2020 Edition with All New Updates / Qs Covered up to Jan

2020

Conceptual Review of
a
Biochemistry
Covering 1300+ MCQs with Explanations, 100+ IBQs &
500 Colored Illustrati
References and Updates from Harper's 31/e, Lehninger's 7 e,
/ Harrison's 20/e,
Lippincott's 7/e, Teitz's 7/e, Devlin's 8 e
/

Recent Qs 2020 - 2012

AIIMS Nov 2019 - 2010
JIPMER May 2019 - 2010
PGI May 2019 - 2010
All India 2011 - 2000
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Biochemistry MICRON
Microbiology Simplified
Conceptual Review of

Pharmacology
for NBE

Iffiri% I
y
.
hi Murugesan

' ' "

.... . .
Forensic Surgery Sixer 4*
Medicine
Nothing Beyond for PGMEE

m is? K Raviraj • VD Agrawal

 Third Edition

Smily Pruthi Pahwa MBBS MD (Biochemistry)

Faculty
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 DISCLAIMER
This book contains questions based on topics asked
in previous years’ State and National Level PG
Entrance and Medical Officer Examinations in India.
Often repeated topics and sub-topics have been
included for students’ benefit. We do not claim that
these questions are exact or similar to questions
asked in recent examinations in India. If any such
similarity is found, it is purely coincidental and by
chance.

ISBN: 978-81-945234-1-3

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Third Edition: 2020

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 Preface
Biochemistry stands out as the most troublesome and unstable subject among the nineteen subjects that one needs to ace to cruise over the
Pre-PG Exams. What makes biochemistry significantly more critical is its basics, which are connected to other subjects, like physiology,
pharmacology and medicine. In Medical Entrance Exams, about 15–20 MCQs are given from biochemistry and in PGI Chandigarh Exam the
number is even more.

Keeping all these facts in mind, the primary aim of this book is to enable you to develop basics with ease. Many self-made formulae are
given in the book to make the subject easy and to help in solving most of the MCQs by using these formulae. The language of the book has
been kept lucid so that all students can understand it easily. In Unit I, Concepts in Biochemistry, emphasizes how the subject is interrelated to
other subjects. This will help you to create a link with other chapters.

The theme of the book is to keep it concise and straightforward with narrative approach. Several Flowcharts, Boxes, Tables and Illustrations
have been added to build a Conceptual Learning. Your last-minute revision has been specially taken care of. For this, the important text has
been highlighted and given in the boxes so that you can revise the subject during eleventh hour. Very few mnemonics are included as the
content of the book has been kept simple and title itself indicates content included under it. Controversies in biochemistry have been carefully
dealt with, wherever needed.

Finally, my suggestion to students is to build an understanding of biochemistry conceptually. Do not cram this interesting and easy
subject. Just mugging up and cramming will not give long-term benefits. Go with the concepts and score well.

Good luck for your examinations! Feel free to get in touch with me for any doubt at my Facebook Group: www.facebook.com/groups/
drsmilybiochem/

Smily Pruthi Pahwa

 Acknowledgements
Although language is the most vital vehicle of human expression, occasionally one finds it difficult to express feelings in appropriate way. It is
not easy to formulate in words, the feeling of gratitude which I have for many persons, who made individual contributions in enabling me to
complete this book.
First and foremost, I thank The Almighty for this precious life and for bestowing His Blessings that helped me carry out this work with
a lot of courage and sheer determination.
Thank you doesn’t seem sufficient for my loving husband Jaspreet Singh whose invaluable presence is constant source of my happiness
and has always endowed me with strength to face challenges in life. Without his relentless support, this book would never had materialized.
Words are insufficient to verbalise my deep sense of gratitude and regards to my parents, who soulfully provided me the sincere
encouragement, inspiration with endless patience, emotional and financial support throughout my work and lifted my morale in this phase
of life. Special and heartiest thanks to my in laws for all the support and encouragement. Especially, I would like to pay homage to my loving
mother-in-law Late Mrs. Jasvinder Kaur, who had been a pillar of support and endless love.
This book would never have been completed without my son Divaahaan’s little contribution, who sacrificed his share of mother’s time.
It is an extraordinary joy to thank my staff Ms Dimple Nagpal, Ms Karamjot Kaur and Ms Taranvir Kaur whose constructive criticism,
deep interest and intellectual acumen were instrumental in accomplishing this task.
Finally, the credit goes to all my students, who have been the actual inspiration for the composition of this particular book.
I would like to thank Mr Satish Kumar Jain (Chairman) and Mr Varun Jain (Managing Director), M/s CBS Publishers and Distributors
Pvt Ltd for providing me the platform in bringing out the book. I have no words to describe the role, efforts, inputs and initiatives undertaken
by Mr Bhupesh Arora, (Vice President – Publishing & Marketing, PGMEE and Nursing Division) for helping and motivating me.

I sincerely thank the entire CBS team for bringing out the book with utmost care and attractive presentation. I thank
Dr Mrinalini Bakshi (Editorial Head & Content Strategist) for her editorial support and Ms Nitasha Arora (Production Head & Content
Strategist), Dr Anju Dhir (Project Manager & Senior Scientific Coordinator), Mr Shivendu Bhushan Pandey (Senior Editor), Mr Ashutosh
Pathak (Senior Proof Reader) and all the production team members, Mr Chaman Lal, Mr Prakash Gaur, Mr Phool Kumar, Mr Bunty
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Mr Vikram Chaudhary, Mr Manoj Malakar and Mr Rahul Negi for devoting laborious hours in designing and typesetting of the book.
 Contents
Preface ----------------------------------------------------------------------------------------------------------------------------------------- v
Acknowledgements -------------------------------------------------------------------------------------------------------------------------- vi
Latest Exam Questions 2020-2019 -------------------------------------------------------------------------------------------------------- ix
Recent Pattern Questions 2020 --------------------------------------------------------------------------------------------------------------------------- x
AIIMS November 2019 ------------------------------------------------------------------------------------------------------------------------------------xi
AIIMS May 2019 -------------------------------------------------------------------------------------------------------------------------------------------xi
JIPMER May 2019 ----------------------------------------------------------------------------------------------------------------------------------------xii
PGI May 2019--------------------------------------------------------------------------------------------------------------------------------------------- xiii

UNIT I    CONCEPTS IN BIOCHEMISTRY

Chapter 1. Concepts in Biochemistry--------------------------------------------------------------------------------------------- 3-26
Theory--------------------------------------------------------------------------------------------------------------------------------------- 3
Multiple Choice Questions--------------------------------------------------------------------------------------------------------------- 18
Answers and Explanations--------------------------------------------------------------------------------------------------------------- 20

UNIT II    C ARBOHYDRATES

Chapter 2. Chemistry of Carbohydrates-----------------------------------------------------------------------------------------29-54

Theory-------------------------------------------------------------------------------------------------------------------------------------- 29
Multiple Choice Questions--------------------------------------------------------------------------------------------------------------- 46
Answers and Explanations--------------------------------------------------------------------------------------------------------------- 49

Chapter 3. Carbohydrate Metabolism------------------------------------------------------------------------------------------ 55-118

Theory-------------------------------------------------------------------------------------------------------------------------------------- 55
Multiple Choice Questions--------------------------------------------------------------------------------------------------------------- 93
Answers and Explanations--------------------------------------------------------------------------------------------------------------103

UNIT III   ENZYMES, AMINO ACIDS AND PROTEINS

Chapter 4. Enzymes-------------------------------------------------------------------------------------------------------------- 119-146

Theory-------------------------------------------------------------------------------------------------------------------------------------121
Multiple Choice Questions--------------------------------------------------------------------------------------------------------------139
Answers and Explanations--------------------------------------------------------------------------------------------------------------142

Chapter 5. Chemistry and Metabolism of Amino Acids-------------------------------------------------------------------- 147-183

Theory-------------------------------------------------------------------------------------------------------------------------------------147
Multiple Choice Questions--------------------------------------------------------------------------------------------------------------174
Answers and Explanations--------------------------------------------------------------------------------------------------------------177

Chapter 6. Urea Cycle------------------------------------------------------------------------------------------------------------ 184-196

Theory-------------------------------------------------------------------------------------------------------------------------------------184
Multiple Choice Questions--------------------------------------------------------------------------------------------------------------192
Answers and Explanations--------------------------------------------------------------------------------------------------------------194
Chapter 7. Proteins--------------------------------------------------------------------------------------------------------------- 197-224
Theory-------------------------------------------------------------------------------------------------------------------------------------197
Multiple Choice Questions--------------------------------------------------------------------------------------------------------------214
Answers and Explanations--------------------------------------------------------------------------------------------------------------218

  UNIT IV    LIPIDS

Chapter 8. Chemistry of Lipids------------------------------------------------------------------------------------------------ 225-242

Theory-------------------------------------------------------------------------------------------------------------------------------------227
Multiple Choice Questions--------------------------------------------------------------------------------------------------------------236
Answers and Explanations--------------------------------------------------------------------------------------------------------------239

Chapter 9. Lipoproteins--------------------------------------------------------------------------------------------------------- 243-255

Theory-------------------------------------------------------------------------------------------------------------------------------------243
Multiple Choice Questions--------------------------------------------------------------------------------------------------------------250
Answers and Explanations--------------------------------------------------------------------------------------------------------------252

Chapter 10. Metabolism of Lipids----------------------------------------------------------------------------------------------- 256-278

Theory-------------------------------------------------------------------------------------------------------------------------------------256
Multiple Choice Questions--------------------------------------------------------------------------------------------------------------270
Answers and Explanations--------------------------------------------------------------------------------------------------------------273

UNIT V    MOLECULAR BIOLOGY

Chapter 11. Chemistry and Metabolism of Nucleotides--------------------------------------------------------------------- 279-293

Theory-------------------------------------------------------------------------------------------------------------------------------------281
Multiple Choice Questions--------------------------------------------------------------------------------------------------------------289
Answers and Explanations--------------------------------------------------------------------------------------------------------------291

Chapter 12. Basics in Genetics--------------------------------------------------------------------------------------------------- 294-321

Theory-------------------------------------------------------------------------------------------------------------------------------------294
Multiple Choice Questions--------------------------------------------------------------------------------------------------------------310
Answers and Explanations--------------------------------------------------------------------------------------------------------------314

Chapter 13. Replication, Transcription, Translation------------------------------------------------------------------------- 322-350

Theory-------------------------------------------------------------------------------------------------------------------------------------322
Multiple Choice Questions--------------------------------------------------------------------------------------------------------------339
Answers and Explanations--------------------------------------------------------------------------------------------------------------343

Chapter 14. Techniques in Molecular Biology--------------------------------------------------------------------------------- 351-374

Theory-------------------------------------------------------------------------------------------------------------------------------------351
Multiple Choice Questions--------------------------------------------------------------------------------------------------------------365
Answers and Explanations--------------------------------------------------------------------------------------------------------------368

  UNIT VI    MISCELLANEOUS

Chapter 15. Miscellaneous------------------------------------------------------------------------------------------------------- 375-412

Theory-------------------------------------------------------------------------------------------------------------------------------------377
Multiple Choice Questions--------------------------------------------------------------------------------------------------------------398
Answers and Explanations--------------------------------------------------------------------------------------------------------------403

Extra Edge Tables ----------------------------------------------------------------------------------------------------413-416

Image–Based Questions---------------------------------------------------------------------------------------------417-456
 Latest Exam Questions
2020-2019

1. Recent Pattern Questions 2020

2. AIIMS November 2019
3. AIIMS May 2019
4. JIPMER May 2019
5. PGI May 2019
 8. Which enzyme is active during low insulin glucagon
RECENT PATTERN QUESTIONS 2020 ratio?
x 1. Which of the following is insoluble in water? a. Hexokinase
a. Lignin b. Glucokinase
b. Inulin c. Glucose 6-phosphatase
c. Amylose d. Phosphofructokinase
CRO BIOCHEMISTRY

d. Chitin 9. Regarding proteins which is true?

2. Nitric oxide is synthesized from: a. Primary, secondary, tertiary structure are destroyed
a. Ornithine by denaturation.
b. Alanine b. Secondary structure shows three dimensions.
c. Aspartic acid c. Secondary structure stabilized by disulfide bridges.
d. L arginine d. Secondary and tertiary structures depends on
3. Given below are the parameters of ABG analysis. pH amino acids
7.2, HCO3 –10 mEq/ L, PCO2 -30 mm Hg. What is your 10. In preterm infants with RDS, what is deficient?
diagnosis? a. Dipalmitoylphosphatidylcholine
a. Metabolic alkalosis b. Phosphatidylethanolamine


b. Metabolic acidosis c. Phosphatidylinositol

c. Respiratory acidosis d. Cardiolipin
d. Respiratory alkalosis 11. A 50-year-old man with paresthesia, Hb = 6.8 gms%,
4. A 50-year-old male presented with tachypnea, peripheral smear–macrocytes and neutrophils with
confusion, blood glucose 350 mg/dl and pH 7.0. What hyper-segmented nuclei and atropic gastritis
is your diagnosis? a. Vitamin B12 deficiency
a. Metabolic alkalosis b. Folate trap
b. Metabolic acidosis c. Riboflavin deficiency
c. Respiratory acidosis d. Vitamin C deficiency
d. Respiratory alkalosis 12. Which of the following binds to the tyrosine kinase
5. Deficiency of which of the following vitamins causes
receptors?
this scenario?
a. Insulin
b. Glucagon
c. Leptin
d. Thyroxine
13. In Non-competitive inhibition:
Ans. a. Vmax decreases, Km remains constant
1. a b. Vmax remains same, Km increases
2. d c. Both Vmax and Km are decreased
3. b d. Km is usually increased, Vmax is reduced
4. b 14. Amino acid which absorbs UV light at 280 nm:
5. c a. Tryptophan
6. a b. Histidine
7. a c. Aspartate
8. c d. Ornithine
a. Riboflavin deficiency
9. d 15. Apo 48 is a remnant of:
b. Vitamin B12 deficiency
10. a a. RNA editing
c. Niacin deficiency
11. a b. RNA interference
d. Vitamin C deficiency
12. a c. RNA splicing
6. Urine turns black on standing in case of:
13. a d. DNA translocation
a. Homogentisate oxidase deficiency – alkaptonuria
14. a 16. Adenosine is 28%, what is the % of cytosine?
15. a
b. Cystathionine beta synthase deficiency –
homocystinuria a. 22%
16. a
c. Tyrosinase deficiency – Albinism b. 26%
17. a
d. Phenylalanine hydroxylase deficiency – c. 44%
phenylketonuria d. 56%
7. Which of the mucopolysaccharide/proteoglycan 17. Which factor in warfarin therapy, have decreased
present in glomerular basement membrane? gamma carboxylation of glutamate residue?
a. Heparan sulfate a. Factor II
b. Chondroitin sulfate b. Factor V
c. Hyaluronic acid c. Factor VIII
d. Keratan sulfate d. Factor III
 24. Which replacement of amino acid does not alter its
AIIMS NOVEMBER 2019 normal function?
18. In urea cycle disorder, which of the following a. Glutamine replaced by arginine xi
substance can be used to reduce the levels of b. Glutamine replaced by alanine
ammonia? c. Glutamine replaced by glutamate
a. L-carnitine d. Glutamine replaced by asparagine

 LATEST EXAM QUESTIONS 2020-2019

b. Phenylbutyrate 25. In CRISPR-Cas 9 system, which repair mechanism is
c. Isoleucine used for genome editing?
d. Glutamate a. Homologous repair
19. The clinical feature given below occurs due to b. Non-homologous end repair
deficiency of: c. Mismatch repair
d. Nucleotide excision repair
26. Assertion: Most common hematological disorder in
chronic lead poisoning is eosinophilia.
Reason: Chronic lead poisoning inhibits the enzyme
ALA dehydratase.
a. Both Assertion and Reason are independently true/
correct statements and the Reason is the correct
explanation for the Assertion
b. Both Assertion and Reason are independently true/
correct statements, but the Reason is not the correct
explanation for the Assertion
c. Assertion is independently a true/correct statement,
but the Reason is independently a false/incorrect
statement
d. Assertion is independently a false/incorrect
statement, but the Reason is independently true/
correct statement
a. Tryptophan e. Both Assertion and Reason are independently false/
b. Phenylamine incorrect statements
c. Cysteine
d. Glutamine
20. Which of the following binds mRNA with ribosome in AIIMS MAY 2019
eukaryotes? 27. Which of the following doesn’t happen in 5’ to 3’ Ans.
a. Poly A tail direction? 18. b
b. tRNA a. Transcription b. DNA replication 19. a
c. 7-methylguanosine cap c. DNA repair d. DNA Editing 20. c
d. Shine-dalgarno sequence 28. Chimeric DNA is used for: 21. a
21. Which of the following glycoprotein has both a. Paternity test 22. d
lubricant and protective action: b. Maternity test 23. a
a. Mucin c. Personal identification 24. d
b. Immunoglobulin d. Organ transplantation 25. b
c. Collagen 29. A bodybuilder starts eating raw eggs for protein. He 26. d
d. Albumin developed fatigue on moderate exercise. The doctor 27. d
22. All of the following are correct about Proteoglycans prescribes a vitamin. Which enzyme is deficient in 28. d
except: him? 29. b
a. Chondroitin sulfate is a proteoglycans a. Glucose 6 Phosphatase 30. a
b. Made of protein and sugar MPS b. Pyruvate Carboxylase
c. Carries negative charge c. PEPCK
d. Doesn’t hold water d. Glycogen Phosphorylase
30. Which of the following shows functional assessment
23. Restriction endonuclease will act on which of the
of B1 deficiency:
following:
a. RBC Transketolase
a. AAGCTT
b. RBC Glutathione reductase
b. AAGAAG
c. Serum Thiamine levels
c. TACGAG
d. RBC Glutathione Peroxidase
d. GAGAGG
 31. Assertion: Central dogma says DNA is transcribed 35. The catalytic efficiency of enzyme is best expressed
into RNA which is subsequently translated into by which of the following kinetic constants:
xii proteins a. Kcat/Km
Reason: Retrovirus show reverse transcriptase b. Km/Kcat
of central dogma where RNA is first converted to c. Km/Ka
DNA which is then transcribed to RNA, which is d. Ka/Km
CRO BIOCHEMISTRY

subsequently translated into proteins 36. Type of mutation seen in sickle cell anemia:
a. Both Assertion and Reasons are independently true a. Insertion
and the Reason is the correct explanation for the b. Deletion
Assertion c. Point mutation
b. Both Assertion and Reasons are independently true, d. Frameshift mutations
but the Reason is not the correct explanation for the 37. In which of the inheritance, if father is affected
Assertion no offspring is affected, but if mother affected, all
c. Assertion is independently true, but the Reasons is offspring affected?
independently a false a. Mitochondrial
d. Assertion is independently a false, but the Reasons b. X linked recessive
is independently a true


c. Autosomal dominant
e. Both Assertion and Reasons are independently false d. Autosomal recessive
32. Which of the following is false regarding reducing 38. Hepcidin decreases iron absorption by inhibiting:
activation energy? a. Divalent metal transporter
a. Decreased entropy b. Hephaestin
b. Desolvation of active site c. Ferroportin
c. Conformational change in the active site of enzyme d. Transferrin
at or after the time of binding of substrate to 39. Which is true regarding the impact of the following
enzyme on cardiac diseases?
d. Complementarity of active site of enzyme and a. Vitamin E deficiency will not cause cardiac
substrate ischemia/problems
33. The graph shown below is the titration curve of a b. Vitamin C deficiency can cause impaired cardiac
biochemical compound. Which of the following health
statement is true? c. Folate has proven role in preventing cardiac illness
d. Vitamin E worsens CAD

Ans. JIPMER MAY 2019

31. b 40. Rapoport Luebering cycle shifts O2 dissociation curve
32. d to right because of:
33. b a. 2, 3 DPG
34. c b. 1, 3 BPG
35. a c. 3 phosphoglycerate
36. c d. 1, 6 fructose bisphosphate
37. a 41. Ketone bodies may be seen in:
38. c
a. Diabetes mellitus
39. b
b. Pancreatitis
40. a
c. Obesity
41. a
d. Myocardial infarction
42. c
42. Arrange according to molecular size:
43. a a. The maximum buffering capacity of the compound a. VLDL > LDL > Chylomicron > HDL
is represented by points A and B. b. HDL > LDL > Chylomicron > VLDL
b. The points A and B represent the range of maximum
c. Chylomicron > VLDL > LDL > HDL
ionization of the carboxyl and amino group
d. VLDL > LDL > Chylomicron > HDL
c. The compound has three ionisable side chains
43. Vitamins playing an important role in citric acid
d. The compound has one ionisable group
cycle:
34. Which of the following TCA cycle intermediate is a
a. Thiamine, riboflavin, niacin, lipoic acid
part of heme metabolism?
b. Thiamine, biotin, riboflavin, pantothenic acid
a. Alpha ketoglutarate
c. Thiamine, pyridoxine, riboflavin, niacin,
b. Fumarate
pantothenic
c. Succinyl CoA
d. Thiamine mecobalamin, pantothenic acid
d. Malate
44. Dimercaprol inhibits: 51. In proteins, the amino acids are folded. These folded
a. ETC complex 1 units are further folded as dimers-Homodimers and
b. ETC complex 2 heterodimer. These dimeric structures are called: xiii
c. ETC complex 3 a. Primary Structure
d. ETC complex 4 b. Secondary Structure
45. Enzyme deficient in Hyperammonemia type 1: c. Tertiary Structure

 LATEST EXAM QUESTIONS 2020-2019

a. Ornithine transcarbamoylase d. Quaternary structure
b. Arginase e. Denatured protein
c. Carbamoyl phosphate synthase I 52. Enzyme(s) of urea cycle is/are:
d. Argininosuccinate synthase a. Glutamate dehydrogenase
46. During Fasting, which of the following is released b. Argininosuccinate lyase
from the muscles? c. Ornithine transcarbamylase
a. Alanine d. Argininosuccinate synthetase
b. Glutamine e. Carbamoyl phosphate synthetase-I
c. Branched keto acid 53. Essential amino acids is/are:
d. Asparagine a. Threonine
47. Ketone bodies not utilized by: b. Phenylalanine
a. Brain c. Alanine
b. RBC d. Methionine
c. Heart e. Cysteine
d. Skeletal muscle 54. Which is/are not seen in hypervitaminosis A:
a. Xerophthalmia
b. Hyperostosis
PGI MAY 2019 c. Desquamation of skin
48. Pentose phosphate pathway occurs in: d. Hirsutism
a. Cytosol e. Liver enlargement
b. Mitochondria 55. Cholesterol is/are absent in:
c. Lysosome a. LDL
d. Golgi apparatus b. HDL
e. Endoplasmic reticulum c. Urine
49. Pellagra is/are caused by: d. Saliva
a. Niacin deficiency e. Exudate and transudate
b. Hartnup disease 56. Which of the following dyads of plasma protein-
function is/are correct: Ans.
c. Tryptophan excess
d. Rifampicin use a. Transferrin- Transport iron 44. c
e. Carcinoid syndrome b. Thyroxin binding globulin- thyroxin binding and 45. c
50. Pyruvate consumption is hampered in which of the transport 46. a
following condition(s): c. Ceruloplasmin- Assisting in iron absorption by 47. b
a. Pellagra transferrin 48. a
b. Beriberi d. Haptoglobin- Heme binding and destruction 49. a,b,e
c. Scurvy e. Hemopexin-Heme binding 50. a,b
d. Rickets 51. d
e. Pernicious anemia 52. b,c,d,e
53. a,b,d
54. a,d
55. d
56. a,b,c,e
 Answers with Explanations
xiv

RECENT PATTERN QUESTIONS 2020 4. Ans. b. Metabolic acidosis

CRO BIOCHEMISTRY

[Ref: Dinesh Puri 3rd ed pg- 16]

1. Ans. a. Lignin
High Blood glucose indicates diabetic condition
[Ref: Lehninger 6th ed pg 257] which will cause ketone body production which is
•• Lignin a complex organic polymer present in cause of decrease in pH due to their acidic nature.
abundant quantities in plant secondary cell wall. A disorder resulting in a decreased pH of arterial plasma
It is a major Insoluble Dietary Fiber. It is not a (<pH 7.35) due to a defective metabolism is called as
Carbohydrate. It is a Poly phenolic compound, a metabolic acidosis.
polymer of Phenyl propane. Due to its non-polar
5. Ans. c. Niacin deficiency


nature it is not soluble in water.

•• Inulin, amylose and chitin are homopolysaccharides [Ref: Vasudevan 6th ed pg 395]
made up of fructose, glucose and N-acetylglucosamine
This is Casal’s necklace shown in picture, which occurs
monomeric subunits respectively. Due to the
in Vitamin B3/Niacin deficiency. Niacin is formed from
presence of polar groups such as –OH, –COOH these
tryptophan amino acid. So, tryptophan deficiency can
are partly soluble in water.
also lead to Casal’s necklace. 60 mg tryptophan gets
2. Ans. d. L arginine converted to 1 mg Niacin.

[Ref: Harper 31st ed pg- 624] 6. Ans. a. Homogentisate oxidase deficiency –

alkaptonuria
NO is also called as Endothelium Derived Relaxing
Factor (EDRF). NO (nitric oxide) is synthesized from [Ref: Harper 31st ed pg - 286]
arginine by enzyme NOS (Nitric oxide synthase) in the
In patients of Alkaptonuria with deficiency of
endothelial cells. The vasodilator – nitroglycerin also
Homogentisate dioxygenase/ oxidase (requires Iron),
enters smooth muscle cells, where its metabolism also
fresh urine is normal in colour; but on standing or
leads to the formation of NO.
on exposed to air urine turns Black. It is because
Extra information: homogentisic acid accumulated in urine gets oxidized
by oxygen in air and gives rise to black colored
No synthase benzoquinone acetic acid.
A •• There are three isoforms of NOS (Nitric oxide
N synthase) 7. Ans. a. Heparan sulfate
S 1. nNOS - neuronal
W 2. iNOS- inducible [Ref: Harper 31st ed pg - 602]
E 3. eNOS- endothelial Heparan sulfate is present in basement membrane of
R •• EC no. 1 (oxidoreductase) glomerulus. Also, it is present on cell surfaces and has a
S •• Monooxygenase (one oxygen is added in substrate) role in retinal cell-cell attachment.
•• Usually belongs to EC no 4 but NO synthase is an
WITH exception. 8. Ans. c. Glucose 6-phosphatase
E 3. Ans. b. Metabolic acidosis [Ref: Harper 31st ed pg – 137-138]
X
P [Ref: Dinesh Puri 3rd ed pg- 16] •• Low insulin glucagon ratio means it is fasting state, as
L in fed state insulin is more. So in fasting state glucose
A disorder resulting from a primary fall in bicarbonate 6-phosphatase will be active as this enzyme is
A level (HCO3) (<22 mEq/L) resulting in a decreased pH
N involved in gluconeogenesis, which occurs during
of arterial plasma (<pH 7.35). This happens because of fasting.
A decreased ratio for bicarbonate concentration to pCO2
T •• Phosphofructokinase is involved in glycolysis. PFK-1
(normal level is 35-45 mm Hg) is called as metabolic is active in any situation and PFK-2 is active only in
I acidosis.
O fed state.
In respiratory acidosis there is increase of both •• Glucokinase phosphorylates glucose in liver and
N bicarbonate level (HCO3) and pCO2 above normal.
S pancreas, active in fed state and is induced by insulin.
 •• Hexokinase also phosphorylates glucose and •• Glucagon binds to G-protein coupled receptors.
is present in all cells of the body. It is active in Leptin binds to cytokine receptor while thyroxine
any situation in body as it is a general enzyme to binds to nuclear receptors. xv
phosphorylate glucose.
13. Ans. a. Vmax decreases, Km remains constant
9. Ans. d. Secondary and tertiary structures depends

 LATEST EXAM QUESTIONS 2020-2019

on amino acids [Ref: Harper 31st ed pg - 78]
•• In Non-Competitive Inhibition: Inhibitor binds at
[Ref: Harper 31st ed pg – 33-38]
regulatory site of the enzyme and induces changes in
•• Disulfide bond is not present in 2° structure, but the active site so that substrate cannot bind and thus
is present in tertiary structure. 1° structure is not decreases the velocity of the reaction. But there is no
destroyed by denaturation. change in Km or Substrate concentration.
•• 2° and 3° structures are based on amino acid •• Km is increased in case of competitive inhibition,
sequence, In fact the overall protein structure where the inhibitor binds at the active site.
depends upon amino acid sequence. Even change
of one amino acid can affect the overall structure &
function of protein.
•• Secondary structure is not the three-dimensional
structure of protein. The overall 3D structure of a
polypeptide is called its tertiary structure.
•• Tertiary structure is due to overall interactions
between the R groups of amino acids. Secondary
structure is due to local interactions between
stretches of a polypeptide chain.

10. Ans. a. Dipalmitoylphosphatidylcholine

[Ref: Harper 31st ed pg - 234]

RDS occurs when there is not enough surfactant in
the lungs. Surfactant is a liquid made by the lungs that
keeps the airways (alveoli) open. Pulmonary surfactant
14. Ans. a. Tryptophan
is a surface-active lipoprotein complex (phospho
lipoprotein) formed by type II alveolar cells. The [Ref: Harper 31st ed pg - 20]
proteins and lipids that make up the surfactant have both
hydrophilic and hydrophobic regions. By adsorbing to Proteins and amino acids absorb UV-light at 280 nm,
the air-water interface of alveoli, with hydrophilic head due to aromatic nature of these amino acids & they A
groups in the water and the hydrophobic tails facing have conjugated double bonds in the rings. Maximum N
towards the air, the main lipid component of surfactant absorption occurs by tryptophan as it has 2 rings in its S
is dipalmitoylphosphatidylcholine (DPPC). side chain. W
E
11. Ans. a. Vitamin B12 deficiency 15. Ans. a. RNA editing
R
[Ref: Harper 31st ed pg - 536] [Ref: Harper 31st ed pg - 238] S
The symptoms presented here are all indicative of In liver cell, Apo B 100 gene gives rise to Apo B 100 WITH
megaloblastic anemia which is mainly caused due to protein. But Apo B 48 protein is made in intestinal cells
from the gene encoding Apo B 100, by the process of E
deficiency of Vitamin B12 or folate.
RNA editing, which is also known as Differential RNA X
12. Ans. a. Insulin processing or chemical modification of RNA. This is a P
post transcriptional modification. L
[Ref: Harper 31st ed pg - 508] •• RNA splicing is also a post transcriptional modi- A
•• The insulin and IGF-I receptors have intrinsic fication but this is removal of introns and not editing. N
ligand-activated tyrosine kinase activity. By binding •• RNA interference is silencing technique where the A
to these receptors, these hormones will activate gene is silenced by not letting the product of gene T
tyrosine kinases inside the cell which in turn formed. I
phosphorylates important enzymes for involved in •• Translocation occurs during translation. This is O
a number of processes such as metabolism, growth movement of ribosome over mRNA. N
and differentiation and also inflammatory response. S
 16. Ans. a. 22%

xvi [Ref: Harper 31st ed pg - 339]

•• By Chargaff’s rule, number of purines equal the
number of pyrimidines in a double stranded DNA.
So, amount of Adenine = Thymine, as they pair with
CRO BIOCHEMISTRY

each other by two hydrogen bonds. As adenosine

given is 28 %, so total A+T is 56%.
•• Now also, amount of Guanine = Cytosine, as they
pair with each other by three hydrogen bonds. Other method to treat urea cycle disorder is Arginine
Remaining 44% equally distributed in 22% each for supplementation (1st line treatment): Arginine is an
guanine and cytosine. essential amino acid. It provides ornithine (required in
2nd step of urea cycle) which is an activator of NAGS.
17. Ans. a. Factor II But it is contraindicated in Arginase deficiency
[Ref: Harper 31st ed pg - 533] 19. Ans. a. Tryptophan


Vitamin K is required for activation of clotting factors

- 2, 7, 9 and 10. Warfarin is a competitive inhibitor of [Ref: Vasudevan 6th ed pg 395]
enzyme epoxide reductase required for vitamin K cycle; This is Casal’s necklace shown in picture, which occurs
hence It will inhibit vitamin k function in turn will affect in Vitamin B3/ Niacin deficiency.
the clotting factor II (prothrombin) activation. •• Niacin is formed from tryptophan amino acid. So,
tryptophan deficiency can also lead to Casal’s
necklace.
•• 60 mg tryptophan gets converted to 1 mg Niacin.

A dditional E dge
Vitamin B3 Deficiency: Pellagra
Characterized by: 4 Ds of Pellagra
•• Dermatitis (Photosensitive Dermatitis)
•• Diarrhoea
•• Dementia
•• Death

20. Ans. c. 7-methylguanosine cap

A
N [Ref: Lehninger 6e/ Pg 1075, 1085]
S In eukaryotic cells, ribosomes recognize most eukaryotic
W mRNAs by binding to the 7-methylguanosine cap at
E their 5´ terminus.
R Shine-Dalgarno sequence (SD sequence) is present at
S AIIMS NOVEMBER 2019 -10 position on mRNA, only in prokaryotes and is rich
in purines. In smaller subunit of ribosome, 16 s rRNA
WITH is present. SD sequence is complementary to 16 s
18. Ans. b. Phenylbutyrate
rRNA, and their binding helps in binding of mRNA with
E
[Ref: Lehninger 6th ed pg- 709] ribosome during initiation of translation in prokaryotes.
X
P In all urea cycle disorders there is accumulation of
21. Ans. a. Mucin
L NH3 (hyperammonemia) which is very toxic and has to
A be removed. [Ref: Harper 31st Ed., p.-547]
N Mucins are a family of high molecular weight and
A Using scavenging agents:
heavily glycosylated proteins.
T •• One of the treatments of urea cycle disorder These large glycoproteins are secreted in mucus, the
I is the use of NH3 scavenging agents. These are viscous fluid that protects and lubricates the epithelium
O Phenylbutyrate and Sodium benzoate. Phenyl of the gastrointestinal, genitourinary, and respiratory
N butyrate reacts with glutamine to get converted into tracts. However, some mucins may be membrane
S phenyl acetyl glutamine which can be excreted. associated and not secreted.
 •• Arginine is basic amino acid, so it is positively
A dditional E dge charged.
•• Glycoproteins are proteins that contain oligosaccharide •• Glutamate is acidic amino acid, so it is negatively xvii
chains (glycans) covalently attached to their polypeptide charged.
backbones. O-linked glycoproteins and N-linked •• Alanine is non-polar
glycoproteins are major classes of glycoproteins.

 LATEST EXAM QUESTIONS 2020-2019

•• O–linked glycoprotein: These are glycoproteins where O-
glycosylation occurs at serine or threonine residues. Serine 25. Ans. b. Non-homologous end repair
is more common.
•• N-linked glycoproteins: These are glycoproteins where N- [Ref: Cell. 2014 Jun 5; 157(6): 1262–1278. https://www.
glycosidic linkage takes place between amide nitrogen of addgene.org/guides/crispr/?utm_source=Addgene+
Asparagine. Ma r c h + 1 4 % 2 C + 2 0 1 3 + Ne w s l e t t e r + Fi n a l & u t m _
campaign = March+2013+ Newsletter&utm_medium=
email]
22. Ans. d. Doesn’t hold water CRISPR-Cas-9 endonuclease system introduces double
stranded breaks in DNA at the specific target site. Now
[Ref: Harper 31st ed., p.601-602] cell tries to repair this double stranded break by two
•• Chondroitin sulfate is a glycosaminoglycan -> mechanism:
option 1 is true •• Nonhomologous end repair or
•• Proteoglycan = core protein (Amino acid) + •• Homologous repair
Glycosaminoglycan (sugar) -> option 3 is true Nonhomologous repair mechanism is always a
•• Sulfate and acetyl groups confer negative charge to possibility as it does not require homologous DNA. So,
the amino acids -> option 4 is true it is the major repair mechanism during CRISPR-Cas
•• As they are negatively charged, they can hold large system.
amount of water, not less -> option 2 is wrong. The homologous repair will only work if the DNA
homologous to the cut DNA is present but is not always
23. Ans. a. AAGCTT present.
[Ref:Robbins 9th/e pg. 176]
26. Ans. d. Assertion is independently a false/incorrect
Restriction endonuclease (also known as Molecular statement, but the Reason is independently true/
Scissor) cuts palindromic sequences. Palindromic correct statement
sequence is that when sequence is same on both the
strands of DNA. [Ref: Harper 31st Ed., p.309-310]
•• A palindromic sequence is a sequence on ds-DNA •• Lead poisoning most commonly cause acquired
that is the same when read from 5' to 3' on one strand porphyria
and 5' to 3' on the other, complementary strand. An •• Lead inhibits two enzymes:
example of a palindromic sequence is 5'-GGATCC-3', A
1. ALA dehydratase
which has a complementary strand, 3'-CCTAGG-5’. N
2. Ferrochelatase
S
W
E
R
S
WITH

•• Option A is a palindrome i.e. 5’ AAGCTT 3’ and other E

complementary strand is 3’ TTCGAA 5’. X
P
24. Ans. d. Glutamine replaced by asparagine L
A
[Ref: Harper 31st Ed., p. 15-16] N
Glutamine and asparagine are uncharged polar amino A
acids. Both have amide group. So exchange of these T
amino acids in a protein will not change the charge of I
the protein. O
N
S
 •• PEPCK is a kinase & is an enzyme in gluconeo-
AIIMS MAY 2019 genesis.
xviii •• Glycogen Phosphorylase is Rate Limiting Enzyme of
27. Ans. d. DNA Editing Glycogenolysis. It requires vitamin B6.
•• Glucose 6 phosphatase is a common enzyme in
[Ref: Harper's 31st Ed./pg.- 238]
gluconeogenesis and glycogenolysis.
CRO BIOCHEMISTRY

•• DNA editing means proofreading. It occurs in 3’→ 5’

direction. 30. Ans. a. RBC Transketolase
•• In DNA Replication whenever synthesis occurs, new
[Ref: Harper's 31st Ed. / pg. 534]
nucleotide is added to 3’end hence replication occur
in 5’to 3’ direction. •• Lab diagnosis of Thiamine deficiency is assessed by
•• In transcription also, RNA is getting synthesized in 5’ Erythrocyte transketolase activity.
to 3’ direction •• Transketolase enzyme is involved in HMP pathway.
•• Hence, DNA replication, removal of primer during •• As HMP pathway occurs in RBCs, Transketolase
replication, repair and transcription occurs in 5’ to 3’ enzyme is abundantly present in RBCs.
direction. •• There is no importance of checking serum thiamine


levels (Option c) instead we check enzyme activity.

28. Ans. d. Organ transplantation •• RBC Glutathione reductase activity is the marker for
vitamin B2 deficiency.
[Ref: http://uagcclinical.arizona.edu/test/chimerism-
testing/, https://www.ncbi.nlm.nih.gov/pmc/articles/ •• Enzyme Transaminase activity in checked for
PMC2662135/] Pyridoxine (Vitamin B6) deficiency.
Note: If Relation between thiamine and energy is asked
More correct option would be Hematopoietic Cell then mark enzyme of Link reaction and TCA.
Transplantation Both link reaction and TCA requires 4 B-complex
•• Chimerism is a condition whereby a person has not vitamins B1, B2, B3 and B5.
one but two complete genomes (sets of DNA) in their
31. Ans. b. Both Assertion and Reasons are
body. One genome is found in one region or organ(s),
independently true, but the Reason is not the correct
while the other genome can be predominant in other
explanation for the Assertion
organs or tissues.
•• Chimerism testing (engraftment analysis) is [Ref: Harper 31st/e pg. 367]
performed for patients who have received a
Both Assertion and Reasons are independently true,
hematopoietic stem cell transplant. The test involves
but the Reason is not the correct explanation for the
identifying the genetic profiles of the recipient and of
Assertion. This can be explained as:
the donor and then evaluating the extent of mixture
•• Assertion: Central dogma of protein synthesis is DNA
A in the recipient’s blood or bone marrow. First, DNA
to RNA and translation from RNA to protein
N is isolated from the recipient and potential donor
DNA → RNA → Protein
S before the transplant and analysis is performed to
•• Reason: Reverse transcriptase converts RNA to DNA
W determine whether the genetic markers unique to
and then normal process of DNA to RNA to proteins
E the donor and the recipient have sufficient power to
happens.
R distinguish the donor from the recipient.
RNA → DNA → RNA → Protein
•• Next, after the transplant takes place, the performance
S •• This reversal of central dogma occurs in retro viruses.
of the transplant engraftment is assessed by
Retro viruses have their genome as RNA and they
WITH evaluating the donor versus recipient contribution
have reverse transcriptase enzyme, which converts
of white blood cells in post-transplant blood or bone
E RNA to DNA.
marrow specimens obtained from the recipient.
X There is no link between these two processes. Both are
P 29. Ans. b. Pyruvate Carboxylase independent phenomenon, but they can occur in same
L cell. For example, in retrovirus both processes happen
A [Ref: Harper's 31st ed. / pg. 538] but have no influence on each other.
N •• Raw eggs contain protein Avidin → tightly binds to
32. Ans. d. Complementarity of active site of enzyme
A Biotin (vit B7) and is excreted along with biotin.
and substrate
T •• Biotin is required for all carboxylases; all
I carboxylases requires ATP also. [Ref: Lehninger 6th ed pg- 195-197]
O •• So, enzyme affected is carboxylase which requires
N The energy derived from enzyme-substrate interaction
Biotin as coenzyme.
S is called binding energy, GB. Binding energy is a major
•• Pyruvate Carboxylase is the first enzyme of
source of free energy used by enzymes to lower the
gluconeogenesis.
 activation energies of reactions. An enzyme completely
complementary to its substrate would be a very poor •• The maximum buffering capacity happens at the point of pK of
enzyme (Option-D) while an enzyme, which is flexible different groups which is midpoint of horizontal part of curve xix
(Option-C) has more chances to make the substrates and not points A and B (option a is fasle)
reach the transition state hence maximizes the binding •• The compound has only one ionizable side chain group as in
energy. histidine (option c is false)

 LATEST EXAM QUESTIONS 2020-2019

Hence statement in option D can be considered as false •• The compound has three ionizable groups (option d is false)
regarding for reducing the activation energy. •• Point A and B represent maximum ionization of carboxyl and
•• Option B: The active sites of enzymes usually occur in amino groups (option b is true)
clefts and crevices in the protein. This design has the
effect of excluding bulk solvent (water), which would
34. Ans. c. Succinyl CoA
otherwise reduce the catalytic activity of the enzyme.
The substrate molecule is desolvated upon binding, [Ref: Harper 31st/e pg. 306]
and shielded from bulk solvent in the enzyme’s active
The first step of heme synthesis uses succinyl CoA
site.
which is an intermediate of TCA.
•• Option A: Entropy reduction: Enzymes bring the
substrate molecules very close and helps to reduce Heme synthesis
the entropy. This facilitates the product formation •• It occurs in Mitochondria + cytoplasm (So, doesn’t
by increasing number of effective collisions between occurs in RBC)
substrate molecules hence helps in catalysis.

33. Ans. b. The points A and B represent the range of

maximum ionization of the carboxyl and amino
group

[Ref: Lehninger 6th ed pg- 84-85]

This curve shown is similar to titration curve of histidine
with three parts

A
N
35. Ans. a. Kcat/Km
S
[Ref: Lehninger 6th edition pg- 205] W
E
•• The kinetic parameters Kcat and Km are useful for the
R
study and comparison of different enzymes, with
simple or complex reaction mechanisms. S
•• Kcat and Km are used as an estimate for the kinetic WITH
efficiency of enzymes, but either parameter alone is
insufficient for this purpose. E
•• For instance, two enzymes catalyzing different X
reactions may have the same Kcat (turnover number), P
yet the rates of the uncatalyzed reactions may be L
different and thus the rate enhancements brought A
about by the enzymes may differ greatly. N
•• Hence, the best way to measure the catalytic A
efficiencies of different enzymes is to compare the T
ratio Kcat/Km for the two reactions. This parameter, is I
called the specificity constant, is the rate constant O
for the conversion of E + S to E + P when [S] << Km. N
S
 36. Ans. c. Point mutation

xx [Ref: Biochemistry by_Satyanarayana and_Chakrapani

pg no: 553]

Sickle-cell anemia
CRO BIOCHEMISTRY

The occurrence of the disease sickle-cell anemia due to

a single base alteration (CTC + CAC in DNA, and CAG +
GUC in RNA) is a classic example of point mutation. The
result is that glutamate at the 6th position of B-chain of
hemoglobin is replaced by valine. This happens since
the altered codon GUG of mRNA codes for valine
instead of glutamate.

37. Ans. a. Mitochondrial

[Ref: Biochemistry by_Satyanarayana and_Chakrapani Other Options



pg no: 739]
•• Divalent metal transporter- absorbs divalent metal
•• In all the offspring, mitochondria is derived only ions from intestine especially Ferrous.
from mother. Hence, any mutation in mother •• Hephaestin – involved in homeostasis of iron &
mitochondria will also be transferred to its offsprings. copper.
So in mitochondrial gene inheritance if mother is
•• Transferrin is a protein which is responsible for the
affected all offspring will be affected.
transport of Iron
Other Options 39. Ans. b. Vitamin C deficiency can cause impaired
•• In case of X-linked disease there is no male to male cardiac health
transmission.
•• In Y linked – father will transmit disease to all his [Ref: Textbook of Medical Biochemistry by Dinesh Puri
sons. 3rd/ed page- 386, 397']
•• In autosomal inheritance, there is equal frequency of •• Option-1 (False) Vit E has anti-atherogenic role.
these disease in male and female Hence Vit E deficiency can cause cardiac problems.
•• Option 4- Vit E due to antioxidant properties
Additional Information:
decreases the risk of CAD. It protects against
•• Mitochondrial disease has high incidence as oxidation of low-density lipoprotein and decreases
mitochondrial DNA repair cannot occur, introns the deposition of atherogenic oxidized low-density
A not present, and there is continuous exposure to the lipoprotein in arterial walls.
N oxygen free radicals of ETC. •• Option 2- Vitamin C also due to its antioxidant role
S •• Due to these few reasons mitochondrial DNA damage prevent heart diseases. So its deficiency can cause
W have more chances of mutations in mitochondrial impaired cardiac health.
E DNA as compared to nuclear DNA. •• Option 3- Folate has main role in the one carbon
R donor THF synthesis which is mainly involved in
38. Ans. c. Ferroportin
S synthesis of nucleic acids and amino acids. It has no
[Ref: Harper 31st/e pg. 523] proven role in cardiac illness. However, it can cause
WITH
anemia.
Hepcidin
E
•• Hepcidin regulates iron metabolism. It mainly
X
regulates iron entry in circulation.
JIPMER MAY 2019
P
L •• By inhibiting ferroportin, hepcidin reduces dietary
A iron absorption. The iron release from macrophages 40. Ans. a. 2, 3 DPG
N is also reduced by ferroportin inhibition. So, increased
hepcidin activity is responsible for reduced iron [Ref: Harper's 31st Ed. / page – 161]
A
T availability. Increased hepcidin will lead to anemia. Shifting the O2 dissociation curve to right means
I Decreased hepcidin leads to iron overload decreasing the oxygen saturation of hemoglobin as
O •• Hepcidin synthesis and secretion by the liver is shown in graph.
N controlled by iron stores within macrophages,
S inflammation, hypoxia, and erythropoiesis.
 42. Ans. c. Chylomicron > VLDL > LDL > HDL

[Ref: Harper's 31st Ed./page - 237] xxi

Lipoprotein can be arranged on the basis of different
parameters as:

 LATEST EXAM QUESTIONS 2020-2019

43. Ans. a. Thiamine, riboflavin, niacin, lipoic acid
[Ref: Lippincott 7th edition page-396]
Role of Vitamins in TCA
Coenzyme Vitamin TCA cycle enzyme

TPP B1 (Thiamine) Alpha - Ketoglutarate

DH, Isocitrate DH
FAD B2 (Riboflavin) Succinate DH
NAD B3 (Niacin) Alpha - Ketoglutarate
DH, Isocitrate DH,
Malate DH
Coenzyme A B5 (Pantothenic Succinyl CoA
acid)
•• 2, 3 DPG is produced as a result of Rapaport Luebering
cycle (RL shunt). This molecule decreases the affinity 44. Ans. c. ETC complex 3
of hemoglobin for oxygen. Hence it will shift the
O2 dissociation curve to the right. Other factors [Ref: Harper's 31st Ed. / page – 123]
which will also shift the curve to right are: Increased Dimercaprol inhibits electron transport via Complex 3
temperature, Carbon monoxide (CO), Low pH by blocking the transfer from Complex 3 to Complex 4.
41. Ans. a. Diabetes mellitus Inhibitors of ETC A
N
[Ref: Harper's 31st Ed. / page - 130] Complexes Inhibitors of ETC S
In diabetes mellitus, glucose utilization is impaired due Complex I Rotenone, Phenobarbitone, W
to absolute or relative insulin deficiency. (NADH-CoQ reductase/ Amobarbital, Piericidin A E
Hence, Fatty acid breakdown occurs to provide energy NADH dehydrogenase) R
and the resultant excessive Acetyl CoA enters the
S
pathway of ketogenesis as shown in the figure. Complex II Malonate (3C), Carboxin (Fungicide),
(Succinate TTFA (Trienoyl Tri fluoro acetone) WITH
CoQ reductase)
E
Complex III Phenformin, Antimycin A, BAL (British X
(Cyt c reductase) Anti Lewisite) or Dimercaprol P
L
Complex IV CO, Cyanide (CN), H2S, Sodium Azide
A
(Cyt c oxidase)
N
Uncouplers of ETC: Dinitrophenol, Thermogenin A
T
I
ADP to ATP conversion is inhibited by: Oligomycin
O
ADP to ATP transfer is inhibited by: Atractyloside N
S
 45. Ans. c. Carbamoyl phosphate synthase I

xxii [Ref: Harper's 31st Ed./page – 277-278]

Hyperammonemia is a metabolic disturbance characterized by an excess of ammonia in the blood. It is a dangerous
condition that may lead to brain injury and death. It may be primary or secondary. This usually occurs in urea cycle
disorders.
CRO BIOCHEMISTRY

•• Deficiency of any of the enzyme of urea cycle leads to ammonia accumulation.

Table: List of diseases due to Urea cycle enzyme deficiency

Disease name Enzyme deficiency Substrate accumulated

} Hyperammonemia Type I CPS-I (Carbamoyl Phosphate synthetase-I) NH3

Most
Hyperammonemia Type II OTC (Ornithine Transcarbamoylase) NH3
severe
(Most common UCD) OMP
UMP
Orotic acid


Citrullinemia Type I Argino-Succinate synthetase NH3 (UCDs)

Citrulline
Least
Hyper
ammo
} Arginosuccinic aciduria

Hyperargininemia
Argino-Succinate Lyase

Arginase
NH3
Arginosuccinic acid
NH3
nemia Arginine

46. Ans. a. Alanine Extra Information:

[Ref: Harper's 31st Ed. / page – 177-178] •• In liver there is absence of thiophorase, first enzyme
of ketone body utilization pathway. So Liver also
Alanine is formed from glucose in muscles during cannot utilize ketone bodies
fasting conditions. This alanine comes out in blood, •• Ketone body synthesis: ketogenesis only occur in liver.
then taken up by liver. In liver it is converted back to Enzyme for ketogenesis present in mitochondrial
glucose by a pathway known as gluconeogenesis. Then matrix
this glucose is again supplied to the fasting muscle. So, a •• Ketone body utilization – ketone body is the main
cycle is formed known as Cahill cycle (see Figure). source of energy to skeletal muscle, renal cortex,
cardiac muscles. During prolonged starvation,
ketone bodies are the major fuel source for the brain
A and other parts of central nervous system.
N
S
W PGI MAY 2019
E
R 48. Ans. a. Cytosol
S [Ref: Harper 31st edition pg 182-183]
WITH
Pathways and their Location
E Mitochondria Cytoplasm Both
X
Fig. Cahill cycle of glucose-allanine in Fasting muscle •• TCA •• Glycolysis •• Urea Cycle
P
L •• ETC •• Glycogen •• Heme synthesis
47. Ans. b. RBC •• Beta oxidation synthesis •• Gluconeogenesis
A
N of Fatty acids •• Glycogen
[Ref: Harper's 31st Ed. / page – 136-137]
•• Ketone body breakdown
A
•• In RBC, there is no mitochondria and ketone body synthesis •• Fatty acid
T
utilization pathway takes place in mitochondria. So, •• Ketone body synthesis
I utilization •• Pentose
RBC cannot utilize ketone bodies
O phosphate
•• Brain, muscle and heart can use ketone bodies during
N pathway
starvation.
S
49. Ans. a. Niacin deficiency; b. Hartnup disease; 52. Ans. b
 . Argininosuccinate lyase;
e. Carcinoid syndrome c. Ornithine transcarbamylase;
d. Argininosuccinate synthetase; xxiii
[Ref: Harper 31st edition pg 288, 535-536]
e. Carbamoyl phosphate synthetase-I
Etiology of Pellagra
•• Dietary deficiency of nicotinic acid or its precursor [Ref: Harper 31st edition pg 275]

 LATEST EXAM QUESTIONS 2020-2019

tryptophan. Enzyme involved in urea cycle are:
ƒƒ Seen in people with staple diet – maize and Jowar. •• Carbamoyl phosphate synthase I
•• Malabsorption •• Ornithine transcarbamylase
•• Drugs like INH, 5FU, and 6-mercaptopurine. •• Arginosuccinate synthase
•• Carcinoid syndrome •• Argininosuccinase
•• Hartnup's disease in which there is Defect in neutral •• Arginase
amino acid transporter (tryptophan) and failure to
absorb tryptophan from intestine and also reabsorb 53. Ans. a. Threonine; b. Phenylalanine; d. Methionine
it from kidneys
[Ref: Harper 31st edition pg 264]
A dditional E dge •• Essential amino acids: Those which cannot be
synthesized in body and are therefore required in
•• Pellagra 4Ds → (Dermatitis, Dementia, Diarrhea, Death)
diet.
•• Non-essential amino acids: Those which can be
50. Ans. a. Pellagra; b. Beriberi synthesized in body and therefore are not required
in diet.
[Ref: Harper 31st edition pg 161-162] •• Semi-essential amino acids: Those which can be
synthesized in body but to some extent. Examples are
Pyruvate is converted to acetyl-CoA by oxidative
Arginine and Histidine.
decarboxylation by enzyme pyruvate dehydrogenase
(PDH) complex
Essential Amino Acids Non-Essential
PDH complex requires five coenzymes – Vitamin B1, Amino Acids
B2, B3, B5 and Lipoic acid
•• Thiamine pyrophosphate prosthetic group (Vit B1) 1. Methionine Sulfur containing 1. Alanine
•• FAD prosthetic group (Vit B2) 2. Threonine –OH containing 2. Asparagine
•• Free NAD+ (Vit B3) 3. Valine 3. Aspartic acid
4. Isoleucine 4. Cysteine
•• CoA (Vit B5)
5. Leucine 5. Proline
•• Lipoic Acid
Branched chain amino acids
Now deficiency of any of the coenzyme will hamper the 6. Lysine 6. Serine
function of PDH and Hence pyruvate consumption will 7. Arginine (Semi essential) 7. Tyrosine
A
be hampered. Basic Amino acids N
•• Pellagra is deficiency of vitamin B3 8.Phenyl Alanine 8. Glycine S
•• Beri beri is deficiency of vitamin B1 9. Tryptophan 9. Glutamic acid W
Hence, both these diseases will affect the pyruvate Aromatic Amino acids E
consumption. 10. Histidine (Essential but semi-essential) 10. Glutamine R
51. Ans. d. Quaternary structure S
54. Ans. a. Xerophthalmia; d. Hirsutism
WITH
[Ref: Harper 31st edition pg 37] [Ref: Harper 31st edition pg 530]
Quaternary structure is the arrangement of two E
Vitamin A toxicity leads to:
or more functional polypeptide chains in three X
•• Alopecia
dimensional complexes e.g. hemoglobin has quaternary P
•• Dry pruritic skin
structure comprising 4 functional polypeptide chains. L
•• Desquamation of skin
Homodimers and heterodimers protein complexes A
•• Hepatomegaly
are, when two same or different polypeptides form N
•• Liver damage
complex with each other respectively and hence they A
•• Thickening of the long bones
are quaternary in nature. T
•• Hypercalcemia
I
O
N
S
 •• Hyperostosis ƒƒ White blood cells,
•• Arthralgia ƒƒ Epithelial cells (from which DNA can be extracted),
xxiv •• Raised intracranial pressure which sometimes mimic ƒƒ Enzymes (such as amylase and lipase),
brain tumor. This is known as benign intracranial ƒƒ Antimicrobial agents such as secretory IgA, and
hypertension (Pseudotumor cerebri). lysozymes.
CRO BIOCHEMISTRY

55. Ans. d. Saliva 56. Ans. a

 . Transferrin- Transport iron;
b. Thyroxin binding globulin- thyroxin binding
[Ref: Harper 31st edition pg 237; https://www.thejpd. and transport;
org/article/S0022-3913(01)54032-9/fulltext] c. Ceruloplasmin- Assisting in iron absorption
•• LDL, HDL has high amounts of cholesterol. by transferrin; e. Hemopexin-Heme binding
•• Urine has relatively very small amount of cholesterol
•• Transudate is fluid pushed through the capillary due [Ref: Harper 31st edition pg 631]
to high pressure within the capillary. Exudate is fluid •• Haptoglobin binds to free hemoglobin, compared
that leaks around the cells of the capillaries caused to hemopexin that binds to free heme released from
by inflammation. They also have minor quantity of erythrocytes.


cholesterol from blood. •• Haptoglobin binds with high affinity and thereby
•• Composition of Saliva inhibits the oxidative activity of hemoglobin.
ƒƒ 99.5% water plus electrolytes, •• The haptoglobin-hemoglobin complex will then be
ƒƒ Mucus, removed by the reticuloendothelial system.

A
N
S
W
E
R
S
WITH

E
X
P
L
A
N
A
T
I
O
N
S
UNIT
I

CONCEPTS IN
BIOCHEMISTRY
Unit Outline
Chapter 1 Concepts in Biochemistry
Concepts in
Biochemistry 1
Overview of Chapter A FORMULA: INSULIN AND GLUCAGON IN
•• Cyclic AMP (cAMP) FED AND FASTING STATE
•• A formula: Insulin and glucagon in fed and fasting state
•• Which are anabolic and catabolic pathways? In fed state, we eat food, which is mainly carbohydrates
•• A formula: Which pathway occurs in which (Glucose). Therefore, this Glucose will cause Hyperglycemia.
compartment? Insulin has to come to decrease this blood glucose. Hence,
•• Sources of blood glucose and main fuel for body if Insulin comes in fed state that means Insulin is anabolic
•• Scene in fasting state
hormone, i.e. mainly, anabolism occurs in body in fed, state.
•• Fuel used in different situations in body
•• Acetyl CoA in fed and fasting states Insulin always causes Dephosphorylation by decreasing
•• Fat and carbohydrate interconversion: Full story cAMP.
Diabetes
•• Understanding plan of nature: How to extract energy
from macromolecules?
•• Atkin’s Diet
•• Concepts of enzymes
•• Cell Organelles
•• Bonds in macromolecules

F undamental B ox
zz Fed state: When we eat food (Within 2 hours of food intake is
called fed state)
zz Fasting state: In between meals, when we are not eating food
(From 12–18 hours after food up to 48 hours is fasting)
zz Starvation: Severe or complete lack of nutrients (since last
2–4 days) Fig. 1.2: Fed state
zz In between meals and night time is called fasting time. That’s
why Breakfast is named → Break the night fast F ormula B ox
Insulin activates all anabolic pathway enzymes (General rule)
CYCLIC AMP (cAMP) zz
zz But Exception → that Insulin activates two catabolic pathways:
This is a second messenger synthesized from ATP, with the 1. Glycolysis (6 C Glucose gets converted into two molecules
help of enzyme Adenyl Cyclase. cAMP activate kinases, of 3 C Pyruvate, so this is breakdown of Glucose, i.e.
which phosphorylate various proteins/enzymes in the body. catabolic pathway)
Basically, cAMP leads to phosphorylation or we can say that 2. Link reaction/Pyruvate Dehydrogenase step (conversion
Adenyl Cyclase leads to phosphorylation, (Fig. 1.1). of 3 C molecule Pyruvate to 2 C molecule Acetyl CoA. So,
cAMP is destroyed by enzyme Phosphodiesterase to this is also breakdown of 3 C compound to a 2 C compound).
5’AMP. So, Phosphodiesterase causes dephosphorylation. So, these 2 things are catabolic, but activated by Insulin.

F ormula B ox
Now, the Opposite Formula for fasting State:
zz In fasting state, Glucagon hormone is released. So, Glucagon is
a catabolic hormone. Glucagon always causes phosphorylation
by increasing cAMP.
zz So, we can conclude that: Glucagon activates all catabolic
pathway enzymes (General rule) EXCEPT: Glycolysis and Link
reaction.
Fig. 1.1: cAMP Synthesis and Breakdown
 WHICH ARE ANABOLIC AND CATABOLIC
4 PATHWAYS?
Anabolic means synthesis occurring in body. Anabolic
pathways are glycogen synthesis (Glycogenesis), HMP
(as HMP synthesizes Ribose-5-P and NADPH), fatty acid
CRO BIOCHEMISTRY

synthesis, cholesterol synthesis and TG synthesis.

Catabolic means breakdown occurring in body. Catabolic
pathways are glycolysis (breakdown of 6C Glucose to two
molecules of 3 C Pyruvate), Pyruvate Dehydrogenase (Link
reaction), glycogen breakdown (Glycogenolysis), Beta
oxidation of fatty acids, Gluconeogenesis, Ketone body
synthesis, Ketone body utilization/breakdown.
Fig. 1.3: Fasting State

NOTE: When I say pathway is anabolic means all enzymes in that



pathway are anabolic.

When I say pathway is catabolic means all enzymes in that
pathway are catabolic.

F undamental B ox
Anabolic Pathways or Catabolic Pathways or
Enzymes Enzymes
•• Glycogenesis •• Glycolysis
•• HMP •• Link reaction
Fig. 1.4 Hormones and cAMP; Insulin is the only hormone •• Fatty Acid Synthesis (Pyruvate Dehydrogenase
which decreases blood Glucose. The hormones which increases •• Cholesterol Synthesis reaction)
blood Glucose are Glucagon, Growth hormone, Glucocorticoids, •• TG Synthesis •• Glycogenolysis
Epinephrine (Epi), Nor-Epinephrine (NE) and Thyroid •• Lipoprotein Lipase •• Beta Oxidation of fatty acids
•• Gluconeogenesis
Q. So, we have so many hormones to increase blood •• Ketone body synthesis
Glucose but only one hormone to decrease blood •• Ketone body utilization/
T Glucose. Why? Breakdown
H A. This is a survival benefit. Because Hypoglycemia is
I •• Hormone sensitive lipase
more dangerous than Hyperglycemia.
N
K
How Gluconeogenesis is Catabolic Pathway?
As Insulin causes dephosphorylation and Phospho-
diesterase enzyme decreases cAMP, so we can conclude The definition is synthesis of Glucose from non-carbohydrate
that insulin activates phosphodiesterase and thus it causes sources but it is not anabolic pathway. Actually, in fasting
dephosphorylation. state, other macromolecules of body (Fats, Proteins, Amino
Acids) are broken down (means catabolic) to produce
All other hormones (Glucagon, Growth Hormone, Glucocor-
Glucose, because Brain and RBCs need Glucose at all times.
ticoids, Epinephrine, Nor-Epinephrine) causes phosphoryla-
So, consider Gluconeogenesis as catabolic pathway.
tion by activating enzyme Adenyl Cyclase.

F ormula B ox How Ketone Body Synthesis and Breakdown –

All anabolic enzymes are active in dephosphorylated state and all both are catabolic?
catabolic enzymes are active in phosphorylated state. Ketone bodies are produced and used in body during
T EXCEPTION: ATP Citrate Lyase is anabolic enzyme involved in Fatty
H starvation only (never in fed state). So, fat (Adipose tissue) is
Acid Synthesis but this enzyme is active in phosphorylated state.
E broken down to release fatty acids, which reaches Liver, gets
broken down to give Acetyl CoA, which synthesizes Ketone
O
R
A dditional E dge Bodies. In short Fats are broken down to produce Ketone
All insulin antagonist hormones (Glucagon, Growth hormone, Bodies. And this ketone body synthesis occurs in Liver.
Y
Glucocorticoids, Epinephrine, Nor-Epinephrine) activate Adenyl Ketone body utilization/breakdown occurs in Heart, Brain
Cyclase BUT Thyroid hormone increases the synthesis of this and Skeletal Muscles to obtain ATP/energy from breakdown
enzyme (by working at the level of gene). of ketone bodies. So, consider ketone body synthesis and
ketone body utilization both are catabolic.
 3 Pathways occurs both in mitochondria and cytoplasm:
Q. Why TCA and ETC are not classified in these lists of
1. Gluconeogenesis
catabolic and anabolic pathways?
T A. TCA and ETC are vital pathways, not affected by 2. Urea cycle
Synthesis of Glucose, Urea
and Haem occur both in 5
H hormones
I 3. Haem Synthesis mitochondria and cytoplasm
• In fact TCA is considered both anabolic and
N
K catabolic (Amphibolic)

CHAPTER 1  CONCEPTS IN BIOCHEMISTRY

• These pathways are always occurring in body Whenever a pathway occurs both in mitochondria and
(whether fed or fasting state). cytoplasm, that pathway starts from mitochondria. So, the
• Anabolic pathways occur in fed state and are first step of these three pathways occurs in mitochondria.
activated by Insulin. Catabolic pathways occur Or first enzyme of these three pathways is always present in
in fasting or starvation and are activated by mitochondria.
Glucagon.
F ormula B ox
 FORMULA: WHICH PATHWAY OCCURS IN
A Pathways Occurring in Mitochondria
WHICH COMPARTMENT? zz All catabolic pathways (except glycolysis and Glycogenolysis),
i.e. Link Reaction, β-oxidation of fatty acids, ketone body syn-
Formula: All catabolic pathways in mitochondria and all thesis, ketone body utilization
anabolic pathways in cytoplasm zz All Vital Pathways – TCA and ETC
Exceptions: zz Replication, Transcription, Translation (for mitochondrial DNA)
zz Glycolysis and Glycogenolysis are catabolic pathways zz Apoptosis
but these occurs in cytoplasm.
Pathways Occurring in cytoplasm:
zz Gluconeogenesis is catabolic but it occurs in both mito- zz All anabolic pathways (HMP, Glycogenesis, fatty acid synthesis,
chondria and cytoplasm. Triglyceride Synthesis, Cholesterol Synthesis)
zz Two catabolic pathways (Glycogenolysis, Glycolysis)
A

SOURCES OF BLOOD GLUCOSE AND MAIN

FUEL FOR BODY

Q. What are the sources of blood Glucose?

A. 1. Food
T 2. Liver glycogen (for 12–18 hrs)
H In sequence
I 3. Gluconeogenesis
N So, the first source of blood glucose is our food but
K when we are not eating food, i.e. in between meals,
the source of blood glucose is Glycogen from Liver,
which can provide Glucose to blood for just 12–18
hours if someone is not eating food for 12–18 hours.
And if lack of food is continued, then next source of
blood glucose is gluconeogenesis pathway. (Do not
B answer fats for this question, as fats can never be
converted to carbohydrates).

A dditional E dge
How Exercise during morning time is beneficial?
During day time, we take three meals. Gap between two meals
is 6–8 hours but gap between dinner and next day breakfast is
12–14 hours. So, when we wake up during the morning time,
then our Glycogen reserves are almost finished. If exercise is
done at this time, then Gluconeogenesis pathway will provide T
blood Glucose, which is a high energy consuming pathway. So, H
this pathway uses energy and in addition exercise will also use E
Fig. 1.5: Pathways occuring in cell energy of body. So, exercise done at this time is much beneficial O
as compared to day time exercise. R
Y
 Q. What is the preferred/main fuel for body? Q. Why Ketone Bodies Synthesized?
6 T
A. • 1st is carbohydrates
T
A. This is a survival benefit that ketone bodies are
synthesized during starvation. They are synthesized
In sequence
H • Next is Fats H for Heart and Brain (vital organs). Because only these
I • Amino acids and proteins I
• The first preferred fuel for body is carbohydrates vital organs (Heart and Brain) can use ketone bodies.
N N
CRO BIOCHEMISTRY

K but if carbohydrates are very less, then body shifts K Other body organs cannot use ketone bodies.* Due to
to fats. And if fats are also finished then body starts this, ketone bodies are always available for these vital
breaking down Proteins of the body. organs as a fuel, so that they can survive this situation
•• So, in fed state, carbohydrates is the main/ of absence of Glucose. So, therefore, Heart and Brain
preferred fuel for body. (Note: Carbohydrate is do not rely on Glucose during starvation. They use
not the only fuel in fed state, it is the main fuel.) ketone bodies.
•• Similarly, Fats is the main/preferred fuel for body •• Ketone bodies appear in urine during starvation
in fasting state. (Note: Fats is not the only fuel •• Muscles can also use Ketone Bodies*, so that use
in fasting, it is the main fuel and in very severe of muscle proteins is delayed.
conditions, body starts using proteins.)

FUEL USED IN DIFFERENT SITUATIONS IN



SCENE IN FASTING STATE BODY

In fasting/starvation, fats is the preferred fuel. So, adipose Table 1.1: Fuel used in different situations in body
tissue is broken down to release fatty Acids in blood. These
Fed Fasting Starvation
fatty Acids reach Liver, where Beta Oxidation is done,
releasing Acetyl CoA, which has three fates. Either this Acetyl Brain Glucose Glucose Ketone Bodies
CoA enters TCA, or enters ketone body Synthesis, or it is used Heart Fatty Acids Fatty Acids Ketone Bodies
for the activation of the first step of Gluconeogenesis (Fig.1.6).
Liver Glucose Fatty Acids Amino Acids
Muscle Glucose Fatty Acids Fatty Acids and KB
Adipose tissue Glucose Fatty Acids Fatty Acids
RBC Glucose Glucose Glucose

zz General thing is that in fed state – fuel is carbohydrates,

i.e. Glucose. In fasting it is fats or fatty acids and in severe
fasting, i.e. starvation, body shifts to ketone bodies for
vital organs, i.e. Heart and Brain.
zz So, in fed state, most organs use Glucose but Heart uses
fatty acids even in Fed state.
zz Fetal Heart uses glucose because during pregnancy,
fetus receives good supply of carbohydrates but less of
fatty acids.
zz Heart (Vital organ) uses fatty acids in fed state and Brain
(Vital organ) uses Glucose in fed state because heart
works 24 hours and requires a fuel which provides more
energy per gram, i.e. Fats.
zz In heart failure, fuel is Glucose because Heart muscle
is starved of oxygen. Carbohydrates (Glucose) use less
oxygen than Fats. So, Glucose becomes the main fuel for
Fig. 1.6: Scene in fasting state Heart during heart failure.
zz In fasting state, most body organs use fatty acids but
Trick for question solving: Diabetic situation is same like Brain cannot use fatty acids because fatty acids cannot
T fasting or starvation as peripheral cells are not receiving
H cross Blood Brain Barrier (BBB) and RBCs cannot use
Glucose so they think that body is in fasting. So, to solve any fatty acids as they don’t have mitochondria.
E question on diabetes, apply the concepts of fasting situation.
O
R
Y
zz In starvation, brain and heart (vital organs) use ketone Propionic Acid (3C) is obtained from oxidation
2. 
bodies. Liver shifts to amino acids and proteins. Muscles of Odd Chain fatty acids. Propionic acid can form
can use fatty acid and or ketone body also. Adipose tissues Succinyl CoA, which is intermediate of TCA cycle. Any 7
use fatty acids only. RBCs still uses glucose because they intermediate of TCA cycle can form Glucose.
don’t have mitochondria.
zz Liver cannot use ketone bodies because the first enzyme
Q. Fats cannot be converted to carbohydrates but

CHAPTER 1  CONCEPTS IN BIOCHEMISTRY

of ketone body utilization (Thiophorase) is absent in
both the breakdown products of TGs are helping in
Liver. T Gluconeogenesis. How?
zz RBCs cannot use ketone bodies because mitochondria H A. Triglyceride is broken down to three fatty acids and
is absent and ketone body utilization occurs in mito- I
one molecule of Glycerol. Fatty acids undergo Beta-
chondria. N
K Oxidation to release Acetyl CoA, which activates the
Q. Heart uses fatty acids during fed and fasting state very first step of Gluconeogenesis. The other product:
and uses ketone bodies during starvation. Then Glycerol is a substrate for Gluconeogenesis. So, both
T what is the need for GLUT-4 in cardiac muscles? fatty acids and Glycerol are helping in this pathway of
H A. Heart does not use Glucose in most of the states but Gluconeogenesis (indirectly).
I But directly Fats cannot be converted to Glucose/
GLUT-4 is a transporter for entry of Glucose which is
N carbohydrates because Link reaction is irreversible.
K present in cardiac muscles because it is present since
fetal life (Fetal heart uses Glucose) and during heart (See Fig. 1.8)
failure also heart needs Glucose as fuel.

Note: R
 BCs always rely on Glucose because they cannot use
Ketone Bodies, fatty acids due to lack of mitochondria

ACETYL CoA IN FED AND FASTING STATES

In fed state, under the effect of Insulin hormone, Pyruvate
Dehydrogenase (link reaction) is activated. So, Pyruvate
is converted to Acetyl CoA, which is used for synthesis of
fats but in fasting state, there is lots of Acetyl CoA (from
Beta Oxidation of fatty acids). So, by product inhibition,
Acetyl CoA itself inhibits Pyruvate Dehydrogenase (Link
reaction enzyme). So, Pyruvate (substrate of link reaction) Fig. 1.7: Acetyl CoA in fasting state
accumulates. This Pyruvate is diverted for the formation of
Oxaloacetate (by enzyme Pyruvate Carboxylase), which is
the first step of Gluconeogenesis. This step is activated by
Acetyl CoA (See Fig. 1.7).

FATS AND CARBOHYDRATES

INTERCONVERSION: FULL STORY
Fats can never be converted to carbohydrates because Link
reaction (Pyruvate to Acetyl CoA) is irreversible. So, Acetyl
CoA can never form Pyruvate, thus can never form Glucose.
Therefore, Acetyl CoA is never Glucogenic, i.e. Acetyl CoA can
never be converted to Glucose.
But two breakdown products of Fats, i.e. Glycerol and Pro- Fig. 1.8: Breakdown products of TGs are helping in
pionic Acid are Glucogenic, i.e. converted to Glucose. Gluconeogenesis
1. Glycerol (3C) is obtained from breakdown of TGs
(triglycerides). Glycerol-3-Phosphate can be converted
T
to DHAP (Di Hydroxy Acetone Phosphate), by enzyme
Glycerol- 3-Phosphate Dehydrogenase. DHAP is inter- H
mediate of Glycolysis, it can be converted to Glucose. E
O
R
Y
 8
CRO BIOCHEMISTRY

Fig. 1.9: Fates of Acetyl CoA

Table 1.2: Fuel used in fasting/starvation

Fasting or Fuel used in Body

Starvation


Early fasting •• Glycogen

(12–18 hours) Fig. 1.10: TCA Suppression in Diabetes
Late fasting •• β-Oxidation of Fatty Acid, TCA suppression in Diabetes occurs due to following
(18–48 hours) Gluconeogenesis (energy for reasons:
Gluconeogenesis is derived from zz Oxaloacetate is used up for Gluconeogenesis, which is
β-Oxidation) not available for TCA
Starvation (>48 •• β-Oxidation, ketone body Synthesis zz Excess Lipolysis leads to increased Beta Oxidation of
hours) (Known as starvation Ketosis) fatty acids which lead to the formation of excess NADH,
•• Substrates of Gluconeogenesis mostly get which further depletes NAD. This suppresses TCA.
depleted during starvation zz Acetyl CoA is diverted to ketone body synthesis

DIABETES Patients have Hyperkalemia. Why?

There is relative or absolute deficiency of anabolic hormone: Insulin is required for K+
Insulin. uptake by cells. Due to
So, Diabetic situation is same like fasting or starvation, i.e. insulin deficiency, K+ uptake
catabolic situation in body. Catabolic pathways or enzymes does not occur. This leads to
will be activated and anabolic pathways or enzymes will be hyperkalemia.
decreased. Also, ketoacidosis increas-
es H+ – K+ Antiport, thus in-
creasing K+ efflux from cells.
Biochemical changes in Diabetes
Q. Why ketoacidosis more common in type I as
• ↑ Hyperglycemia • ↑ K B Synthesis
compared to type II diabetes?
•• ↓ Lipoprotein Lipase • ↑ VLDL and TG in blood T A. In type I patients, the onset of disease is abrupt. So,
H body switches abruptly from glucose to fatty acids as
•• ↑ Gluconeogenesis • ↑ Beta oxidation of FA I
the main fuel. Breakdown of fatty acids forms excess
N
•• Insulin dependent GLUT-4 not working, So, blood Glucose K Acetyl CoA which leads to the formation of ketone
cannot enter Peripheral tissues bodies.
•• Plasma Free Fatty Acid levels will be raised as catabolic
enzyme- Hormone Sensitive Lipase (HSL) will be active in
UNDERSTANDING PLAN OF NATURE:
adipose tissues, which breaks down TGs into fatty acid and HOW TO EXTRACT ENERGY FROM
glycerol. Fatty acids travel to blood and to Liver, where Beta MACROMOLECULES?
Oxidation of fatty acids takes place. This leads to excess
T Acetyl CoA which is taken for TG and thus VLDL synthesis. An underlying theme occurs to extract energy from macro-
H (Only one anabolic mechanism activated in diabetes). molecules:
E zz This extraction of energy is like a business done. For any
O business, we have to first invest, later we can gain out of
R it. Any good business is that where we invest less and
Y obtain more.
zz These macromolecules (made up of C, H, O, N) don’t Note: Keto diet is used for weight reduction, (i.e. very low or
have energy in them as such. Energy can neither be absence of carbohydrates in diet) so that body is forced to use fats
created, nor be destroyed. It can just be converted from and ketone bodies for energy.
9
one form to another. So, a plan is made by nature to
convert one form of energy to another.
Q. A person on fat free, Carbohydrate rich diet continues
zz First a high energy bond (like phosphate or CoA) is added to grow obese. Which lipoprotein is increased?

CHAPTER 1  CONCEPTS IN BIOCHEMISTRY

in a macromolecule, making it activated macromolecule. A. VLDL.
Then this high energy bond is broken energy is released, Explanation: This person is not taking any exogenous fat
which is entrapped in the form of ATP. but taking excess carbohydrates, which will be converted
zz Any pathway that is giving energy (ATP) in the end, is a to fat in the body. This fat is endogenous fat, transported
catabolic pathway. Because mostly macromolecules are in the form of VLDL.
broken down to give energy in body. So, the first step of
any catabolic pathway is the activation of macromolecule
(like investment phase of any business) and second
H igh R eturn
phase is breakage of that high energy bond to obtain Thermogenic effect of food → Energy required to digest,
energy. E.g. Glycolysis, Beta Oxidation of fatty acid (See absorb, transport and metabolize the food material is known as
Fig. 1.11) Thermogenic effect of food. It is maximum for proteins, then
zz For carbohydrates, mostly this activation is done by a carbohydrates and least for fats. So more proteins in diet will use
high energy phosphate and for Fats this activation is lots of energy of body. Thermogenic effect of food is also known
mostly by Coenzyme A. as Thermic effect of food or SDA (specific dynamic action) or DIT
(diet induced thermogenesis). It leads to the production of heat
in the body, e.g. in Brown fat.

CONCEPTS OF ENZYMES
Enzymes are divided into 6 categories, also known as Enzyme
Commission Number/Code Number – or simply EC Number:
1. Oxidoreductases 2. Transferases
3. Hydrolases 4. Lyases
5. Isomerases 6. Ligases

F undamental B ox
Fig. 1.11: Plan of nature to extract ATP from any macromolecule is Basics
a rule of business zz To break a bond, water is added by Hydrolases
zz To make a bond, energy from ATP is used by Ligases.

A dditional E dge zz Lyase category can make a bond or break a bond, but neither
uses water nor ATP.
Whenever Phosphate or CoA (high energy bond) is added, then
ATP is used. zz Oxidoreductase: Enzymes which carry Oxidation, Re-
Exception: duction reactions
1. 
Glyceraldehyde-3-P Dehydrogenase (enzyme in Glycolysis) → zz Transferase: Enzymes which transfer a group from one
adds phosphate, without using ATP molecule to another.
Thiophorase (enzyme in ketone body utilization) → adds CoA
2.  zz Isomerase: Enzymes which interconvert various isomers
in Acetoacetate, without using ATP into each other.

ATKIN’S DIET H igh R eturn

Difference between Transferase and Isomerase:
Atkin’s Diet is Low Calorie, Low Carbohydrate diet. zz If Molecular formula is changed then it is Transferase.
In diet, 60–70% is carbohydrates, 15–20% is Lipids (also known zz If Molecular formula is not changed then it is Isomerase.
as exogenous Fat, transported in the form of Chylomicrons)
and rest are proteins zz Enzyme Mutase is for intramolecular transfer but EC No.
Major function of carbohydrates is to provide energy but is 5 because molecular formula not changed. T
in a sedentary lifestyle, only 50% of these carbohydrates are H
used to give energy, 10% is converted to Glycogen, which is F undamental B ox E
used in between meals and rest 40% is converted to fats. This zz If Oxygen is added → known as Oxidation O
Fat is known as endogenous fat (as it is formed in body from zz If hydrogen is added → known as Reduction R
carbohydrates). Endogenous fat is transported in the form of zz If electron is added → known as Reduction Y
VLDL. zz In biochemistry, consider hydrogen atom is equivalent to
electron or reducing equivalent
zz Hydrogen ion (H+) → equivalent to Proton
 Dehydrogenases
10 Removal of hydrogen is done, oxidation occurs. The hydrogen
which is removed can be taken by either NAD or FAD or
NADP, forming respectively NADH, FADH2 and NADPH.
NADH enters ETC to give 2.5 ATPs. FADH2enters ETC to give
CRO BIOCHEMISTRY

1.5 ATPs. NADPH has no role in ATP formation, instead it has

a role in reductive biosynthesis.
F undamental B ox
zz Dehydrogenases are named after the substrate which gets
oxidized
zz Reductases are named after the substrate which gets reduced.
EC Number:
� Dehydrogenase → 1 � Reductase → 1
� Kinases → 2 � Phosphorylase → 2
� Phosphatase → 3


Table 1.3: Difference between Kinase and Phosphorylase

Kinase Phosphorylase
•• Transfer Phosphate •• Transfer Phosphate
•• They transfer organic •• They transfer inorganic
Fig. 1.12: Dehydrogenases phosphate, i.e. ATP/ADP phosphate, i.e. Phosphate
present free in medium (Pi)

NADPH Phosphatase belongs to EC no. 3, because this enzyme

removes phosphate by adding water. The phosphate then is
NADPH produced from NADPH used in released into the medium as inorganic phosphate.
 MP (major source of 1. Reductive Biosynthesis e.g.
1. H
NADPH) •• Fatty Acid Synthesis
H igh R eturn
2. Malic enzyme or NADP •• Cholesterol Synthesis zz Cofactor for Kinases: Mg
Malate Dehydrogenase •• Steroid Synthesis zz But Pyruvate Kinase requires K (potassium) more than Mg
3. Cytoplasmic Isocitrate- •• Deoxyribonucleoside
Dehydrogenase Diphosphate Synthesis Whenever Kinases work, either ATP is used or ATP is produced.
2. Synthesis of Reduced ATP used means ATP is converted to ADP. ATP produced
Glutathione (GSH) in RBC means ADP is converted to ATP (known as Substrate Level
3. Cytochrome P450 system Phosphorylation)
4. Detoxification of reactive
oxygen intermediates Table 1.4: Difference between SLP and ETC

Substrate Level Oxidative Phosphorylation

Phosphorylation (SLP)
•• Enzyme is Kinase. ATP •• Electron Transport Chain
produced when substrate is (ETC) is known as Oxidative
converted to product Phosphorylation
•• Few ATPs derived •• Most ATPs derived via this
route
H igh R eturn •• Easy way of ATP formation •• Very complex ETC involved
zz Coenzyme for cytoplasmic Isocitrate Dehydrogenase → NADP (only single step)
zz Coenzyme for mitochondrial Isocitrate Dehydrogenase → NAD
T Table 1.5: Difference between Synthase and Synthetase
H
Synthase Synthetase
E Reductive Biosynthesis
O •• Bond formed but ATP not •• Bond formed and ATP used
In anabolic pathways, Reductase enzyme is present. This used
R enzyme needs H2 for Reduction. This H2 is derived from •• EC No. = 4 •• EC No. = 6
Y
NADPH. So, NADPH has a role in reductive biosynthesis, e.g.
Thioredoxin Reductase
 Q. Why kinases like hexokinase, PFK-1 are not ligases, Note: A
ll these are mainly amino acids which undergo
as kinase also use ATP? decarboxylation to produce Amine.
T A. Both kinase and synthetase (a ligase) use ATP 11
H •• If ATP used for transfer of Phosphate → Kinase
I
enzyme name is used
N
K •• If ATP used for making a bond →Synthetase

CHAPTER 1  CONCEPTS IN BIOCHEMISTRY

enzyme named is used.

Carboxylation and Decarboxylation

For addition of CO2,enzyme is Carboxylase. It requires ATP,
Biotin (Vitamin B7), CO2 and Mg+2 ions. As this enzyme is
making a bond by adding CO2 to a compound, so it belongs Q. Why Decarboxylation of Glutamate and Lysine is
to enzyme category – Ligase (EC No. 6). not forming amine?

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N
ABC of Carboxylases (All Carboxylases use A, B, C) K
zz A → ATP required
zz B → Biotin required
zz C → CO2 required

F undamental B ox
zz All Kinases and Carboxylases use Magnesium (Mg) because zz In Oxidative Decarboxylation, Enzyme is Dehydrogenase, so
ATP is involved EC Number is 1
zz Whenever ATP is used then Mg is used to stabilize the outgoing zz In Simple Decarboxylation, EC Number is 4 because this carbon
phosphate. Manganese (Mn) can also be used instead of bond is broken without using water, i.e. it is a Lyase
Magnesium.

Decarboxylation
Decarboxylation is removal of CO2. This is of two types:
Oxidative Decarboxylation and Simple Decarboxylation.
1. Oxidative Decarboxylation involves a reaction where
enzyme is a Dehydrogenase and CO2 is getting removed.
As the name of enzyme is Dehydrogenase, so EC No. is
1. There are many oxidative decarboxylation reactions
in the body. They require Vitamin B1 (Thiamine) as
coenzyme. All oxidative decarboxylation enzymes are
multi enzyme complexes, present in mitochondria and
require 5 coenzymes (Lipoic acid, Vitamin B1, Vitamin
B2, Vitamin B3 and Vitamin B5. Fig. 1.13 Carboxylases and Decarboxylases
2. Simple Decarboxylation requires B6 (PLP-Pyridoxal
Phosphate). EC Number is 4 (Lyase), as CO2 is removed Summary
without using water. Examples of reactions of Simple
zz For Substrate Level Phosphorylation, enzyme is a Kinase
Decarboxylation are:
zz For Oxidative Decarboxylation, enzyme is a Dehydrogenase
●● Histidine → Histamine (Vasodilator) zz Oxidative Decarboxylation requires Vitamin B1
●● Tryptophan → Tryptamine zz Simple Decarboxylation requires Vitamin B6
●● Tyrosine → Tyramine zz Carboxylation requires Vitamin B7
T
●● DOPA → Dopamine
Energy Saving by Body H
●● Serine → Ethanolamine E
●● Cysteine → Beta Mercaptoethanol Synthesis of proteins requires lots of energy, so body saves its O
●● Glutamate →  GABA (Gamma Amino Butyric energy by using enzymes of Glycolysis in many pathways e.g. R
Acid) 1. Gluconeogenesis Y
2. Fructose Metabolism
●● Lysine → Cadaverine (a foul-smelling
3. Galactose Metabolism
diamine)
12 m nemonic
zz FAD → Vitamin B2
Riboflavin (B2) is a constituent of FAD, FMN
NAD → Vitamin B3
CRO BIOCHEMISTRY

zz
Niacin (B3) is a constituent of NAD, NADP
zz CoA → Vitamin B5
Pantothenic Acid (B5) is a constituent of CoA

Rossmann Fold
Nucleus
Rossmann fold/ βαβ fold- a super-secondary structure (con- zz Contain genetic material DNA, organized into chromo-
served during evolution), characterized by an alternating βαβ somes. There are two nuclear membranes, i.e. outer
(beta strand-alpha helix-beta strand). The β-strands are hy- and inner nuclear membrane. Outer membrane is
drogen bonded forming a β-sheet. The βαβ fold structure is


continuous with the endoplasmic reticulum. At several

commonly observed in enzymes that have dinucleotide co- places, inner and outer membrane are fused to form
enzymes, such as FAD, SAM, NAD and NADP. The associa- pores known as nuclear pores.
tion between a Dehydrogenase & NAD or NADP is relatively
loose. So, the coenzyme can quickly diffuse from one enzyme
to another, acting as a water-soluble carrier of electrons from
one metabolite to another. The enzymes which have Ross-
mann fold are known as Rossmann enzymes e.g. Glyceralde-
hyde-3-Phosphate Dehydrogenase, Alcohol Dehydrogenase
Isocitrate Dehydrogenase, Lactate Dehydrogenase, Phospho-
glycerate Dehydrogenase, Glutathione Re-ductase, D-Amino
Acid Oxidase

zz Nucleolus: This is rich in rRNA. It disappears during cell

division.
Mitochondria
zz Power house of cells as they are responsible for ATP
production. There are multiple mitochondria in each cell
zz They have their own mitochondrial DNA, RNA and ribo-
somes which synthesize around 10% of the mitochondrial
proteins and 20% proteins of ETC.
zz Mitochondria control the levels of calcium in cytoplasm.

T
CELL ORGANELLES
H Cell is the structural and functional unit of life. Eukaryotic
E cells (animal cell, plant cell, fungi, protozoa) have many Enzymes in various regions of mitochondria:
O organelles. Organelles are not found in prokaryotic cell. zz OMM has enzymes for lipid metabolism
R zz Organelle: Any membrane bound structure within a cell zz Inter membrane space has enzymes for nucleotide
Y is known as an organelle Exception: Ribosome. metabolism
zz Ribosomes are not organelles as they are non-membra- zz IMM has all ETC components
nous
zz Mitochondrial matrix has enzymes of beta oxidation zz H2O2 formed
of fatty acids, TCA cycle and enzyme Glutamate Dehy- zz Contain oxidative enzymes e.g. Catalase and Peroxidase
drogenase of oxidative deamination which can destroy Hydrogen Peroxide 13
Function of peroxisomes is neutralization of peroxides,
A dditional E dge
zz
oxidation of very long chain fatty acids and alpha
Evolution of Mitochondria and Chloroplast: oxidation of fatty acids, i.e. lipid oxidation without ATP

CHAPTER 1  CONCEPTS IN BIOCHEMISTRY

Aerobic bacteria during evolution have developed a symbiotic formation.
relation with primordial anaerobic eukaryotic cells that ultimately zz Peroxisome Biogenesis Disorders (PBD) are rare dis-
lead to the arrival of aerobic eukaryotic cells. Therefore, orders which affect peroxisomes.
mitochondria and chloroplast resemble prokaryotic cells. Most severe PBD is Zellweger syndrome (Cerebro-Hepato-
Renal syndrome) in which there is absence of peroxisomes
Lysosomes (Digestive Organelles): in all tissues. So, there is defect in oxidation of very long chain
zz They are of intermediate size between mitochondria and fatty acids (VLCFA).
endoplasmic reticulum
zz Contain many Hydrolase enzymes with optimum pH < 5 Endoplasmic Reticulum
zz Lipofuscin/Residual bodies are formed when indigestible zz This is a system of membranes with a network of vesicular
material gets accumulated in lysosomes spaces. These membranes run parallel to each other
zz They are mainly responsible for digestion and creating spaces known as Cisternae. It is well connected
destruction of various biomolecules. If there is some with perinuclear space and extracellular space.
inherited deficiency of some lysosomal enzyme, then
that particular macromolecule will be accumulated
in lysosome, collectively these diseases are known as
Lysosomal Storage Diseases (a type of inborn error of
metabolism). Organs mostly affected are Brain, Bone,
Cartilage, Viscera.
Lysosomal Storage Diseases (LSDs)
1.   MPS (Mucopolysaccharidosis) → ↑ Mucopolysaccharides
(covered in carbohydrate chemistry)
2.   Pompe’s disease (Only Glycogen Storage Disease which
is a LSD) → Defect in minor pathway of Glycogenolysis
(covered in Glycogen storage diseases)
3.   Sphingolipidosis or lipid storage disease (Most common
LSD is Gaucher’s disease) → ↑ Sphingolipids (covered in
lipid chemistry)
4.   Wolman’s disease → Defect in Lysosomal Acid Lipase
(covered in lipid chemistry)
5.   Cystinosis → Defect in Cystine Transporter (covered in
Proteins)
6.   Mucolipidosis → I cell disease (covered in carbohydrate
chemistry)
7.   Glycoproteinoses → e.g. a and β Mannosidosis, Sialido-
sis.

LSD with ERT (Enzyme Replacement Therapy)

•• MPS – I, II, VI
•• Pompe’s disease: α-Glucosidase
•• Sphingolipidosis:
ƒƒ Gaucher’s disease –β-Glucosidase
ƒƒ Niemann Pick disease
ƒƒ Fabry’s– α-Galactosidase A T
•• Wolman’s disease H
Fig. 1.14: Different Types of ER E
Peroxisomes/Microbodies/Glyoxysomes zz During cell fractionation, RER is disrupted to form small O
zz Single membrane organelle
vesicles known as microsomes. R
zz No ATP formed
zz Microsomes do not exist in a cell. Y
 Golgi Apparatus or Dictyosomes PROTEIN TARGETTING DISORDERS
These are system of membranes with many vesicles. [Diseases due to defective protein targeting]
14 zz
They are rich in lipids. These are active in cells which
•• Zellweger Syndrome (↑↑ VLCFA) (covered in lipid metabolism)
produce proteins for export
zz They receive protein from RER which enter Golgi •• Cystic Fibrosis Defect in CFTR gene, which forms a protein or
channel i.e. ATP gated Cl– channel which secrets Cl– in lungs
CRO BIOCHEMISTRY

apparatus, where they are stored, post translationally

modified, assembled, segregated or packaged and and GIT and reabsorb Cl– in sweat glands.
supplied to the extracellular side. So they are also known
Ribosomes
as packaging bodies.
zz Functions of Golgi apparatus: zz Free ribosomes or Cytoplasmic ribosomes: Ribo-
1. Protein sorting/packaging somes are not considered organelles as they are non
2. Protein glycosylation (O-glycosylation) membranous. They are spherical bodies made up of rRNA
and proteins. rRNA is more in amount than proteins. So,
3. Processing oligosaccharide chain of glycoproteins
rRNA is considered as marker for this organelle. They
4. Membrane synthesis of other organelles e.g. Peroxi- synthesize proteins which are to be used by cell e.g. for
somes, Lysosomes cell metabolism, proteins used in cytoplasm, proteins


N-glycosylation is done in endoplasmic reticulum or enzymes destined for nucleus, mitochondria and
peroxisomes.

Technique for Separation of Various Organelles:

Sucrose Density Gradient Centrifuge
zz A centrifuge tube is filled with a solution (density of which
increases from top to bottom) e.g. a solute- Sucrose is
dissolved at different concentrations to produce the
density gradient.
zz Mixture of organelles is layered on top of this density
gradient.
zz Centrifuge the tube at high speed.
zz Individual organelles sediment until their buoyant
Fig. 1.15: Subcellular fractions density exactly matched that in the gradient.
zz Each layer is collected separately.
Protein Targeting
Protein sorting or targeting means correct localization of
protein in the cell. It can occur co-translationally or post
translationally.

PROTEIN TARGETTING DISORDERS

[Diseases due to defective protein targeting]
•• I – cell disease or Inclusion body disease (A Mucolipidoses)
(covered in carbohydrate chemistry)
•• Primary Hyperoxaluria (defect in Glycine Metabolism)
(covered in amino acids)
•• Familial Hypercholesterolemia (Type II (a)
Hyperlipoproteinemia)(covered in lipoproteins) Fig. 1.16: Sucrose density gradient centrifuge

Contd…

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 This way purified organelle can be isolated. The isolation of an organelle enriched in certain enzyme is often the first step
in the purification of that enzyme.
15

CHAPTER 1  CONCEPTS IN BIOCHEMISTRY

Fig. 1.17: Biochemical markers of various cell organelles

BONDS IN MACROMOLECULES zz Amide bond is formed when a carboxy group joins with
amino group
zz If Amide bond is in protein then it is known as peptide
bond
zz Strongest covalent bond is peptide bond
zz Disulphide bond (S-S)- formed when two sulfhydryl
groups (–SH) are joined
zz Weakest bond – Van der Waals interactions (They arise
from the rapid movement of electrons of all neutral
atoms)
zz Hydrophobic interactions – tendency of nonpolar
compounds to self-associate in an aqueous medium.
These are not hydrophobic bonds, they are hydrophobic
interactions.
zz Electrostatic/ionic bonds are also known as salt bridges/
linkages, which occur between oppositely charged
groups.
zz Strongest Bond: Covalent bond
zz Weakest Bond: Van der Waals forces
zz Covalent > Ionic > Hydrogen > Hydrophobic > Van der Waals

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 Summary of Formulae
16
Smile Formula 1: Which are anabolic, Which are catabolic pathways?
zz Anabolic pathways/enzymes are: HMP, Glycogenesis, FA synthesis, Cholesterol synthesis, TG synthesis, Lipoprotein Lipase enzyme.

zz Catabolic pathways/enzymes are: Glycolysis, Link reaction, Glycogenolysis, beta oxidation of fatty acids, Gluconeogenesis, Ketone
CRO BIOCHEMISTRY

body synthesis, Ketone body utilization.

Smile Formula 2: Insulin & Glucagon activates which pathway or enzymes?
zz Insulin activates all anabolic pathways enzymes, i.e. HMP, Glycogenesis, FA synthesis, Cholesterol synthesis, TG synthesis, Lipoprotein
Lipase enzyme.
zz Also, Insulin activates two catabolic pathway enzymes, i.e. Glycolysis & Link reaction.

zz Glucagon activates all catabolic pathway enzymes, i.e. Glycogenolysis, beta oxidation of fatty acids, Gluconeogenesis, Ketone body
synthesis, Ketone body utilization.
zz But Glucagon does not activate two catabolic pathway enzymes, i.e. Glycolysis & Link reaction, as these are activated by Insulin.

Smile Formula 3: Which hormone causes phosphorylation & which causes dephosphorylation?


zz Enzymes which are activated by Insulin (anabolic enzymes) are always active in dephosphorylated state. Enzymes which are activated
by Glucagon (catabolic enzymes) are always active in phosphorylated state but One Exception: ATP Citrate Lyase is anabolic enzyme
but it is active in phosphorylated state.
Smile Formula 4: Which pathway occurs in which compartment of the cell?
zz In cytoplasm, anabolic pathways occur, i.e. HMP, Glycogenesis, FA synthesis, Cholesterol synthesis, TG synthesis.
zz But two catabolic pathways also occur in cytoplasm, i.e. Glycolysis & Glycogenolysis.

zz In mitochondria, catabolic pathways occur (beta oxidation of fatty acids, ketone body synthesis, ketone body utilization, link reaction),
Vital pathways, i.e. TCA and ETC and Replication, Transcription, Translation (for mitochondrial DNA) and Apoptosis.
zz Gluconeogenesis is catabolic but it occurs in both mitochondria and cytoplasm.

SITE OF PATHWAYS
•• CYTOPLASM
� Glycolysis � Glycogenesis
� Glycogenolysis � Nucleotide Synthesis
� HMP � Fatty Acid Synthesis
� Cholesterol Synthesis � Steroid Synthesis
� Activation of Fatty Acid for β-Oxidation
•• MITOCHONDRIA
ƒƒ Link reaction/PDH complex
ƒƒ TCA
ƒƒ ETC
ƒƒ β-Oxidation of Fatty Acid
ƒƒ Ketone Body Synthesis
ƒƒ Ketone Body Utilization/Breakdown
•• BOTH CYTOPLASM AND MITOCHONDRIA
ƒƒ Urea Cycle
ƒƒ Gluconeogenesis
ƒƒ Haem Synthesis
•• ENDOPLASMIC RETICULUM (ER)
ƒƒ Desaturation of Fatty Acid
ƒƒ Elongation of Fatty Acid
T ƒƒ ω-oxidation
H ƒƒ Protein synthesis (Rough ER)
E
•• PEROXISOMES
O
ƒƒ α-oxidation
R ƒƒ Oxidation of Very Long Chain fatty acids (VLCFA)
Y
 Pearls of the Chapter
17
zz Acetyl CoA is never Glucogenic, i.e. Acetyl CoA can never form Glucose but Acetyl CoA activates the very first step of Gluconeogenesis,
i.e. Pyruvate Carboxylase
zz In fed state, Acetyl CoA is used for the synthesis of Fats
In starvation, Acetyl CoA has three fates – TCA or ketone body Synthesis or Gluconeogenesis

CHAPTER 1  CONCEPTS IN BIOCHEMISTRY

zz

zz Fats can never be converted to carbohydrates. Exception: Two breakdown products of Fats, i.e. Glycerol and Propionic acid can be
converted to carbohydrates
Three pathways which occur both in mitochondria and cytoplasm are Gluconeogenesis, Urea Cycle and Haem Synthesis
zz Sources of blood Glucose are: Food, Liver Glycogen (for 12–18 hrs) and Gluconeogenesis.
zz Lipoprotein Lipase is activated by Insulin, but only in the capillary beds of Adipose tissues.
zz Hormone Sensitive Lipase is inhibited by Insulin.
zz Brain cannot use fatty acids as fatty acids cannot cross Blood Brain Barrier (BBB).
zz Ketone Bodies are formed during starvation for vital organs – Heart and Brain.
zz Atkin’s diet is low calorie, low carbohydrate diet.
zz Fructose is the Most Lipogenic Sugar.
zz Low Insulin: Glucagon Ratio means Catabolic State.
zz High Insulin: Glucagon Ratio means Anabolic State.
zz Diabetic situation is almost same like fasting and starvation
zz Most severe PBD is Zellweger syndrome.
zz Strongest covalent bond is peptide bond and weakest bond is Van Der Waals interactions

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 18 Multiple Choice Questions
Formula Questions 11. Pathway which occurs both in fed and fasting state:
a. TCA b. Glycolysis
(Insulin, Glucagon, Compartment, Phosphorylated
c. HMP d. Glycogenesis
State and Dephosphorylated State) 12. Pathway which occurs both in cytoplasm and mito-
1. Which of the following does not occur in mito- chondria is/are:
chondria?  (AIIMS Nov 2016) a. Urea cycle
a. Beta-oxidation b. DNA synthesis b. Gluconeogenesis
c. Fatty acid synthesis d. Protein synthesis c. Haem synthesis
2. Insulin promotes lipogenesis by all EXCEPT:  d. All
 (PGI May 2017) 13. Effect of glucagon:  (FMGE June 2018)
a. Decreasing cAMP a. Retard Glycogenolysis
b. Increase Glucose uptake b. Retards Ketogenesis
c. Inhibiting Pyruvate Dehydrogenase c. Promote Gluconeogenesis
d. Increasing Acetyl CoA d. Decrease plasma amino acids
3. Mitochondria are involved in all of the following 14. Which of the following enzyme activity decreases in
EXCEPT: (AIIMS Nov 2015) fasting? (AIIMS May 2018)
a. ATP production a. Hormone Sensitive Lipase
b. Apoptosis b. Glycogen Phosphorylase
c. Tri-Carboxylic Acid cycle c. Acetyl CoA Carboxylase
d. Cholesterol Synthesis d. Pyruvate Carboxylase
4. Hormone Sensitive Lipase is not activated by:  15. Which of the following is active in dephosphorylated
 (PGI Nov 2017) state? (PGI May 2017)
a. Insulin b. Glucagon a. Glycogen Synthase
c. Catecholamines d. T4 b. Pyruvate Carboxylase
M 5. Which of the following is not seen in low insulin c. Glycogen Phosphorylase
C glucagon ratio? (PGI Nov 2017) d. Acetyl CoA Carboxylase
Qs a. Gluconeogenesis b. Glycogen Breakdown e. Pyruvate Dehydrogenase
Ans. c. Ketogenesis d. Glycogen Storage 16. What is effect of Cortisol on metabolism:
6. Which of the following is active in dephosphorylated  (PGI Nov 2008)
1. c state?  (AIIMS May 2017) a. ↑ Gluconeogenesis
2. c a. Glycogen Synthase b. ↑ Lipogenesis
3. d b. Pyruvate Carboxylase c. ↑ Proteolysis
4. a c. Glycogen Phosphorylase d. ↑ Export of amino acid to liver
5. d d. PEPCK e. ↑ Glycolysis
6. a 7. All occur in mitochondria EXCEPT: (PGMEE 2015)
7. a a. Glycolysis b. TCA cycle Diabetes
8. a c. ETC d. Ketogenesis 17. The activity of which of the following enzymes is
9. d 8. The biosynthesis of the enzyme Pyruvate Carboxylase increased in Diabetes Mellitus?
10. a is repressed by: (PGMEE 2012, 13) a. CPT-I
11. a a. Insulin b. Cortisol b. Phosphoenol Pyruvate Carboxykinase
12. d c. Glucagon d. Epinephrine c. Glucose-6-Phosphatase
13. c 9. The enzyme activated with low Insulin: Glucagon d. All
14. c ratio is:  (AIIMS Nov 2013) 18. Starvation and diabetes mellitus can lead to
15. a,d,e a. Glucose-6- phosphate dehydrogenase ketosis. Which of the following features are in
16. a,c,d b. Glucokinase favor of ketosis in diabetes mellitus?
17. d c. Pyruvate Kinase a. Increase in glucagon/insulin ratio, increased cAMP
18. a d. Glucose 6 Phosphatase and increased blood glucose
10. The gene expression of which of the following b. Decreased insulin, increased free fatty acid which is
enzymes are not increased by insulin? equivalent to blood glucose
a. Pyruvate Carboxylase c. Decreased insulin, increased free fatty acid which is
b. Acetyl CoA Carboxylase not equivalent to blood glucose
c. Phosphofructokinase-I d. Elevated insulin and free fatty acid, equivalent to
d. Pyruvate Dehydrogenase blood glucose
 19. Patient with Type I Diabetes mellitus, with complains 31. Which of the following biochemical reaction is
of polyuria. Which of the following will occur involved in the conversion of histidine to histamine?
normally in his body? (AIIMS Nov 2018)  (PGMEE 2015) 19
a. Glycogenesis in muscle a. Decarboxylation b. Carboxylation
b. Increased protein synthesis c. Amination d. Oxidation
c. Increased conversion of fatty acid to Acetyl CoA 32. Which of the following is NOT the source of cytosolic
d. Decreased in Cholesterol synthesis NADPH? (Recent Pattern Jan 2019 Q)
20. In type I diabetes, which of the following is true? a. Isocitrate Dehydrogenase
 (Recent Pattern Jan 2019 Q) b. ATP Citrate Lyase
a. Increased lipolysis c. Malic enzyme
b. Decreased protein catabolism d. G6PD
c. Decreased hepatic glucose output 33. NADPH is produced by:  (PGI May 2014)
d. Increase glucose uptake a. Pyruvate Dehydrogenase
b. Isocitrate Dehydrogenase
Fuel for body and Acetyl CoA c. a-ketoglutaryl-Dehydrogenase
21. Preferred fuel for body in fasting state?  d. Succinate Dehydrogenase
 (AIIMS May 2017) e. Malate Dehydrogenase
a. Carbohydrates b. Fats
c. Proteins d. Amino acids Organelles and Bonds in Macromolecules
22. Acetyl CoA cannot be converted to: 34. Protein segregation occurs in:  (PGMEE 2012, 13)
a. Fatty acids b. Glucose a. Golgi apparatus b. ER
c. Ketone bodies d. Cholesterol
c. Peroxisomes d. Mitochondria
23. Myocardium normally utilizes: (FMGE June 2018)
a. Glucose b. Lactose 35. Oxidation of drugs mainly takes place in:
c. Fatty acid d. Glycogen  (PGMEE 2015)
24. During 3rd day to 2nd week of starvation, brain a. Cytoplasm b. Rough ER
depends on which of the following substance as fuel? c. Nucleus d. Smooth ER
a. Ketone bodies b. Glucose 36. The cellular component of protein synthesis is:
c. Fatty acid d. Amino Acid a. Smooth endoplasmic reticulum
25. Main source of energy is derived from (By human b. Rough endoplasmic reticulum
body): (PGI Nov 2008) c. Ribosomes M
a. Fat b. Glycogen d. Mitochondria C
c. Lactate d. Ketone 37. Which of the following is seen in association with Qs
e. Acetone membrane raft? (AIIMS Nov 2012)
Ans.
a. Mannose binding protein
Glycemic Index b. GTP associated receptor 19. c
26. Which of the following has highest glycemic index? c. GPI anchored protein 20. a
d. Spectrin associated protein 21. b
a. Glucose c. Sucrose 38. Plasma Membrane marker(s) is/are: (PGI Nov 2011) 22. b
b. Fructose d. Sorbitol a. 5’-Nucleotidase b. Galactosyltransferase 23. c
27. Low glycemic index food is: (FMGE Nov 2018 ) c. ATP Synthetase d. Adenyl cyclase 24. a
a. Easily digestible
e. Na+-K+ ATPase 25. b
b. Increase plasma glucose
c. Has slower absorption 26. a
Miscellaneous 27. c
d. Increase glycogen deposits 39. Entropy is a measure of the: (PGMEE 2007) 28. c
Phosphocreatine a. Reversibility of reaction 29. c
b. Randomness in a system 30. b
28. During exercise, most rapid way to synthesize ATP is: c. Exothermicity
 (AIIMS Nov 2018) 31. a
d. Free energy for an enzymatic reaction 32. b
a. Glycogenolysis b. Glycolysis
40. Storage form of free energy in the cell:  33. b,e
c. Phosphocreatine d. TCA Cycle
 (PGMEE 2015) 34. a
29. Source of energy for a running race athlete in the
a. NADH b. ATP 35. d
initial 3 minutes of running (PGMEE 2013)
c. G-6-P d. Creatine Phosphate 36. c>>b
a. Free fatty acid b. Creatine Phosphate
41. Glowing of firefly is due to(PGMEE 2008, 2009, 2010) 37. c
c. Muscle glycogen d. Blood Glucose
a. ATP b. NADH 38. a,d,e
Enzyme Basics c. GTP d. Phosphocreatinine 39. b
30. Which is required in anabolic reactions? 42. How many calories are supplied per gram of dietary 40. b
 (AIIMS May 2017) fiber? 41. a
a. NAD b. NADP a. 2 kilocalories b. 4 kilocalories 42. a
c. FAD d. FADP c. 9 kilocalories d. 7 calories
 43. Silver staining is done for:  (PGI) 44. Themogenic food is which of the following: 
a. DNA b. RNA  (AIIMS May 2017)
20 c. Protein d. Karyotyping analysis a. High protein diet
e. Collagen b. High Carbohydrate diet
c. High fat diet
d. It does not depend on the macronutrients

Answers with Explanations

1. Ans. (c) Fatty acid synthesis 5. Ans. (d) Glycogen Storage
[Ref: Harper 30th/e pg. 233] [Ref: Harper 30th/e pg. 176]
•• Fatty acid synthesis is anabolic pathway, so it occurs •• Low insulin means catabolic situation in body. So,
in cytoplasm. β-oxidation (option a) is catabolic question is that which of the following is not seen
pathway, so it occurs in mitochondria. DNA synthesis in catabolic state. So, answer is Glycogen Storage,
(Replication) and protein synthesis (Translation) of which is anabolic. Rest three pathways given are
mitochondrial DNA occurs in mitochondria (option catabolic – Gluconeogenesis, Glycogen Breakdown
b and d) and Ketogenesis.

2. Ans. (c) Inhibiting Pyruvate Dehydrogenase 6. Ans. (a) Glycogen Synthase

[Ref: Harper 30th/e pg. 182]
[Ref: Harper 30th/e pg. 188]
•• Glycogen Synthase is enzyme of glycogen synthesis,
•• Insulin activates link reaction (enzyme Pyruvate
which is anabolic pathway. So, this enzyme is active
Dehydrogenase) and thus increases Acetyl CoA.
in dephosphorylated state. Rest all enzymes are
M •• Insulin decreases cAMP, by activating enzyme involved in catabolic pathways, which are active in
C Phosphodiesterase (option a). phosphorylated state.
Qs •• Insulin increases glucose uptake via activating GLUT- •• Pyruvate carboxylase and PEPCK are in Gluconeo-
Ans. 4 (present on peripheral tissues) (option b). genesis. Glycogen Phosphorylase is in glycogenoly-
43. all sis.
44. a 3. Ans. (d) Cholesterol Synthesis
7. Ans. (a) Glycolysis
[Ref: Harper 30th/e pg. 267]
•• The biochemical processes taking place in mito- [Ref: Harper 30th/e pg. 168]
chondria are all catabolic pathways, Vital pathways •• Glycolysis occurs in cytoplasm (Also see explanation
i.e. TCA and ETC, Replication, Trans-cription and of Q3)
Translation for mitochondrial DNA and apoptosis
•• Cholesterol synthesis is anabolic pathway, so it 8. Ans. (a) Insulin
occurs in cytoplasm (option d) [Ref: Harper 30th/e pg. 188]
4. Ans. (a) Insulin •• Pyruvate Carboxylase is enzyme of gluconeogenesis
(catabolic pathway). So, it is repressed by Insulin
[Ref: Harper 30th/e pg. 262] (anabolic hormone).
•• Hormone Sensitive Lipase (HSL) is a catabolic
9. Ans. (d) Glucose 6 Phosphatase
enzyme (enzyme which breaks down the triglycerides
of adipose tissue). So, it is inhibited by Insulin. [Ref: Harper 30th/e pg. 176]
Insulin activates all anabolic pathway enzymes. Rest •• Glucokinase and Pyruvate Kinase are enzymes of
all hormones like Glucagon, Thyroid, Epinephrine Glycolysis, so these are active in fed state (i.e. High
activates HSL.
 Insulin Glucagon ratio). G6PD is enzyme of HMP, Phosphorylase (Glycogenolysis) and Pyruvate Carbo-
which is activated by insulin. Glucose 6 Phosphatase xylase (Gluconeogenesis) is increased in Fasting.
is enzyme of gluconeogenesis, so it is activated in 21
fasting state (i.e. Low insulin glucagon ratio). 15. Ans. (a); (d); (e)

10. Ans. (a) Pyruvate Carboxylase [Ref: Harper 30th/e pg. 188]

CHAPTER 1  CONCEPTS IN BIOCHEMISTRY

•• Dephosphorylation is done by hormone, insulin. Insulin
[Ref: Lehninger 5th /e p591]
activates all anabolic enzymes in fed state.
•• Enzymes where gene expression is increased ƒƒ (Option a): Glycogen Synthase, enzyme of glycogen
by Insulin: (usually enzymes of glycolysis, Link synthesis, so it is anabolic.
reaction, HMP, fatty acid synthesis) are Hexokinase- ƒƒ (Option b): Pyruvate carboxylase, involved in
II, Hexokinase IV, phosphofructokinase-I (PFK-I), gluconeogenesis, so it is catabolic.
PFK-II, Pyruvate Kinase, Glucose-6-Phosphate ƒƒ (Option c): Glycogen Phosphorylase, enzyme of
Dehydrogenase (G6PD), Pyruvate Dehydrogenase, glycogen break-down, so it is catabolic.
Acetyl CoA Carboxylase, Malic enzyme, ATP Citrate ƒƒ (Option d): Acetyl CoA Carboxylase, involved in
Lyase, FA synthase complex fatty acid synthesis, so, it is anabolic.
•• Enzymes where gene expression is decreased by ƒƒ (Option e): Pyruvate Dehydrogenase, enzyme of link
Insulin: (usually enzymes of gluconeogenesis) Pho- reaction. Link reaction is catabolic, but it is activated
sphoenolpyruvate Carboxykinase (PEPCK), Glucose- by insulin (EXCEPTION).
6-Phosphatase
16. Ans. (a); (c) (d)
11. Ans. (a) TCA
[Ref: Lehninger 7th/e pg. 937, 938]
[Ref: Harper 30th/e pg. 125, 162]
•• Cortisol is a catabolic hormone like Glucagon. It acts
•• Answer is TCA & ETC. They are vital pathways. These primarily on muscle, liver and adipose tissues. A variety
do not depend on Hormones. of stressors release Cortisol from adrenal cortex. This is
a relatively slow acting hormone. Metabolically, it will
12. Ans. (d) All cause increase in Glycogenolysis, Gluconeogenesis,
[Ref: Harper 30th/e pg. 188, 189] Glycolysis, Beta Oxidation of Fatty Acid.
•• There is increased proteolysis and increased export of
•• Pathways taking place in two compartments, i.e. both amino acids to liver for gluconeogenesis.
cytoplasm and mitochondria are Haem synthesis,
Urea Cycle and Gluconeogenesis. 17. Ans. (d) All

13. Ans. (c) Promote Gluconeogenesis [Ref: Lehninger 5th /e p591]

[Ref: Harper 30th/e pg. 171] •• In Diabetes Mellitus, the activity of catabolic enzymes A
is increased. N
•• Glucagon activates all catabolic pathway enzymes •• Carnitine Palmitoyl Transferase-I (CPT-I) is an S
(EXCEPT: Glycolysis and Link reaction) enzyme of β-Oxidation which is a catabolic pathway.
•• It increases, gluconeogenesis glycogenolysis, W
Therefore, is increased in diabetes
lipolysis, ketogeresis and proteolysis leading to E
•• Phosphoenol Pyruvate Carboxykinase and Glucose
increase plasma amino acids. R
-6-Phosphatase are enzymes of gluconeogenesis,
which is a catabolic pathway and hence also S
14. Ans. (c) Acetyl CoA carboxylase increased in Diabetes. WITH
[Ref: Harper’s Illustrated Biochemistry, 30th ed., pg. 188, •• Only one arabolic mechanism increased in diabetes
Lippincott’s illustrated reviews, 6th ed., pg. 107] is TG and VLDL synthesis, which is due to excess E
Acetyl CoA from b-oxidation. X
•• Anabolic pathways and its enzymes are decreased in P
fasting e.g. Glycogenesis, Lipogenesis, Cholesterol 18. Ans. (a) Increase in glucagon/insulin ratio, increased L
synthesis, Fatty acid synthesis and TG synthesis. cAMP and increased blood glucose A
•• Also, glycolysis & link reaction is decreased in fasting. N
Acetyl CoA Carboxylase is an enzyme of FA synthesis. [Ref: Satyanarayana, 3rd/e p 296]
A
So, its activity is decreased in fasting. •• Diabetic situation is same like fasting or starvation. T
•• Catabolic pathways and its enzymes are increased in There is relative or absolute deficiency of Insulin. I
fasting: Gluconeogenesis, Glycogenolysis, Lipolysis This leads to increase in Glucagon, cAMP & Blood O
(Hormone Sensitive Lipase). Activity of Glycogen Glucose. N
S
 19. Ans. (c) Increased conversion of fatty acid to Acetyl 24. Ans. (a) Ketone bodies
CoA
22 [Ref: Lippincott 4th/e pg.327]
[Ref: Lippincott 4th/e pg. 338, 339] Glucose is the main/preferred fuel for the brain in fed
•• Diabetic situation is same like fasting, i.e. breakdown as well as fasting state whereas during starvation, brain
or catabolism is increased and anabolism or synthesis depends upon Ketone Bodies as the fuel.
CRO BIOCHEMISTRY

is decreased.
25. Ans. (b) Glycogen
•• So in diabetes, fats (TG) are broken down in adipose
tissue to give fatty acids. These fatty acids go in blood [Ref: Harper 30th/e pg. 178]
and then to liver. In liver, these fatty acids are broken
down by beta oxidation of fatty acids to give Acetyl In fed state, all body cells prefer carbohydrates, i.e.
CoA. Glucose or Glycogen (whenever question does not
•• This excess Acetyl CoA obtained from fatty acid mention fed or fasting state, then consider fed state).
breakdown is used for formation of fats e.g. fatty Fat is the main fuel in fasting or starvation. Ketone
acids, endogenous TGs, VLDL & cholesterol. bodies are fuel for Heart and Brain during starvation.
•• Body is mainly in catabolism, not anabolism [Option Acetone is a ketone body which does not act as a fuel in


a and b are wrong] body. Lactate is used for gluconeogenesis which mainly
•• There is increased cholesterol synthesis [Option d is occurs in fasting.
wrong]
26. Ans. (a) Glucose
20. Ans. (a) Increased lipolysis [Ref: Lippincott’s Illustrated Reviews 4th/e pg. 366]
[Ref: Harper 30 /e pg. 149]
th
•• Glycemic Index (GI)- This is the increase in blood
•• In diabetes, body is in catabolic situation. catabolic glucose after the test dose of a carbohydrate compared
pathways are increased and anabolic are decreased. with that of an equivalent amount of glucose. GI is a
So, there is increased lipolysis, increased protein value assigned to foods based on how slowly or how
catabolism, increased hepatic glucose output due to quickly those foods cause an increase in blood glucose
Gluconeogenesis. Due to insulin deficiency, there is levels.
decreased glucose uptake by peripheral cells. •• The highest glycemic index is for glucose and galactose
(also lactose, maltose, trehalose and isomaltose get
21. Ans. (b) Fats converted to glucose, so they also have high GI).
•• GI of fructose is less than that of glucose.
[Ref: Harper 30th/e pg. 230]
•• Fructose and sugar alcohols have less glycemic index
•• In fasting state, preferred fuel is Fats and Adipose tissues because they are not absorbed completely.
are broken down to give energy. •• Sucrose also has low glycemic index than glucose as it
A •• Preferred fuel for body in fed state is carbohydrates, i.e. gets split to glucose and fructose.
N Glucose.
S 27. Ans. (c) Has slower absorption
22. Ans. (b) Glucose
W
[Ref: Harper 30th/e pg. 156]
E [Ref: Harper 30th/e p172]
R •• Acetyl CoA is never a substrate for Gluconeogenesis •• A food with high GI raises blood glucose more than a
S because Acetyl CoA can never be converted back to food with low GI. (Also see Q.26)
Pyruvate as Link reaction (Pyruvate Dehydrogenase) is •• Foods with a low-GI value are the preferred choice as
WITH
irreversible. they are slowly digested and absorbed, causing a slower
•• Acetyl CoA is a starting material for Fatty Acid and smaller rise in blood sugar levels.
E
Synthesis, Cholesterol Synthesis and ketone body •• On the other hand, foods with a high GI value should
X
Synthesis (Option a, c, d). be limited since they are quickly digested and absorbed,
P
resulting in a rapid rise and fall of blood sugar levels.
L
23. Ans. (c) Fatty acid
A Low GI (55 or less) Fructose, beans, grains, walnuts
N [Ref: Harper 30th/e pg. 150]
A Medium GI (56-69) Table sugar or sucrose, bread,
•• Heart uses fatty acids in fed and fasting state. It uses juice, raisins
T Ketone bodies during starvation. Fetal heart uses
I High GI (70 or higher) High fructose corn syrup, glucose,
Glucose. In heart failure, fuel is Glucose. sweets, rice, maltose
O
N
S
 28. Ans. (c) Phosphocreatine 29. Ans. (c) Muscle Glycogen
[Ref: Harper 30th/e pg. 135] [Ref: Harper 30th/e pg. 135] 23
Immediate source of energy for Muscles is Creatine •• For explanation, refer Ques No. 28
Phosphate for first 8–10 seconds. This is known as •• Muscle glycogen gives glucose in muscles. Do not
Phosphagen system.

CHAPTER 1  CONCEPTS IN BIOCHEMISTRY

mark blood glucose (Option d)
Reasons for use of Phosphocreatine:
•• Does not uses O2 30. Ans. (b) NADP
•• Does not produce lactate
•• Quickest source of energy [Ref: Lehninger 7th/e pg. 312]
•• Most direct form of energy production
•• NADPH is required for reductive biosynthesis (Fatty
But this is of limited supply in muscles, so depleted
Acid Synthesis, Cholesterol Synthesis, Steroid Hormone
quickly.
Synthesis, in RBC for formation of Reduced Gluta-
•• Then second system used is Anaerobic Glycolysis
thione), i.e. mainly anabolic reactions
or Lactate system. This system derives ATPs using
Glucose, which is mainly derived from Muscle •• NADPH is synthesized mainly in HMP pathway. Minor
Glycogen and this system give energy for 1-3 minutes sources of NADPH are Malic enzyme and cytoplasmic
of exercise or running. Isocitrate Dehy-drogenase.
•• After 3 minutes of exercise, the body starts to supply •• Here the best answer is NADP (If NADPH given, mark
working muscles with oxygen & now Aerobic System that)
(or Mitochondrial Respiration) is used which
can use carbohydrates, Fats, or Proteins to produce 31. Ans. (a) Decarboxylation
energy. This system is slowest in providing energy, [Ref: Harper 30th/e pg. 299]
but more efficient than the other two systems as it
provides more amount of energy. This system uses •• Histidine to Histamine is simple decarboxylation
mainly TCA & ETC. step, requiring Vitamin B6 as coenzyme. (See other
examples of simple decarboxylation in theory)
Muscle Power (Rate Capacity Fuel uses
energy of ATP (Total ATP 32. Ans. (b) ATP Citrate Lyase
systems production) produced)
[Ref: Harper 30th/e pg. 197]
Phosphagen Very high Very low Creatine-
system Phosphate •• NADPH is synthesized by HMP (G-6PD), Malic enzyme
or Phospho- & cytoplasmic Isocitrate Dehydrogenase.
creatine •• NADPH is used in reductive biosynthesis (anabolic
Anaerobic High Low Muscle pathways).
system/ glycogen
In Sequence

Lactate
33. Ans. (b); (e) A
system N
[Ref: Harper 30th/e pg. 197]
Aerobic Low Very high Muscle
S
For explanation, Refer to ques No. 30 W
system/ glycogen,
Mito- blood E
34. Ans. (a) Golgi apparatus
chondrial glucose, R
respiration adipose [Ref: Harper 30th/e pg. 555] S
tissue and •• Golgi apparatus receives proteins from Rough
intramuscular ER they are stored, post translationally modified,
WITH
fat
assembled, segregated, packaged and supplied to E
the extracellular side. So, they are also known as X
packaging bodies. P
L
35. Ans. (d) Smooth ER A
[Ref: Harper 30th/e pg. 121] N
A
•• SER has a role in oxidative metabolism and detoxi- T
fication of drugs in hepatocytes. I
O
N
S
 36. Ans. (c) Ribosomes elevated cholesterol and sphingolipid content. They
are more rigid than rest of the membrane.
24 [Ref: Harper 30th/e pg. 121] •• Proteins associated with lipid rafts are:
•• Ribosomes >> Rough endoplasmic reticulum 1. GPI anchored proteins
•• Rough ER is that ER on which ribosomes are attached 2. Src family kinase
and protein synthesis occurs in ribosomes. So, best •• Many proteins get attached to lipid membranes
CRO BIOCHEMISTRY

answer here is c. via a post-translational lipid modification, the

Glycosylphosphatidyl Inositol (GPI) anchor & are
37. Ans. (c) GPI anchored protein known as GPI anchored proteins. They are found in
almost all cells. GPI-anchor has molecular interaction
[Ref: Harper 30th/e pg. 484] with lipid rafts and are associated with protein/lipid
•• Lipid rafts are specialized regions of the plasma sorting, signaling mechanisms and with human
membrane, made up of proteins and lipids, arranged disease.
in an ordered and tightly packed pattern. They have


A 41. Ans. (a) ATP

38. Ans. (a); (d) ; (e)
N
S [Ref: Harper 30th/e pg. 479] [Ref: Harper 30th/e pg. 120]
W •• Fireflies produce light due to a chemical reaction and
For explanation refer Figure 1.16
E this process is known as Bioluminescence.
R 39. Ans. (b) Randomness in a system
S [Ref Harper’s 30th /ed pg. 114 Lippincort’s 5th/e pg. 69]
WITH •• Entropy is degree of disorder or randomness in a
system, i.e. movement or vibration of molecules in a
E
system.
X
P 40. Ans. (b) ATP
L
A [Ref: Harper 30th/e pg. 120]
N •• ATP is the storage form of free energy in most of the
A cells of the body.
T •• Creatine phosphate is the immediate storage form in
I muscles. 42. Ans. (a) 2 kilocalories
O •• High energy compounds in biological systems are: •• Energy obtained per gram of macromolecules is as
N ATP, GTP, UTP, Creatine Phosphate, Phosphoenol- follows:
S pyruvate, Coenzyme A, 1,3-bisphospho glycerate and ƒƒ Dietary fibers: 2 kcal/gm
carbamoyl phosphate.
 ƒƒ Carbohydrates: 4 kcal/gm
ƒƒ Soluble fibers are broken by intestinal bacteria to
produce short chair fatty acids that provide energy. 25
Insoluble fibers do not provide energy.
ƒƒ Proteins: 4.2 kcal/gm
ƒƒ Fats: 9 kcal/gm

CHAPTER 1  CONCEPTS IN BIOCHEMISTRY

ƒƒ Alcohol: 7 kcal/gm (empty calories)

43. Ans. (a); (b); (c); (d); (e)

•• Silver staining is highly sensitive method (can detect
in low nanogram range) which is used for visualiza-
tion of intracellular and extracellular components
such as DNA, RNA, Proteins e.g. collagen and retic-
ulin fibres. •• Coomassie blue stain – is for proteins but it is less
•• Method is simple, equipments and chemicals are sensitive than silver stain.
cheap.
•• Silver ions (from silver nitrate in the staining reagent) 44. Ans. (a) High protein diet
interact and bind with certain functional groups. [Ref: Harper 30th/e pg. 254]
•• Themogenic or thermic effect of food is the amount
of energy required to digest, absorb, transport and
metabolize that food. This is maximum for proteins.

A
N
S
W
E
R
S
WITH

E
X
P
L
A
N
A
T
I
O
N
S
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________________________________________________________________________________________________________________
UNIT
II

CARBOHYDRATES
Unit Outline
Chapter 2 Chemistry of Carbohydrates
Chapter 3 Carbohydrate Metabolism
Chemistry of
Carbohydrates
2
Overview of Chapter
•• Definition of Carbohydrates
•• Isomerism in Carbohydrates
•• Classification of Carbohydrates
•• Glucose Transport

DEFINITION OF CARBOHYDRATES
Fig. 2.1: Aldehyde and Keto Group
zz Definition → Polyhydroxy aldehydes or ketones
zz Polyhydroxy → (Poly means many, Hydroxy means
‘OH’) means carbohydrates have many ‘OH’ groups Functional Carbon is Symmetric
Functional carbon (Ald or Keto) is C=O, i.e. 2 valencies are
General Points about Hydroxy Group occupied by same atom, i.e. Oxygen. So, functional carbon
zz Any compound having –OH group is Polar. (Polar is Symmetric, but only in linear configuration. (In cyclic
compounds are soluble in water) configuration, functional carbon becomes Asymmetric. See-
zz Suffix for –OH is ‘ol’ e.g. GlycerOL, AlcohOL Anomerism)
CholesterOL means cholesterol has polar component.
H igh R eturn
zz
But we know that cholesterol is a lipid having non-polar
component also. So, cholesterol is Amphipathic (very zz Molecular Formula for Carbohydrates (CH2O)n
clear from the name). Where n = Number of Total Carbons
zz OH group always has high tendency to bind phosphate. zz Number of total isomers possible for a compound is given by
E.g. Glucose gets converted to Glucose-6-P immediately the formula = 2n
when Glucose enters the cell, as phosphate has high Where n = Number of Asymmetric Carbons
tendency to bind OH. (Also –OH containing amino acids NOTE: Both formulas contain ‘n’ But this n is different.
have maximum tendency to bind phosphate).
zz In case of Carbohydrates, number of OH groups are one
less the number of Carbons. E.g. Glucose has 6 C and 5 Asymmetric Carbon (or Chiral Carbon)
–OH; Ribose has 5 C and 4 –OH groups. Valency of carbon is 4. If all these 4 valencies are occupied
by different atoms or group of atoms, then the carbon is
What is Aldehyde and Keto? asymmetric.
These are the functional groups of Carbohydrates. Either
Carbohydrate has Aldehyde group or Keto group. This is
carbonyl group present at C1 or C2. (Do not consider ‘OH’ as
functional group in carbohydrates)

F undamental B ox
CARBONYL GROUP (C=O)
zz If C = O is present at C1, it is Aldehyde group
zz If C = O is present at C2, it is Keto group

Contd…
 Symmetric Carbon
If any two, three or four valencies are occupied by same atoms
30 or group of atoms, then the carbon is Symmetric.
CRO BIOCHEMISTRY

Fig. 2.3: 3 Carbon Functional Isomers

H igh R eturn


zz Parent alcohol in Carbohydrates: Glycerol

zz Parent carbohydrate which gives rise to other carbohydrates
is D: Glyceraldehyde
Fig. 2.2: Symmetric and Asymmetric Carbons zz Simplest Carbohydrate is D: Glyceraldehyde
zz Minimum number of carbons possible in a carbohydrate: 3
zz Minimum number of –OH group possible in a carbohydrate:
2 (Recall: Number of OH is one less the number of carbons)
F undamental B ox zz Minimum number of Functional group possible in a carbo-
“Any compound having asymmetric carbon shows both structural hydrate: 1
and optical isomerism”. zz Monosaccharide with zero Asymmetric Carbon: Dihydroxy-
Carbohydrates and Amino Acids have asymmetric carbon, so both acetone (DHA)
structural and optical isomerism present. zz Number of asymmetric carbons in Ketoses are one less the
number in Aldoses

ISOMERISM IN CARBOHYDRATES
Compounds having same Molecular formula but different
structure are known as Isomers e.g. Glucose, Fructose,
Galactose, Mannose have same Molecular formula, i.e.
C6H12O6, but different structure.

Fig. 2.4: 6C Functional Isomers

Glucose and Fructose are Functional isomers of each other.

H igh R eturn
Functional Isomerism Number of Asymmetric carbons in Glucose = Four (C2, C3, C4, C5)
T In this isomerism, Molecular Formula is same but functional C1 and C6 are symmetric (C1 is C=O) (C6 is CH2OH)
H groups are different, i.e. one is aldehyde, other is keto form So, total no of isomers possible = 2n
E e.g. Glyceraldehyde and Dihydroxy-Acetone (DHA) are func- Where n = Number of asymmetric carbons
O tional isomers of each other (Fig. 2.3) So, there are 16 isomers possible for Glucose (24 = 16)
R
Y
Enantiomerism (Also known as D and L Isomerism)
They are also known as mirror images of each other. 31
TRICK for Q Solving: If mirror images written in some
Question of Isomerism, you are very sure it is Enantiomerism
Definition: Different –H and –OH orientation around the

CHAPTER 2  CHEMISTRY OF CARBOHYDRATES

penultimate carbon or 2nd last carbon. This carbon is also
known as Reference carbon. e.g. D-and L-Glyceraldehyde
(Fig. 2.5)

H igh R eturn
zz Which enantiomer of carbohydrate is abundant in body/
nature /plasma/cell? → D
zz All carbohydrates in our body are in D-form. Exception:
L-Fucose is present in glycoproteins. e.g. Blood group antigen
zz Which enantiomer of amino acid is abundant in proteins? → L
Fig. 2.5: 3 Carbon Enantiomers zz Which enantiomer of a free amino acid is present? → Both
D and L
zz Amino acid is present in body either in proteins or free amino
acid is present in amino acid pool of body. This free amino acid
may be present in D-form of L-form. But L-form is abundant.
E.g. of D-amino acids: D-Serine and D-Asparate are found in
brain and act as neurotransmitters.
(Note: That we have mentioned 'D' here to write the name of a 'D'
amino acid. But whenever the name of any amino acid is written
without mentioning 'D' or 'L' that means obviously it is 'L')
zz Which enantiomer of amino acid is present in body? → Both
D and L
zz The amino acids which we can synthesize in our body
are always in 'L'- form. So the source of 'D' amino acids is
exogenous i.e. from diet.

Diastereomers
Fig. 2.6: 6 Carbon Enantiomers: Now in case of Glucose,
second last carbon is C5. So, at C5, if OH is on right side, then it is Diastereomers are structural isomers having different H and
D-Glucose. And at C5, if OH is on left side, then it is L-Glucose OH orientation around C2, C3 and C4. But unlike Enantiomers,
they are not mirror images of each other E.g. Galactose and
A dditional E dge Mannose.
zz THINK: To make mirror image, we have to alter –H and
–OH orientation around all asymmetric carbons. But then
Epimerism
why definition of Enantiomerism mentions that –H and –OH Definition: Different –H and –OH orientation around only
orientation changed only around one carbon, i.e. 2nd last one carbon, other than the penultimate carbon. E.g. Mannose
carbon is epimer of Glucose at C2. Galactose is epimer of Glucose
 In case of Glyceraldehyde, there is only one Asymmetric
at C4.
carbon, i.e. C2
But in case of Glucose, there are four Asymmetric carbons.
So, when we make Enantiomer of Glucose, –H and –OH
orientation is changed at around four carbons (Asymmetric)
i.e. C2, C3, C4, C5
T
zz CONCEPT: Actually Enantiomerism is different –H and –OH
orientation around all asymmetric carbons. But the definition H
mentions only one carbon, which means that out of all E
asymmetric carbons, only one carbon is taken as Reference O
carbon where we have to look H and OH orientation to tell R
which is D and which is L isomer. So, this is known as Reference
Y
carbon.
Contd… Contd…
 also becomes Asymmetric. As isomerism is because of
Asymmetric carbon, so one more isomerism possible in
32 cyclic structures, i.e. Anomerism
Two types of cyclic structures are formed:
1. Pyranose– a 6 membered ring with one Oxygen and 5
CRO BIOCHEMISTRY

Carbons.

Anomerism 2. 
Furanose – a 5 membered ring with one Oxygen and 4


Definition: Different –H and –OH orientation around the Carbons.

Functional carbon.
zz Anomerism exist only in cyclic structures, which exist in
H igh R eturn
solutions Q. How many Carbons are present in the Pyranose ring?
zz In powder form → Carbohydrates are in their linear Answer is 5 (not 6). Because 5 Carbons and one is Oxygen
configuration. But in solution form, this linear structure Q. How many Carbons are present in the Furanose ring?
gets converted to cyclic configuration. Answer is 4 (not 5). Because 4 Carbons and one is Oxygen
zz To convert Linear structure to Cyclic structure, two
carbons in linear structure make a bond with oxygen in Glucose is mainly Pyranose, i.e. 99% Pyranose but 1%
between them. Furanose also exist.
zz Always functional carbon makes a bond with second last NOTE: Glucose is not always Pyranose. Glucose is mainly Pyranose.
carbon (to bring ald/keto group in close proximity to the Similarly, Fructose is mainly Furanose, i.e. 99% Furanose but
2nd last carbon’s –OH group). 1% Pyranose also exist.
NOTE: Fructose is not always Furanose. Fructose is mainly Furanose.

F undamental B ox
zz Hexoses can exist both as Pyranose and Furanose
zz But Pentoses (5C) can exist only as Furanose

For making Ring/Cyclic structure, minimum number of

carbons present should be 4.

zz Always functional carbon makes a bond with second last

carbon.
T
zz In case of Glucose, it’s C1 (functional carbon) and C5
H (2nd last). So, C6 is out of the ring.
E zz In case of Fructose, it’s C2 (functional carbon) and C5
O Functional carbon is Symmetric in case of linear (2nd last). So, C1 and C6 are out of the ring.
R configuration. But in Cyclic structures, functional carbon zz In case of Ribose, it’s C1 (functional carbon) and C4
Y (2nd last). So, C5 is out of the ring.
Two types of Anomers are → Alpha (α) and Beta (β) NOTE: D and L (in capitals) are Structural Isomers (Enantiomers)
1.   α means at Functional carbon, –OH is downwards But d and l (in small) are Optical Isomers.
2.   β means at Functional carbon, –OH is upwards 33
H igh R eturn
zz Glucose is always dextrorotatory (d)
Always

CHAPTER 2  CHEMISTRY OF CARBOHYDRATES

zz Fructose is always levorotatory (l)

Racemic Mixture
Equal d and l isomers present (better to use the word ‘equal’
not ‘both’). As they are present in equal amount, so Racemic
Summary of Structural Isomerism mixture is optically inactive.
1. Functional Functional group different (ald or keto)
isomerism Racemase Enzyme (Misnomer Name)
2. Enantiomerism Different –H and –OH orientation around zz Enzyme which interconverts D and L isomers into
the penultimate carbon each other (not small d and l). The name Racemase is a
3. Epimerism Different–H and –OH orientation misnomer.
around only one carbon, other than the
penultimate carbon H igh R eturn
4. Anomerism Different –H and –OH orientation around zz Structural Isomerism: Compounds with same Molecular
the functional carbon. formula but different structure
zz Optical Isomerism: Compounds with same Molecular formula
but different optical properties.
NOTE: F unctional Isomerism is different functional groups – one
is Aldehyde, other is Keto. But Anomerism is different –H What is Primary, Secondary and Tertiary Alcohol?
and –OH orientation around the Functional carbon.
zz Primary alcohol (1°) → OH is attached to that carbon,
which is further attached to one carbon.
Optical Isomerism zz Secondary alcohol (2°) → OH is attached to that carbon,
When a plane polarized light is passed through a Carbohydrate which is further attached to two carbons.
solution, then this light gets rotated either towards right or zz Tertiary alcohol (3°) → OH is attached to that carbon,
left. which is further attached to three carbons.
zz If the light gets rotated towards right, then the
carbohydrate present in the solution is known as dextro-
rotatory, also represented with d/ (+) sign.
zz If the light gets rotated towards left, then the carbohydrate
present in the solution is known as levorotatory, also
represented with l/ (–) sign.

Fig. 2.7: Primary, Secondary, Tertiary alcohol

CLASSIFICATION OF CARBOHYDRATES
Carbohydrates are also known as Saccharides. So, they
are classified as Monosaccharides, Disaccharides, Oligo- T
saccharides and Polysaccharides. H
E
O
R
Y
34
CRO BIOCHEMISTRY


Monosaccharides–Simplest Unit of Carbohydrates

Table 2.1: Monosaccharides

No of C Generic Example (Ald) Example (Keto)

atoms name
3 Trioses Glyceraldehyde Dihydroxyacetone
4 Tetroses Erythrose Erythrulose
5 Pentoses Ribose Ribulose
Xylose (epimer of Xylulose
Ribose) Fig. 2.8: Sialic Acid Structure
Arabinose
Reactions of Monosaccharides
6 Hexoses Glucose Fructose
Oxidation
Galactose
Mannose Monosaccharides on oxidation forms Acids.

7 Heptoses Glucoheptose Sedoheptulose Table 2.2: Oxidation of Monosaccharides

9 Nanoses Sialic acid (NANA) -------------- Oxidation at Carbon Acid

Oxidation at C1 Aldonic acid
H igh R eturn Oxidation at C6 (or Oxidation at Primary alcohol) Uronic acid
Glucose Oxidation at Both C1 and C6 Saccharic acid
zz Solution of Glucose is known as Dextrose
zz Glucose is the main energy source for Brain, RBCs, Retina, # In case of Glucose, C6 is primary alcohol (see Glucose structure)
Cornea, Renal Medulla, Testis
zz RBCs can only use Glucose (in Fed/Fasting/Starvation) Reduction
zz Glucose is the universal fuel for Fetus. On reduction, carbohydrates forms their Alcohols.
T
H All these alcohols are hygroscopic in nature. Hygroscopic
Nanoses means they absorb water. So, if this hygroscopic alcohol is
E
One example is Aldehyde form, i.e. Sialic acid, which is a present in excess in a cell, it will lead to cell swelling. e.g.
O
modified 9 C sugar. Sialic acid is rich in functional groups 1. Glucose forms Sorbitol which gives rise to Snow Flake
R
(one N-acetyl group, three Secondary alcohol groups, a Cataract
Y
Primary alcohol group, and a Carboxylic acid group), which 2. Galactose forms Galactitol/Dulcitol which gives rise to
gives the possibility of various binding sites. Sialic acid is often Oil Drop Cataract
incorporated in Oligosaccharides as a terminal residue. It is a 3. Mannose forms Mannitol which is used to reduce intra
constituent of Glycoproteins and Gangliosides (Glycolipids). cranial pressure
 H igh R eturn
zz Type of Cataract in Diabetic patient: Snow Flake Cataract 35
zz Type of Cataract in Galactosemia patient: Oil Drop Cataract

F undamental B ox

CHAPTER 2  CHEMISTRY OF CARBOHYDRATES

Q. Sorbitol – also known as Polyol. Why?
A. Because it has 6 C, 6 OH groups
(Recall: Glucose has 6C, 5 OH groups, so Sorbitol has 6C
and same number of –OH groups)

Fig. 2.9: Structure of Disaccharides

Disaccharides
Table 2.4: Tests Given by Monosaccharides and Disaccharides
Means 2 monosaccharides joined by a bond. That bond is
known as Glycosidic bond. Test Features
1. Molisch test General test for Carbohydrates. But one
Table 2.3: Disaccharides
condition is number of carbons should be 5
Disaccharide Carbohydrate Bond Reducing or more, only then Molisch test is positive
units nature 2. Benedict’s test Given positive by all Reducing Sugars
Maltose Glu + Glu α (1 → 4) Reducing 3. Barfoed’s test Distinguish between Monosaccharides
and Disaccharides. It is given positive by
Isomaltose Glu + Glu α (1 → 6) Reducing
Monosaccharides.
Trehalose Glu + Glu α(1 → 1) Non-Reducing
4. Seliwanoff’s Distinguish between Aldehyde and Keto
Sucrose Glu + Fruc α(1 → 2) Non-Reducing test sugar. It is given positive by Keto sugars.
Lactose Gal + Glu β(1 → 4) Reducing

NOTE: A
 ll Monosaccharides are Reducing (Reducing means func-
tional group is free)

T
H
E
O
R
Fig. 2.10: Tests for Carbohydrates Y
zz Osazones: These are crystals & all reducing sugars form
Osazones.
 Table 2.5: Shapes of Osazone crystals Oligosaccharides
36 Carbohydrate Shape of Osazone crystal They do not exist as such in nature. They are just breakdown
products of polysaccharides. e.g. Maltotriose – a trisaccharide
Glucose, Fructose, Needle shaped or broom stick like
Mannose appearance
[made up of 3 Glucose units, joined together by alpha (1 → 4)
glycosidic bonds]
Galactose Rhombic plate
CRO BIOCHEMISTRY

Maltose Sunflower shaped

Lactose Powder puff/ Hedgehog appearance


Homopolysaccharides
1. Starch
Major carbohydrate in diet

2. Glycogen
Storage form of carbohydrate in Liver and Muscle

Table 2.7: Difference between Starch and Glycogen

Homo Present in Branching Branching

polysaccharide point comes
after
Starch Plants Less 24–30 Glucose
(Storage form) branched residues
Fig. 2.11: Osazones
Glycogen Animals More 8–12 Glucose
GOD-POD Test (Storage form) branched residues
An enzymatic quantitative test, specific for Glucose. This
is a cheap and accurate method as it is enzymatic method. Similarities between Starch and Glycogen
(Enzymes are very specific for their substrates). So, this is the
routinely used method for Blood Glucose estimation. zz Both are made up of a-Glucose residues.
zz Bonds in straight chain is alpha (1 → 4) bonds

NOTE: Only the branch point is alpha (1 → 6). Not the full branch
is alpha (1 → 6).

In this test, Glucose in the sample is first acted by enzyme

– Glucose Oxidase (GOD), which oxidizes C1, making Aldonic
acid (in case of Glucose, known as Gluconic acid) plus H2O2
produced. This H2O2 is acted by enzyme POD (Peroxidase),
which will produce some colored compound. By using
T principle of colorimetry, we can estimate the amount of
H Glucose from this colored compound. This way quantitative
E estimation of Glucose can be done with very accurate results.
O Table 2.6: Methods of Estimation of Glucose
R Reductometric Methods Enzymatic Method
Y
• Nelson’s Somogyi method • Hexokinase method (Gold
• Folin-Wu method Standard)
• Ortho- Toluidine method • GOD-POD test
 → 6) bonds. So, after the combined action of Amylase and
Isomaltase, we get lots of Disaccharides – Maltose. Maltose
is digested by Maltase enzyme, which breaks the remaining 37
alpha (1 → 4) bonds, releasing many Monosaccharides, i.e.
Glucose.

CHAPTER 2  CHEMISTRY OF CARBOHYDRATES

Fig. 2.12: Digestion of Starch in Intestine

3. Dextran
zz Complex HMW (high molecular weight) homopolysac-
charide made up of a-Glucose
zz Highly branched with many bonds, which are not broken
by human enzymes.
Starch is made up of:
zz Amylose: 20%, unbranched Uses
zz Amylopectin: 80%, branched
zz If given i/v (intravenously) then it is not broken by any
As only Amylopectin is branched. So, when we compare
enzyme plus it is high molecular weight, so cannot leave
Glycogen with Starch, we use the word Amylopectin. (We will
intravascular compartment. So, it is used as a plasma
use this concept in Glycogen Storage Diseases).
volume expander in patients of hypovolemic shock.
Iodine test: Positive for Polysaccharides which is zz Dental plaques contain network of Dextran
indicated by formation of blue/brown/red colour. This color zz The gel used in Gel Filtration Chromatography is Dextran
is due to formation of a complex between Iodine and the (commercial name is Sephadex)
Polysaccharide due to adsorption.
4. Cellulose
Most Homopolysaccharides are branched but Cellulose is
unbranched.
Cellulose is made up of β Glucose.
“Beta bond is difficult to be broken”. So, cellulose having
beta bond is not broken in our body. So, it acts as fiber in the
diet. It is an insoluble fiber. (Humans lack enzyme Cellulase,
which breaks beta bond in Cellulose)

Table 2.8: Difference between Soluble and Insoluble Fiber

Insoluble fiber Soluble fiber

Digestion of Starch e.g. Cellulose, Hemi- e.g. Pectins, Gums, Psyllium, Inulin T
cellulose, Lignin H
Amylase starts breakdown of Starch in intestine. Amylase
Excreted unchanged from They absorb water, get converted E
can only break alpha (1 → 4) bonds. So, after the action intestine, in its original to gel form and get excreted
of Amylase, a structure is left having lots of alpha (1 → 6)
O
form R
bonds, which is known as Limit Dextrins. Limit Dextrins are
then broken by enzyme Isomaltase, which breaks alpha (1 Prevent constipation Better in preventing constipation Y
 5. Inulin Properties of GAGs
Made up of β-Fructose. Beta bond is difficult to be broken. So, Unbranched, synthesized in ER and Golgi
38 Inulin is not broken in our body
zz
zz Highly sulfated
Uses of Inulin zz Tandem repeats of amino sugar and uronic acid
zz Attached to proteins making proteoglycan
Ideal substance for the measurement for GFR
CRO BIOCHEMISTRY

zz
zz Carboxy, Sulfate and Acetyl groups gives too much of
zz Act as a Prebiotic (not Probiotic). negative charge, which repels, giving them slimy and
A dditional E dge slippery nature. That’s why they are present in mucus
secretions. (act as lubricant, or as shock absorber)
Prebiotic is like a food for the bacteria (Probiotic), which is given zz Amino group in amino sugars comes from Glutamine
for the healthy growth of the bacteria. E.g. Inulin has beta bond, so
cannot be broken in humans, so when Inulin is not broken in small
intestine, this will reach large intestine where bacteria present
can break this beta bond and they can use this carbohydrate
(Fructose) as their food.


H igh R eturn
“Beta bond is difficult to be broken”
Exceptions:
If beta bond is present on the side of Galactose, then beta bond
becomes easy to be broken. Any such enzyme therefore is known
as Beta Galactosidase e.g.
1. Lactase: deficient in lactose intolerance (Lactase breaks
lactose, which is galactose joined to glucose by beta bond)
2. Beta Galactosyl Ceramidase: deficient in Krabbe’s disease
(this enzyme breaks Beta-Galactosyl ceramide, a Glycolipid in
which Galactose is joined to Ceramide by beta bond).

6. Chitin
Chitin is made up of N-Acetyl Glucosamine residues in
β (1 → 4) linkage. It is a structural polysaccharide in the
exoskeleton of insects.
Table 2.9: Homopolysaccharides
Fig. 2.13: Structure of Proteoglycan (Bottle Brush appearance)
Polysaccharide Bond Constituent
Amylose a (1 → 4) Glucose Structure of Proteoglycan (Figure 2.13)
Starch
Amylopectin a (1 → 4), a (1 → 6) Glucose GAGs are attached to proteins, therefore called Proteoglycans.
Glycogen a (1 → 4), a (1 → 6) Glucose On Hyaluronic acid backbone, many core proteins and link
Dextran a (1 → 4), a (1 → 6) Glucose proteins are attached. Linear GAG chains of Amino sugars
a (1 → 2), a (1 → 3) and Uronic acid (tandem repeats) are covalently attached
to core proteins. This linear chain of GAG is linked to core
Cellulose b (1 → 4) Glucose
protein by a core trisaccharide (Galactose-Galactose-Xylose).
Inulin b (2 → 1) Fructose
Chitin b (1 → 4) N-Acetyl No need to No need to
learn these. learn these.
Glucosamine
Just read once Just read once
Heteropolysaccharides/GAGs/Mucopolysaccharides GAGs Amino sugar Uronic acid Location
Hyaluronic N-Acetyl D-Glucuronic Synovial fluid,
T
H
F undamental B ox Acid Glucosamine Acid Vitreous Humor,
Let's understand the names: (longest) Role in Wound
E
zz Heteropolysaccharides: Because they are made up of different Healing and cell
O Migration during
Carbohydrate units
R Morphogenesis,
zz GAGs (Glycosaminoglycans): Because they have lots of amino
Y groups (Note: Usually Carbohydrates do not contain amino Tumor cell
group. Amino acids and Proteins contain amino group) migration
zz Mucopolysaccharides: Because they are found in mucus Contd…
secretions
 No need to No need to Role in retinal cell attachment: Heparan Sulfate
learn these. learn these. Cell-cell adhesion: Heparan Sulfate
Just read once Just read once 39
Longest GAG: Hyaluronic Acid
GAGs Amino sugar Uronic acid Location zz All GAGs are covalently attached to Proteins, EXCEPT: Hya-
luronic Acid which is noncovalently attached to other Proteo-
Chondroitin N-Acetyl D-Glucuronic Cartilage, glycans in ECM.

CHAPTER 2  CHEMISTRY OF CARBOHYDRATES

Sulfate Galactosamine Acid Bone, Tendon, zz GAG synthesized mainly by arterial Smooth Muscle Cells:
(most 4-Sulfate Ligament, Dermatan Sulfate
abundant) Cornea zz Dermatan Sulfate binds LDL and has role in atherosclerosis
Keratan N-Acetyl D-Galactose Cornea (Type I zz Cornea has mainly Keratan Sulphate, but also some Chon-
Sulfate Glucosamine Keratin Sulfate), droitin Sulfate
6-Sulfate Cartilage (Type II zz Mostly GAGs are extracellular but Heparin is intracellular.
Keratin Sulfate) zz Mucin Clot Test/ Rope Test → If few drops of synovial fluid

Dermatan N-Acetyl L-Iduronic Acid Skin, Blood is added to Acetic acid, clot is formed due to polymerization
Sulphate Galactosamine Vessels, Valves of Hyaluronic Acid. Poor clot formation occurs in Rheumatoid
(Wide 4-Sulfate Arthritis, Septic Arthritis, Gouty Arthritis (inflammatory
Distribution) conditions). But it is a nonspecific test.

Heparin N-Sulfo L-Iduronic acid Anticoagulant Table 2.10: Difference between Proteoglycan and
Glucosamine (Mast cells and Glycoprotein
Liver)
Heparan N-Sulfo D-Glucuronic Cell surfaces Proteoglycan Glycoprotein
sulfate Glucosamine Acid (role in Retinal Carbohydrate >>>> Protein Protein >>>> Carbohydrate
cell-cell
Carbohydrate portion is always Carbohydrate portion is
attachment),
a Heteropolysaccharide either a Monosaccharide or
in Basement
Oligosaccharides (Never a
Membrane of
Polysaccharide)
Glomerulus
Carbohydrate is long and linear Carbohydrate is short and
H igh R eturn highly branched
e.g. Aggrecan, Syndecan e.g. Collagen
Hyaluronic Acid
zz In Extracellular Matrix (ECM)
zz Present in both bacteria and animals
Mucopolysaccharidosis (MPS)
zz No Sulfate in Hyaluronic Acid These are Autosomal Recessive diseases. They come under
zz Role in Cell Migration during Morphogenesis Lysosomal Storage Diseases as the lysosomal enzyme is
zz Role in Wound Healing absent which normally breaks down Mucopolysaccharides.
So, Mucopolysaccharides starts getting accumulated in
Heparin lysosomes.
More sulfated than Heparan Sulfate. Has highest negative
charge (because of Sulfate). This too much of negative charge Clinical Feature
chelates the positive charge of calcium, leading to anti- zz Coarse facial features, depressed nasal bridge, frontal
coagulation. Heparin binds Anti-thrombin, Factor IX and bossing
Factor XI. zz Copious nasal discharge
zz Short stature due to growth retardation
H igh R eturn zz Protuberant abdomen due to Umbilical Hernia or
Most widely distributed GAG: Dermatan Sulfate (a relatively Hepatosplenomegaly
smaller GAG) zz Corneal Clouding/Opacity due to direct infiltration of
Most abundant GAG: Chondroitin Sulfate (a very large mole-cule) GAG in cornea
zz Most heterogeneous GAG: Keratan Sulfate I and II because zz Clawing of hands
they contain additional monosaccharides such as L-Fucose, zz Thickening of cardiac valve T
Mannose and NANA. zz Hearing impairment H
Highest Negative Charge: Heparin Skeletal abnormalities- Dysostosis Multiplex due to
zz E
zz No sulfate: Hyaluronic Acid defective bone formation, bullet shaped middle phalanx.
zz No Uronic Acid: Keratan Sulfate
O
zz Corneal transparency: Keratan Sulfate
R
zz Major component of cartilage: Chondroitin Sulfate Y

Contd…
40
CRO BIOCHEMISTRY

Fig. 2.14: Bullet shaped middle phalanges in Mucopolysaccharidosis



zz Presence of Mucopolysaccharides in urine Table 2.11: Mucopolysaccharidosis

zz Reilly Body Inclusions or Alder-Reilly bodies in
Leukocytes (specially granulocytes). These are large, Type Disease Enzyme Special GAG
name defect features accumulated
coarse, dark purple granules or inclusions in cytoplasm.
Specially these inclusions are seen in Hurler disease MPS Hurler Alpha-L- Inguinal DS + HS
I–H disease Iduronidase hernias
(AR) often
present
MPS I–S Scheie Alpha-L- No mental DS
(AR) disease Iduronidase retardation
MPS II Hunter Iduronate No corneal DS + HS
(XR) disease Sulfatase clouding,
(Mild Exclusively
Hurler + males
Aggressive affected
behaviour)
MPS VI Maroteaux Aryl No mental DS
(AR) Lamy Sulfatase B retardation
Syndrome
• DS – Dermatan Sulfate
• HS – Heparan Sulfate
Fig. 2.15: Reilly bodies Table 2.12: Enzyme Replacement Therapy for Mucopoly-
saccharidosis
H igh R eturn Disease Name Enzyme Used
zz All diseases are Autosomal Recessive, EXCEPT Hunter disease, TYPE - I Hurler disease Aldurazyme
which is X-linked recessive (so exclusively in males)
zz Accumulation of Heparan Sulfate is responsible for Mental TYPE - II Scheie’s disease Elaprase
retardation TYPE - VI Hunter’s disease Naglazyme
zz Accumulation of Dermatan Sulfate is responsible for
Atherosclerosis For Type-III (Sanfilippo) and Type-IV (Morquio), Enzyme
zz Most common MPS → Sanfilippo (Type III) > Hunter (Type II) > Replacement Therapy is currently under clinical trial stage.
T
Hurler (Type I)
H I-cell Disease or Inclusion Body Disease
zz MPS with no corneal clouding → Hunter and Sanfilippo
E zz Alcian Blue Spot test is screening test for MPS (detects urinary zz a Mucolipidosis
O GAGs) zz a Lysosomal Protein Targeting Disorder
R zz Not a Mucopolysaccharidosis but clinical features are
Y same like MPS specially Hurler disease
zz But clinical features are more severe in I-cell Disease
than MPS.
zz Normally Mannose is present on Hydrolases and this 41
Mannose is phosphorylated to Mannose-6-Phosphate by
enzyme N-Acetyl Glucosamine Phospho Transferase
(present in Golgi Apparatus). Phosphorylation is

CHAPTER 2  CHEMISTRY OF CARBOHYDRATES

required so that these Hydrolases reach lysosomes and
take part in digestion of Mucolipids inside lysosomes.
zz In I-cell disease, this enzyme is defective. So, Hydrolases
do not reach lysosomes and undigested material
accumulated in lysosomes, known as Inclusion Bodies.
Fig. 2.18: Secondary Active transport
GLUCOSE TRANSPORT
Glucose is polar (polyhydroxy) but membrane is lipid bilayer, Active Transport/Na+ Glucose Symport is present in:
1. Intestine 2. PCT (Kidneys)
i.e. Non polar. So, how polar glucose will cross the cell
membrane and enters the cells as glucose is the main fuel for
the cells.
H igh R eturn
So, we need a carrier in the membrane which acts like a zz SGLT-1 → Small intestine and Kidneys → for Glucose and
channel in the membrane for the transport of Glucose. Galactose transport
zz SGLT-2 → in Kidneys → only for Glucose

GLUTs (Glucose Transporters)/Facilitative

Diffusion/Sodium Independent Glucose Transport
Active Transport is present only in Intestine and Kidneys. So,
mainly for Glucose Transport, GLUTs are present. GLUTs are
bidirectional.

Q. Whenever Glucose enters a cell, quickly it is

converted to Glu-6-P. Why?
Fig. 2.16: Glucose transport in cell T A. Because if it is Glucose, it can escape the cell via
H GLUT as GLUTs are bidirectional. But if it is Glu-6-P,
I
then it cannot escape the cell via GLUT.
N
K So, we can say that phosphorylation of Glucose is
done for the entrapment of Glucose inside the cells.

Table 2.13: GLUTs (Glucose Transporters)

GLUT Tissue Location Function

GLUT-1 Brain, Placenta, Kidney, Basal Glucose uptake
Fig. 2.17: Glucose transport is carrier mediated RBC
GLUT-2 Liver, Pancreas, Intestine- absorption,
Active Transport Intestine Liver-Glycogen formation,
Also known as Sodium Glucose Symport or Sodium Pancreas-Insulin secretion
Dependent Glucose Transport (SGLT)
GLUT-3 Brain (Neuronal), Basal Glucose uptake
Q. Why it is known as Secondary Active Transport or Placenta, Kidneys
indirectly Active Transport?
T A. Explanation: ‘Symport’ means same direction, i.e. GLUT-4 Skeletal muscles, Insulin stimulated Glucose
H both Glucose and Na are entering inside the cell (in
Cardiac muscles , uptake after meals
I Adipose tissues
T
same direction). No energy used when glucose and
N GLUT-5 Small intestine, Testis Fructose transporter
K Na+ enters the cell. H
But Na+ has to be thrown out of cell otherwise water (Sperms) E
will enter and cell will burst and die. So, Na+ is thrown O
GLUT-6 WBC, Spleen Not known
out with the help of Na+/K+ ATPase pump, which R
requires energy. So, energy is not used when glucose GLUT-7 Liver, Endoplasmic Glucose transporter in ER Y
enters. Energy is used indirectly for Glucose transport Reticulum
via Na+/K+ ATPase pump (Fig 2.19).
Contd…
 GLUT Tissue Location Function Q. GLUT 1 and 3 dependent on Insulin?
42 GLUT-8 Blastocyst, Testis, Brain Insulin dependent, T A. No
can transport Glucose, H Exp: GLUT 1 and 3 are active in Fasting state. Insulin
Fructose and Galactose I always comes in Fed state. So, there is no relation
GLUT-9 Liver, Kidney Urate transporter
N between them.
CRO BIOCHEMISTRY

K Q. GLUT-2 dependent on Insulin?

GLUT-11 Heart, Skeletal muscles Fructose transporter A. No
GLUT-2 works before Insulin. So, GLUT-2 is not
dependent on Insulin.
Q. GLUT-4 dependent on Insulin?
A. Yes
ONLY GLUT-4 is dependent on Insulin. Insulin
recruits GLUT-4 on the membrane of peripheral cells.

A dditional E dge


Q. Why we suggest Diabetic patient to do little exercise or

a brisk walk after each meal?
A. Because exercise can also activate GLUT-4, and this effect
does not requires insulin. This effect requires 5’ AMP
Fig. 2.19: Glucose entry via GLUT-2 releases Insulin from Pancreatic cells activated kinase.

Insulin and GLUT-4

H igh R eturn
Once Insulin is released in blood, it will make GLUT-4 active.
GLUT (MCQs)
GLUT-4 is present in Peripheral tissues (Skeletal muscles,
zz Most widely distributed GLUT → GLUT-1
Cardiac muscles, Adipose tissues) so that Glucose enters the
zz Most abundant in RBCs → GLUT-1
Peripheral tissues and this way Blood Glucose is decreased, zz Major GLUT in brain and placenta → GLUT-1
leading to hypoglycemic action of insulin. zz Major GLUT in neurons → GLUT-3
When Glucose enters muscles, it is converted to Glycogen. zz GLUT-1 and GLUT-3 both are in Brain
When Glucose enters Adipose tissues, it is converted to Fats. zz GLUT-3 is in neurons but not GLUT-1

NOTE: Carbohydrates can be converted to Fats, But Fats cannot

be converted to Carbohydrates.

H igh R eturn
zz Facilitative Transport is also known as Sodium Independent
Glucose Transport (GLUT)
zz Secondary Active Transport is also known as Sodium Depen-
dent Glucose Transport (SGLT)

T Fig. 2.21: Glucose Transport in Intestine

H
We have both Active transport and Facilitative transport
E
present in Intestine. Type of active transport present is
O
SGLT-1 (at the Apical membrane, which is towards Luminal
R side). Type of GLUT present is GLUT-2 (in the Basolateral
Y membrane, which is towards the interstitial fluid side)

Fig. 2.20: Sequence of Events in Fed State

Table 2.14: Mutation and the Disease Caused Procedure of GTT Test

Mutation Disease caused

A fasting blood and urine sample is 43
collected. Then a Glucose load of 50 gm
SGLT-1 Glucose-Galactose Malabsorption is given to the patient. Blood and urine
samples are collected at hourly interval
SGLT-2 Familial Renal Glucosuria

CHAPTER 2  CHEMISTRY OF CARBOHYDRATES

for next 3 hours. Glucose is estimated in
GLUT-2 Fanconi-Bickel Syndrome all these samples and a curve is plotted with blood glucose
levels on y-axis and time of collection on X-axis.
GTT – Glucose Tolerance Test Indications of GTT- Glucose Tolerance Test
A normal body is able to tolerate glucose load as glucose zz Asymptomatic persons with sustained or transient
can enter the peripheral cells via GLUT-4 which is insulin glucosuria
dependent. But Glucose tolerance can be decreased or zz Person with symptoms of DM, but no hyperglycemia or
increased in various conditions. glucosuria
zz A person with or without symptoms of DM, with one
H igh R eturn abnormal blood finding
Decreased Glucose tolerance seen in conditions causing Hyper- zz In patient with neuropathy or retinopathy of unknown
glycemia: origin
zz Diabetes Mellitus zz Women with history of having delivered a large baby
zz Hyperactivity of Anterior Pituitary, Thyroid, Adrenal Cortex zz Gestational DM
zz Stress
Increased Glucose tolerance seen in conditions causing Hypo- Instructions for GTT
glycaemia:
zz Decreased function of Pituitary, Thyroid, Adrenal Cortex
zz Normal carbohydrate diet 3 days prior to test
zz Increased Insulin
zz Avoidance of drugs which affect blood glucose levels
zz Decreased gastrointestinal absorption e.g. Celiac disease zz No smoking and heavy exercise on the day of test

Fig. 2.22: Normal GTT and GTT in diabetic patient

T
H
Normal GTT: Fasting blood glucose is well within normal levels falls due to entry of blood glucose into the cells via E
range. There is rise in blood glucose after glucose load and GLUT-4. Then normal levels are reached within 150 minutes, O
peak is observed at 1 hour, due to absorption of blood glucose i.e. after 2.5 hours. Glucosuria never occurs – all urine samples R
from intestine. Then there is release of insulin and glucose are negative for Glucose.
Y
 Diabetic Curve HbA1c/ Glycated or Glycosylated Hemoglobin
44 Fasting blood glucose is higher than normal. The highest There occurs irreversible binding of Glucose to Hb non
value is reached at 1 hour or 1.5 hour. This highest value enzymatically. It is a ratio (Glycated Hb/Total Hb). It has no
exceed renal threshold. Glucose is found in almost all the units. Around 5% fraction of total Hb is glycosylated when
urine samples. Blood glucose does not return to normal blood glucose is within normal limits.
CRO BIOCHEMISTRY

levels even after 2.5 to 3 hours of taking glucose.

zz Impaired Glucose Tolerance: Blood Glucose levels are Benefits
above normal but less than diabetic values. zz Tells Glucose control over past 2-3 months (as the RBC
zz Alimentary Glycosuria: A temporary condition which lifespan is 120 days)
occurs due to excessive ingestion of Glucose. Excessive zz Does not require a fasting sample
Glucose appears in urine producing Glycosuria.
zz Renal Glycosuria: Blood Glucose is normal but urine HbA1c levels Significance
sugar is present due to decreased renal threshold 3.8–6.4% Normal
(Normal is 180 mg/dl). Mostly it occurs due to Mutation 5.7–6.4% Pre-diabetes
in SGLT-2.


≥ 6.5% Diabetes Mellitus

Pearls of the Chapter

zz Hydroxy group always gives polarity, has high tendency to bind Phosphate.
zz Aldehyde group always present at C1
zz Keto group always present at C2
zz Functional Isomerism → Functional group different (Ald or Keto)
zz Enantiomerism → Different–H and –OH orientation around the Penultimate carbon
zz Epimerism → Different –H and –OH orientation around only one carbon, other than the Penultimate carbon
zz Anomerism → Different –H and –OH orientation around the Functional carbon.
zz Functional carbon is Symmetric in linear configuration, but Asymmetric in cyclic configuration.
zz In Molecular formula of Carbohydrates, n =Number of Total carbons.
zz In Isomeric formula of Carbohydrates, n = Number of Asymmetric carbons.
zz If all the four valencies of carbon are occupied by different atoms or group of atoms, then the carbon is Asymmetric.
zz If any two, three or four valencies are occupied by same atoms or group of atoms, then the carbon is Symmetric.
zz Any compound having Asymmetric carbon will show both optical and structural Isomerism
zz GLUT -1 is in Brain, Placenta, Kidneys and also in RBC
zz GLUT-3 is in Brain (neuronal), Placenta, Kidneys
zz Phosphorylation of Glucose is done for the entrapment of Glucose inside the cells
zz Affinity is inversely proportional to Substrate concentration
zz Hypoglycemic action of Insulin is via GLUT-4
zz Enantiomerism, also known as -D and -L Isomerism, also known as mirror images of each other
zz Pentoses (5C) can exist only as Furanose
zz Anomerism exist in Cyclic structures or in Solutions
zz In cyclic structures, Functional Carbon makes a bond with second last carbon
zz Mannose and Galactose are not Epimers of each other, as they differ in two carbons
zz Monosaccharides on oxidation forms Acids
T zz Monosaccharides on reduction forms Alcohols
H zz Molisch Test: General test for Carbohydrates. But one condition → number of carbons should be 5 or more, only then Molisch test is
E positive
O zz Benedict’s Test: Given positive by all Reducing sugars
R zz Barfoed’s Test: Distinguish between Monosaccharides and Disaccharides. It is given positive by Monosaccharides.
Y
Contd…
zz All Monosaccharides are Reducing
Seliwanoff’s Test: Distinguish between Aldehyde and Keto sugar. It is given positive by Keto sugars.
45
zz

zz Heteropolysaccharides and Cellulose are unbranched

zz Glycogen is more branched than Starch
zz Cellulose is not broken due to beta anomerism at C1 and Inulin is not broken due to beta anomerism at C2

CHAPTER 2  CHEMISTRY OF CARBOHYDRATES

zz Beta bond is difficult to be broken but if beta bond is on one side of Galactose then it can be broken. Any such enzyme is known as Beta
Galactosidase. E.g. Lactase and Beta Galactosyl Ceramidase
zz Cellulose is most abundant polysaccharide in nature. Chitin is second most abundant.
zz Pectin is a heteropolysaccharide made up of D-Galacturonic acid in α(1 → 4) linkages. It is a soluble dietary fiber in fruits. Softening of
ripe tomato occurs by enzyme Poly Galacturonase. Gene knock down of enzyme’s gene helps to preserve tomatoes.
zz Heparan Sulfate in excess is responsible for Mental Retardation
zz Dermatan Sulfate in excess is responsible for Atherosclerosis
zz No sulfate in Hyaluronic Acid
zz Hyaluronic Acid is not attached to Proteins
zz Heparan has highest negative charge
zz Keratan Sulfate does not contain Uronic acid
zz GAGs are slimy and slippery because of negative charge given by Carboxy, Sulfate and Acetate.
zz Malt contains β-Amylase which hydrolyses Starch into Maltose by sequential removal of disaccharide units from the Non-reducing
ends.
zz Blood Group Antigens (A,B,O) are Glycosphingolipids in RBCs and Glycoproteins in secretions, where Carbohydrate is Oligosaccharide.
zz D-Lyxose is a Pentose sugar and is a constituent of ‘Lyxoflavin’ isolated from human heart muscle.
zz Bial’s test: is specific for Pentoses (bluish color). Hexoses generally react to give green, red or brown products.
zz Inositol – A polyhydroxy alcohol, not a Vitamin

T
H
E
O
R
Y
 46 Multiple Choice Questions
Isomerism 11. A young man finds that every time he eats dairy
1. Which of the following is NOT correct? products, he feels very uncomfortable. His stomach
 (Recent Question 2018) becomes distended. He develops gas and diarrhoea
a. Parent carbohydrate which gives rise to other frequently. These symptoms do not appear when he
carbohydrates is Glycerol eats food other than dairy products. Which of the
b. Minimum number of carbons possible in a following is most likely enzyme in which this young
carbohydrate is 3 man is deficient: (Recent Question 2016)
c. Minimum number of ‘OH’ group possible in a a. Alpha amylase b. Beta galactosidase
carbohydrate is 2 c. Alpha glucosidase
d. Minimum number of functional group possible in a d. Sucrase
carbohydrate is 1 12. A male child presented with coarse facies, protu-
2. Which of the following statement about isomerism is berant abdomen, frontal head enlargement,
NOT correct? (Recent Question 2016) thickening of cardiac valve, hepatosplenomegaly,
a. Racemic mixture is equal d and l isomers present aggresive behaviour, clear vision and hearing
b. Racemic mixture is optically inactive impairment. What is the most probable diagnosis?
c. Racemase enzyme interconverts d and l isomers  (AIIMS Nov 2018)
into each other a. Hurler’s disease
d. Enantiomerism is also known as D and L-Isomerism b. Hunter’s disease
3. Dextrose is:  (Recent Question 2017) c. Fragile X syndrome
a. D + Glucose b. D - Glucose d. Tay-Sachs disease
c. L + Glucose d. L – Glucose 13. All of the following should be avoided by a patient
4. Number of isomers possible for Glucose are:  with lactose intolerance, EXCEPT (AIIMS May 2018)
 (Recent Question 2017) a. Condensed milk b. Ice-cream
a. 32 b. 64 c. Skimmed milk d. Yoghurt
M c. 16 d. 8 14. Which of the following does not exist in cyclic
C 5. Parent alcohol in carbohydrates is: structure?
Qs  (Recent Question 2016) a. Glyceraldehyde b. Glucose
Ans. a. Glycerol b. Ethanol c. Erythrose d. Ribose
c. Methanol d. Cholesterol 15. Which of the following enzyme helps in catalyzing
1. a
6. Which form of Glucose and Fructose is predominant? conversion of aldose sugars to ketose sugars?
2. c
a. a b. β  (PGMEE 2010)
3. a
c. Both d. Variable a. Oxidoreductase b. Aldolase
4. c
7. Cellulose is not broken due to beta anomerism at: c. Decarboxylase d. Isomerase
5. a
 (Recent Question 2018) 16. Which test is given positive by Glyceraldehyde?
6. b
a. C1 b. C2  (Recent Question 2016)
7. a
c. C5 d. C6 a. Benedicts test b. Molisch test
8. a,d
8. Which of the following are epimers: (PGI Nov 2011) c. Seliwanoff’s test d. Gerhard’s test
9. c,d,e
a. D-Galactose and D-Glucose 17. Which of the following test CANNOT be done for
10. b
b. D-Galactose and L-Glucose Glucose estimation?
11. b
c. D-Mannose and L-Mannose a. Glucose oxidase
12. b
d. D-Mannose and L-Glucose b. Dextrostix
13. d
e. D-Glucose and L Glucose c. Ferric chloride test
14. d
9. Which of the following is a keto sugar? (PGI Nov 2017) d. Nelson somogyi method
15. d
a. Glucose b. Sorbitol 18. Glycosaminoglycans present in cornea:
16. a
c. Fructose d. Sedoheptulose  (PGMEE 2015)
17. c
a. Dermatan sulfate b. Chondroitin Sulfate
18. d e. Ribulose
c. Hyaluronic acid d. Keratan Sulfate
19. c
19. Sucrose is hydrolyzed by (PGMEE 2009)
20. a
Classification of Carbohydrates a. Saccharase b. Sucrose Phosphorylase
10. Which of the following is a component of poly- c. Invertase d. Amylase
saccharide Chitin?  (PGMEE 2009) 20. Side chain linkage in Proteoglycans: (PGMEE 2015)
a. Ascorbic acid b. Glucosamine a. Covalent b. Hydrogen bond
c. Synovium d. Glucuronic acid c. Van der Waals force d. Electrostatic bond
21. Heparin is a: (PGMEE 2015) 35. Saccharic acid is produced as a result of:
a. Glycosaminoglycan b. Carbohydrate a. Oxidation of C1 of Glucose
c. Proteoglycan d. Polysaccharide b. Oxidation of C6 of Glucose 47
22. Reilly bodies are seen in? (PGMEE 2012) c. Reduction of C1 of Glucose
a. Behçet’s disease b. Gangliosidosis d. Oxidation of both C1 and C6 of Glucose
c. Gaucher’s disease d. Hurler disease 36. Which of the following is an ideal substance to check
23. Which disaccharide is NOT broken down in GIT? Glomerular Filtration Rate (GFR)?
 (PGMEE 2013) a. Inulin b. Phenol red
a. Lactulose b. Maltose c. Creatinine d. Cr51 EDTA
c. Sucrose d. Lactose 37. Which of the following test is positive for poly-
24. Glucosamines used in following condition: saccharides only?
 (AIIMS Nov 2017) a. Benedicts test b. Seliwanoff’s test
a. Arthritis b. Niemann pick disease c. Iodine test d. Molisch’s test
c. Alzheimer’s disease d. Cancer 38. Which of the following is not fermented by Gastroin-
25. D-Xylose test is used in diagnosis of testinal microorganisms?
 (PGMEE 2015, 2011) a. Lignin b. Pectin
a. Zinc deficiency c. Cellulose d. Hemicellulose
b. Malabsorption syndrome 39. All are true about glycosaminoglycans EXCEPT:
c. Coeliac sprue  (PGI May 2017)
d. Bacterial overgrowth syndrome a. Protein associated with glycosaminoglycans is
26. Excess of which of the following can result in cata- called core proteins
ract? (PGMEE 2015) b. May be associated with connective tissues
a. Sugar alcohol b. Glucose c. Highly positively charged
c. Sugar amines d. Galactose d. Negatively charged
27. Number of –OH groups in Ribose? e. Component of ECM
a. 4 b. 5 40. Which of the following is not a dietary fiber? 
c. 6 d. 2  (Recent Pattern Jan 2019)
28. Which of the following is branched: a. Inulin b. Cellulose
a. Starch c. Pectin d. Gum
b. Cellulose 41. Which of the following helps in wound healing? M
c. Heteropolysaccharide  (FMGE Nov 2018) C
d. All a. Keratan sulfate
29. All are functions of Glycosaminoglycans EXCEPT: Qs
b. Dermatan sulfate Ans.
 (PGMEE 2015) c. Hyaluronic acid
a. Anticoagulant b. Wound healing d. Chondroitin sulfate 21. a
c. Lubrication d. Transport of lipids 42. MPS in eye: (Recent Pattern June 2018) 22. d
30. Which of the following has NO asymmetric carbon? a. Keratan Sulfate – I and Chondroitin Sulfate 23. a
b. Keratan Sulfate – I and Heparan Sulfate 24. a
a. Glucose 6 Phosphate c. Chondroitin Sulfate and Keratan Sulfate – II 25. b
b. Glyceraldehyde 3 Phosphate d. Chondroitin Sulfate and Dermatan Sulfate 26. a
c. Dihydroxyacetone Phosphate 43. Hyaluronic acid is composed of: (PGI Nov 2014) 27. a
d. Fructose a. Longest Glycosaminoglycan 28. a
31. Which is the major energy providing carbohydrate cons- b. N-Acetyl Galactosamine 29. d
tituent of Vegan diet? c. Has Glucuronic acid 30. c
a. Amylose b. Lactose d. N-acetyl neuramic acid 31. a
c. Cellulose d. Glycogen e. Iduronic acid 32. a
32. Which of the following is a test to distinguish between 44. In benedict test, red colour is/are produced by: 33. a
monosaccharides and disaccharides?  (PGI Nov 2014) 34. d
a. Barfoed’s test b. Bial’s Test a. Sucrose b. Inositol 35. d
c. Seliwanoff’s test d. Hydrolysis test c. Fructose d. Lactose 36. a
33. The oxidation of galactose with strong oxidizing e. Maltose 37. c
agent produces: 38. a
45. Mucopolysaccharidosis, which is a lysosomal storage
a. Mucic Acid b. Gluconic Acid 39. c
disease, occurs due to abnormality in:
c. Galacturonic acid d. Saccharic Acid 40. d
a. Hydrolase enzyme  (PGI May 2015)
34. Which of the following is a lectin? 41. c
b. Dehydrogenase enzyme
a. Ricin 42. a
c. Lipase enzyme
b. Hemagglutinin 43. a,c
d. Phosphatase
c. Cholera toxin 44. c,
e. Acetyl CoA Carboxylase
d. All are lectins d,e
45. a
 46. Danaparoid contains: (PGI Nov 2009) 56. GLUT (Glucose transporter) present in neurons is:
a. Keratin sulfate 
48 b. Chitin a. GLUT-1 b. GLUT-2
c. Dermatan sulfate c. GLUT-3 d. GLUT-4
d. Heparan sulfate 57. Mutation in GLUT-2 causes - (PGMEE 2012,13)
e. Keratin a. Menke's disease
b. Fanconi-Bickel syndrome
Glucose Transporters c. Beckwith syndrome
47. The monosaccharide with maximum rate of absor- d. Dandy walker syndrome
ption in intestine is:  (AIIMS Nov 2017) 58. Glucose transporter present in erythrocytes (RBCs):-
a. Glucose b. Galactose (PGMEE 2015, 16, 17 )
c. Mannose d. Fructose a. GLUT– 1
48. The rate of absorption of sugars by the small intestine b. GLUT – 2
is highest for (PGMEE 2012) c. GLUT – 3
a. Polysaccharides b. Disaccharides d. GLUT – 4
c. Hexoses d. Pentoses 59. GLUT responsible for secretion of insulin from beta
49. Glucose is reabsorbed in which part?  cells of pancreas – (PGMEE 2012, 13)
 (FMGE Nov 2018) a. 4 b. 2
a. Early PCT c. 3 d. 1
b. Henle loop 60. Which of the following is NOT correct?
c. Collecting duct a. Sodium dependent Glucose transporter (SGLT) is
unidirectional
d. Distal convoluted tubule
b. SGLT-2 is in kidneys only for Glucose transport,
50. How many ATPs are used during transport of glucose
SGLT-1 is in kidneys and intestine for Glucose and
via active transport?
galactose transport
a. 1 b. 2
c. This Sodium-Glucose symport carries 2 Na+for each
c. 4 d. 0
Glucose
51. GLUT-5 is transporter for- (PGMEE 2012,13)
d. This Sodium-Glucose symport carries 3 Na+for each
a. Galactose b. Fructose
Glucose
c. Mannose d. Glucose 61. After an overnight fast, GLUTs are reduced in :
M 52. Defect in renal glucosuria: (PGMEE 2015) a. Brain b. RBC
C a. GLUT-1 b. GLUT-2 c. Kidney d. Adipose Tissues
Qs c. SGLT-1 d. SGLT-2 62. Which of the following does not depend on insulin for
Ans. 53. Identify the correct statement: Glucose uptake? (PGMEE 2012, 13)
a. GLUT-2 responsible for Glucose and fructose a. Brain
46. a,c,d
absorption b. Cardiac muscles
47. b
b. Glucose is absorbed independent of Na c. Skeletal muscles
48. c
c. Fructose requires sodium for absorption d. Adipose tissue
49. a
d. SGLT-2 is specific for Glucose 63. Which of the following is Insulin dependent?
50. d
54. Method of transport of glucose in the intestine is a. GLUT-1
51. b
 (AIIMS Nov 2018) b. GLUT-2
52. d
a. Primary active transport c. GLUT-3
53. d
b. Secondary active transport d. GLUT-4
54. b
c. Simple diffusion 64. Which of the following depends totally on Glucose?
55. b
56. c
d. Counter transport a. Liver b. Renal cortex
57. b
55. Active uptake of glucose is inhibited by: c. Renal medulla d. Brain
58. a
 (PGMEE 2013) 65. HbA1c (glycated hemoglobin) is glycosylated with
59. b
a. Insulin b. Phlorizin which amino acid? (Recent Question 2017)
60. d
c. Indoacetate d. Fluoride a. Arginine b. Glutamate
61. d c. Valine d. Leucine
62. a
63. d
64. d
65. c
 Answers with Explanations
49
1. Ans. (a) Parent carbohydrate which gives rise to •• Parent carbohydrate, and simplest carbohydrate is
other carbohydrates is Glycerol D-Glyceraldehyde

CHAPTER 2  CHEMISTRY OF CARBOHYDRATES

[Ref: Lehninger7th/e pg. 242] 6. Ans. (b) β
Parent carbohydrate which gives rise to other carbo-
[Ref: Harper 30th/e pg. 154]
hydrates is D-Glyceraldehyde (not Glycerol) and this
is also the simplest carbohydrate which exists. So, β−Glucose and β−Fructose are more predominant than
Options b,c,d are correct as the simplest carbohydrate is their α forms
Glyceraldehyde with 3 Carbons, 2 ‘OH’ groups and one
7. Ans. (a) C1
functional group i.e. Aldehyde.
[Ref: Harper 30th/e pg. 153]
2. Ans. (c) Racemase enzyme interconverts d and l Anomerism is different –H and –OH orientation around
isomers into each other. the functional carbon. β-anomers cannot be broken
[Ref: Harper 30th/e pg. 154; Fig.15–2] in body. Cellulose is made up of Glucose. Glucose
functional carbon is at C1 (Aldehyde). So, Cellulose is
Racemase enzyme interconverts D- and L- isomers
not broken due to β-anomerism at C1
(Enantiomers) into each other in which D means OH is
•• Inulin (Homopolysaccharide of β-Fructose) is
on right side of penultimate carbon. L means OH is on
not broken due to β- anomerism at C2 because in
left side of penultimate carbon. The name Racemase is
fructose- (a keto sugar), functional carbon is at C2
a misnomer.
•• d and l (small d and l) are Optical isomers. 8. Ans. (a); (d)
•• d(dextrorotatory)(+) – means rotation of plane
polarized light towards right side. [Ref: Harper 30th/e pg. 153]
•• l(levorotatory)(–) –means rotation of plane In Epimerism, there is different –H and –OH orientation
polarized light towards left side. around only one carbon, other than the penultimate
•• Racemic mixture is equal d and l isomers present carbon. Mannose is epimer of Glucose at C2. Galactose
(option a). Racemic mixture is optically inactive is epimer of Glucose at C4. Galactose and Mannose are
(option b). Enantiomerism is also known as D and not epimers of each other.
L-isomerism. (option d).
9. Ans. (c); (d); (e)
3. Ans. (a) D + Glucose [Ref: Harper 30th/e pg. 153]
[Ref: Harper 30th/e pg. 154] Glucose is aldehyde. Sorbitol is a sugar formed from A
Solution of Glucose is known as Dextrose. ‘D’ means reduction of Glucose. Ribulose is a 5C keto sugar; N
structural isomer, i.e. Enantiomer. Abundant form of Fructose is a 6C keto sugar and Sedoheptulose is a 7C S
carbohydrate is always ‘D’. keto sugar. W
(+) means dextrorotatory (d). Glucose is always dextro- E
10. Ans. (b) Glucosamine R
rotatory (d) (+). Fructose is always levorotatory (l) (–).
[Ref: Harper 30th/e pg. 159] S
4. Ans. (c) 16
Chitin is a linear homopolysaccharide which consists WITH
[Ref: Harper 30th/e pg. 154] of β (1 → 4) linkage between N-Acetyl-Glucosamine
•• Number of isomers possible for a compound is given residues. E
It is present in cell wall of fungi and is a principal X
by formula = 2n, where n= number of asymmetric
component of hard exoskeletons of arthropods. P
carbons.
L
•• In case of Glucose, number of asymmetric carbons
11. Ans. (b) Beta Galactosidase A
are 4 (C2, C3, C4, C5). C1 is symmetric as it is C=O. C6
[Ref: Harper 30th/e pg. 155] N
is CH2OH, so symmetric. Hence, in case of Glucose,
A
isomers are 16. Patient has abdominal discomfort and diarrhoea after T
taking milk. Diagnosis is Lactose Intolerance. Lactase is I
5. Ans. (a) Glycerol also known as Beta-Galactosidase. Alpha Glucosidase or O
[Ref: Lehninger7th/e pg. 242] Acid Maltase is minor pathway of Glycogen breakdown N
in lysosomes [Option c], Alpha Amylase digests Starch S
•• Parent alcohol in carbohydrates is Glycerol.
 [Option a]. If these symptoms appear after taking sugar •• Folin Wu method
or sugarcane juice, then enzyme deficient is Sucrase •• Dextrostix test strips
50 [Option d].
18. Ans. (d) Keratan Sulfate
12. Ans. (b) Hunter’s disease
[Ref: Harper 30th/e pg. 637]
CRO BIOCHEMISTRY

[Ref: Harper 30th/e pg. 179] Keratin Sulfate has no Uronic acid and their sulfate
Coarse facial features is a typical feature of content is variable. They are found in cornea, connective
Mucopolysaccharidosis (MPS). In Hunter disease (MPS tissue and also a variety of horny structures formed of
Type II), Iduronate Sulfatase is deficient. There is no dead cells: horn, hair, hoofs, nails and claws. Keratin
corneal clouding. This is X-linked recessive, so occurs Sulfate is responsible for corneal transparency.
exclusively in Males. Dermatan Sulfate and Heparin
Sulfate accumulates. 19. Ans. (c) Invertase

13. Ans. (d) Yoghurt [Ref Harper 30th/e pg. 156]

Milk and dairy products are avoided in Lactose •• Invertase is a Sucrase which breaks Sucrose into


Intolerance. Yogurt can be taken as in curd, bacteria its constituent parts, i.e. Glucose and Fructose. It
produce Lactase which breaks Lactose. is derived from yeast Saccharomyces cerevisiae for
industrial use.
14. Ans. (a) Glyceraldehyde •• It is also synthesized by bees to produce honey from
[Ref: Lehninger 7th/e pg. 241] nectar.
Monosaccharides of four or more carbons tend to have
20. Ans. (a) Covalent
cyclic structures. But erythrose (4C, Keto doest not has
cyclic structure) [Ref: Harper 30th/e pg. 634]
Structure of Proteoglycan: On Hyaluronic Acid back-
15. Ans. (d) Isomerase
bone, many core proteins and link proteins are attached.
[Ref: Harper 30th/e pg. 196] •• Linear GAG chains (of amino sugars and Uronic
Aldo: Keto interconversion is done by Isomerase. acid) are covalently attached to core Proteins. Linear
Aldolase [Option b] is a lyase. Aldolase A is in Glycolysis. chain of GAG is linked to Core Protein by a core
Aldolase B is in Fructose metabolism. Trisaccharide (Galactose-Galactose-Xylose).

16. Ans. (a) Benedicts test 21. Ans. (a) Glycosaminoglycan (GAG)>> Polysaccha-
ride
[Ref: Rafi Practical pg 4]
•• Glyceraldehyde is a Monosaccharide. All mono- [Ref: Lehninger7th/e pg. 261]
A saccharides are reducing. All reducing sugars gives Heparin, an anticoagulant is a Glycosaminoglycan
N positive Benedict’s test. (GAG). This is sulfated, negatively charged GAG, releas-
S •• Molisch test is a general test for all the carbohydrates, ed from mast cells and liver cells.
W But it is positive if number of carbons are 5 or more
E [Option b]. Glyceraldehyde contains only 3 carbons 22. Ans. (d) Hurler disease
R •• Seliwanoff’s test is positive for keto sugars [Option
[Ref: Pediatric bone Marrow By Lila Penchnask pg. 32]
S c]. Glyceraldehyde is an aldehyde. Gerhard’s test and
Rothera’s test are positive for ketone bodies[Option Reilly Bodies, also known as Alder-Reilly bodies are
WITH d]. found in many Mucopolysaccharidosis, e.g. Hurler
disease and Maroteaux-Lamy Syndrome. These are
E 17. Ans. (c) Ferric chloride test metachromatic granules, surrounded by a clear zone.
X These are present in the cytoplasm of all leukocytes
P [Ref: Rafi Practical pg 6]
including Neutrophils. (See picture) in theory.
L Ferric Chloride urine test is done for the diagnosis of
A Phenylketonuria, Alkaptonuria, Tyrosinemia, MSUD. 23. Ans. (a) Lactulose
N This test detects phenols and branched chair acids.
A Glucose estimation methods are: [Ref: Harper 30th/e pg. 157]
T •• Enzymatic methods → Hexokinase, GOD-POD Lactulose is a synthetic disaccharide made up of
I •• Reduction methods (e.g. Ferricyanide, Copper Galactose and Fructose, joined by beta bond. Due
O Sulfate) to beta bond, it is not broken in intestine when it is
N •• Nelson somogyi method ingested So, it is used in the treatment of constipation.
S (Lactose is made up of Galactose and Glucose)
24. Ans. (a) Arthritis Dihydroxyacetone Phosphate is the only carbohydrate
which has no asymmetric carbon (Option c)
[Ref: Lehninger 7th/e pg. 261] 51
Glucosamine is amino sugar. Amino sugars are consti- 31. Ans. (a) Amylose
tuents of GAGs (Glycosaminoglycans) like Hyaluronic [Ref: Lehninger 7th/e pg. 253]
acid, which has a very important function of joint

CHAPTER 2  CHEMISTRY OF CARBOHYDRATES

lubrication. So, used in arthritis. Starch is the main constituent in Vegan diet. It is made
up of Amylose and Amylopectin.
25. Ans. (b) Malabsorption syndrome
32. Ans. (a) Barfoed’s test
[Ref: Varley 6th/e pg. 699]
[Raf: Practical pg 4-6]
Xylose is a pentose sugar, which is incompletely
metabolized in body and therefore 50% of it is excreted •• Barfoed’s test is primarily used to distinguish between
in urine. So, this test is a general screen test for Monosaccharides and Reducing Disaccharides.
malabsorption. The reduction of Cu++ in acidic Barfoed’s reaction is
slower than that of (Alkaline) Benedict’s reaction.
26. Ans. (a) Sugar alcohol •• Monosaccharides are able to reduce Cu++ (cuprous) to
Cu+ (cupric) within 3 minutes whereas disaccharides
[Ref: Harper 30th/e pg. 205]
take a longer time (up to 10 minutes). If heating is
•• Sugar alcohol >> glucose or galactose prolonged in this test, then disaccharides also give
•• Reduction of Monosaccharides forms sugar alcohols, this test positive.
which are hygroscopic in nature i.e. they absorb
water. So, if this sugar alcohol is present in excess 33. Ans. (a) Mucic Acid
in cells, it will cause cell swelling. Especially it gets
accumulated in lens of the eye causing cataract. [Ref: Harper 30th/e pg. 191]
Galactitol causes oil drop cataract. Sorbitol causes In the presence of strong oxidizing agent, both first and
snow flake cataract. last carbon of sugar is oxidized to produce Saccharic
Acid.
27. Ans. (a) 4 •• Glucose is oxidized to Glucosaccharic acid
[Ref: Lehninger 7th/e pg. 242] •• Mannose to Mannaric acid
•• Galactose to Mucic Acid. Mucic Acid forms insoluble
Number of –OH groups are always one less than the
crystals. In mild oxidation conditions, either C1 is
number of carbons, in case of carbohydrates. So,
oxidized or C6 is oxidized.
Glucose (6C) has 5–OH groups and Ribose (5C) has 4
–OH groups. 34. Ans. (d) All are lectins
28. Ans. (a) Starch [Ref: Lehninger 7th/e pg. 268]
A
[Ref: Lehninger7th/e pg. 253] •• Lectins are proteins that bind carbohydrates with
N
high affinity and specificity. Lectins serve in a
•• Starch is branched (less branched than Glycogen). S
wide variety of cell-cell recognition, signaling and
•• Cellulose (Option b) is unbranched homopolysac- W
adhesion processes and in intracellular targeting of
cha-ride (made up of beta Glucose). Heteropolysac- E
newly synthesized proteins.
charides (Option c) are linear, unbranched, tandem R
•• In the laboratory, purified lectins are useful reagents
repeats of Amino sugar and Uronic acid. S
for detecting and separating glycoproteins with
29. Ans. (d) Transport of lipids different oligosaccharide moieties. WITH
[Ref: Harper 30th/e pg. 638] 35. Ans. (d) Oxidation of both C1 and C6 of Glucose E
GAGs have role in lubrication, cushioning effect in [Ref: Harper 30th/e pg. 155] X
tissues, wound healing, anticoagulation, cell adhesion P
and transparency of cornea. Transport of lipids in blood Aldonic acid is produced as a result of oxidation at L
is done by lipoproteins e.g. VLDL, LDL C1.Oxidation at C6 or Primary alcohol produces Uronic A
Acid. Oxidation at both C1 and C6 produces Saccharic N
30. Ans. (c) Dihydroxyacetone Phosphate acid A
[Ref: Harper 30th/e pg. 154] T
36. Ans. (a) Inulin
I
Glucose 6 phosphate has 4 chiral/asymmetric
[Ref: Harper 30th/e pg. 191] O
carbons (Option a). Glyceraldehyde-3-Phosphate
•• For the estimation of Glomerular Filtration rate, N
has 1 chiral/asymmetric carbon (Option b). Fructose
criteria for a component to be used is that it should S
has 3 chiral/asymmetric carbons (Option d). While
neither be excreted nor be absorbed from the tubules.
 •• Inulin is an ideal substance to check glomerular 41. Ans. (c) Hyaluronic acid
filtration rate as per above criteria.
52 •• Creatinine provides an easier method for GFR deter-
[Ref: Harper 30th/e pg. 423]
mination as it is an endogenous metabolite •• Hyaluronic acid has a role in cell migration during
•• Cr51 EDTA has also been used to determine GFR morphogenesis and also in wound healing.
•• Phenol red is 94% excreted by renal tubules and only
CRO BIOCHEMISTRY

42. Ans. (a) Keratan Sulfate – I and Chondroitin Sulfate

6% by the glomeruli, hence provides a test for the
tubular function known as PSP excretion test. [Ref: Lippincott 4th/e pg. 159]
•• Keratan sulfate-I is present in cornea while Keratan
37. Ans. (c) Iodine test sulfate-II is present in cartilages.
[Raf: Practical pg 9] •• Chondroitin sulfate is present in cartilages and eyes.
•• Heparan sulfate is present on cell surfaces and is
•• Iodine test is a test for starch which is a polysaccharide. required for cell adhesion.
•• Starch forms adsorption complex with iodine to give •• Dermatan sulfate is present in skin, blood vessels and
blue color. Deep blue color disappears on heating due valves.
to the breaking of the complex and then reappears on


cooling due to reformation of complex. 43. Ans. (a) ; (c)

[Ref: Harper 30th ed pg. 423]
38. Ans. (a) Lignin
•• Hyaluronic acid is longest GAG. It does not contain
[Ref: Harper 30th ed pg. 157] any sulfate. It is not attached to proteins. The
•• Lignin resists attack by microorganisms, and repeating disaccharides in Hyaluronic acid is:
anaerobic processes tend not to attack the aromatic Glucuronic Acid (β1 → 3) NAc-Glucosamine. It is a
rings at all. Aerobic breakdown of lignin is slow and major GAG of extracellular matrix.
may take many days. Lignin is an insoluble fiber so it •• Hyaluronates form clear, highly viscous solutions
is not fermented. that serve as lubricants in the synovial fluid of joints
•• Pectin is soluble fiber. Soluble fiber is acted upon and gives the vitreous humor of the vertebrate eye its
by the microflora and fermented to produce short jelly like consistency.
chain volatile fatty acid, which improves the colonic 44. Ans. (c); (d); (e)
environment, regulates immune response, involved
in synthesizing some vitamins, helps to lower down [Ref: Rafi Practical pg. 4]
the colonization of pathogenic bacteria. •• Benedict’s test is positive by reducing sugars. All
monosaccharides are reducing. Nonreducing dis-
39. Ans. (c) Highly positively charged accharides are Sucrose and Trehalose.
[Ref: Harper 30th ed pg. 423] •• Benedict’s test is also positive by :Uric acid,Ascorbic
A acid, Glutathione, Glucuronic acid, Homogentisic
N •• GAGs are covalently attached to a protein called acid and Salicylates
S core protein to form hybrid molecules called
W Proteoglycans. 45. Ans. (a) Hydrolase enzyme
E •• GAGs are negatively charged due to carboxy, sulfate
[Ref: Harper 30th/e pg. 68]
R and acetyl group. GAG is found in ECM. GAGs can
hold a lot of water, so hygroscopic in nature. Lysosomes contain Hydrolase enzyme for breaking
S down Mucopolysaccharides.
WITH
40. Ans. (d) Gum
46. Ans. (a); (c); (d)
[Ref: Harper 30th ed pg. 157]
E [Ref: Harper 30th ed pg. 423]
X •• This is a tricky question, all these are fibers but
question is asking about dietary fiber. Danaparoid is a low molecular weight compound
P
consisting of Heparin, Dermatan Sulfate and Chon-
L •• Pectin and cellulose are present in fruits and
droitin Sulfate. It has antithrombotic activity. It has a
A vegetables
very low tendency to cause bleeding and has minimal
N •• Inulin is found in a wide variety of fruits, vegetables,
effects on the fibrinolytic system.
A wheat, onions, bananas.
T •• Gums are used as food thickeners e.g. alginin, guar 47. Ans. (b) Galactose
I gum, locust bean gum, and xanthan gum. They are
O normally not taken in diet as fiber. [Ref: Harper 30th/e pg. 191]
N •• The absorption rate in intestine is:
S Galactose> Glucose > Fructose
 •• Glucose and Galactose are absorbed by SGLT-1 55. Ans. (b) Phlorizin
(Sodium dependent Glucose transport), against the
concentration gradient.
[Ref: Harper 30th/e pg. 191] 53
•• Fructose is absorbed by GLUT-5, down the concen- Phlorizin is a competitive inhibitor of secondary active
tration gradient. transport of Glucose, i.e. Sodium Glucose Symport
•• All the monosaccharides exit from intestinal cells (SGLT-1 and 2). It competes with D-Glucose for binding

CHAPTER 2  CHEMISTRY OF CARBOHYDRATES

into blood via GLUT-2. to the carrier, decreasing renal glucose reabsorption,
leading to decreased blood glucose. It is a normally
48. Ans. (c) Hexoses occurring substance found in the bark of pear, apple,
[Ref: Harper 30th/e pg. 191] cherry and other fruit trees.
•• Hexose absorption is twice as fast as pentose 56. Ans. (c) GLUT-3
absorption.
[Ref: Harper 30th/e pg. 191]
49. Ans. (a) Early PCT •• GLUT-1 and GLUT-3 both are in Brain
[Ref: Harper 30th/e pg. 191] •• Major GLUT in Brain – GLUT-1
•• Major GLUT in neurons – GLUT-3
•• 100% Glucose is reabsorbed from PCT by secondary
•• GLUT-3 is in neurons but not GLUT-1
active transport, i.e. SGLT-2
•• Also, 100% amino acids gets reabsorbed from PCT. 57. Ans. (b) Fanconi-Bickel syndrome
50. Ans. (d) 0 [Ref: https://www.ncbi.nlm.nih.gov/pubmed/22865906]
[Ref: Harper 30th/e pg. 191] Fanconi-Bickel syndrome and Autosomal Recessive
Proximal Tubulopathy with hypercalciuria are allelic
•• No ATP is used when glucose is getting transported.
•• Energy is in fact used indirectly by sodium potassium variants caused by GLUT-2 mutations.
ATPase pump. That's why this transport is called 58. Ans. (a) GLUT– 1
secondary active or indirect active transport.
[Ref: Harper 30th/e pg. 191]
51. Ans. (b) Fructose In RBCs, only GLUT-1 is present, which is responsible
[Ref: Harper 30th/e pg. 191] for basal glucose uptake and is not dependent on
GLUT-5 is for the transport of Fructose. insulin.

52. Ans. (d) SGLT 2 59. Ans. (b) 2

[Ref: Harper 30th/e pg. 191] [Ref: Harper 30th/e pg. 191]
Renal Glycosuria is a condition in which Blood Glucose GLUT-2 is present in Pancreas, Intestine and Liver and
A
is normal but still Glucose appears in urine. This is is active in Fed state. It is responsible for secretion of
N
because of defect in SGLT-2, which is a transporter insulin from beta cells of Pancreas
S
present in kidneys for the transport of Glucose. W
60. Ans. (d) This Sodium-Glucose symport carries 3 Na+
53. Ans. (d) SGLT-2 is specific for Glucose for each Glucose E
R
[Ref: Harper 30th/e pg. 191] [Ref: Harper 30th/e pg. 191]
S
•• GLUT-2 is only for Glucose (in liver and pancreas). •• Secondary active transport of Glucose, also known
•• Glucose is absorbed from intestine by Na dependent as Sodium dependent Glucose transporter (SGLT) is WITH
secondary active transport. unidirectional.
E
•• Fructose is absorbed by GLUT-5, which does not •• SGLT-2 is in kidneys only for Glucose transport,
X
requires sodium. SGLT-1 is in kidneys and intestine for Glucose and
P
•• SGLT-1 is for Glucose and Galactose absorption (in galactose transport.
L
intestine and renal tubules) •• This Sodium-Glucose symport carries 2 Na+ for each
A
•• SGLT-2 is for Glucose absorption only (in renal Glucose (Not 3).
N
tubules)
61. Ans. (d) Adipose Tissues A
54. Ans. (b) Secondary active transport T
[Ref: Harper 30th/e pg. 191] I
[Ref: Harper 30th/e pg. 191] GLUT are open during fed state in Skeletal muscles, O
Method of transport of Glucose in the intestine is Cardiac muscles and Adipose tissues. Therefore, after N
Secondary active transport or sodium glucose symport. an overnight fast, GLUTs are reduced in these tissues. S
 62. Ans. (a) Brain 64. Ans. (d) Brain

54 [Ref: Harper 30th/e pg. 191] [Ref: Harper 30th/e pg. 191]
Glucose uptake occurs in muscles (Cardiac and Brain>>Renal Medulla
Skeletal) and Adipose tissue via GLUT-4, which is Glucose is the main energy source for Brain, RBCs,
Insulin dependent. In Brain, GLUT-1 and GLUT-3 are Retina, Cornea, Renal Medulla, Testis.
CRO BIOCHEMISTRY

present which are not dependent on Insulin.

65. Ans. (c) Valine
63. Ans. (d) GLUT-4
•• HbA1c is formed non-enzymatically by irreversible,
[Ref: Harper 30th/e pg. 191] covalent glycosylation of human haemoglobin, taking
Only GLUT-4 is Insulin dependent. Insulin recruits place under physiological conditions, at a specific
GLUT-4. GLUT-4 is present on the membranes of site on protein. There is an attachment of Glucose to
peripheral tissues (Skeletal muscles, Cardiac muscles N- terminal amino acid valine of a beta globin chain,
and Adipose tissues). It helps in Insulin dependent though some HbA1c molecules are glycated at the
Glucose uptake after meals. N-terminal valine residues of two beta chains.


A
N
S
W
E
R
S
WITH

E
X
P
L
A
N
A
T
I
O
N
S
Carbohydrate
Metabolism
3
Overview of Chapter zz Step 3: Conversion of fructose-6-P to fructose-1,
6-Bisphosphate phosphate is added to first carbon on
•• Glycolysis fructose (this cannot be done on glucose because C1 of
•• Link Reaction glucose does not contain OH and phosphate has tendency
to bind with OH). So for adding phosphate (high energy
•• TCA Cycle
bond), we require ATP, which gets converted to ADP.
•• Shuttles Whenever ATP is used for adding phosphate, enzyme is
•• Electron Transport Chain Kinase. Here it is phosphofructokinase-I (PFK-I).
•• Gluconeogenesis
•• Glycogen H igh R eturn
•• Hexose Monophosphate Pathway PFK-I
•• Uronic Acid Pathway zz Rate limiting enzyme of glycolysis
zz 1st committed step of glycolysis
•• Galactose and fructose Metabolism
zz Most important control point
zz Bottle neck of glycolysis

GLYCOLYSIS
Table 3.1: Flux generating step v/s Rate limiting step
This is major pathway for the oxidation of glucose. Purpose
of glycolysis in cells is to make ATP. Also known as Embden Flux generating step Rate limiting step
Meyerhoff Pathway/EMP Pathway
Example: Hexokinase in Example: PFK-I in glycolysis
H igh R eturn glycolysis
Usually have low Km Usually have high Km
zz Nature of Pathway: Catabolic
zz Occurs in Fed state Speed of reaction is high Very slow speed of reaction
zz Activated by hormone: Insulin (usually anabolic Pathways are All substrates get converted to Not all substrates are
activated by Insulin. But this catabolic Pathway is activated by products converted to products
Insulin)
zz Organelle: Cytoplasm (Any anabolic Pathway occurs in
cytoplasm. But this catabolic pathway occurs in cytoplasm)
zz Organ/cell: In all the cells of the body
zz Only Pathway which occurs in both aerobic and anaerobic
conditions
zz Glucose is the only molecule which can produce ATP without
O2

EXPLANATION OF 10 STEPS OF GLYCOLYSIS

(FIG. 3.1)
zz Step 1: Phosphorylation of glucose to Glu-6-P
The first step for any monosaccharide is phosphorylation.
This is for the entrapment inside the cells. So for adding
phosphate (high energy bond), we require ATP, which
gets converted to ADP. Whenever ATP is used for adding Step 4: Conversion of fructose 1,6 Bisphosphate to
zz
phosphate, enzyme is Kinase. Here it is Hexokinase or Glyceraldehyde-3-P and DHAP
Glucokinase. This is flux generating step of glycolysis. 
Aldolase A breaks fructose 1,6- Bisphosphate into two 3 C
zz Step 2: Conversion of Glu-6-P to Fruc-6-P molecules:
The enzyme is isomerase (Phospho Gluco Isomerase or 1. 3 C (ald) with a phosphate i.e. Glyceraldehyde-3-P
Phospho Hexose Isomerase) as glucose and fructose are
functional isomers of each other.
 2. 3C (keto) with a phosphate i.e. Dihydroxy Acetone category because this is intramolecular transfer so
phosphate (DHAP) molecular formula remains same here.
56 This enzyme breaks this bond without using water. So it’s zz Step 9: Conversion of 2-Phospho Glycerate to
a Lyase. Phospho-Enol-pyruvate
 is step occurs to attach last phosphate with a high
Th
Remember: Aldolase B (the hepatic isoform) is in fructose
energy bond i.e. Enol bond. Enzyme required is Enolase,
CRO BIOCHEMISTRY

Metabolism
which belongs to Lyase category. (Enolase is inhibited by
zz Step 5: Conversion of DHAP into Glyceraldehyde-3-P Fluoride, which is used in Blood glucose estimation. Also
 nzyme is Isomerase (Triose phosphate Isomerase) as
E water fluoridation reduces dental caries as it decreases
DHAP and Glyceraldehyde-3-P are Functional Isomers lactate production by mouth bacteria).
of each other. So we get 2 molecules of Glyceraldehyde- zz Step 10: Conversion of Phospho-Enol-pyruvate to
3-P, which will enter the next steps. pyruvate
 ere this last phosphate bond is broken and pyruvate
H
Remember: DHAP can also be used for TG synthesis. is formed. The phosphate removed is taken by ADP and
gets converted to ATP. Formation of ATP at substrate level
 teps 1 to 5 constitute Phase I, also known as Energy
S is known as Substrate Level Phosphorylation (SLP). The


Utilizing Phase. Its like investment phase of business,

enzyme is pyruvate Kinase (always enzyme of SLP is a
where 2 ATPs are used
Kinase). Pyruvate is the end product in aerobic glycolysis.
Steps 6 to 10 constitute Phase II, also known as Energy
zz Step 11: Only in case of anaerobic conditions: Conver-
Producing Phase.
sion of pyruvate to lactate:
Understand the plan: For any good business, ATP used
 naerobic glycolysis is useful in cells mitochondria (e.g.
A
must be less and ATP produced should be more. Only
RBCs) or tissues which are poorly vascularized (Kidney
then these reactions are of use. So the plan is that in
Phase I, only one molecule is entering i.e. one molecule of Medulla, Lens and Cornea of eye) and during Hypoxia.
glucose entering. So that each time when we use energy This step occurs for the regeneration of NAD (From
in Phase I, then we use it only for one molecule. But Phase glucose to pyruvate conversion, NAD is used). Lactate is
II is energy producing phase and two molecules are formed as the end product. Enzyme used is LDH– lactate
entering Phase II i.e two molecules of Glyceraldehyde- Dehydrogenase
3-P are entering. So that all the energetics in Phase II is (EC No. = 1)
simply multiplied by 2. Lactate is also regarded as the dead end of glycolysis.
Step 6: Conversion of Glyceraldehyde-3-P to 1, 3-Bis
A dditional E dge
zz
Phospho Glycerate
 ere one phosphate is added to position number 1.
H Q. How NAD is regenerated from NADH in case of aerobic
But ATP is not used. This phosphate is taken from conditions?
inorganic phosphate. So this enzyme Glyceraldehyde- By Substrate Shuttles (Malate and Glycerol phosphate
3-P Dehydrogenase is the only enzyme of glycolysis, Shuttle) which transfer NADH from cytoplasm to mito-
chondria. During the Shuttle reaction in cytoplasm NADH
which can use inorganic phosphate. Here, oxidation
gets converted to NAD. This NADH is delivered into the
occurring by Dehydrogenase (removal of Hydrogen). That mitochondria, where it enters ETC. Starting material for ETC
Hydrogen removed is taken by NAD, which is converted is NADH, which gets converted to NAD. ETC requires O2 so
to NADH. This step is the first Oxidation-Reduction step we can say that O2 is required for the regeneration of this
of glycolysis. NAD. And this occurs in case of aerobic glycolysis.
From two molecules of Glyceraldehyde-3-P, 2 NADH So there are two methods for the conversion of NADH to
obtained, which will enter ETC to give 5 ATPs. NAD:
So this is a special step, where phosphate added but ATP 1. Lactate Dehydrogenase (pyruvate to lactate conver-
sion)
not used. 2. ETC (starts mostly from NADH)
zz Step 7: Conversion of 1, 3-Bis Phospho Glycerate to
3-Phospho Glycerate
O  ne of the high energy phosphate bond is broken
Summary Box
here so that 1, 3-Bis Phospho Glycerate gets converted zz Enzymes used in Phase I (Energy Utilizing Phase)
to 3-Phospho Glycerate. The phosphate removed is 1. Hexokinase (EC No. = 2)
T taken by ADP and gets converted to ATP. Formation 2. Phospho Hexose Isomerase (EC No. = 5)
H of ATP at Substrate level is known as Substrate level 3. Phosphofructokinase-1 (EC No. = 2)
E 4. Aldolase A (EC No. = 4)
Phosphorylation (SLP). The enzyme is Phosphoglycerate
5. Triose phosphate Isomerase (EC No. = 5)
O Kinase (always enzyme of SLP is a Kinase) zz Enzymes used in Phase II (Energy Producing Phase)
R zz Step 8: Conversion of 3-Phospho Glycerate to 2-Phos-  lyceraldehyde-3-phosphate Dehydrogenase (EC No. = 1)
6. G
Y pho Glycerate 7. Phosphoglycerate Kinase (EC No. = 2)
H  ere intramolecular transfer of phosphate is occurring 8. Phosphoglycerate Mutase (EC No. = 5)
from position number 3 to position number 2. Enzyme is 9. Enolase (EC No. = 4)
Phospho Glycerate Mutase. Mutase belongs to Isomerase 10. Pyruvate Kinase (EC No. = 2)
Table 3.2: Kinases and Dehydrogenases of Glycolysis
Kinases of Glycolysis Dehydrogenase of Glycolysis
1. Hexokinase/Glucokinase Only one i.e
57
2. Phosphofructokinase-1 Glyceraldehyde-3-phosphate Dehydrogenase
3. Phosphoglycerate Kinase
4. Pyruvate Kinase

CHAPTER 3  CARBOHYDRATE METABOLISM

All SLP steps are reversible. Only pyruvate Kinase is irreversible.

T
H
Fig. 3.1: Glycolysis E
O
Table 3.3: Irreversible/ Regulatory steps and SLP steps of glycolysis
R
Irreversible/Regulatory steps Substrate level Phosphorylation (SLP) steps Y
1. HexoKinase/Glucokinase 1. Phosphoglycerate Kinase
2. Phosphofructokinase-1 2. Pyruvate Kinase
3. Pyruvate Kinase
 zz Glycolysis is irreversible But in case of anaerobic conditions, pyruvate gets
zz Glycolysis is not complete breakdown of glucose converted to lactate, by enzyme lactate Dehydrogenase. In
58 zz Complete breakdown of glucose leads to production of this reaction NADH gets converted to NAD. This extra step of
CO2. But in glycolysis no CO2 is produced anaerobic glycolysis occurs to replenish NAD because NAD is
C6H12O6 + 6O2—→ 6CO2 + 6H2O + Energy (Complete required for the conversion of glucose to pyruvate.
breakdown of glucose)
E nergetics B ox
CRO BIOCHEMISTRY

Table 3.4: Pyruvate Kinase and G-6-PD Deficiency Energetics of anaerobic glycolysis
Pyruvate Kinase deficiency G-6-PD deficiency NADH is used in step 11 (lactate Dehydrogenase). So 5 ATPs which
are derived from NADH in aerobic condition, are not obtained in
Second most common human First most common human anaerobic conditions. So energetics:
enzyme deficiency enzyme deficiency zz 2 Substrate Level Phosphorylations + 4 ATPs
zz HexoKinase and PFK-1 uses ATPs – 2 ATPs
This is enzyme of EMP This is enzyme of HMP pathway
pathway Total 4–2 = 2 ATPs
So 2 ATPs are obtained in case of anaerobic glycolysis
Hemolysis occurs because Hemolysis occurs due to
RBCs rely only on glucose. oxidative stress. (No NADPH Lactate: Dead end of Glycolysis


When glucose cannot give and Reduced Glutathione to

Lactate is regarded as the dead end of glycolysis. This means
ATPs, then Na+ K+ ATPase scavenge H2O2) (See HMP)
that lactate cannot be converted to some other thing, in the
pump of RBCs is affected
cell where lactate is produced. But in body it can be converted
Heinz bodies absent Heinz bodies present to glucose by gluconeogenesis. This is easily understood by
Enzyme deficiencies of glycolysis and HMP lead to Hemolytic Cori’s cycle.
Anemia
Cori’s Cycle
Energetics of Glycolysis In case of exercising muscles, anaerobic conditions are
created. So here glucose gets converted to lactate. lactate
When one molecule of glucose gets converted to pyruvate,
is the dead end. So in this muscle cell (where lactate is
10 steps are involved. 2 ATPs are used (one at Hexokinase
formed), lactate cannot be converted to glucose or any other
and other at PFK-1). Two steps of Substrate Level
compound. So this lactate has to come out of muscle cell
Phosphorylation (SLP) occur → one at Phosphoglycerate
and enters blood. From blood it is taken up by liver. In liver
Kinase and other at pyruvate Kinase. SLP steps occur in
lactate forms pyruvate (by lactate dehydrogenase, which is
Phase II. All energetics of Phase II is multiplied by two (as two
reversible) and pyruvate forms glucose (by gluconeogenesis).
molecules of Glyceraldehyde-3-P enters Phase II). Enzyme
This glucose is again supplied to the exercising muscles. This
Glyceraldehyde-3-P Dehydrogenase converts NAD to NADH.
cycle formed is known as Cori’s cycle.
This also occurs in Phase II so multiplied by two. So 2 NADH
produced. These 2 NADH will go into ETC to give 5 ATPs
(each NADH gives 2.5 ATPs).

Fig. 3.3: Cori’s Cycle

Fig. 3.2: Regeneration of NAD by LDH step
E nergetics B ox
T E nergetics B ox Q. Number of ATP produced in RBC in Fed state?
H Q. Number of ATP produced in RBC in aerobic state?
Energetics of aerobic glycolysis
E Answer to both questions is = 2 ATPs
2 NADH + 5 ATPs Because in RBCs there is no mitochondria. NADH can give
O 2 Substrate level Phosphorylations + 4 ATPs ATPs after entering ETC, which occurs in mitochondria. So,
R Hexokinase and PFK-1 uses ATPs – 2 ATPs NADH can never give ATPs in RBCs. So in RBC energetics is 4
Y Total: 9–2 = 7 ATPs (ATPs from Substrate Level Phosphorylation) – 2 ATPs (used by
So 7 ATPs are obtained in case of aerobic glycolysis Hexokinase and PFK-1) = 2 ATPs
RAPAPORT LEUBERING SHUNT (RL-SHUNT)
RL shunt occurs only in RBCs 59

CHAPTER 3  CARBOHYDRATE METABOLISM

Fig. 3.5: Number of ATPs in RBCs can never be more than two, but
it can be less than two if it is RL shunt

Role of 2, 3 BPG
Fig. 3.4: RL Shunt 2, 3 BPG helps in the release of oxygen from HbA. It binds
10% of glucose which is entering RBCs , enters RL shunt. to beta globin chain of Haemoglobin and releases oxygen.
In RL shunt 1, 3-Bis Phospho Glycerate (1, 3 BPG) gets It does not affect any other Haemoglobin (like HbA2, HbF)
converted to 2,3 Bis Phospho Glycerate (2,3 BPG). Then 2,3 because there is no beta chain in these Hb.
BPG gets converted to 3-Phospho Glycerate and glycolysis A dditional E dge
continues. So Phospho Glycerate Kinase (one of the Substrate
zz 2, 3 BPG is the most abundant organic phosphate in RBC
Level Phosphorylation) step is bypassed, which produces zz It binds to Deoxyhemoglobin (not Oxyhemoglobin)
2 ATPs in glycolysis. So in RL shunt, these 2 ATPs are not zz It binds to a pocket formed by two beta-globin chains of
produced in RBCs. So energetics are: 2 ATPs (Substrate Level Deoxyhaemoglobin. This pocket contains positive charged
Phosphorylation by pyruvate Kinase) – 2 ATPs (used by amino acids that form negative charge with the phosphates
Hexokinase and PFK-1) = 0 (zero) ATPs. in 2, 3 BPG
zz HbO2 (Oxyhemoglobin) + 2,3 BPG → Hb–2,3 BPG (Deoxy-he-
moglobin) + O2

Tissues that mainly depend on glucose as a fuel are RBCs, brain, GIT, renal medulla, sperm, many cancer cells, White fibres of Skeletal Muscles

Inhibitors of Glycolysis
Table 3.5 Inhibitors of Glycolysis
Inhibitor Enzyme Inhibited
Iodoacetate Glyceraldehyde-3-P Dehydrogenase
Arsenate Glyceraldehyde-3-P Dehydrogenase
Sodium fluoride Enolase
Oxamate Lactate Dehydrogenase
Sodium Fluoride is used in Blood glucose estimation, to obtain true glucose value.

•• ARSENATE (Pentavalent Arsenic ) – inhibits Glyceraldehyde-3- phosphate Dehydrogenase of glycolysis.

NORMAL STEPS IN GLYCOLYSIS IF ARSENIC IS PRESENT

T
H
E
O
R
Y
Note: So, glycolysis can continue in the presence of Arsenic, but there is loss of 7 ATPs
•• Arsenite (Trivalent Arsenic): inhibits the Sulhydryl group of PDH complex and α- Ketoglutarate Dehydrogenase
Fig. 3.6: Arsenic as Inhibitor
 Regulation of Glycolysis PFK-I PFK-II
60 PFK-I is regulated by allosteric activation and inhibition. •• Has no covalent •• Covalently activated by insulin
Inhibitors are ATP and Citrate. Activators are fructose 2,6 Bis regulation (Insulin dephosphorylate and
phosphate and ADP. activates this enzyme)
CRO BIOCHEMISTRY

Fates of glucose-6-P
zz HMP
zz Glycogenesis
zz Gluconeogenesis
Fig. 3.7: Regulation of PFK-I

A dditional E dge
zz Homotropic Modulator: When substrate or substrates like


substance act as regulator. E.g. ATP and fructose 2,6 Bis

phosphate are Homotropic modulators of glycolysis
zz Heterotrophic Modulator: When molecule other than sub-
strate act as regulator E.g. Citrate in glycolysis

Linking of PFK-1 and Pyruvate Kinase

Pyruvate Kinase is activated by fructose 1,6 Bis phosphate
(produced by PFK-1). So this way two Kinases are linked.
Increased PFK-1 increases fructose 1,6 Bis phosphate, which
activates pyruvate Kinase.
Also pyruvate Kinase has covalent modification. It is active
in dephosphorylated state (done by insulin in Fed state).
It is inactive in phosphorylated state (done by Glucagon in Fig. 3.8: Fates of pyruvate
Fasting state).
LINK REACTION (CONVERSION OF PYRUVATE
H igh R eturn TO ACETYL CoA)
zz Presence of ATP → indicates high energy state in cell
Presence of ADP → indicates low energy state in cell
zz
So presence of ATP will inhibit the rate limiting enzyme of H igh R eturn
glycolysis and presence of ADP will activate. Link between Glycolysis and TCA
zz Inhibition by Citrate directs glucose for glycogen synthesis. •• Activated by hormone – Insulin
zz Fructose 2,6 Bis phosphate is the most potent activator of •• Occurs in mitochondria
PFK-1. It can activate even in high ATP concentrations. This is •• Occurs in fed state
formed from fructose-6-phosphate by enzyme PFK-II (Phos-
phofructokinase-II). Pyruvate is formed in glycolysis in cytoplasm. But link
zz Insulin activates Glucokinase, PFK-1 and pyruvate Kinase reaction occurs in mitochondria. So pyruvate must cross
Inner mitochondrial Membrane (IMM). Pyruvate enters
Table 3.6: Difference between PFK-I and PFK-II IMM with the help of pyruvate-Proton symport. (Fig. 3.10)
PFK-I PFK-II Pyruvate (3C) gets converted to acetyl CoA (2C), so
removal of CO2 occurs. But name of enzyme is pyruvate
•• Converts fructose-6- •• Converts fructose-6-phosphate
phosphate to fructose to fructose 2, 6–Bis phosphate Dehydrogenase complex. Dehydrogenase means oxidation
1, 6–Bis phosphate occurring. So this is oxidative decarboxylation (removal of
hydrogen and removal of CO2). Oxidative Decarboxylation
T •• Not a bifunctional •• A bifunctional enzyme i.e. having
step always requires vitamin B1. Therefore B1 deficiency,
enzyme both enzymatic activities– PFK-II
H Beriberi, causes Lactic acidosis. Because in B1 deficiency,
and fructose 2, 6-bisphosphatase
E pyruvate accumulates, reaches cytoplasm and gets converted
O to lactate. So, Lactic Acidosis occurs. In this step, NAD forms
R NADH (Fig. 3.9).
Y
 61

CHAPTER 3  CARBOHYDRATE METABOLISM

Fig. 3.9: Link reaction occurring in mitochondria. Pyruvate enters mitochondria with the help of pyruvate-Proton symport. Malate and
Aspartate can cross IMM. NADH and Oxaloacetate cannot cross IMM
How pyruvate enters mitochondria?

Fig. 3.11 PDH reaction occurs in mitochondrial matrix but enzyme

is present in IMM
Fig. 3.10: Pyruvate-Proton Symport present in Inner mitochondrial
Membrane (IMM)

3 Enzymes: pyruvate Dehydrogenase Complex

(PDH Complex) H igh R eturn T
5 coenzymes required are: H
PDH complex is a multienzyme complex having three 1. TPP (Thiamine Pyrophosphate or Vit B1) E
components- E1, E2, E3. 2. FAD (or vitamin B2) All these 5 also
required in TCA
O
zz E1–pyruvate Dehydrogenase or pyruvate Carboxylase 3. NAD (or vitamin B3)
R
zz E2–Dihydro Lipoyl Transacetylase 4. Coenzyme A (or vitamin B5)
5. Lipoic acid Y
zz E3–Dihydrolipoyl Dehydrogenase
 Deficiency of these vitamins (Vitamin B1, B2, B3, B5) lead
to decreased energy production and CNS problems because
62 Brain cells depend on this aerobic reaction.
Link Reaction is Irreversible
Because link reaction is irreversible, therefore pyruvate can
CRO BIOCHEMISTRY

form acetyl CoA but acetyl CoA can never form pyruvate.
Pyruvate is mainly derived from carbohydrates and acetyl
CoA mainly form fats (TGs, Fatty Acids, Cholesterol) in the
body. So we can conclude that carbohydrates (in excess) can H igh R eturn
form fats but fats can never be converted to carbohydrates zz All enzymes of TCA lies in the mitochondrial matrix
because link reaction is irreversible.
Two exceptions to this rule: Glycerol and Propionic Acid
Exception:
are derived from fats, but they can be converted to glucose.
zz Succinate Dehydrogenase – is in IMM (Inner mitochondrial

H igh R eturn Membrane)



Regulation of PDH Amphibolic role of TCA

zz End Product Inhibition/allosteric inhibition by:
TCA cycle is amphibolic pathway i.e. it is both catabolic as
� Acetyl CoA � NADH � ATP
well as anabolic.
zz Covalent Modification
zz Catabolic role of TCA: All macromolecules finally forms
 Active in dephosphorylated state (done by insulin)
acetyl CoA, which then enters TCA.
In muscles, this enzyme is activated by calcium during zz Anabolic role of TCA: Many intermediates of TCA cycle
muscle contraction. synthesize important compounds.
Deficiency of PDH Complex
zz X-linked dominant
zz Rare but most common biochemical cause of congenital
Lactic Acidosis
zz Brain is very sensitive to acidosis. Therefore it leads to
neurodegeneration, muscle spasticity and in neonatal
onset may lead to early death.
zz Treatment: Dietary restriction of carbohydrate and
Thiamine supplementation can reduce symptoms. No Fig. 3.12: Catabolic role of TCA
specific treatment.
TCA CYCLE
H igh R eturn
zz Nature of Pathway: catabolic and anabolic (Amphibolic)
zz Neither activated by Insulin nor Glucagon
zz Organelle: Mitochondria
zz Organ/cell: In all the cells of the body (where mitochondria is
present)
zz A Vital Pathway for cells (vital pathways occur in mitochondria)
zz Occurs only in aerobic conditions
zz Occurs both in Fed and Fasting state
zz TCA is called a cycle, not a pathway because it begins and ends
with oxaloacetate

Fig. 3.13: Anabolic role of TCA

T Names
H
E zz TCA–Tricarboxylic Acid Cycle because citrate and Anaplerotic Reactions
O isocitrate are tricarboxylic Acids
zz Kreb’s cycle- by the name of the scientist- Hans Adolf Those reactions which synthesize intermediates of TCA cycle
R also known as ‘Filling Up-reaction’
Krebs
Y 1. Pyruvate → OAA (Oxaloacetate)
zz Citric Acid cycle because the first compound formed is
2. Propionyl CoA to Succinyl CoA
citrate
3. From amino acids e.g.
 Aspartate and Asparagine → OAA (Oxaloacetate) F undamental B ox
 Glutamate and Glutamine → Alpha-Ketoglutarate Q. TCA is activated by: 63
a. Insulin b. Glucagon
c. Both d. None
A. (d) None

CHAPTER 3  CARBOHYDRATE METABOLISM

• TCA has no hormonal control. It is a vital cycle
(Enzyme deficiency of TCA not known)

TCA only depends on 2 things

Energy status of the cell: If a cell has enough energy,
1. 
then TCA will not occur. If energy of the cell is less,
then TCA will occur.
Availability of Oxaloacetate (not acetyl CoA):
2. 
Number of Carbons in some Compounds Therefore Oxaloacetate is regarded as the first substrate
zz Alpha-ketoglutarate = 5 C of TCA cycle or the carrier of TCA cycle.
Glutamate = 5 C
H igh R eturn
zz
zz Glutamine = 5 C
zz Oxaloacetate (OAA) 4 C zz Oxaloacetate is regarded as:
zz Aspartate = 4 C  Carrier of TCA cycle
 1st substrate of TCA
zz Asparagine = 4 C
 Has a catalytic role in TCA
zz Succinyl CoA/Succinate = 4 C zz Sources of Oxaloacetate: Produced from pyruvate or Aspartate
zz Malate = 4C
zz Malonate/Malonyl CoA = 3C

Fig. 3.14: TCA Cycle

Explanation of the 8 Steps of TCA Cycle

T
zz Step 1: Acetyl CoA joins oxaloacetate to form citrate (known as condensation step) H
 cetyl CoA (2C) joins oxaloacetate (4C) to form citrate (6C). Enzyme is Citrate Synthase. This is irreversible step. Citrate
A E
synthase EC No. is 2 i.e. Transferase category (earlier it was 4: Lyase). Citrate inhibits glycolysis (by inhibiting PFK-1) and O
activates fatty acid synthesis (by activating acetyl Coa Carboxylase) R
zz Step 2: Conversion of Citrate to Isocitrate Y
 itrate (6C) gets converted to its isomer (6C) -Isocitrate by enzyme Aconitase, also known as Aconitate Hydratase, which
C
belongs to Lyase category. Aconitase is an iron-sulfur protein.
 zz Step 3: Conversion of Isocitrate (6C) to Alpha-Keto- Ligase/Succinyl–CoA Synthetase (EC No. = 6). This is
glutarate (5C) the only substrate level phosphorylation step of TCA
64  6-C compound gets converted to 5 C compound, so
A cycle.
decarboxylation occurs. But the name of enzyme is
Isocitrate Dehydrogenase. Dehydrogenase means H igh R eturn
oxidation occurring. So this is oxidative decarboxylation Mostly succinate thiokinase produces ATP. But in liver and kidney
CRO BIOCHEMISTRY

(removal of hydrogen and removal of CO2). Oxidative during starvation, this enzyme produces GTP, for PEPCK (Phospho
decarboxylation step always requires vitamin B1. In Enol pyruvate Carboxy Kinase) enzyme of Gluconeogenesis.
this step, NAD forms NADH. This is the first oxidative
decarboxylation step of TCA cycle. zz Step 6: Conversion of Succinate (4C) to Fumarate (4C)
zz Step 4: Conversion of Alpha-Ketoglutarate (5C) to  nzyme is Succinate Dehydrogenase. FAD gets con-
E
Succinyl CoA (4C) verted to FADH2 in this step.
 5-C compound gets converted to 4 C compound, so
A
decarboxylation occurs. But the name of enzyme is H igh R eturn
Alpha-Ketoglutarate Dehydrogenase. Dehydrogenase Succinate Dehydrogenase:
means oxidation occurring. So this is oxidative •• Present in IMM


decarboxylation (removal of hydrogen and removal of •• Part of Complex II

CO2). Oxidative decarboxylation step always requires •• Inhibited by Malonate (3C)
vitamin B1. In this step, NAD forms NADH. This is •• FAD is the Hydrogen Acceptor
the second oxidative decarboxylation step of TCA zz Step 7: Conversion of Fumarate (4C) to Malate (4C)
cycle. Alpha-Ketoglutarate Dehydrogenase enzyme is a (Hydration step):
multienzyme complex similar to PDH.
 nzyme is Fumarase or Fumarate Hydratase. This is a
E
Table 3.7: PDH complex and α-Ketoglutarate Dehydrogenase Lyase
zz Step 8: Conversion of Malate (4C) to Oxaloacetate (4C)
pyruvate Dehydrogenase Alpha-Ketoglutarate
 e enzyme is Malate Dehydrogenase. In this step, NAD
Th
complex (PDH) Dehydrogenase complex
gets converted to NADH.
•• A multienzyme complex •• A multienzyme complex
•• Requires 5 coenzymes– •• Requires 5 coenzymes–
H igh R eturn
Lipoic acid, TPP, FAD, NAD, Lipoic acid, TPP, FAD, NAD, zz Intermediates of TCA are: Oxaloacetate, Citrate, Isocitrate,
Coenzyme A Coenzyme A Alpha-Ketoglutarate, Succinyl CoA, Succinate, Fumarate,
•• Regulated by •• Not regulated by Malate
Phosphorylation and Phosphorylation and zz Acetyl CoA is not the intermediate of TCA
Dephosphorylation Dephosphorylation
EC Numbers of Enzymes of TCA
zz Step 5: Conversion of succinyl CoA (4C) to succinate zz Citrate Synthase– EC No.2 (earlier it was 4)
(4C) zz Aconitase– EC No.4
 uccinyl CoA is a high energy thioester similar to acetyl
S zz Isocitrate Dehydrogenase– EC No. 1
CoA. In this step, there is removal of this high energy CoA zz Alpha Ketoglutarate Dehydrogenase– EC No. 1
bond. When this high energy bond is broken, energy is zz Succinate Thiokinase– EC No.6
released which is entrapped in the form of ATP or GTP. zz Succinate Dehydrogenase– EC No.1
So this is known as substrate level phosphorylation zz Fumarase– EC No.4
step. Enzyme is Succinate Thiokinase/Succinate –CoA zz Malate Dehydrogenase– EC No.1

T
H
E
O
R
Y
 65

CHAPTER 3  CARBOHYDRATE METABOLISM

Fig. 3.15: Energetics of TCA cycle

ENERGETICS OF TCA CYCLE PDH is also considered regulatory step for TCA
PDH regulates TCA, mainly in brain, where major source of
acetyl CoA is carbohydrates.

H igh R eturn
Irreversible Steps of TCA
1. Citrate Synthase
2. Alpha-Ketoglutarate Dehydrogenase
If both given in question, then Citrate Synthase is best option to
be marked.

TCA Regulation
zz Inhibited by ATP, NADH and succinyl CoA. These
compounds indicate high energy state of cell.
zz Activated by ADP and NAD, which indicate low energy
state of cell
 Five coenzymes required for link reaction and TCA
Fig. 3.16: Energetics from one glucose are –lipoic Acid, Vitamin B1, Vitamin B2, Vitamin B3 and
Vitamin B5
There is not ‘One’ Rate Limiting Enzyme of TCA
Cycle Q. 
Why we take multivitamins (specially B-complex T
vitamins) when we feel lethargic? H
3 enzymes can be rate limiting, depending on various T A. Because 4 vitamins from B-complex group are E
complex conditions in the body: H required in link reaction and TCA. If these vitamins O
1. Citrate Synthase I
N are deficient then energy from link reaction and TCA R
2. Isocitrate Dehydrogenase is not released. So lethargy occurs.
3. Alpha-Ketoglutarate Dehydrogenase K Y
 Role of Vitamins in TCA
66 Coenzyme Vitamin TCA cycle enzyme
TPP B1 (Thiamine) •• Alpha-Ketoglutarate DH
•• Isocitrate DH
NAD B3 (Niacin) •• Alpha-Ketoglutarate DH
CRO BIOCHEMISTRY

•• Isocitrate DH
•• Malate DH
FAD B2 (Riboflavin) Succinate DH
Coenzyme A B5 As part of acetyl CoA,
(Pantothenic acid) Succinyl CoA

Q. Why TCA cannot occur in anaerobic conditions?

A. This is because of regulation from incoreased NADH
T


and FADH2.
H
I Fig. 3.17: Mechanism of action of Fluoroacetate in inhibiting
N Aconitase (Fluoroacetate is a plant toxin, used in pesticide)
K
H igh R eturn
zz Acetyl CoA is not the carrier of TCA cycle
zz Acetyl CoA is not the 1st substrate of TCA cycle
zz Acetyl CoA is not the intermediate of TCA cycle
zz Acetyl CoA is never Glucogenic

Summary Box
Energetics
zz Anaerobic glycolysis = 2 ATPs
zz Aerobic glycolysis = 7 ATPs
zz Complete breakdown of glucose = 32 ATPs
zz One acetyl CoA/TCA cycle = 10 ATPs
zz RBC (Aerobic/Anaerobic state) = 2 ATPs
Inhibitors of TCA
zz RBC (Fed/Fasting state) = 2 ATPs
zz RL shunt = 0 ATPs
Inhibitor Enzyme inhibited
zz Cancerous cells = 2 ATPs
1. Fluoro citrate Aconitase zz If aerobic glycolysis uses Glycerol phosphate shuttle = 5 ATPs

2. Fluoro acetate Aconitase

3. Arsenite Alpha-Ketoglutarate DH
H igh R eturn
Type of glycolysis in cancerous Cells
4. Malonate Succinate DH zz Pasteur’s effect: This effect occurs in any normal cell of body.
It says that whenever oxygen is present, then anaerobic
Malonate (3C) is inhibitor of: glycolysis is inhibited (When excess O2 present, then more ATP
1. TCA (Succinate Dehydrogenase) and Citrate gets formed, which inhibits PFK-I)
zz Warburg’s effect: This effect occurs in cancerous cells. This is
2. ETC (Complex II)
3. 
Beta-oxidation of Fatty Acids (Carnitine Palmitoyl a paradox from normal. It says that even if oxygen is present;
T Transferase I or CPT-I) glucose gets converted to lactate. This is known as aerobic
H glycolysis. So, end product of this aerobic glycolysis is lactate.
E A dditional E dge Number of ATPs in this aerobic glycolysis is 2 ATPs (as lactate
O is formed).
zz Fluoroacetate is non-competitive inhibitor of Aconitase.
 Glycolysis in cancerous cells = aerobic glycolysis
R zz Fluorocitrate is competitive inhibitor of Aconitase.
 No. of ATPs from each glucose in cancerous cells = 2 ATPs
Y
 End product of glycolysis in –Cancerous cells = lactate
 of NH2 group. So in short we can say that malate shuttle
takes NADH from cytoplasm and delivers NADH in the
mitochondria. 67
This is also known as Malate-Aspartate Shuttle as Malate
and Aspartate cross IMM in this shuttle.

CHAPTER 3  CARBOHYDRATE METABOLISM

Glycerol-Phosphate Shuttle

Fig. 3.18: Warburg effect

SHUTTLES
NADH cannot cross IMM (inner mitochondrial membrane)
but NADH is the starting material for ETC which occurs in
mitochondria. So to transport NADH from cytoplasm to
mitochondria, we need the help of shuttles. There are two
shuttles:
1. Malate Shuttle Fig. 3.20: Glycerol-Phosphate Shuttle
2. Glycerol phosphate Shuttle zz This shuttle is more important in brain and skeletal
Malate Shuttle muscles. In cytoplasm, DHAP (Dihydroxy Acetone
phosphate) gets converted to Glycerol-3-phosphate by
enzyme cytoplasmic Glycerol-3-phosphate Dehydro-
genase. NADH gets converted to NAD.
zz This Glycerol-3-phosphate formed can cross IMM and
reaches mitochondria. In mitochondria, Glycerol-3-
phosphate gets converted to DHAP by enzyme mito-
chondrial Glycerol-3-phosphate Dehydrogenase. This
reaction produces FADH2, which can enter ETC and
gives 1.5 ATPs.
zz So in short we can say that glycerol-3-phosphate shuttle
takes NADH from cytoplasm and delivers FADH2 in the
mitochondria.

A dditional E dge
Why G-3-P shuttle is present in brain, although it forms less ATP?
It is a shorter shuttle, so is a quicker source of ATP (Although less,
Fig. 3.19: Malate Shuttle but gives ATP quickly). Brain has to act rapidly so this shuttle is
This shuttle is more important in liver and heart. In present in brain
cytoplasm, oxaloacetate gets converted to malate (this zz Muscle is involved in emwgmiris, so this shuttle is present in
is opposite of TCA cycle) by enzyme cytoplasmic malate muscles also.
Dehydrogenase. NADH gets converted to NAD. This malate
formed can cross IMM and reaches mitochondria. In T
Two more shuttles in body H
mitochondria, malate gets converted to oxaloacetate by
enzyme mitochondrial Malate Dehydrogenase. This reaction Citrate Shuttle E
produces NADH, which enters ETC and gives 2.5 ATPs. To
This shuttle is used in fatty acid synthesis. Acetyl CoA is
O
complete the cycle, oxaloacetate must reach cytoplasm again R
the starting material for fatty acid synthesis. Fatty acid
but oxaloacetate cannot cross IMM, so oxaloacetate gets Y
converted to aspartate (by addition of NH2 group). Aspartate synthesis occurs in cytoplasm but acetyl CoA is produced in
can cross IMM and reaches cytoplasm. In cytoplasm, mitochondria (via link reaction). So this shuttle is required to
aspartate again gets converted to oxaloacetate by removal transport acetyl CoA from mitochondria to cytoplasm.
 ETC (FIG. 3.23)
68 H igh R eturn
zz Organelle: mitochondria
zz Organ/cell: In all the cells of the body (where mitochondria is
present)
CRO BIOCHEMISTRY

zz A vital pathway for cells (vital pathways occur in mitochondria)

zz Occurs only in aerobic conditions (requires O2)
zz Occurs both in fed and fasting state
zz ETC occurs in IMM. All components lie in IMM (Inner mitoch-
ondrial Membrane).

ETC Components
zz 5 protein Complexes – I, II, III, IV, V
Fig. 3.21: Citrate Shuttle: In mitochondria, Oxaloacetate and zz Coenzyme Q (also known as Ubiquinone)
acetyl CoA combines to form citrate (enzyme Citrate Synthase). zz Cytochrome c – a Peripheral Membrane protein


Citrate crosses IMM and reaches cytoplasm. In cytoplasm, Citrate

is broken down (by enzyme Citrate Lyase) into again Oxaloacetate
and acetyl CoA. Acetyl CoA in cytoplasm is used for the synthesis H igh R eturn
of Fatty Acid zz Coenzyme Q is the only non-protein member of ETC. It is a
lipid soluble Quinone. It is also an anti-oxidant
Creatine-Phosphate Shuttle
zz Cytochrome c is the only member of ETC, not lying in IMM.
To transport ATP from mitochondria to cytoplasm. (Fig. 3.22) zz Complex IV is the only electron carrier in which Haem iron has
an available coordination site that can react directly with O2.
Protein Complex I to IV are involved in electron transport

F undamental B ox
zz Hydrogen Atom → Equivalent to Electron
zz H+ Ion → Equivalent to Proton
Fig. 3.22: Creatine Phosphate Shuttle: ADP-ATP Translocase is
present in IMM, which transfers ATP out into the intermembrane Table 3.8: ETC Complexes
space and ADP is transferred inside into the mitochondrial Matrix.
In intermembrane space, enzyme Creatine Kinase-Mitochondrial ETC Complex Components
(CKm) converts ATP to ADP. On the other hand, this phosphate
Complex I FMN + Fe-S Complex
is used to convert creatine to creatine-phosphate. Creatine
phosphate reaches cytoplasm and again gets converted to Creatine Complex II FAD + Fe-S Complex
and phosphate used to form ATP. This ATP in muscles is used in
Complex III Cyt b, Cyt c, Fe-S Complex
muscle contraction.
Complex IV Cyt aa3, Cu

T
H
E
O
R
Y
 69

CHAPTER 3  CARBOHYDRATE METABOLISM

Fig. 3.23: Electron transport chain. mitochondria shown here with two membranes— outer and inner, separated by Intermembrane
space. Most interior is a gel like solution, known as mitochondrial Matrix.

Electron Flow Occurs from NADH to O2

Complex I takes electrons from NADH and gives it to
coenzyme Q. So, NADH gets oxidized (removal of hydrogen
or electron) and CoQ gets reduced. So name of complex I
is NADH-CoQ Reductase. Complex III takes electrons from
CoQ and gives it to Cytochrome c. So name of Complex III
is CoQ-Cytochrome c Reductase (or simply Cytochrome c
Reductase also known as cytochrome b/c1). Complex IV
takes electrons from Cytochrome c and gives it to oxygen. So
cytochrome c gets oxidised (removal of hydrogen or electron).
So name of Complex IV is cytochrome c oxidase (also known
as cytochrome a/a3) and it requires Cu as prosthetic group. T
Electrons ultimately combine with O2 and H+ to form H2O. H
This requirement of O2 is the greatest portion of O2 use for E
body, therefore ETC is also known as respiratory chain. O
(Fig. 3.24) R
Y

Fig. 3.24: Electron flow occurs from NADH to O2

 can gain electron from an electron donor and can donate
A dditional E dge these electrons to the next acceptor in the chain. In ETC,
70 zz ATP Synthase- also known as F1/F0-ATPase because it can also always next molecule has more Redox Potential. So in ETC,
catalyse the hydrolysis of ATP to ADP and Pi NADH has minimum R.P. (a Strong Electron Donor) and
oxygen has maximum R.P. (a Strong Electron Acceptor)
zz Electron flow occurs from NADH to O2. This energy of e–
CRO BIOCHEMISTRY

flow is used by complexes (I, III and IV). Using this energy
of electron flow, these complexes transfer some protons
out from mitochondrial matrix into the intermembrane
space. So these extra protons in intermembrane space
creates osmotic effect, creating a proton gradient
(chemiosmotic theory). So protons have high tendency
to go back again into the matrix. These protons can
go back via complex V, which is having ATP Synthase
activity. So ATP is produced.


Fig. 3.25: Sequence of events occurring in oxidative

phosphorylation (ETC)

Protons and ATPs Produced

One molecule of NADH gives 2.5 ATPs. When one NADH
enters ETC, then 3 complexes are involved i.e. Complex I, III
and IV. Complex I transfers 4 H+, Complex III transfers 4 H+
and Complex IV transfers 2 H+. So total 10 H+ are responsible Fig. 3.26: A flowchart showing redox pairs with increasing redox
for providing 2.5 ATPs. That means complex I gives 1 ATP. potential. Redox Pair consists of two components- one is oxidized
Complex III also gives 1 ATP and Complex IV gives 0.5 ATP. and other is reduced during electron transfer.

H igh R eturn Complex II (Fig. 3.27)

Protons (H+) FADH2 gives electrons to succinate dehydrogenase, which
zz Complex I → 4 H+ → 1 ATP produced further gives to complex II and then to coenzyme Q. After
zz Complex III → 4 H+ → 1 ATP produced CoQ, electron transport chain is same as discussed earlier. So
zz Complex IV → 2 H+ → 0.5 ATP produced Complex II takes electrons from Succinate Dehydrogenase
and gives to CoQ. So name of complex II is Succinate-CoQ
Redox Potential (Fig. 3.26) Reductase. This complex cannot transfer any proton from
matrix into intermembrane space. So it will not form any
T Electron Flow occurs due to Redox Potential (R.P.). Redox ATP. (As ATP formation depends on how many protons are
H Potential is tendency to gain electrons. Each carrier in ETC transferred)
E
O
R
Y
 transfer in such a case. So, ATP formation (phosphorylation)
does not occur. But there is no problem in oxidation i.e.
electron flow. So energy is not used for ATP formation, instead 71
it is diverted for heat generation. This is known as Non-
shivering thermogenesis.

Oligomycin

CHAPTER 3  CARBOHYDRATE METABOLISM

zz Inhibits Complex V
zz It is not an uncoupler
zz Inhibits both oxidation and phosphorylation

Translocase (Fig. 3.29)

zz It is a protein present in IMM. It helps in the transfer
of molecules across the IMM. They have Flip Flop
Mechanism.
zz In ETC, ADP-ATP Translocase is present in IMM. It
transfers ATP out from mitochondria into intermembrane
space and on the other hand it transfers ADP from
Intermembrane space into the mitochondria.
zz Atractyloside inhibits ADP-ATP Translocase

Fig. 3.27: Sequence of events when ETC starts from FADH2

ETC Couples (Fig. 3.28)

In ETC, oxidation (flow of electrons) and phosphorylation
(ATP formation) are like couples i.e. they always occur
together.
Uncoupling: Oxidation is occurring, but phosphorylation
does not occur. A substance which creates a hole in the
IMM, acts as an uncoupler. E.g. of uncouplers are → Drugs
acting as uncouplers (2, 4-Dinitrophenol) and natural Fig. 3.29: Flip Flop Mechanism: Translocase– transfers molecules
uncouplers [Thermogenin (also known as uncoupling across IMM. E.g. It binds molecule X on mitochondrial side and
protein-1–UCP–1)] present in brown fat is responsible for molecule Y on the cytoplasmic side. Then it flip flop. So that
non-shivering thermogenesis. So, brown fat is involved in molecule X comes on cytoplasmic side and molecule Y comes to
energy expenditure. mitochondrial side.

H igh R eturn
zz ADP to ATP conversion is inhibited by - Oligomycin
zz ADP to ATP transfer is inhibited by – Atractyloside
zz ATP formation is also inhibited by uncouplers but uncouplers
indirectly inhibit this. Oligomycin directly inhibit Complex V
and inhibit ADP to ATP conversion

Table 3.9: Inhibitors of ETC

Complex Inhibitors
Fig. 3.28: ETC Uncouplers Complex I Rotenone, Phenobarbitone, Amobarbital T
Piericidin A H
H igh R eturn Complex II Malonate (3C) E
zz Non-Shivering Thermogenesis: Thermogenin acts as Carboxin (fungicide) O
uncoupler. This is a protein present in brown fat (in Neonates TTFA (Trienoyl Tri Fluoro Acetone) R
and Hibernating animals), which is like a H+ channel. So when
themogenin is present, then protons move through this Complex III Phenformin Y
thermogenin channel. Complex V is not used for proton Antimycin A
BAL {British Anti Lewistie} or Dimercaprol
Contd…
Contd…
 Complex Inhibitors GLUCONEOGENESIS
72 Complex IV CO, CN, H2S, Sodium Azide H igh R eturn
•• Uncouplers of ETC: Dinitrophenol, Thermogenin zz Nature of pathway: Catabolic
•• ADP to ATP conversion is inhibited by: Oligomycin
zz Activated by hormone: Glucagon
•• ADP to ATP transfer is inhibited by: Atractyloside
CRO BIOCHEMISTRY

zz Occurs in fasting state

zz Organelle: Both mitochondria and cytoplasm
H igh R eturn zz Organ/cell: Liver and kidney
Phenformin
zz Oral Hypoglycemic agent
zz Inhibits ETC so NADH accumulate
zz This leads to increased conversion of pyruvate to lactate,
leading to Lactic acidosis.
zz Lactic Acidosis leads to Hyperuricemia because lactic acid
competes with uric acid for excretion by kidneys. If excess
lactic acid is present, kidneys keep excreting lactic acid, leaving


uric acid in body. This leads to Hyperuricemia.

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Fig. 3.30: Gluconeogenesis Pathway
Fasting/Starvation State (Refer Fig. 3.31) NOTE: This is not malate shuttle. Because the purpose of malate
In fasting/starvation, fat is the preferred fuel. So adipose shuttle is to take NADH from cytoplasm to mitochondria. But here
Tissue is broken down to release fatty acids in blood. These NADH is getting transported from mitochondria to cytoplasm 73
fatty acids reache liver, where beta oxidation is done, releasing (opposite to the purpose of malate shuttle). In gluconeogenesis,
acetyl CoA, which has 3 fates. Either this acetyl CoA enters we need to transport OAA from mitochondria to cytoplasm. This
NADH generated in cytoplasm is used in gluconeogenesis in the

CHAPTER 3  CARBOHYDRATE METABOLISM

TCA, or enters ketone body synthesis, or it is used for the
activation of the first step of Gluconeogenesis. conversion of 1,3 Bisphospho Glycerate to Glyceraldehyde-3-P by
enzyme Glyceraldehyde-3-P Dehydrogenase.
NOTE: Acetyl CoA can never form glucose. Acetyl CoA activates
the very first step of Gluconeogenesis i.e. Pyruvate Carboxylase.

Rule of 2 for Gluconeogenesis

zz 2 compartments–mitochondria and cytoplasm
zz It starts from mitochondria (Any pathway which occurs
both in mitochondria and cytoplasm starts from
mitochondria. Other examples–Urea Cycle and Haem
Synthesis)
zz 2 organs: Liver and Kidney –90% in Liver, 10% in Kidneys.
Small Intestine can also make glucose.
zz 2 situations: zz Next step is conversion of OAA to Phosphoenol pyruvate
1. Fasting/Starvation (not eating food) (by decarboxylation and phosphorylation). Enzyme is
2. Diabetes Mellitus Phospho-Enol-pyruvate Carboxy Kinase (PEPCK). This
enzyme is a Lyase. GTP is used for this phosphorylation
Gluconeogenesis Pathway (Fig. 3.31) step.
It starts from mitochondria. In fasting state, pyruvate zz The combined action of pyruvate Carboxylase and
is diverted towards the formation of oxaloacetate in PEPCK provide energy favourable for the conversion of
pyruvate to PEP (here pyruvate Kinase, (irreversible),
mitochondria. Enzyme used is pyruvate carboxylase, which
cannot be used).
requires biotin and ATP [any carboxylase requires biotin-
zz Then Phosphoenol pyruvate (PEP) gets converted to
(Vit B7 and ATP)]. This is the first step of Gluconeogenesis.
fructose 1,6 Bisphosphate by same enzymes which
This is activated by acetyl CoA (When OAA is used for
are used in glycolysis i.e. Enolase, Phospho Glycero
gluconeogenesis in fasting state, then acetyl CoA is not used
Mutase, Phospho Glycerate Kinase, Glyceraldehyde-3-P-
up in TCA. So acetyl CoA accumulates. This acetyl CoA is Dehydrogenase and Aldolase A. These enzymes catalyze
very important allosteric activator of pyruvate Carboxylase). reversible reactions.
This reaction is also the most important anaplerotic reaction zz Then fructose 1,6 Bisphosphate gets converted to fructose-
(replenish TCA intermediate OAA) (Fig. 3.32). 6-P by enzyme fructose 1,6 Bisphosphatase (In glycolysis
opposite occuring i.e. conversion of fructose 6-phosphate
to fructose 1,6 Bisphosphate by enzyme PFK-I).
zz Then fructose-6-phosphate gets converted to Glucose-
6-P by enzyme Phospho Hexose-Isomerase (same
enzyme used in glycolysis).
zz Then the last step is conversion of glucose-6-P to
glucose by enzyme glucose-6-Phosphatase (opposite
to this occur in glycolysis i.e. conversion of glucose
to glucose-6-P by enzyme Hexokinase). This last step
of gluconeogenesis does not occur in cytoplasm,
instead enzyme glucose-6-Phospatase is present in the
Fig. 3.31: Acetyl CoA during Fasting membrane of ER (Endoplasmic Reticulum)
zz Gluconeogenesis is not just the reversal of glycolysis as
zz So this reaction produces oxaloacetate (OAA) in
three enzymes are different in gluconeogenesis
mitochondria. Now next steps of gluconeogenesis
T
occurs in cytoplasm. So this oxaloacetate has to come
H
out in cytoplasm but OAA cannot cross IMM. OAA gets
converted to Malate by enzyme mitochondrial Malate E
Dehydrogenase. Malate formed can cross IMM and it O
reaches cytoplasm, where it again gets converted to OAA R
by enzyme cytoplasmic Malate Dehydrogenase Y
 Table 3.10: Irreversible enzymes of glycolysis shown and the fructose1, 6 -Bisphosphatase is used in Gluconeogenesis.
counterpart enzymes in gluconeogenesis. Instead of Hexokinase, glucose-6 Phosphatase is used in
74 Gluconeogenesis.
Glycolysis Gluconeogenesis
Pyruvate Kinase •• Pyruvate Carboxylase H igh R eturn
•• PEPCK
CRO BIOCHEMISTRY

Q. Why glucose-6-phosphate of hepatocyte is not acted

PFK–1 Fructose 1, 6 Bis Phosphatase upon by glucose-6-Phosphatase as soon as it is formed?
A. Because glucose-6-Phosphatase is present in the mem-
Hexokinase Glucose-6-Phosphatase brane of ER, not in cytoplasm (Fig. 3.32). This enzyme
Instead of pyruvate Kinase, enzymes used in cannot be kept in cytoplasm because it will break glucose
gluconeogenesis are pyruvate Carboxylase and PEPCK -6- phosphate of cytoplasm which should enter into
(Phosphoenol-pyruvate Carboxy Kinase). Instead of PFK-1, glycolysis and should not be broken down by glucose -6-
Phosphatase


Fig. 3.32: Glucose -6- Phosphatase enzyme present in membrane of ER (Endoplasmic Reticulum). In fasting when glucose-6-phosphate
is formed in cytoplasm via gluconeogenesis, then this Glu-6-P enters ER (via T1 transporter) and gets broken down by Glucose-6-
Phosphatase. Products formed are glucose and inorganic phosphate. Glucose exits ER via T2 transporter and inorganic phosphate exits ER
via T3 transporter.
Rate Limiting Enzymes
There is no ‘one’ rate limiting enzyme in gluconeogenesis.
Here three enzymes can be rate limiting, depending on
various complex situations in body. They are:
1. Pyruvate Carboxylase
2. PEPCK (Phospho Enol pyruvate Carboxy Kinase)
3. fructose 1, 6-Bisphosphatase

H igh R eturn
Enzyme Commission Number of Enzymes of
Substrates of Gluconeogenesis: Precursors which can be
Gluconeogenesis
converted to glucose. Mostly 3 C compounds are good substrates
T
Enzyme EC No. of Gluconeogenesis.
H 1. Pyruvate-(3C) (Source of pyruvate in Fasting: Link Reaction not
E pyruvate Carboxylase 6 occurring so pyruvate accumulates)
O 2. Lactate: 3C (lactate from anaerobic glycolysis gets converted
PEPCK 4 to pyruvate by enzyme lactate Dehydrogenase, which is
R
fructose 1, 6-Bisphosphatase 3 reversible enzyme)
Y
3. Glycerol: 3C (Glycerol converted to Glycerol-3-P by Glycerol
glucose-6-Phosphatase 3 Kinase in Liver and then Glycerol-3-P forms DHAP (an inter-
mediate of glycolysis) by enzyme Glycerol-3-P dehydrogenase)

Contd…
 4. P ropionic Acid: 3C (gets converted to Succinyl CoA, which is
intermediate of TCA)
5. Any TCA intermediate can form glucose as they can form OAA, 75
which can be converted to glucose
6. Amino Acids (See Fig. 3.33)
Fats cannot be converted to carbohydrates. But Glycerol and

CHAPTER 3  CARBOHYDRATE METABOLISM

Propionic Acid are two breakdown products of fats which can be
converted to Carbohydrates.

H igh R eturn
zz Purely Ketogenic amino acids = 2
zz Both Ketogenic and Glucogenic amino acids = 5
zz Purely Glucogenic amino acids = 13
zz So in total, Ketogenic amino acids = 7 (2 + 5)
zz So in total, Glucogenic amino acids = 18 (13 + 5)
zz So only 2 amino acids can never form glucose. Rest 18 can
form glucose Fig. 3.33: Amino Acids can be Glucogenic, Ketogenic or both.
Alanine (3C) is most glucogenic amino acid

Energetics of Gluconeogenesis (Fig. 3.34)

zz 2 Pyruvate → Glucose (6 ATP used)
zz 2 Lactate→ Glucose (6 ATP used)
zz 2 Alanine → Glucose (6 ATP used)

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Fig. 3.34: Two pyruvate (3C) gets converted to one molecule of glucose (6C) and this requires use of 6 high energy phosphate bonds and
oxidation of 2 NADH molecules. Same energetics is with lactate or alanine.
76
CRO BIOCHEMISTRY


Fig. 3.35: Energetics from Lactate (Gluconeogenesis): Involves replenishment of NADH

Reciprocal regulation of glycolysis and As fructose 2,6 Bisphosphate is formed in fed state. So
gluconeogenesis (Fig. 3.36) fructose 2,6 Bisphosphate acts as an intracellular signal of
glucose abundance.
In body, reciprocal regulation occurs always for two Hormonal role: Glucagon activates gluconeogenesis in
opposite pathways e.g. glycolysis (glucose to pyruvate) fasting state. Insulin inhibits gluconeogenesis in fed state.
and gluconeogenesis (pyruvate to glucose). In fed state,
glycolysis occurs and gluconeogenesis is inhibited. But in
fasting state, opposite occurs i.e. glycolysis is inhibited and H igh R eturn
gluconeogenesis is activated. zz Fructose 1, 6-Bisphosphate – Glycolysis
zz Fructose 1, 6-Bisphosphatase – Gluconeogenesis
zz Fructose 2, 6-Bisphosphate – Reciprocal Regulator (produced
in fed state)
zz Fructose 2, 6-Bisphosphatase – Active in cancerous mutation

T Q. If fructose 2, 6-Bisphosphatase is active, what will

H happen to glycolysis ?
I A. Glycolysis decreases
N Explanation: If fructose 2, 6 Bisphosphatase is active,
K then fructose 2,6 Bisphosphate gets converted to
fructose-6-P. So fructose 2,6 Bisphosphate decreases.
Fructose 2,6 Bisphosphate usually activates
glycolysis. So if fructose 2, 6 Bisphosphate is not
present, then glycolysis decreases.
Fig. 3.36: Reciprocal regulation of glycolysis and gluconeogenesis.
In fed state, enzyme PFK-II (Phosphofructokinase–II) is activated.
This enzyme produces fructose 2, 6 Bisphosphate. This molecule
TIGAR (TP-53 Induced Glycolysis and Apoptosis
acts as reciprocal regulator i.e. it activates glycolysis (by activating Regulator) (Fig. 3.37)
the rate limiting enzyme of glycolysis) and on the other hand, it
inhibits gluconeogenesis (by inhibiting the enzyme fructose-1, TIGAR regulates glycolysis and Apoptosis in the Cancerous
T cell. TIGAR is a protein which is induced by TP-53 protein.
6-Bisphosphatase).
H
E In fasting state, opposite will occur. Fructose-2,6 Bisphos-
O phate is not formed. So, glycolysis decreases and on the other
R hand, gluconeogenesis is activated.
Y
 77

CHAPTER 3  CARBOHYDRATE METABOLISM

Fig. 3.37: TIGAR. PFK-I enzyme is the rate limiting enzyme of glycolysis. PFK-II enzyme is active only in fed state, producing fructose 2, 6
Bisphosphate. This fructose 2, 6 Bisphosphate activates PFK-I and thus activates glycolysis. Fructose 2, 6 Bisphosphatase enzyme is active
in cancerous mutation. If a cancerous mutation occurs in some cell, then p-53 gene (Tumor Suppressor gene) is activated, producing its
protein TP-53 . This protein induces another protein – TIGAR. TIGAR has enzymatic activity of fructose 2,6 Bisphosphatase. This enzyme
decreases glycolysis in that cancerous cell. The benefit of decreasing glycolysis is that further replication will not occur so that another
cancerous cell is not formed. And when glycolysis is decreased, then repair of the cancerous mutation can occur. If repair occurs then
TIGAR return back. But if repair does not occur, then TIGAR will induce apoptosis of this cancerous cell. So TIGAR regulates glycolysis and
apoptosis in the cancerous cell. Hence full form is TP-53 induced Glycolysis and Apoptosis Regulator.

GLYCOGEN
It is the storage form of carbohydrates or energy in the body.
We all know that fats gives more energy than carbohydrates
plus fats have more abundant stores in body but glycogen
reserves are limited. Then why body stores energy in the form
of carbohydrates (glycogen) and not in fats?
The reason is:
zz Fats cannot be metabolized anaerobically. During
exercise, we have anaerobic conditions so only glucose
can be used, which is obtained from glycogen.
zz Mobilization of glycogen is faster than fats
zz Fats can never be converted to glucose so fats cannot
maintain blood glucose levels.
Fig. 3.38: Glycogenin in centre of a glycogen granule
zz Also fatty acids cannot cross Blood Brain Barrier (BBB)
but glucose can.
Glycogen Metabolism
Glycogenin zz Synthesis of glycogen from glucose is known as
zz Biochemically, it is a glycoprotein Glycogenesis T
zz Acts as a primer in glycogen synthesis zz Breakdown of glycogen to produce glucose is known as H
zz Acts as auto-catalyst (in attaching glucose on glycogenin) Glycogenolysis E
zz Glucose gets attached to –OH group of Tyrosine on zz Glyconeogenesis: Synthesis of glycogen from Non- O
glycogenin carbohydrate sources. This is almost same like gluconoe- R
zz Glycogenin is present in the core of a full glycogen genesis. But the last step (glucose-6-P to glucose) is not
Y
structure, known as glycogen granule (Fig. 3.38). occurring. Instead, glucose-6-P directly goes for the
formation of glycogen.
 In liver, the function of glycogen is to maintain normal
H igh R eturn blood glucose levels. So when glycogenolysis occurs in liver,
78 zz Compartment: Both glycogenesis and glycogenolysis occurs in the end product (i.e. Glucose) should come out of the Liver
cytoplasm cell. (GLUTs are bidirectional but only glucose can cross the
zz Both rate limiting enzymes of glycogen metabolism are cell membrane, Glu-6-P cannot cross via GLUT). So the end
transferases product is free glucose.
CRO BIOCHEMISTRY

Glycogen gets stored in liver and muscle. Minor amount In muscles, the function of glycogen is to give energy for
stored in brain also. But because very little glycogen is present muscle activity/ contraction. So when glycogenolysis occurs
in brain, that’s why brain largely depends on blood glucose. in muscles, then glucose should not come out of muscle cells
because muscle cell has to use it for its own purpose. So the
F undamental B ox end product is glucose-6-phosphate and not glucose.

zz By weight, glycogen is more in liver (means if we take same Linkages in Glycogen (Fig. 3.39 (a) and (b))
weight of liver and muscle, then glycogen is more in Liver)
zz By percentage, glycogen is more in muscle (means if we Straight chains have α 1 → 4 bonds. And the branching point
calculate total body’s percentage of glycogen, then it is more has α 1 → 6 bond.


in muscles because muscle mass is more than liver mass)

(a)

Table 3.11: Difference between Liver glycogen and Muscle

(b) Glycogen

Liver Glycogen Muscle Glycogen

Function– Blood glucose Function–Muscle contraction
maintainence
End product– Free glucose End product: Glucose-6-phosphate
(non-phosphorylated)
Stores increased in fed Stores not affected in short period
state and depleted in of fasting and is only moderately
fasting decreased in prolonged fasting
glucose-6-Phosphatase glucose-6-Phosphatase absent
present

Glycogen Synthesis (Fig. 3.40 & 3.41)

H igh R eturn
T zz Also known as Glycogenesis
H Fig. 3.39: Reducing and non-reducing ends of Glycogen shown. If zz Nature of Pathway: Anabolic
E C1 (functional carbon) of glucose is free, then glucose is reducing. zz Occurs in Fed state
O If C1 (functional carbon) of glucose is involved in bond, then zz Activated by hormone: Insulin (all Anabolic pathways are
R glucose is non-reducing activated by Insulin)
Y zz Organelle: Cytoplasm (any Anabolic pathway occurs in
cytoplasm)
zz Organ/cell: Liver and Muscle
zz Glucose gets converted to glucose-6-phosphate by enzyme Hexokinase/Glucokinase.
zz Then glucose-6-phosphate is converted to glucose-1-phosphate by enzyme mutase (intramolecular transfer of phosphate
occurs). 79
zz Glucose-1-phosphate is converted to UDP-glucose by enzyme UDP glucose Pyro-Phosphorylase. Pyrophosphate, released
by this reaction is hydrolysed to two inorganic phosphates by pyro phosphatase.
zz Then UDP-glucose is used to add glucose residues to non- reducing end of glycogenin (primer required for glycogen

CHAPTER 3  CARBOHYDRATE METABOLISM

Synthesis). This is done by the rate limiting enzyme –Glycogen synthase, which adds α (1 → 4) bonds i.e. only straight
chains are synthesized by this enzyme.
zz Another enzyme Branching enzyme (also known as amylo α (1 → 4) → α (1 → 6) transglycosylase or simply 4:6 Trans-
ferase) creates branch points i.e. α (1 → 6) bonds.
zz So, this way glycogen is formed having α (1 → 4) bonds and α (1→ 6) bonds.

Fig. 3.40: Biosynthesis of glycogen

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Fig. 3.41: Glycogen metabolism (synthesis and breakdown)

 Glycogen Breakdown
80
H igh R eturn
zz Also known as Glycogenolysis
zz Nature of pathway: Catabolic
CRO BIOCHEMISTRY

zz Occurs in fasting or in between meals

zz Activated by hormone: Glucagon (all Catabolic pathways are
activated by Glucagon)
zz Organelle: Cytoplasm (both glycogen synthesis and
breakdown occurs in cytoplasm)
zz Organ/cell: Liver and Muscle

Minor Pathway of Glycogenolysis: Only this glycogen

breakdown (1-3%) occurs in lysosomes. Minor pathway
enzyme is Acid Maltase (also known as Acid α (1 → 4)
Glucosidase). Deficiency of this enzyme is Pompe’s disease


(Type II Glycogen Storage Disease).

Major pathway of Glycogenolysis: Occurs in cytoplasm.
Enzymes involved are (Fig. 3.42):
1. Glycogen Phosphorylase: Rate Limiting and the
first enzyme which starts breakdown of glycogen.
This enzyme breaks a (1 → 4) bonds i.e. only straight
chains are broken. And it starts breaking down glucose
residues from the non-reducing end of every branch.
This glucose is released in the form of glucose-1-
phosphate. Glycogen Phosphorylase can break glucose
residues in a chain only till 4 glucose residues are left in
the branched chain Phosphorylase breaks only a(1→
4) bonds and not a (1 → 6) bonds. The structure left
is known as Limit Dextrin. This enzyme uses PLP –
Pyridoxal phosphate (Vit B6) as a coenzyme. PLP acts
as a phosphate donor here. Phosphorylase enzyme
transfers phosphate to glucose from PLP (known as
Phosphorolysis).
2. Glucan Transferase: Transfers 3 glucose residues
from one chain to the neighbouring chain. Full name
of this enzyme is Oligo a (1 → 4) → a (1→ 4) Glucan
Transferase or simply 4:4 Transferase.
3. Debranching enzyme: Breaks a (1→ 6) bond, which
is present at the starting of a branched chain. This
glucose is released in the form of free glucose. This
enzyme is also known as Amylo a (1→ 6) Glucosidase
activity

A dditional E dge
Debranching enzyme is a bifunctional enzyme (two enzymatic
activities on a single protein i.e. Glucan Transferase and Debran-
T ching enzyme).
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Fig. 3.42: Glycogen breakdown
Y
 zz Muscle glycogen function is to provide glucose for muscle
contraction so this glucose should not come out of muscle cell.
Therefore end product is glucose-6-P and not free glucose. 81
zz Pompe’s disease is the only Glycogen Storage Disease, which is
a Lysosomal Storage Disease
zz Enzyme common b/w glycolysis and glycogenesis- Hexokinase

CHAPTER 3  CARBOHYDRATE METABOLISM

/Glucokinase
zz Enzyme common b/w glycogenesis and glycogenolysis- Phos-
phoglucomutase
Fig. 3.43: Summary of glycogen breakdown zz Enzyme common between glycogenolysis and gluconoe-
genesis- glucose-6-Phosphatase
Isomerization of Glucose-1-P to glucose-6-P
This is done by enzyme Mutase (or Phosphoglucomutase). Glycogen Regulation (Figs. 3.44 & 3.45)
This enzyme is common to both glycogenesis and In body, reciprocal regulation occurs always for two
glycogenolysis. Next glucose-6-P is converted to free glucose opposite pathways e.g. Glycogenesis (Glycogen synthesis)
by enzyme glucose-6-Phosphatase (same enzyme is used and Glycogenolysis (Glycogen breakdown). In fed state,
in gluconoegenesis-last step). This last step occurs only in glycogenesis occurs and glycogenolysis is inhibited. But in
liver because this free glucose in liver is used to maintain fasting state, opposite occurs i.e. glycogenesis is inhibited
blood glucose levels. But this last step is not needed in and glycogenolysis is activated.
muscles because directly glucose-6-P is entering glycolysis to Basics: Glycogen Synthase is anabolic (involved in
provide energy to muscle cell for contraction. So glucose-6- anabolic pathway i.e. glycogen synthesis) enzyme. Any
Phosphatase is absent in muscles. anabolic enzyme is activated by insulin and is active in
dephosphorylated state. Glycogen phosphorylase is a
H igh R eturn catabolic enzyme (involved in glycogen breakdown). Any
zz Free glucose can cross cell membrane via GLUTs but phos- catabolic enzyme is activated by glucagon and is active
phorylated glucose (glucose-6-P) is entrapped inside the cells. in phosphorylated state. So we conclude that glycogen
synthase is active in dephosphorylated state and glycogen
Contd… phosphorylase is active in phosphorylated state.

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Fig. 3.44: In fasting state, Glucagon hormone is released. Glucagon always causes phosphorylation by increasing cAMP. So both RLE are R
phosphorylated. When Glycogen Synthase is phosphorylated, it is inactive and when Glycogen Phosphorylase is phosphorylated, it is
Y
active. This leads to activation of glycogenolysis and inactivation of glycogen synthesis
82
CRO BIOCHEMISTRY


Fig. 3.45: In fed state, insulin hormone is released. Insulin always causes dephosphorylation by decreasing cAMP. So both RLE are
dephosphorylated. When Glycogen Synthase is dephosphorylated, it is active and when Glycogen Phosphorylase is dephosphorylated, it
becomes inactive. This leads to activation of glycogen synthesis and inactivation of glycogenolysis
Forms of Glycogen Phosphorylase (Fig. 3.46)
Glycogen Phosphorylase (RLE of glycogen breakdown) has
two forms: a and b. 'a' is active form. 'b' is inactive form. 'b'
gets converted to 'a' form by phosphorylation (Any catabolic
enzyme is active in phosphorylated state)

Fig. 3.47: In response to Hypoglycemia (Glucagon released) and

stress (Epinephrine released), blood glucose needed, so liver
glycogenolysis is activated. During muscle contraction, there
is urgent and rapid need of ATP, so muscle glycogenolysis is
activated. Glucagon can act only on liver. Epinephrine can act on
both liver and muscle. Both Glucagon and Epinephrine increase
glycogenolysis by increasing cAMP and causing phosphorylation.

Glycogen Storage Diseases (GSD)

T zz Most diseases affect glycogenolysis

H zz That’s why known as Glycogen Storage Diseases i.e.
Fig. 3.46: Activators and inhibitors of Glycogen Phosphorylase glycogen is not broken down and thus it gets stored.
E
O A dditional E dge zz Liver: If GSD affects liver, then patient presents with
Hypoglycemia and hepatomegaly because function of
R
zz Muscle Phosphorylase has a binding site for 5’AMP. It activates liver glycogen is to give glucose to blood.
Y this enzyme without phosphorylation zz Muscle: If GSD affects muscles, then patient presents
zz Epinephrine acts in muscle and liner with muscle cramps and exercise intolerance because
zz In muscle and liver, there is cAMP independent activation of function of muscle glycogen is muscle contraction.
glycogenolysis with Ca-Calmodulin sensitive Phosphorylase Kinase
Table 3.12: Glycogen Storage Diseases (GSD)– Type-I to Type-VI 3. Cori’s disease also known as Limit Dextrinosis
because in this disease debranching enzyme not
Type Disease Enzyme Substrate Organ present. So α (1 → 6) bonds are not broken, therefore a 83
Accumu- structure is left having lots of α (1 → 6) bonds , which is
lated known as Limit Dextrins.
I Von Gierke’s Glucose-6- Glucose-6- Liver Debranching Deficiency

CHAPTER 3  CARBOHYDRATE METABOLISM

Phosphatase phosphate↑  Kidneys not enlarged
II Pompe’s Acid Maltase Glycogen in  No lactic acidosis and hyperuricemia
Lysosomes 4. Anderson’s disease also known as Amylopectinosis.
L iver, Due to lack of branching enzyme, a less branched
III Cori’s/Limit Debranching Limit
Muscle (or no branches at all) structure is formed known
Dextrinosis Enzyme Dextrins
and as amylopectin (a form of starch and starch is less
IV Anderson/ Branching Amylopectin Brain branched than glycogen).
Amylopectinosis Enzyme 5.  Mc Ardle’s disease: The characteristic diagnostic
V Mc Ardle’s Muscle Muscle Muscle feature is normal lactate after exercise because the
Phosphorylase Glycogen first enzyme of glycogen breakdown of muscles is not
VI Her’s Liver Liver Liver present. So glycogen is not broken down in muscles,
Phosphorylase Glycogen glucose not released and therefore lactate is not
formed. Therefore lactate is normal after exercise
1. Von Gierke’s disease: This is Type I glycogen storage because cells other than muscles can produce lactate.
disease and it is also the most common GSD (Fig. 3.48): After exercise, raised levels of lactate is because of
2. Pompe's disease is the only GSD which is a Lysosomal muscle glycogen breakdown (This cannot occur in Mc
Storage Disease. Patient has hypertrophic cardio- Ardle’s disease).
myopathy, progressive skeletal myopathy. Death 6.  Her’s disease: Only liver affected like Type I GSD but
occurs within 2 years due to cardiac failure. hypoglycemia is more severe in Type I as compared to
Type VI.

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Fig. 3.48: Von Gierke's disease clinical features Y
 Cause of Hyperuricemia in Von Gierke’s Disease
A dditional E dge
1. Lactic Acidosis
84 2. HMP
Other pathways which do not produce ATP
� HMP
3. Sequesteration of Pi (Phosphate is bound in the form � α-oxidation
of Glu-6-phosphate. So ATP is not formed to AMP � Uronic acid pathway
CRO BIOCHEMISTRY

and ADP are degraded leading to increased uric acid � Oxidation of VLCFA [Very Long Chain Fatty Acids]
formation) � RL shunt
Table 3.13: Comparison of Type I and Type VI
Table 3.14: Differences between HMP and EMP (Glycolysis)
Type I (Von Gierke’s disease) Type VI (Her’s disease)
EMP HMP
Deficiency of glucose-6-Phosphatase Deficiency of Glycogen
(this enzyme is common to both Phosphorylase Major pathway for Glucose Minor pathway for Glucose
glycogenolysis and gluconeogenesis) Catabolic pathway Anabolic pathway
Only Liver affected Only Liver affected No CO2 produced CO2 produced
No Glycogenolysis No Glycogenolysis ATP used and produced No ATP used and produced


Severe Hypoglycaemia Mild Hypoglycemia Occurs in all cells Special sites

Ketosis occurs Ketosis does not occur Dehydrogenase —NAD Dehydrogenase —NADP
Starting point - glucose Starting point — glucose-6-P
HMP
HMP is more complex than glycolysis
H igh R eturn
zz Nature of pathway: Anabolic HMP Pathway – Oxidative Phase (Fig. 3.49)
zz Occurs in fed state 1. First reaction is conversion of glucose-6-P to 6-Phospho
zz Activated by hormone: Insulin (usually anabolic pathways are
Gluconate by enzyme G-6-PD (glucose-6-phosphate
activated by Insulin)
zz Organelle: Cytoplasm (any anabolic pathway occurs in cyto- Dehydrogenase). This is oxidation step and NADPH is
plasm) formed. G6PD is the rate limiting step,
zz Organ/cell: Liver, Adipose tissue, Lactating Mammary glands, 2. Oxidation step by enzyme 6-Phospho Gluconate
Adrenal Cortex, Gonads and RBCs Hydrolase converts 6-Phospho Gluconate to 3-Keto
zz Alternate pathway for oxidation of glucose 6-Phospho Gluconate. NADPH produced at this step
zz Major source for NADPH also.
zz No ATP generated but CO2 is produced

zz HMP→ Hexose Mono phosphate Pathway: This name

given because glucose-6-phosphate is the starting
material (not glucose)
zz PPP→ Pentose phosphate Pathway: This name given
because Ribose-5-phosphate is synthesized by this path-
way
The only source of Ribose-5-phosphate (synthesize
DNA and RNA) is HMP. But NADPH comes from 2 other minor
sources also, i.e. Malic enzyme (NADP dependent Malate
Dehydrogenase) and cytoplasmic Isocitrate Dehydrogenase

F undamental B ox
zz HMP has 2 phases:
1. Oxidative Phase: synthesize NADPH, irreversible Fig. 3.49: Oxidative phase (3 steps) of HMP
2. Non-Oxidative Phase: synthesize Ribose-5-P, reversible
Cell with greater need for NADPH than Ribose-5-P: Oxidative 3. Spontaneous Decarboxylation → No enzyme required
T zz
and non oxidative phase both occurs. Oxidative phase here. The product formed is Ribulose-5-P.
H
E produces NADPH here and non-oxidative phase reactions Summary Box
O occur and finally produce glycolysis intermediates, which can
enter glycolysis. •• So, in Oxidative Phase, 2 NADPH, 1 CO2 and one Ribulose-
R 5-P is formed per glucose-6-P molecule oxidized.
zz Cell with greater need for Ribose-5-P than NADPH: Only non-
Y oxidative phase occurs in this case because it is reversible. Non-
•• 3 molecules of glucose-6-P actually enters HMP, so in total
6 NADPH, 3 CO2 and 3 molecules of Ribulose-5-phosphate
oxidative phase can provide Ribose-5-P from Glyceraldehyde-
produced.
3-P and fructose-6-P.
HMP Pathway – Non-Oxidative Phase: (Fig. 3.50) Q. Molecule which is intermediate as well as end
zz This phase uses three molecules of Ribulose-5-P and product in HMP?
T A. Glyceraldehyde-3-phosphate 85
various rearrangement reactions occur. H Q. Molecule which is substrate as well as end product
zz 2 molecules of Ribulose-5-P produce Xylulose-5-P I
in HMP?
(by enzyme Epimerase) and Ribose-5-P (by enzyme N
A.

CHAPTER 3  CARBOHYDRATE METABOLISM

Isomerase). Transketolase enzyme transfers 2 carbon K Glucose-6-phosphate
units from Xylulose-5-P to Ribose-5-P to produce Q. Is fructose-6-phosphate an end product of HMP
Sedoheptulose-7-phosphate and Glyceraldehyde-3-P. pathway?
Next is Transaldolase reaction which transfers 3 carbon A. Fructose-6-phosphate is formed in HMP, but it gets
unit and form fructose-6-P and Erythrose-4-P. converted to glucose-6-phosphate. So it is not an end
product of HMP.
zz Now third molecule of Ribulose-5-P is used to again
form Xylulose-5-P (by enzyme Epimerase). Then second
Transketolase reactions occur and 2-C unit transferred Table 3.15: Sites of HMP
from Xylulose-5-P to Erythrose-4-P producing fructose Uses of NADPH Sites of Pathway
6 – phosphate and Glyceraldehyde–3–phosphate.
zz So we have got 2 molecules of fructose–6–P and one 1. Fatty Acid Liver, Adipose tissue,
Synthesis Lactating Mammary Glands All these
molecule of Glyceraldehyde–3–phosphate. Fructose-6-P
sites are sites
forms glucose-6-P. Glyceral dehyde–3–phosphate and 2. Cholesterol Liver, Adipose tissue
of HMP as
glucose-6-P can then enter glycolysis. 3. Steroid Adrenal cortex, gonads NADPH is
(Testis and ovaries), produced by
Placenta HMP
4. To convert RBCs
GSSG to GSH

H igh R eturn
zz Tissues which are never the site of HMP are: Non Lactating
Mammary Glands and Skin
zz HMP activity is also low in Skeletal Muscles and brain, where
almost all the glucose is used by glycolysis

Function of HMP (Fig. 3.51)

To convert reduced glutathione to oxidized glutathione.

Glutathione
Glutathione is a tripeptide made up of three amino acids
—Glutamate, Cysteine and Glycine. It is a reducing agent.
Reducing property is due to –SH (Sulfhydryl) group in
Cysteine.

Fig. 3.50: Non-oxidative phase of HMP

Summary Box
So complete reaction of HMP can be written like this:
3 G–6–P + 6 NADP _→ 2 G–6-P + Glyceraldehyde–3–P + 3CO2 + 6 NADPH
T
E asy to L earn H
H igh R eturn zz Glutathione = Glutamate + Cysteine + Glycine
E
Creatine = Arginine + Methionine + Glycine O
zz Transketolase → requires TPP (derivative of Vit B1) and Mg zz

zz Transketolase transfers 2 carbon units zz Carnitine = Lysine + Methionine R

zz Transaldolase transfers 3 carbon units zz SAM formed by methionine is used actually as methyl donor in Y
both creatine and carnitine.
 URONIC ACID PATHWAY (FIG. 3.52)
86
H igh R eturn
Similarities with HMP:
zz Minor pathway for Oxidation of glucose
CRO BIOCHEMISTRY

zz Starts from glucose-6-phosphate

zz No ATP produced
zz Site – Cytoplasm
zz Organ – Liver

Fig. 3.51: In RBCs, HMP occurs and forms NADPH. When it

gets oxidized (removal of Hydrogen) to NADP, on the other
hand, glutathione gets reduced. Enzyme involved is Glutathione


Reductase, which requires FAD (or Vit B2). So function of HMP

in RBCs is to produce this Reduced Glutathione. This is used to
destroy H2O2. Now glutathione gets oxidized and H2O2 is reduced
to water. Enzyme involved is Glutathione Peroxidase, which is
a selenoprotein (requires Selenocysteine-21st amino acid at its
active site). In G6PD deficiency, H2O2 accumulates leading to
hemolytic anemia.

H igh R eturn
zz Marker for B1 deficiency – Transketolase activity
zz Marker for B2 deficiency – Glutathione Reductase activity
zz Marker for B6 deficiency – Transaminase activity

G-6-PD enzyme
zz Rate limiting enzyme
zz First committed step
zz Regulated step of HMP
zz NADPH is a potent competitive inhibitor of G6PD
zz Insulin upregulates the expression of gene for G6PD. Fig. 3.52: Uronic Acid Pathway
zz Uronic Acid Pathway synthesize Glucuronic Acid,
G-6-PD Deficiency Pentoses and Vit C
zz Most common human enzyme defect zz Humans cannot synthesize Vit C due to deficiency of enzyme –
zz XR (X-linked recessive) L-Gulonolactone Oxidase.
zz Haemolytic anemia, Heinz bodies present (denatured
zz Uses of Glucuronic acid:
Hb).
 Incorporated into proteoglycans (Glucuronate used)
zz Neonatal jaundice 1-4 days after birth (due to unconju-
zz Acts as a conjugating agent (Phase II conjugation
gated bilirubin)
reactions like bilirubin conjugation)
zz Shortened lifespan due to chronic hemolysis complica-
tions Essential Pentosuria
zz Increased resistance to plasmodium falciparum malaria
T (Sickle cell trait and Thalassemias also provide resistance zz Deficiency of Xylulose Reductase.
H to malaria) zz L-Xylulose in urine, which gives positive benedict’s test
E (but glucose Oxidase test strip negative)
O Q. Why only RBCs affected in G6PD deficiency? zz One of the condition of Garrod’s Tetrad (Pentosuria,
A. There are 2 Reasons: Albinism, Alkaptonuria, Cystinuria) (Table 3.18)
R T 1. HMP is the only means of generating NADPH in zz It can also occur after consumption of large amount of
Y H RBCs (in other cells, Malic enzyme is present).
I fruits that are rich in pentoses
2. RBCs lack nucleus and ribosomes. So, they cannot
N zz Harmless condition but should be differentiated from
K renew the supply of this enzyme. diabetes.
Table 3.16: Garrod’s Tetrad four diseases described by scientist monosaccharides like Galactose and fructose, body will not
Edward Garrod, which are all inherited defects in metabolic waste its energy in making all 30-40 enzymes different for
pathways them. Just few initial changes will be done in case of Galactose 87
and fructose metabolism and finally same enzymes of
Mnemonic Disease Defect glycolysis will be used

CHAPTER 3  CARBOHYDRATE METABOLISM

C Cystinuria Defect in dibasic amino acid NOTE:
transporter which is for absorption •• Galactose also uses the enzymes of glycolysis
and reabsorption of basic amino •• Fructose also uses the enzymes of glycolysis In order to
acid and cysteine •• Gluconeogenesis also uses the enzymes of save energy
A Alkaptonuria Defect in Homogentisate Oxidase glycolysis

A Albinism Defect in Tyrosinase

Galactose Metabolism
P Pentosuria Defect in Xylulose Reductase
H igh R eturn
GALACTOSE AND FRUCTOSE METABOLISM zz Source: Milk and milk products is Lactose which contains
glucose and galactose. Also galactose obtained by lysosomal
For glucose metabolism, we have 30-40 enzymes (in degradation of glycoproteins and glycolipids.
glycolysis, link reaction, TCA, ETC). Now to metabolize other zz Organ: Liver exclusively
zz Rate limiting enzyme: Galactose-1-P Uridyltransferase (GALT)Q

Fig. 3.53: Galactose metabolism

Galactose Metabolism Occurs in 3 Steps (Fig. 3.53) Galactose is not Essential in Diet
1. Phosphorylation: Galactose is converted to Galactose-
The Epimerase reaction is reversible so UDP-Galactose can
1-P by enzyme Galactokinase. ATP is used at this step.
be obtained from UDP-glucose also. See diagram below:
2. Synthesis of UDP-Galactose: This is an exchange
reaction. UDP is taken from UDP-glucose and
phosphate is transferred from Galactose-1-P to
glucose. Products are UDP-galactose and glucose-1-P.
Enzyme is Galactose-1-P Uridyl Transferase (GALT). T
Glucose -1-P can be used for glycolysis. H
3. Conversion of UDP Galactose to UDP: Glucose– E
Enzyme is Epimerase (UDP Galactose-4- Epimerase O
or UDP-Hexose-4-epimerase). UDP-glucose can be R
used in glycogen synthesis and this reaction will not Y
occur if directly UDP-Galactose is required for Lactose
synthesis.
 GALACTOSE ENERGETICS (See Fig. 3.54) Galactosemia (Fig. 3.55)
88 zz Galactose energetics is same like glucose. Developed countries screen each newborn for Galactosemia
zz One ATP is used at Galactokinase step, finally forming (specially GALT deficiency)
glucose-6-phosphate which enters glycolysis. In case of zz Deficiency of Galactokinase: known as Minor Type
glucose, 1 ATP is used at Hexokinase step. Galactosemia. Excess Galactose gets converted to
CRO BIOCHEMISTRY

zz So, starting with either glucose or Galactose, there is a Galactitol/Dulcitol with the help of enzyme Aldose
net investment or loss of 1 ATP in arriving at the G6P step. Reductase (same enzyme which converts glucose to
zz So complete breakdown of Galactose = 32 ATPs. sorbitol). Galactitol is hygroscopic in nature. In excess it
will cause oil drop cataract.
zz Deficiency of GALT: this is known as Classical Type
Galactosemia. Excess Galactose-1-P accumulates
in liver and brain leading to jaundice and mental
retardation. Excess Galactose-1-P also inhibits enzyme
Galactokinase by product inhibition. If this patient still
continue to take galactose in diet then excess galactose
is converted to galactitol leading to oil drop cataract.


zz Deficiency of Epimerase is rare

Diagnosis
zz Urine
 Benedict’s test positive
 Glucose Oxidase test negative
zz Chromatography for presence of galactose in urine
zz Confirmatory diagnosis: Direct enzyme assay using ery-
throcytes

Treatment
zz Breast milk avoided
Fig. 3.54: Galactose Energetics (Same like glucose) zz Lactose free diet till 4-5 years of age.
zz Galactose-1-P Pyrophosphorylase becomes active by 4-5
years of age, so after 5 years Lactose can be given as this
enzyme uses Galactose-1-phosphate and reduces the
level of Galactose-1-phosphate.

T
H
E
O
R
Y

Fig. 3.55: Galactosemia

Lactose Synthesis in Mammary Glands DHAP and Glyceraldehyde-3-P both are glycolytic inter-
mediates, so they enter glycolysis. Rest metabolism is same
zz Lactose synthesis occurs in golgi by enzyme lactose
Synthase (also known as UDP-Galactose: glucose like phase II of glycolysis. Therefore, the energetics of fructose 89
Galactosyl Transferase) which transfers Galactose from is same like glucose i.e. 32 ATPs obtained per molecule.
UDP-Galactose to glucose, releasing UDP. Because in phase I of glycolysis, 2 ATPs are used and in
fructose metabolism also 2 ATPs are used.

CHAPTER 3  CARBOHYDRATE METABOLISM

zz Lactose is made up of galactose joined to glucose by
β(1 → 4) linkage.
Table 3.17: Differences between Aldolase A, B and C

Aldolase A Aldolase B Aldolase C

Location Most tissues Liver, kidneys, Brain
Small Intestine
Reaction Cleavage of Cleavage of Cleavage of
catalyzed fructose 1,6 fructose-6-P fructose 1,6
Fructose Metabolism Bisphosphate into DHAP and Bisphosphate
into DHAP and Glyceraldehyde into DHAP and
H igh R eturn Glyceraldehyde-
3-P
Glyceraldehyde-
3-P
zz Organ: Liver mainly
zz Source: Disaccharide – sucrose (contains equal amount of
glucose and fructose), in monosaccharide form i.e. Fructose in Fructose as Most Lipogenic Sugar
Fruits, Honey and high fructose corn syrup (55% fructose, 45%
glucose), Sugarcane
zz Fructose entry into cells is insulin independent
zz Fructose ingestion does not leads to insulin release
zz It is the major source of energy for sperms (present in seminal
fluid)

Fructose metabolism occurs in 3 steps (Fig. 3.56)

1. Phosphorylation: Fructokinase/Hexokinase enzyme
converts fructose to Fruc-1-P. Hexokinase has high
Km, means more fructose is required for this enzyme
to act. But Fructokinase is the primary enzyme for
Phosphorylation of fructose and it has low Km (means
even if less fructose is present, then also this enzyme
can do the Phosphorylation). One ATP is used at this
step. Fig. 3.57: Fructose is rapidly metabolized as compared to
2. Action of Aldolase B (also known as fructose-1-P glucose because it bypasses PFK-1 step (Rate Limiting Enzyme of
Aldolase): Cleavage of fructose-1-P into DHAP (Di glycolysis). So fructose forms acetyl CoA and thus fats. So fructose
Hydroxy Acetone phosphate) and Glyceraldehyde is regarded as most lipogenic sugar.
3. Conversion of Glyceraldehyde to Glyceraldehyde-
3-P: This phosphorylation step also needs one ATP. H igh R eturn
zz Fructose energetics is same as Glucose zz Energetics of glucose and fructose is same. So, complete
breakdown of fructose = 32 ATPs
zz Fructose gets rapidly metabolized because it bypasses PFK-I
step. So it is most lipogenic sugar.
zz Unlike Glucokinase, Fructokinase’s activity is not affected
by Insulin. So, fructose is metabolized normally in a diabetic
patient.

Essential Fructosuria/Benign Fructosuria T

zz Deficiency of Fructokinase H
zz Accidental finding, when urine benedict’s test is positive E
(but glucose Oxidase test strip negative) O
zz Increased fructose in blood R
zz Fructose excreted in urine because of lack of renal Y
threshold for fructose

Fig. 3.56: Fructose Metabolism

 Diagnosis

90 zz Benedict’s test positive (shows some reducing sugar

present)
zz Seliwanoff’s test positive (shows some keto sugar pres-
ent)
CRO BIOCHEMISTRY

zz Urine chromatography for fructose

A dditional E dge
Why not fructosemia?
Fructose not raised in blood:
zz Hexokinase (a non specific enzyme) can phosphorylate fructose
as well as other sugars but it has high km (low affinity) for fructose
zz So fructose is always not elevated
zz Glucose is the true substrate for this enzyme
zz Fructose 6-phosphate - the end product of Hexokinase


reaction can enter glycolytic pathway to be utilized further, so

it does not accumulate to produce the toxic effects
Fructose excreted in urine:
zz Because of lack of renal threshold for fructose
zz Therefore it is called Essential Fructosuria/Benign Fruc-tosuria
Fig. 3.58: Hereditary fructose Intolerance
and not fructosemia
Q.1. A newborn begin to vomit 3 days after birth,
Hereditary Fructose Intolerance (HFI) (Fig. 3.58) after breastfeeding. After 3 weeks, he develops
T Jaundice. On examination, he has abdominal
zz Deficiency of Aldolase B H distension with Liver enlarged. Urine dipstick
zz AR (Autosomal Recessive) I test specific for glucose was negative but
N urine benedict's test was positive. What is the
zz Asymptomatic but fructose ingestion causes symptoms K
(typically child presents after weaning when fruit juices diagnosis?
are started in the diet) A. Galactosemia
Q.2. A 7 month old baby girl with severe jaundice
zz Hypoglycemia, jaundice, hepatomegaly, vomiting, seiz-
presents to hospital. The child is irritable from
ures, lethargy, irritability
the time weaning was started. Which enzyme is
zz If not treated → liver and kidney failure can oocur. defective ?
zz No cataract as fructose is not the substrate for Aldose A. Fructose intolerance
Reductase. Lets compare these 2 questions. In question-1, age of
zz Hyperuricemia because of sequestration of phosphate child is 3 days after birth (or before weaning when
zz This hypoglycemia is known as fructose induced milk is the only food). In Q2, age of child is 7 months
hypoglycemia despite the presence of high glycogen (i.e. after weaning when fruit juices are started).
reserves Both patients have jaundice (Liver is affected in
both Galactosemia and fructose intolerance). In
both cases, urine benedicts test will be positive but
Diagnosis glucose Oxidase test strip will be negative.
zz Fructose in urine and confirmatory diagnosis by doing A.1. 
Diagnosis is Galactosemia (GALT deficiency is
Enzyme assay in Liver more common). Liver affected, jaundice occurs.
Galactose present in urine so benedict’s test positive
(Galactose- a monosac-charide is a reducing sugar).
Treatment
But glucose Oxidase test is specific for glucose so it
zz Complete exclusion of Fructose, Sorbitol and Sucrose in is negative.
the diet A. 2. 
Diagnosis is fructose intolerance. Weaning with
fruit juices contains fructose so child presents
after weaning. Liver is affected so jaundice occurs.
T Fructose present in urine so benedict’s test positive
H (fructose- a monosaccharide is a reducing sugar).
E But glucose Oxidase test is specific for glucose so it
O is negative.
R
Y
Sorbitol Pathway Sorbitol Pathway in Diabetes
This is a short pathway to synthesize fructose in body via
sorbitol. First glucose is reduced to its alcohol i.e. Sorbitol (or
In diabetic patients, glucose cannot enter peripheral cells 91
like muscles & adipose tissues, But excess glucose entry
Glucitol) by enzyme Aldose Reductase (same enzyme which occurs specially in insulin insensitive tissues and leads to
reduce Galactose to Galactitol). Then sorbitol is oxidized to the formation of sorbitol. But Sorbitol Dehydrogenase is not

CHAPTER 3  CARBOHYDRATE METABOLISM

fructose by enzyme Sorbitol Dehydrogenase. available in most of these cells. So sorbitol accumulation
This synthesis of fructose in body is specially useful in occurs. Sorbitol is hygroscopic in nature. So, in excess it will
seminal vesicles for sperms as they mainly use fructose as the cause cell swelling. Excess sorbitol in lens leads to snow flake
source of energy. cataract.
Insulin insensitive tissues: i.e. tissues in which Insulin not required
for glucose entry are Peripheral Nerves, Renal Glomeruli, Lens
and Retina.
Therefore sorbitol pathway is considered to be partly
responsible for retinopathy, neuropathy and nephropathy
Fig. 3.59: Sorbitol pathway in diabetic patients. In addition to this, NADPH is used
by Aldose Reductase. This leads to decreased NADPH,
so decreased glutathione (antioxidant). This also adds to
diabetic complications.

Fig. 3.60: Diabetic Cataract – Snowflake on breadcrumb cataract

Pearls of the Chapter

zz Number of ATPs in aerobic glycolysis = 7 ATPs
zz In RBCs always anaerobic glycolysis occurs
zz Link reaction enzyme – Pyruvate Dehydrogenase complex. It requires B1, B2, B3, B5 vitamins, Lipoic acid
zz Link reaction gives 5 ATPs from two pyruvate
zz Oxaloacetate → 1st substrate, carrier of TCA and has Catalytic role in TCA
zz Acetyl CoA is not the intermediate of TCA
zz TCA has no hormonal control and is both anabolic and catabolic
zz Anaplerotic reactions are those reactions which replenish TCA intermediates
zz Mostly 3 C compounds are good substrates of Gluconeogenesis
zz Glucose-6-Phosphatase enzyme is present in the membrane of ER
zz Gluconeogenesis occurs during Fasting/ Starvation/ Diabetes Mellitus
zz Acetyl CoA activates pyruvate Carboxylase of Gluconeogenesis T
zz Coenzyme Q is the only non-protein member of ETC H
zz NADH has minimum Redox potential O2 has maximum Redox potential E
zz Malonate (3C) is inhibitor of complex II of ETC, Succinate Dehydrogenase of TCA cycle and CPT-I of Beta Oxidation of Fatty Acid O
zz Increased NADH leads to Lactic Acidosis and Hyperuricemia R
zz Complex I and Complex III transfers 4 H+, so they are responsible for the production of 1 ATP each Y
 zz Complex IV transfers 2 H+ , so it is responsible for the production of 0.5 ATP
Malate and Glycerol-P Shuttle transfers NADH from cytoplasm to mitochondria
92 zz
zz Citrate shuttle transfers acetyl CoA from mitochondria to cytoplasm
zz Creatine-phosphate Shuttle transport ATP from mitochondria to cytoplasm
zz Both rate limiting enzymes of glycogen metabolism are transferases
End product of liver glycogenolysis is free glucose
CRO BIOCHEMISTRY

zz
zz End product of muscle glycogenolysis is glucose-6-phosphate
zz Pompe’s disease is the only Glycogen Storage Disease, which is a Lysosomal Storage Disease
zz Most common GSD- Von Gierke’s disease
zz Epinephrine acts in Muscle and Liver but Glucagon acts only in Liver
zz HMP is a minor pathway for oxidation of glucose but a major source for NADPH
zz No ATP generated but CO2 is produced in HMP
zz Pathways which do not produce ATP are HMP, Uronic acid pathway and RL shunt
zz NADPH is Produced from: HMP (major source), Malic enzyme and Cytosolic Isocitrate Dehydrogenase
zz Tissues which are never the site of HMP are non lactating mammary glands and skin
zz G-6-PD deficiency is the most common human enzyme deficiency. Pyruvate Kinase deficiency is the second most common human


enzyme deficiency.
zz HMP is the only means of generating NADPH in RBCs (in other cells, Malic enzyme is present).
zz Humans cannot synthesize Vit C due to deficiency of enzyme – L-Gulono Lactone Oxidase
zz Garrod’s Tetrad (Pentosuria, Albinism, Alkaptonuria, Cystinuria)

T
H
E
O
R
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 Multiple Choice Questions 93

Glycolysis 12. The number of ATPs produced by Rapaport Leubering

1. The major metabolic product produced under cycle in RBC from glucose? (PGMEE 2015)
normal circumstances by erythrocytes and by muscle a. 1 b. 2
cells during intense exercise is recycled through liver c. 3 d. 4
in Cori’s cycle. The metabolite is: 13. ATP yield via substrate level phosphorylation in
 (Recent Question 2018) glycolysis: (PGMEE 2013)
a. Oxaloacetate b. Alanine a. 5 b. 6
c. Glycerol d. lactate c. 4 d. 3
2. Rate limiting enzyme (RLE) in Glycolysis is PFK-I. 14. Post prandial utilization of glucose is by which
Which among the following is the most potent enzyme? (PGMEE 2012)
a. Fructokinase b. Glucokinase
allosteric activator of PFK-I? 
c. Hexokinase d. All of above
a. Low pH (PGMEE 2013, 14)
15. Inhibition of glycolysis by increase supply of O2 is
b. Citrate
called – (PGMEE 2013)
c. ATP
a. Carbtree effect
d. fructose 2, 6 Bisphosphate
b. Pasteur effect
3. Which of the following decreases affinity of oxygen
c. Lewis effect
with hemoglobin?  (JIPMER May 2018)
d. None
a. Decreased H+ ions
16. Immediate metabolic products during conversion
b. 2,3 BPG
of fructose 1-6 bisphosphate to 2 molecules of pyr-
c. Increase in temperature uvate: (PGMEE 2015)
d. Decreased sorbitol a. 3-Phosphoglycerate and 1,3-Bisphosglycerate
4. How many ATPs are used in energy investment phase b. Glyceraldehyde -3-phosphate and 1, 3
of glycolysis? (Recent Question 2017) Bisphosphoglycerate
a. 2 b. 3 c. Dihydroxyacetone phosphate and Dihydroxy- M
c. 4 d. ZERO acetone phosphate C
5. In anaerobic glycolysis, end product is- d. Glyceraldehyde-3-phosphate and Dihydroxy- Qs
a. 2 ATP + 2 NAD b. 2 ATP acetone phosphate Ans.
c. 2 ATP + 2 NADH d. 4 ATP + 2 FADH2 17. The purpose of extra step of anaerobic glycolysis is :  1. d
6. In anaerobic glycolysis, there is gain of – a. Production of 2 lactate (JIPMER 2016) 2. d
a. 2 ATP + 2 NAD b. 2 ATP b. Production of one lactate 3. b
c. 2 ATP + 2 NADH d. 4 ATP + 2 FADH2 c. Replenishment of NAD 4. a
7. All tissues convert glucose to predominantly lactate d. Replenishment of NADH 5. a
EXCEPT: (Recent Question 2017) 18. Zero ATP in RBC in glycolysis occurs in: 6. b
a. Brain b. Cornea a. Arsenic poisoning b. RL shunt 7. c
c. Lens d. RBCs c. Both a and b d. None 8. a
8. Which is a negative heterotropic allosteric modulator 19. How many ATPs are produced in glycolysis : 9. c
of glycolysis? (Recent Question 2017) a. 7 b. 9 10. c
a. Citrate b. ATP c. 3 d. 0 11. b
c. ADP d. AMP 20. 2, 3-BPG binds to ___ site(s) of hemoglobin and ____ 12. b
9. Which of the following is TRUE about glycolysis? the affinity for oxygen? (AIIMS May 2014) 13. c
a. Occurs in mitochondria a. 4, decreases b. 1, decreases 14. b
b. Complete breakdown of glucose c. 4, increases d. 1, increases 15. b
c. Conversion of glucose to 3 C pyruvate 21. Which of the following enzymes catalyze the irrever- 16. d
d. 3 ATPs produced in anaerobic glycolysis sible step of glycolysis? (AIIMS May 2013) 17. c
10. What activate Kinase of glycolysis? (PGMEE 2015) a. Glucokinase, Phosphofructokinase, pyruvate Carbo- 18. c
a. ATP b. cAMP xylase 19. a
c. Insulin d. Glucagon b. Hexokinase, fructose 1, 6 Bisphosphatase, pyruvate 20. b
11. In glycolysis which of the ion is most important? Kinase 21. c
 (PGMEE 2009) c. Glucokinase, Phosphofructokinase, pyruvate Kinase
a. Zn b. Mg d. Enolase, fructose 1, 6 Biphospahatase,
c. Cu d. Ca Phosphofructokinase
 22. Glycolytic enzyme (s) inhibited by Fluoride is/are: 31. Which of the following is/are INCORRECT? 
 (PGI Nov 2008)  (PGI Nov 2017)
94 a. Hexokinase b. Aldolase a. Fats can be converted to carbohydrates
c. Enolase d. Pyruvate Kinase b. Carbohydrates can be converted to fats
e. Phosphofructokinase c. Glycerol can be converted to glucose
23. Which of the following is INCORRECT about RBCs? d. Beri-Beri leads to lactic acidosis
 (PGI Nov 2017) e. Link reaction is irreversible
a. RBCs cannot use fatty acids, amino acids and ketone 32. Pyruvate Dehydrogenase complex has all enzyme
bodies for energy components EXCEPT: (PGI Nov 2017)
b. RBCs does not contain enzyme Isocitrate Dehy- a. Decarboxylase b. Dehydrogenase
drogenase c. Carboxylase d. Transacetylase
c. Lactate dehydrogenase is absent in RBCs e. Phosphatase
d. Production of 2,3 BPG does not yield any ATP 33. True about acetyl CoA: (PGI Nov 2011)
e. ATP Synthase is present in RBCs a. Precursor for synthesis of cholesterol and other
24. In traumatic brain injury, changes in brain steroids.
metabolism are seen. All are true EXCEPT:  b. Form ketone bodies
 (AIIMS November 2014) c. Starting material for synthesis of fatty acid
a. There is shut down of pyruvate Dehydrogenase d. Arise from glycolysis
activity e. Can never be converted to glucose
b. There is accumulation of lactate in brain
c. There is increased lactate uptake from circulation TCA Cycle
d. Increased CSF lactate is associated with good 34. Which is not the intermediate of TCA cycle?
prognosis a. Acetyl CoA b. Oxaloacetate
Link Reaction c. Alpha- Ketoglutarate d. Succinyl CoA
35. Cyanide taken up by child. First one to be affected in
25. Which of the following statement about link reaction Kreb’s cycle is:
is CORRECT? (Recent Question 2016) a. Aconitase b. NAD
a. This is a link between TCA and ETC c. Citrate d. Acetyl CoA
b. This is oxidative deamination of pyruvate
36. Thiokinase of TCA produces:
c. This is oxidative decarboxylation of acetyl CoA
M a. ATP b. GTP
d. This reaction requires lipoic acid and four
C c. Both a and b d. NADH
B-complex vitamins
Qs 37. Which among the following controls is an allosteric
26. Major source of acetyl CoA is/are:
Ans. inhibitor of TCA cycle?
a. Triglycerides b. Fatty acids
a. Isocitrate Dehydrogenase
22. c
c. Pyruvate d. Alanine
b. Malate Dehydrogenase
23. c,e
27. Thiamine deficiency results in decrease energy
c. Ketoglutarate Dehydrogenase
24. d
production, because TPP:  (AIIMS May 2017)
d. Pyruvate Dehydrogenase
25. d
a. Interferes with alcohol metabolism
b. Interferes with transketolase activity 38. Which of the following is anaplerotic reaction?
26. c  (AIIMS May 2017)
27. c
c. Is cofactor for pyruvate dehydrogenase and Alpha
Ketoglutarate dehydrogenase a. Conversion of pyruvate to Lactic acid
28. c b. Conversion of pyruvate to Oxaloacetate
29. c
d. Interferes with energy production from amino acids
28. Which of the following is reversible enzyme? c. Conversion of pyruvate to Acetyl CoA
30. c d. Conversion of pyruvate to Acetaldehyde
31. a
a. Pyruvate Kinase
b. Pyruvate Dehydrogenase 39. Why TCA cycle is called amphibolic cycle?
32. a,c,e a. It can proceed both in forward and backward
33. a,b,
c. Lactate Dehydrogenase
d. Hexokinase direction
c,d,e b. It is both endothermic and exothermic
34. a
29. Congenital lactic acidosis may occur due to defect in
c. Metabolites are used in both amino acid and ketone
35. b
a. Pyruvate Carboxylase body synthesis
36. c
b. Pyruvate Decarboxylase d. Same enzyme can be used in reverse direction
37. a
c. Pyruvate Dehydrogenase 40. Succinate Dehydrogenase is inhibited by: 
38. b
d. Transketolase  (PGMEE 2013)
39. b
30. A baby is hypotonic and shows that pyruvate cannot a. Fluoroacetate b. Arsenite
40. d
form acetyl CoA in fibroblasts. Also lactic acidosis is c. Cyanide d. Malonate
41. a
found. Administration of which of the following can 41. What is liberated when citrate converted to cis-
revert this situation?  (JIPMER May 2018) aconitate? (PGMEE 2015)
a. Biotin b. Pyridoxal phosphate a. H2O b. CO2
c. Thiamine d. Pyruvate c. H2O2 d. H2
42. In TCA cycle, which is first formed? (PGMEE 2009) 54. The type of enzyme inhibition in which Succinate
a. Succinate b. Citrate Dehydrogenase reaction is inhibited by malonate is
c. Isocitrate d. None an example of: 95
43. The net ATP yield when one molecule of pyruvate is a. Noncompetitive b. Uncompetitive
completely oxidized to CO2 and H2O is: c. Competitive d. Allosteric
 (PGMEE 2013) 55. TRUE statement regarding Lactate Dehydrogenase
a. 12.5 b. 12 deficiency: (PGI Nov 2014)
c. 15 d. 30 a. Fumarate level increases
44. All of the following are correct EXCEPT: b. Exercise intolerance
c. Muscle cramps may occur
a. Fluorocitrate is competitive inhibitor of Aconitase
d. It operate in anaerobic condition
b. Fluoroacetate is non-competitive inhibitor of Aconi-
e. It is key enzyme of Kreb cycle
tase 56. NAD acts as a cofactor for: (PGI Nov 2011)
c. Malonate is competitive inhibitor of Succinate a. Citrate Synthase
Dehydrogenase b. Isocitrate Dehydrogenase
d. Iodoacetate inhibits Glycerol-3-phosphate c. a-Ketoglutarate Dehydrogenase
Dehydrogenase d. Malate Dehydrogenase
45. Enzyme responsible for complete oxidation of e. Succinyl Thiokinase
glucose to CO2 and H2O is present in : 57. In TCA, CO2 is released by: (PGI May 2017)
a. Cytosol b. Lysosomes a. Citrate Synthase
c. Mitochondria d. Endoplasmic reticulum b. Alpha-Ketoglutarate Dehydrogenase
46. Two carbon atoms which leave in the form of CO2 in c. Citrate Dehydrogenase
TCA, are derived from: (PGI May 2017) d. Isocitrate Dehydrogenase
a. Acetyl CoA b. Oxaloacetate e. Succinate Thiokinase
c. CO2 d. Citrate 58. First substrate of Kreb’s cycle is: 
e. Pyruvate  (PGI Nov 2017)
47. Source of energy in TCA is: a. Glucose b. Glycine
a. NAD b. NADH c. Citrate d. Acetyl CoA
59. Unaltered final product of TCA is: 
c. FAD d. NADPH
 (PGI May 2017)
48. Which of the following is not the dehydrogenase of
a. Acetyl CoA b. Oxaloacetate
TCA : c. CO2 d. Pyruvate M
a. Succinate Dehydrogenase 60. Fluoroacetate inhibits which metabolic pathway? C
b. Pyruvate Dehydrogenase  (AIIMS May 2018) Qs
c. Malate Dehydrogenase a. TCA cycle Ans.
d. Isocitrate Dehydrogenase b. Glycolytic pathway
49. Rate limiting step of TCA is/are: c. Oxidative phosphorylation 42. b
a. Citrate synthase d. ETC 43. a
b. Isocitrate dehydrogenase 44. d
c. Alpha-ketoglutarate dehydrogenase Shuttles 45. c
d. All 61. Malate shuttle is required for: 46. b
50. Which enzyme of TCA is present in Inner Mito- a. Glycolysis (Recent Question 2018) 47. b
chondrial Membrane (IMM)? b. Pyruvate Dehydrogenase complex 48. b
a. Alpha-Ketoglutarate Dehydrogenase c. TCA 49. all
b. Malate Dehydrogenase 50. d
d. All
c. Fumarate Dehydrogenase 51. b
62. If aerobic glycolysis uses Glycerol-3-phosphate
d. Succinate Dehydrogenase 52. b
shuttle, how many ATPs are produced? 
51. TCA cycle depends on: 53. a
 (Recent Question 2018)
a. Availability of acetyl CoA 54. c
a. 2 ATP b. 5 ATP
b. Availability of Oxaloacetate 55. b,c,d
c. 7 ATP d. 3 ATP
c. Availability of Insulin 56. b,c,d
63. NADPH via Glycerol phosphate shuttle gives how
d. Availability of Glucagon 57. b,d
many ATPs? (Recent Question 2018)
52. Oxalo-acetate + Acetyl-Co-A → Citrate + Co-ASH 58. d
a. 2.5 b. 1.5 59. b
this reaction is: c. 3 d. Zero
a. Reversible b. Irreversible 60. a
64. Reason of presence of less ATP forming glycerol-P- 61. a
c. Endergonic d. None shuttle in brain are all EXCEPT: 62. b
53. Thiamine requirement increases in excessive intake a. This is a shorter shuttle (Recent Question 2017) 63. d
of: (AIIMS May 2009) b. It is a quick source of ATP 64. c
a. Carbohydrates b. Fats c. After going in brain in ETC, it gives high energy
c. Proteins d. None d. Brain needs a quick source of ATP
 ETC 77. Enzyme involved in oxidative phosphorylation
65. Which is the only non-protein member of ETC?  (PGMEE 2012)
96  (Recent Question 2017) a. Succinyl CoA Thiokinase
a. Cytochrome c b. Coenzyme Q b. Pyruvate Kinase
c. Complex V d. Complex II c. NADH Dehydrogenase
d. Pyruvate Dehydrogenase
66. Cytochrome c Oxidase requires:
78. Which of the component of respiratory chain reacts
 (Recent Question 2016)
directly with molecular oxygen? (PGMEE 2013)
a. Cu b. Mg
a. Cyt b b. CoQ
c. Ca d. Zn
c. Cyt c d. Cyt aa3
67. ATPs given by complex IV of ETC are: 
79. Last electron acceptor in ETC is (PGMEE 2014)
 (Recent Question 2016)
a. Oxygen
a. 1 b. 2
b. Tetrachloroethylene
c. 3 d. 0.5
c. Nitrate
68. ETC is located in: (Recent Question 2015)
d. Iron
a. Inner mitochondrial Membrane 80. Atractyloside act as-  (PGMEE 2013)
b. Outer mitochondrial Membrane a. Inhibitor of complex III of ETC
c. Mitochondrial matrix b. Inhibitor of oxidative phosphorylation
d. Inter membrane space c. Uncoupler
69. Which of the following is NOT TRUE regarding ETC? d. Inhibitor of glycolysis
 (Recent Question 2015) 81. Creatinine is the breakdown product of-
a. Coupling of oxidation and phosphorylation occurs  (PGMEE 2015)
b. Occurs in mitochondrial matrix a. Adenosine triphosphate
c. Known as chemiosmotic theory b. Purine nucleotides
d. ADP + Pi → ATP c. Pyrimidine nucleotides
70. Mitochondrial membrane contains a protein which d. Creatine phosphate
is transporter of: (Recent Question 2015) 82. Cyanide affects respiratory chain by: (PGMEE 2013)
a. Oxaloacetate b. Acetyl CoA a. Non-competitive reversible inhibition
c. NADH d. ATP b. Competitive reversible inhibition
71. Which couple has minimum redox potential c. Suicide irreversible inhibition
M  (PGMEE 2012) d. Non-competitive irreversible inhibition
C a. NADP+/NADPH b. CoQ-CoQ H2 83. True about 2, 4- Dinitrophenol is? (PGMEE 2012)
Qs c. FAD/FADH2 d. NAD+/NADH a. Prevents ATP synthesis and electron transport chain
Ans. 72. Most important source of ATP? b. Prevents ATP synthesis and electron transport chain
a. Oxidative phosphorylation is increased
65. b
b. Substrate level phosphorylation c. Blocks electron transport chain but ATP synthesis is
66. a
c. Aerobic glycolysis normal
67. d
d. TCA d. Blocks ATP synthesis but electron transport chain is
68. a
73. Mechanism of action of uncouplers: (PGMEE 2015) normal
69. b
a. Inhibition of ATP synthesis only not ETC 84. Uncouplers of oxidative phosphorylation include:
70. d
b. Inhibition of both ATP synthesis and ETC  (PGI May 2018)
71. d
c. Inhibition of only ETC not ATP synthesis a. 2,4 – DNP b. H2S
72. a
d. None of the above c. Cyanide d. Thermogenin
73. a
74. Barbiturates act on which step of mitochondrial e. Carboxin
74. a
respiratory chain? (PGMEE 2013) 85. Electrons in electron transport chain travel from:
75. d
a. Complex I to Coenzyme Q  (PGMEE 2013)
76. c
b. Coenzyme Q to Complex III a. One way irrespective of the potential
77. c
c. Complex II to Coenzyme Q b. Low to high potential
78. d
d. Cytochrome C to Complex IV c. Two way
79. a
75. CO binds with which complex of the electron trans- d. High to low potential
80. b
port chain? (PGMEE 2013) 86. Which vitamin is used in ETC?
81. d
a. Complex I b. Complex III a. Thiamine b. Biotin
82. d
c. Complex II d. Complex IV c. Nicotinic acid d. Pyridoxal phosphate
83. b
76. ATP is generated in ETC by- (PGMEE 2013) 87. Which of the following vitamin is a component of
84. a,d
a. ADP kinase ETC? (PGMEE 2009, 2007)
85. b
b. Na+ Cl ATPase a. Vitamin B12
86. c
c. Fo–F1 ATPase b. Riboflavin
87. b
d. Na+ K+ ATPase c. Nicotinic acid
d. Thiamine
88. Oxidative phosphorylation is inhibited by all 98. Which is NOT glucogenic? (Recent Question 2018)
EXCEPT: (FMGE Nov 2018) a. Acetyl CoA b. OAA
a. CO b. Antimycin A c. Pyruvate d. Lactate 97
c. Malonate d. Thermogenin 99. Which of the following substrates CANNOT contribute
89. MELAS inhibit all ETC Complexes EXCEPT:  to gluconeogenesis in mammalian liver? 
 (PGMEE 2015)  (PGI May 2017)
a. I b. II a. Alanine b. Glutamate
c. III d. IV c. Palmitate d. Pyruvate
90. Oxidative phosphorylation is NOT inhibited by: e. Odd chain fatty acids
 (PGI May 2016) 100. A 15-year-old male presents with increased thirst,
a. Fluoride hunger, urination, and weight loss. His fasting blood
b. 2, 4-dinitrophenol (DNP) glucose level is 400 mg/dl and is diagnosed with type 1
c. Oligomycin diabetes mellitus. What is the reason for this patient's
d. Carboxin inability to maintain a normal blood glucose level?
e. Ouabain  (Recent Question 2017)
91. ETC is regulated by: (PGI Nov 2011) a. Increased ketone body production
a. NADH Co-Q Reductase b. Abnormal response to glucagon
b. Cytochrome C Oxidase c. Decreased glucagon to insulin ratio
c. Glutathione Reductase d. Decreased uptake of glucose by peripheral cells
d. Isocitrate Dehydrogenase 101. Which of the following is the sequence of compart-
e. Co-Q Cytochrome C Reductase ments of gluconeogenesis? (Recent Question 2017)
92. Which of the following is high energy phosphate a. Mitochondria → Cytoplasm → ER
bond (produce ATP on hydrolysis): (PGI Nov 2011) b. Cytoplasm → ER → mitochondria
a. Fructose-6-phosphate c. ER → mitochondria → Cytoplasm
b. Creatine phosphate d. Only in mitochondria and Cytoplasm
c. Carbamoyl phosphate 102. Which of the following is most effective for gluco-
neogenesis: (Recent Question 2017)
d. Glucose-1-phosphate
a. Fructose 2,6 bisphosphate inhibits fructose 1,6
e. Glucose-6-phosphate
Bisphosphatase
93. Number of ATPs produced in adipose tissue from
b. Acetyl CoA activates Pyruvate Carboxylase
1 NADH (NAD+/NADH) through respiratory chain: M
c. Citrate stimulates Acetyl CoA Carboxylase
 (PGI Nov 2011)
d. Citrate inhibit.s acetyl CoA Carboxylase C
a. 0 ATP b. 1 ATP
103. A child having hypoglycemia is unable to use both Qs
c. 2 ATP d. 2.6 ATP
glycogenolysis and gluconeogenesis pathways. Ans.
e. 3 ATP Which of the following enzyme is affected?
94. Which component transfers four protons :  (Recent Question 2016) 88. d
 (PGI Nov 2011) a. Glucokinase 89. b
a. NADH-CoQ Oxidoreductase b. Phospho-Fructo Kinase -1 90. a,d,e
b. Cytochrome c Oxidase c. Glucose-6-Phosphatase 91. a,b,e
c. CoQ Cytochrome c Reductase d. Transketolase 92. b,c
d. Isocitrate Dehydrogenase 104. Which of the following is/are substrate (s) for glucon- 93. d
e. Succinate CoQ Reductase eogenesis?  (PGI May 2018) 94. a,c
95. Which of the following releases/provide energy : a. Glycerol b. Fatty acids 95. b,d,e
a. Conversion of ADP to ATP c. Alanine d. Lysine 96. c
b. Breaking of high energy bond to low energy bond e. Leucine 97. c
c. Conversion of pyruvate to lactate 105. Pyruvate can be converted directly into all of the 98. a
d. Electrical gradient across inner and outer side of following EXCEPT: (Recent Question 2016) 99. c
mitochondrial membrane a. Phosphoenol pyruvate 100. d
e. Passage of electron through FADH2 in ETC b. Alanine 101. a
c. Acetyl CoA 102. b
Gluconeogenesis d. Lactate 103. c
96. All are substrates of gluconeogenesis EXCEPT: 106. Which pathway can use propionic acid?  104. c
 (Recent Question 2018)  (Recent Question 2016) 105. a
a. Lactate b. Alanine a. Glycolysis b. Gluconeogenesis 106. b
c. Leucine d. Lysine c. Glycogenolysis d. Glycogenesis 107. a
97. Glucose can be synthesized from all EXCEPT: 107. Glucose may be synthesized from:
 (FMGE June, 2018 )  (Recent Question 2016)
a. Amino acids b. Glycerol a. Glycerol b. Adenine
c. Acetoacetate d. Lactic acid c. Palmitic acid d. Guanosine
 108. Amino acid which cannot be used for glycogen syn- 119. Which of the following reactions takes place in two
thesis: (PGI May 2017) compartments? (PGMEE 2015)
98 a. Alanine b. Threonine a. Glycogenesis b. Gluconeogenesis
c. Phenylalanine d. Leucine c. Glycolysis d. Glycogenolysis
109. Gluconeogenesis occurs in: (Recent Question 2015) 120. Step of Gluconeogenesis is: (PGMEE 2015)
a. Muscles b. Kidney a. Fructose 6 phosphate to glucose 6 phosphate
c. Liver d. Intestine b. Pyruvate to lactate
110. Conversion of lactate to glucose requires all EXCEPT: c. Phosphoenol pyruvate to Oxaloacetate
a. Pyruvate Carboxylase d. Pyruvate to acetyl CoA
b. PFK-1 121. Which of the following metabolites is involoved in
c. Enolase glycogenolysis, glycolysis and gluconeogenesis?
d. Glucose-6-Phosphatase a. Fructose – 6- phosphate  (PGMEE 2016, 17)
111. Regulatory enzymes in gluconeogenesis are all b. Glucose – 6 – phosphate
EXCEPT: (Recent Question 2014) c. Uridine diphospho glucose
a. Pyruvate Carboxylase d. Galactose- 1- phosphate
b. Aldolase B 122. Substrate for gluconeogenesis- (PGMEE 2015)
c. PEP carboxykinase a. Fatty acid
d. Glucose-6-Phosphatase b. Acetyl-CoA
112. Enzymes involved in gluconeogenesis are all c. Pyruvic acid (pyruvate)
EXCEPT: (Recent Question 2014) d. All of the above
a. Phosphoglycerate Kinase 123. Which of the following hormones can cause
b. Fructose 1,6 bisphosphatase hyperglycemia without known effects on glycogen?
c. Phosphoglucomutase  (PGMEE 2016, 17)
d. Pyruvate Carboxylase a. Epinephrine b. Nor epinephrine
113. Glyconeogenesis is: (Recent Question 2013) c. Thyroxine d. Glucagon
a. Synthesis of galactose from non-carbohydrate 124. Gluconeogenesis is favoured in fasting state by:
sources  (PGI-May 2017)
b. Synthesis of glycogen from glucose a. Activation of pyruvate Carboxylase by acetyl CoA
c. Synthesis of glucose from glycerol b. Increased conversion of phosphoenol pyruvate to
d. Synthesis of glycogen from non-carbohydrate pyruvate by activation of pyruvate Kinase
M sources
C c. Increased fatty acid oxidation in liver
114. A genetic disorder renders fructose 1,6 bisphos-
Qs d. Inhibition of PFK-II
phatase in liver less sensitive to regulation by fructose
Ans. e. Increase release of alanine from muscles to liver
2,6- bisphosphate. All of the following metabolic
108. d changes occur EXCEPT: (Recent Question 2014) Glycogen
109. c a. Level of fructose 1,6 bisphosphate is higher than
125. Which vitamin is required for Glycogen Phosphory-
110. b normal
lase?  (AIIMS Nov 2017, PGMEE 2015, 2018)
111. b b. Level of fructose 1,6 bisphosphate is lower than
a. TPP (Thiamine Pyrophosphate)
112. c normal
b. PLP (Pyridoxal phosphate)
113. d c. Less pyruvate formed
d. Less ATP formed c. Riboflavin
114. a
115. During gluconeogenesis, oxaloacetate is transported d. Lipoic acid
115. a
from mitochondria to cytoplasm by:  126. Glycogen Phosphorylase is regulated by all EXCEPT:
116. a
 (Recent Question 2013) a. Protein Kinase b. Calmodulin
117. a
a. Malate b. Pyruvate c. cAMP d. Glycogenin
118. c
c. Glutamate d. Phosphoenol pyruvate 127. Allosteric stimulator of glycogen synthase:
119. b
116. Malate shuttle is important in: a. Insulin
120. a
a. Glycogenesis b. Glycolysis b. Glucose 6 phosphate
121. b
c. Gluconeogenesis d. Glycogenolysis c. Glucagon
122. c
117. During prolonged starvation, rate of gluconeogenesis d. Fructose 1,6 bisphosphate
123. c
depends on:  (Recent Question 2013) 128. A 28-year-old professional cyclist has been training
124. a,c,
a. Increased alanine levels in liver for an opportunity to go for a long race. His coach
d,e
b. Decreased cGMP levels in liver strongly suggests the intake of carbohydrates after
125. b
c. ADP in liver the work out to ensure muscle glycogen storage.
126. d
d. Decreased essential fatty acids in liver The activity of muscle glycogen synthase in resting
127. b
128. c 118. All of the following amino acids forms Acetyl CoA via muscles is increased by the action of which of the
pyruvate Dehydrogenase EXCEPT: following? (Recent Question 2017)
a. Glycine b. Hydroxyproline a. Epinephrine b. Glucagon
c. Tyrosine d. Alanine c. Insulin d. Phosphorylation
129. Muscle CANNOT maintain blood glucose because of 140. During the breakdown of glycogen, free glucose is
deficiency of :  (Recent Question 2017) formed from which of the following?
a. Glucose-6-phosphatase a. Glucose residues in a-1,4 glycosidic linkages 99
b. Glycogen Phosphorylase b. The reducing end
c. Hexokinase c. The non reducing end
d. Phospho-gluco-mutase d. Glucose residues in a -1,6 glycosidic linkages
130. Muscle CANNOT make use of glycogen because of 141. Glycogen catabolism is best described by which of
deficiency of : (Recent Question 2017) the following statements- (Recent Question 2015)
a. Glucose-6-Phosphatase a. In brain, it yields glucose for skeletal muscle cons-
b. Glycogen Phosphorylase umption
c. Hexokinase b. It requires a debranching enzyme in the erythrocytes
d. Phospho-gluco-Mutase c. It is not a major pathway in brain
131. Major carbohydrate store in the body is-  d. It uses Phosphorylase for glucose residue cleavage
 (PGMEE 2015) from the reducing end of glycogen in liver
a. Hepatic Glycogen 142. The degradation of glycogen normally produces
b. Blood glucose which of the following: (Recent Question 2015)
c. Glycogen in adipose tissue a. More glucose than glucose-1-P
d. None of the above b. More glucose-1-P than glucose
132. A 15-year-old type I diabetic patient faints after c. Equal amount of glucose and glucose-1-P
injecting himself with insulin. He is administered d. Neither glucose nor glucose-1-P
glucagon and rapidly recovers consciousness. 143. The energy for glycogenesis is derived from :
Glucagon induces activity of: (Recent Question 2017) a. GTP b. ATP
a. Glycogen synthase b. Glycogen phosphorylase c. UDP d. UTP
c. Glucokinase d. Hexokinase 144. UDP-glucose is not used in: (Recent Question 2015)
133. Glycogenin is a: (Recent Question 2016) a. HMP
a. Lipid b. Galactose metabolism
b. Polypeptide c. Glycogen synthesis
c. Polysaccharide d. Uronic acid pathway
d. Glycosa amino glycans (GAGs) 145. Which is branching enzyme? (FMGE June 2018)
134. If muscle glycogen is used for anaerobic glycolysis, a. Glycogen Synthetase
b. glucose-6 Phosphatase M
how many ATPs are formed? C
c. Amylo (1 → 4), (1 → 6) Transglycosylase
a. 2 b. 7 Qs
d. Glycogen Phosphorylase
c. 3 d. ZERO Ans.
146. In starvation how many hours needed for depletion
135. Which of the following yields 3 molecules of ATP
of Glycogen: (PGMEE 2015) 129. a
under anaerboic metabolism? (AIIMS May 2017)
a. 9 b. 18 130. a
a. Glucose b. Galactose
c. 24 d. 48 131. a
c. Glycogen d. Amino Acid
147. Glycogen is released from muscle because of increa- 132. b
136. All are sources of glucose EXCEPT:
sed cAMP due to: 133. b
a. Liver Glycogen b. Gluconoegenesis
a. Glucagon b. Insulin 134. c
c. Muscle Glycogen d. Alanine c. Epinephrine d. Growth hormone 135. c
137. A 30-year-old presents with intractable vomiting 148. Alpha amylase secreted by pancreas digest starch 136. c
and inability to eat or drink for the past 3 days. His into which of the following major products? 137. a
blood glucose level is normal. Which of the following a. Amylose, Amylopectin, and Maltose 138. b
is most important for maintenance of Blood glucose? b. Glucose, Galactose, and fructose 139. c
a. Liver b. Heart c. Glucose, Sucrose, and Maltotriose 140. d
c. Skeletal Muscle d. Lysosome d. Limit Dextrins, Maltose, and Maltotriose 141. c
138. Glycogen phosphorylase degrades glycogen to 149. Glycogen synthesis and breakdown takes place in 142. b
produce: (Recent Question 2015) the same cell, having enzymes necessary for both 143. d
a. Glucose b. Glucose-1-P pathways. Why is glucose-6-phosphate produced 144. a
c. Glucose-6-P d. UDP glucose during glycogenesis in the cytoplasm of liver cells, 145. c
139. Glycogenolysis is best described by which of the not acted upon by glucose-6-phosphatase enzyme? 146. b
following statements? (Recent Question 2015)  (AIIMS Nov 2015) 147. c
a. It involves enzymes cleaving beta (1-4) glycosidic a. Steric inhibition of phosphatase by albumin 148. d
linkage b. Glucose-6-phosphatase is present in endoplasmic 149. b
b. Requires activation of glycogen synthase reticulum while Glycogen is in cytoplasm
c. Requires a bifunctional enzyme (debranching and c. It is thermodynamically viable only when Gluco-
transferase) noegenesis has stated
e. Requires inactivation of phosphorylase kinase d. Require protein kinase for activation
 150. Enzyme involved in both glycogenesis and glycoge- 158. Enzyme deficient in Her's disease- (PGMEE 2015)
nolysis is? (AIIMS May 2015, 2014) a. Muscle Phosphorylase
100 a. Glycogen synthase b. Phosphoglucomutase b. Acid maltase
c. Phosphorylase d. Glycogen Transferase c. Liver Phosphorylase
151. In glycogen metabolism, some metabolically active d. Debranching enzyme
important enzymes found in the liver are converted 159. Glycogen storage disease which presents as lyso-
from their inactive dephosphorylated state to active somal storage disease: 
phosphorylated state. Which of the following is true? a. Andersen’s disease (PGMEE 2012, 2013, 2015)
 (AIIMS November 2012) b. Pompe’s disease
a. Always activates the enzyme c. Mc Ardle’s disease
b. Catecholamines directly stimulate it d. Von gierke’s disease
c. More commonly seen in fasting state than in fed 160. Hypoglycemia is more severe in Type I Glycogen
state Storage Disease as compared to Type VI Glycogen
d. Always activated by cAMP dependent protein kinase Storage Disease because: (AIIMS May 2014)
a. No Gluconoegenesis in type I disease
Glycogen Storage Diseases b. No Gluconoegenesis in type VI disease
152. A 10 year old boy rapidly develops hypoglycemia after c. Both
moderate activity. Blood examination reveals raised d. Type I disease affects muscles and liver both
levels of ketone bodies, lactic acid and triglycerides. 161. Baby has hypoglycemia, specially early morning
On examination, liver and kidneys were enlarged. hypoglycemia. Glucagon given. It raises blood
Histopathology of liver shows deposits of glycogen in glucose if given after meals But does not raises blood
excess amount. What is the diagnosis? glucose if given during fasting. Liver biopsy shows
 (AIIMS Nov 2017) increased glycogen deposits. Enzyme defect is? 
a. Von Gierke’s disease b. Cori’s disease a. Muscle Phosphorylase (AIIMS May 2016)
c. Mc Ardle’s disease d. Pompe’s disease b. Glucose-6-phosphatase
153. An adolescent male patient came with pain in calf c. Branching enzyme
muscles on exercise. On biopsy excessive amount d. Debranching enzyme
of glycogen present was found to be present in the 162. In Von Gierke’s disease, the levels of ketone bodies
muscle. What is the most likely enzyme deficiency? are increased due to all EXCEPT: 
 (AIIMS May 2015)
M  (AIIMS May 2018)
a. The patients have hypoglycaemia
C a. Muscle debranching enzyme
b. The patients have low blood glucose
Qs b. Phosphofructokinase I
c. Less mobilization of fats
Ans. c. Glucose 6 phosphatase
d. OAA is required for Gluconoegenesis
d. Phosphorylase enzyme
150. b 163. Glycogen storage disorder (s) is/are:
154. A 3-month-old infant presents with hepatosple-
151. c a. Niemann-Pick disease  (PGI Nov 2014)
nomegaly and failure to thrive. A liver biopsy
152. a b. Gaucher disease
reveals glycogen with an abnormal, amylopectin
153. d c. Tay-Sachs Disease
like structure with long outer chains and missing
154. b d. Pompe’s disease
branches. Which of the following enzymes would e. McArdles disease
155. a
most likely be deficient?
156. a
157. a
a. Alpha Amylase b. Branching enzyme HMP Pathway, Uronic Acid Pathway and Sorbitol
c. Debranching enzyme d. Glycogen Phosphorylase Pathway
158. c
155. A 30-year-old male presents with severe muscle
159. b 164. Products of HMP shunt are all EXCEPT: 
cramps. His blood lactate levels did not increase after
160. a  (Recent Question 2018)
exercise. His blood glucose by GOD-POD levels was
161. d a. Glyceraldehyde-3-P b. Glycerol-3- P
found to be normal. He has :
162. d c. 2 NADPH d. 3 NADPH
a. Mc Ardle's disease
163. d,e 165. HMP is the only source for : (Recent Question 2018)
164. b
b. Glycogen storage disease type III
a. NADPH b. NADH
165. c
c. Von Gierke’s disease
c. Ribose-5-P d. CO2
166. c
d. Glycogen storage disease type VI
166. Which vitamin is required for glucose -6- phosphate
167. d
156. All of the following are associated with non-ketotic
Dehydrogenase?  (Recent Pattern June 2018)
hypoglycemia, EXCEPT:
a. Riboflavin b. Thiamine
a. Von Gierke’s disease b. Insulinoma
c. Niacin d. Biotin
c. Carnitine deficiency d. MCAD deficiency 167. NADPH is produced from: 
157. Increased uric acid levels are seen in which glycogen a. HMP  (Recent Pattern 2018)
storage disease?  (Recent Question) b. Malic enzyme
a. Type I b. Type II c. Cytoplasmic isocitrate dehydrogenase
c. Type III d. Type IV d. All
168. Which pathway DOES NOT generate ATP?  180. Essential pentosuria is due to deficiency of- 
 (Recent Question 2017)  (PGMEE 2013-12)
a. Glycolysis b. HMP a. Fructokinase 101
c. TCA d. Fatty acid oxidation b. Phosphoglucomutase
169. Severe thiamine deficiency is associated with : c. Xylulose Reductase
 (Recent Question 2017) d. Gulonolactone Oxidase
a. Increased clotting time 181. All are true about hexose monophosphate pathway
b. Decreased RBC transketolase activity (HMP) EXCEPT:  (PGI May 2017)
c. Decreased RBC Glutathione activity a. Produce NADPH in oxidative phase of pathway
d. Increased Xanthic acid excretion b. Doesn’t produce ATP
170. Which of the following metabolic pathway in carbo- c. Occurs in testes, ovaries, placenta and adrenal
hydrate metabolism is required for nucleic acid cortex
synthesis? (Recent Question 2017) d. Produces ribose 5-phosphate in oxidative phase of
a. Glycolysis b. Glycogenesis pathway
c. HMP d. Gluconeogenesis e. Glucose 6-phosphatase dehydrogenase enzyme is
171. HMP shunt occurs in all organs EXCEPT : involved
 (Recent Question 2016) 182. UDP glucose is used for: (PGI Nov 2017)
a. Liver a. Glycogen synthesis
b. Non lactating mammary glands b. Galactose metabolism
c. Adipose tissues c. Heparin synthesis
d. RBCs d. Bilirubin metabolism
172. Glutathione is a: (PGMEE 2006) e. Gangliosides synthesis
a. Dipeptide b. Polypeptide
c. Tripeptide d. Oligopeptide
Galactose and Fructose Metabolism
173. Reduced NADPH is produced by:  183. A breast-fed infant began to vomit frequently and lose
 (PGMEE 2016-17, 2015) weight. Several days later she developed jaundice,
a. Krebs cycle hepatomegaly and bilateral cataract. What is the
b. Hexose monophosphate pathway (HMP shunt) possible cause for these symptoms?
c. Uronic acid pathway  (Recent Question 2016)
d. Anerobic glycolysis a. Galactosemia
M
174. Dehydrogenases of HMP shunt are specific for b. Von-Gierke's disease
c. Juvenile diabetes Mellitus
C
 (PGMEE 2009) Qs
a. TPP b. NADP+ d. Hereditary fructose intolerance
184. Most common enzyme deficient in galactosemias: Ans.
c. FMN d. FAD
 (Recent Pattern Jan 2018; FMGE June 2018) 168. b
175. NADPH is generated in the reaction catalysed by:
a. Galactose-1-phosphate Uridyl Transferase/ GALT 169. b
 (PGMEE 2013, 2005) 170. c
a. LDH b. G6PD b. Galactosidase
c. UDP galactose epimerase 171. b
c. G3PD d. Alcohol Dehydrogenase 172. c
d. Galactokinase
176. NADPH in extra mitochondrial site helps in the 173. b
185. All are true about galactosemia EXCEPT: 
production of: (PGMEE 2013, 2012) 174. b
 (PGI Nov 2013)
a. Ketone bodies b. Steroids 175. b
a. Deficiency of galactokinase
c. Glycogen d. None 176. b
b. Disease manifest only at adolescence
177. Products of uronic acid pathway in human beings are 177. a
c. Accumulation of Galactose-1-phosphate
all EXCEPT: (PGMEE 2015) 178. b
d. Accumulation of galactitol
a. Vitamin C b. Pentoses 179. c
e. Deficiency of enzyme Galactose-1-phosphate
c. NADH d. Glucuronic acid 180. c
uridyltrans­ferase
178. Glucose is converted to glucuronic acid by- 181. d
186. Reducing sugar in urine is seen in: 
a. Oxidation of aldehyde group (PGMEE 2013-12)  (Recent Question 2015) 182. all
b. Oxidation of terminal alcohol a. Galactosemia 183. a
c. Oxidation of both b. Lactose intolerance 184. a
d. None c. Phenylketonuria 185. b
179. Due to which of the following enzyme deficiency, d. Alkaptonuria 186. a
vitamin C CANNOT be synthesised in humans? 187. Oil drop cataract is produced because of the activity 187. a
 (AIIMS May 2018) of which enzyme? 
a. L-Glucuronic acid oxidase a. Aldose Reductase (Recent Question 2015, 2014)
b. L-Gulonic acid reductase b. Galactose Reductase
c. L-Gulonolactone oxidase c. Fructose Dehydrogenase
d. L-Gulonolactone reductase d. Sorbitol Dehydrogenase
 188. A child presents with hepatomegaly and bilateral 194. Which of the following is NOT metabolised in our
lenticular opacities. Deficiency of which of the body?
102 following enzyme will not cause such features? a. Glucose b. Fructose
 (Recent Question 2014) c. Sucrose d. Sorbitol
a. UDP-Galactose-4-epimerase 195. An enzyme involved in fructose metabolism is :
b. Galactokinase a. Glucokinase
c. Glucokinase b. Glyceraldehyde-3-P Dehydrogenase
d. Gal-1-P uridyltransferase c. Aldolase A
189. E. coli sepsis is commonly seen in: d. PFK-1
 (Recent Question 2013) 196. Essential Fructosuria occurs due to deficiency of
a. Urea cycle disorder  (PGMEE 2013)
b. Glycogen storage diseases a. Aldolase A
c. Galactosemia b. Aldolase B
d. Fructose intolerance c. Fructokinase
190. Enzyme deficiency in Galactosemia- (PGMEE 2015) d. Enolase
a. Aldolase – B 197. A patient has blood glucose levels by GOD–POD
b. Galactokinase method to be normal. But urine shows positive
c. Glucokinase Benedict’s test. The reason is: (AIIMS Nov 2016)
d. All of the above a. False positive
191. Familial Fructokinase deficiency causes no symp- b. Fructosemia
toms because:  (Recent Question 2012) c. Galactosemia
a. Hexokinase can phosphorylate fructose d. Glucose intolerance
b. LiverAldolase can metabolize it 198. Snow flake cataract is produced because of which
c. Excess fructose does not escape in to urine enzyme?  (Recent Question 2013)
d. Excess fructose is excreted through feces a. Aldose Reductase
e. Excess fructose is converted to glucose b. Galactose Reductase
192. Which will cause post-prandial hypoglycemia? c. Fructose Dehydrogenase
a. Fructose d. Sorbitol Dehydrogenase
b. Galactose 199. What can be prevented in a diabetic patient by giving
c. Glucose drugs which are aldose reductase inhibitors?
M d. Sorbitol a. Diabetic retinopathy
C 193. Fructose intolerance is due to deficiency of:  b. Cataract
Qs  (PGMEE 2013-12) c. Neuropathy
Ans. a. Aldolase B b. Triokinase d. Deafness
188. c
c. Fructokinase d. Aldolase A
189. c
190. b
191. a
192. a
193. a
194. c
195. b
196. c
197. c
198. a
199. b
 Answers with Explanations
103
1. Ans. (d) lactate •• But Q-6 is asking what is the net gain in anaerobic
glycolysis. Gain is only of 2 ATP as 2 NAD is used also

CHAPTER 3  CARBOHYDRATE METABOLISM

[Ref: Harper 30th/e pg. 171] (Glyceraldehyde-3-phosphate Dehydrogenase step)
•• RBCs lack mitochondria. So, the end product of and 2 NAD produced also (LDH step). So, net gain of
glycolysis is always lactate. During exercise, anae- NAD is zero.
robic conditions are created in muscles. So, the end
product is lactate. 7. Ans. (c) Brain
•• From muscles it travels via blood to liver where
[Ref: Harper 30th/e pg. 173]
lactate undergoes gluconeogenesis to form glucose.
This glucose is supplied again to muscles. This cycle •• In brain, mostly aerobic glycolysis occurs because
formed is known as Cori's cycle or Glucose-Lactate brain is highly aerobic tissue. So, glucose is not
cycle. (Refer Fig. 3.3) converted to lactate in brain. Whereas cornea, lens,
RBCs convert glucose to predominantly lactate.
2. Ans. (d) Fructose 2, 6 Bisphosphate
8. Ans. (a) Citrate
[Ref: Harper 30th/e pg.171]
•• Homotropic Modulator: When Substrate or
•• Fructose 2, 6 Bisphosphate is the most potent Substarate like substance act as regulator. E.g. ATP
allosteric activator of PFK-1. It activates glycolysis
and fructose 2, 6 Bis phosphate are Homotropic
and on the other hand, it inhibits gluconeogenesis.
modulators of glycolysis
•• ATP and citrate are allosteric inhibitors of PFK-1.
•• Heterotrophic Modulator: When molecule other
(Refer to figure 3.7)
than substrate act as regulator, E.g. Citrate is a negative
3. Ans. (b) 2, 3 BPG heterotropic allosteric modulator of glycolysis.
2, 3 Bis Phospho Glycerate (2,3-BPG) is produced only 9. Ans. (c) Conversion of glucose to 3 C pyruvate
in RBCs from RL shunt. It decreases the affinity of Hb for
oxygen, therefore it helps in the release of oxygen from [Ref: Harper 30th/e pg. 168]
Hb. It binds to β-chain of Hb (a2β2)
•• Glycolysis occurs in cytoplasm. Glycolysis is not
HbO2 (Oxyhemoglobin) + 2,3 BPG β Hb–2,3 BPG
complete breakdown of glucose. It is conversion of
(Deoxyhemoglobin) + O2.
glucose to 3C pyruvate. There is net gain of 2 ATPs in
4. Ans. (a) 2 anaerobic glycolysis.
A
[Ref: Harper 30th/e pg. 169, table 17-1] 10. Ans. (c) Insulin
N
•• The first phase of glycolysis, also known as energy [Ref: Harper 30th/e pg. 188 Table 19-1] S
Investment or energy utilizing phase uses 2 ATPs W
– one at Hexokinase/Glucokinase step and other at •• Insulin activates glycolysis and link reaction. Insulin
E
PFK-I (Phospho Fructo Kinase-I) step. activates Kinases of glycolysis and PDH of link
R
•• Second phase of glycolysis (Energy producing phase) reaction.
S
produces 9 ATP. So, energetics are 9–2 = 7 ATP
11. Ans. (b) Mg
(Aerobic glycolysis) WITH
[Ref: Harper 30th/e pg. 170] E
5. Ans. (a) 2 ATP + 2 NAD
•• Mg is very important for Kinases. There are four X
6. Ans. (b) 2 ATP Kinases in glycolysis (Hexokinase, Phospho- P
fructokinase, Phosphoglycerate Kinase and pyruvate L
[Ref: Harper 30th/e pg. 169, figure 17-1] Kinase). So, Mg is the most important ion in A
•• Q5, 6 These are two tricky questions asked on glycolysis. N
anaerobic glycolysis. Be careful, these are different A
questions. In Q-5, they are asking end product. In 12. Ans. (b) 2 T
the end of anaerobic glycolysis, 2 ATP and 2 NAD are I
[Ref: Harper 30th/e pg. 173, 174] O
obtained.
N
S
 •• In RBCs, there is no mitochondria. So, NADH cannot of NAD as NAD gets used by Glyceraldehyde
travel into mitochondria to give 5 ATPs in ETC. So, -3-phosphate Dehydrogenase, So, it must be
104 only 2 ATP are produced in RBCs. replenished in the cell for the continuation of
•• In RL shunt, one SLP (Phosphoglycerate kinase) step glycolysis. (Refer to fig 3.2)
is not occurring. So, number of ATPs produced in
RBCs during RL shunt are 2 ATPs (Substrate Level 18. Ans. (c) Both a and b
CRO BIOCHEMISTRY

Phosphorylation by pyruvate Kinase only), but num-

[Ref: Harper 30th/e pg. 172]
ber of ATPs used are also 2 (one at Hexokinase/
Glucokinase step and other at PFK-I step). So net gain Refer Q.12 for explanation
is zero. •• Arsenite inhibits Glyceraldehyde-3-P dehydrogenase
•• But as zero option is not given, so we can mark 2 enzyme. In the presence of arsenite, glycolysis
because number of ATPs produced are 2. continues. But zero ATP are formed. (Also Refer Table
3.6).
13. Ans. (c) 4
[Ref: Harper 30th/e pg. 192] 19. Ans. (a) 7 (Mark 7, not 9 )


•• There are two substrate level phosphorylation steps [Ref: Harper 30th/e pg. 169, table 17-1]
in glycolysis i.e. by enzyme Phosphoglycerate Kinase
•• By default, all questions on ATP ask you net gain.
and Pyruvate Kinase. These steps occur in Phase II
•• ATP produced in glycolysis is net 7
of glycolysis, So, the number of ATPs produced are
•• If 7 is not an option, then mark 9.
multiplied by 2, i.e. total 2 × 2 = 4 ATPs.
•• If options doesn't contain any answer according to
•• Net ATP yield in glycolysis from oxidative
net gain, then mark that answer which tells total ATP
phosphorylation (ETC) is 5 (Glyceraldehyde-3-
produced.
Phosphate Dehydrogenase step).
20. Ans. (b) 1, decreases
14. Ans. (b) Glucokinase
[Ref: Harper 30th/e pg. 172]
[Ref: Harper 30th/e pg. 192]
Post prandial utilization of glucose i.e. in fed state. •• 2,3-BPG is produced only in RBCs from RL shunt. It
It is done by Glucokinase enzyme, which is active in decreases the affinity of Hb for oxygen, therefore it
fed state. Glucokinase has high km for glucose i.e. helps in the release of oxygen from Hb. It binds to one
more amount of glucose is required for this enzyme site of Hb. i.e. β chain of HbA (a2 β2).
to work and more glucose is present during fed state.
21. Ans. (c) Glucokinase, Phosphofructokinase,
Glucokinase is activated by insulin. Hexokinase has low
pyruvate Kinase
Km and feedback inhibition from substrate glucose -6-
phosphate. [Ref: Harper 30th/e pg. 170]
A
N 15. Ans. (b) Pasteur effect •• Irreversible/regulatory steps of glycolysis are Hexo-
S kinase, Phosphofructokinase-1 (PFK-I) and pyruvate
W [Ref: Harper 30th/e pg. 171] Kinase. Rate limiting step is PFK-I. Flux generating
E •• Pasteur effect occurs in any normal cell of the body. It step is Hexokinase.
R says that if oxygen is present then anaerobic glycolysis
is inhibited. 22. Ans. (c) Enolase
S
16. Ans. (d) Glyceraldehyde-3-phosphate and Dihy- [Ref: Harper 30th/e pg. 161]
WITH
droxyacetone phosphate •• Sodium Fluoride inhibits Enolase of glycolysis. It is
E used in blood glucose estimation.
X [Ref: Harper 30th/e pg. 202]
P •• This reaction of glycolysis is catalysed by 23. Ans. (c) ; (e)
L enzyme Aldolase A, which converts fructose 1-6
A [Ref: Harper 30th/e pg. 691, table 53-2]
bisphosphate to glyceraldehyde-3-phosphate and
N dihydroxyacetone phosphate (DHAP). •• RBCs can only use glucose as a fuel (Fatty acids,
A amino acids, ketone bodies cannot be used). Lactate
T 17. Ans. (c) Replenishment of NAD Dehydrogenase is present in RBCs. There is no
I mitochondria in RBCs. Therefore, no mitochondrial
O [Ref: Harper 30th/e pg. 170] enzyme is not present in RBCs. Production of 2, 3
N •• Purpose of the extra step of anaerobic glycolysis is not BPG (RL shunt) does not yield any ATP in RBCs. ATP
S lactate formation, but the purpose is replenishment Synthase is absent in RBCs.
24. Ans. (d) Increased CSF lactate is associated with 30. Ans. (c) Thiamine
good prognosis
[Ref: Harper 30th/e pg. 174] 105
[Ref: Harper 30th/e pg. 170] •• Q 30, 31 Congenital lactic acidosis may occur due
•• In brain injury, anaerobic condition prevail. So, to defect in thiamine which is required for pyruvate
lactate levels are increased which are associated with dehydrogenase which converts pyruvate to acetyl

CHAPTER 3  CARBOHYDRATE METABOLISM

poor prognosis. Pyruvate dehydrogenase activity CoA. If this reaction is defective then pyruvate
is decreased. Also, increase lactate uptake from accumulates and leads to the formation of lactate.
circulation is seen. As brain cells start using lactate as This leads to lactic acidosis. Also this enzyme requires
fuel because glucose uptake. vitamins B1, B2, B3, B5 and Lipoic acid. Option (a) is
vitamin B7- Biotin. Option (b) is PLP- vitamin B6.
25. Ans. (d) This reaction requires lipoic acid and four Option (d) is Pyruvate which is coming from food
B-complex vitamins (carbohydrates).

[Ref: Harper 30th/e pg. 172] 31. Ans. (a) Fats can be converted to carbohydrates
•• Link reaction is a link between glycolysis and TCA [Ref: Harper 30th/e pg. 173,174]
(not TCA and ETC). It is oxidative decarboxylation
•• Fats cannot be converted to carbohydrates because
(not oxidative deamination) of Pyruvate not Acetyl
link reaction is irreversible. Excess carbohydrates
CoA vis diminished following injury.
can be converted to fats. Glycerol and Propionic
•• This reaction requires 5 coenzymes i.e. Lipoic acid
acid are two breakdown products of fats which
and four B-complex vitamins (B1, B2, B3, B5). can be converted to glucose. Beri-Beri (vitamin B1
•• This reaction is irreversible. Therefore Fats cannot be deficiency) leads to lactic acidosis.
converted to carbohydrates.
32. Ans. (a) ; (c) ; (e)
26. Ans. (c) Pyruvate
[Ref: Harper 30th/e pg. 172]
[Ref: Harper 30th/e pg. 172] •• Pyruvate Dehydrogenase complex has three enzyme
•• Acetyl CoA is derived from carbohydrates, fats and components:
proteins also. But major source is carbohydrates. ƒƒ E1: Pyruvate Dehydrogenase
Glucose forms pyruvate and then it forms acetyl CoA. ƒƒ E2: Dihydrolipoyl Transacetylase
So, best answer here is pyruvate. ƒƒ E3: Dihydrolipoyl Dehydrogenase.

27. Ans. (c) Is cofactor for pyruvate dehydrogenase and 33. Ans. (a); (b); (c); (d); (e)
alpha ketoglutarate dehydrogenase [Ref: Harper 30th/e pg. 203]
•• Acetyl CoA is the precursor for the synthesis of
[Ref: Harper 30th/e pg. 174] A
cholesterol, steroids, fatty acids and ketone bodies. It
•• Link reaction and TCA cycle requires 5 coenzymes is derived from glycolysis and link reaction (glucose N
i.e. Lipoic acid and four B-complex vitamins (B1, B2, → Pyruvate → Acetyl CoA). It is never glucogenic S
B3, B5). Due to this reason, Thiamine (Vitamin B1) (i.e. can never be converted to glucose). W
deficiency results in decrease energy production as E
TCA and link reaction both are responsible for energy 34. Ans. (a) Acetyl CoA R
production in cells. [Ref: Harper 30th/e pg. 162] S
28. Ans. (c) Lactate dehydrogenase •• Acetyl CoA is not the intermediate of TCA cycle. WITH
•• Citrate, Isocitrate, Alpha-ketoglutarate, Succinyl
[Ref: Harper 30th/e pg. 172] CoA, Succinate, Fumarate, Malate and Oxaloacetate E
are intermediates of TCA cycle. X
•• Pyruvate Dehydrogenase complex is enzyme of P
link reaction, which is irreversible. 3 enzymes of 35. Ans. (b) NAD L
glycolysis are irreversible – Hexokinase/Glucokinase, A
Phosphofructokinase-1 and pyruvate Kinase. But [Ref: Harper 30th/e pg. 119]
N
lactate dehydrogenase (pyruvate to lactate) is •• Cyanide inhibits complex IV of ETC. So, ETC is A
involved in anaerobic glycolysis, is reversible. inhibited and NADH accumulates. If NADH accu- T
mulates, means NAD depleted. So, TCA affected I
29. Ans. (c) Pyruvate Dehydrogenase because each TCA cycle requires 3 NAD (for Isocitrate O
Dehydrogenase, alpha-ketoglutarate dehydrogenase N
and Malate Dehydrogenase). S
 36. Ans. (c) Both a and b 42. Ans. (b) Citrate

106 [Ref: Harper 30th/e pg. 163] [Ref: Harper 30th/e pg. 163]
•• Thiokinase of TCA produces ATP most of the times. •• Citrate is the first compound of TCA cycle therefore
But in liver and kidney, during starvation it produces this cycle is known as citric acid cycle.
GTP, for PEPCK enzyme of gluconeogenesis.
CRO BIOCHEMISTRY

43. Ans. (a) 12.5

37. Ans. (a) Isocitrate Dehydrogenase
[Ref: Harper’s 30th/e pg. 169-170]
[Ref: Harper 30th/e pg. 166]
•• When one pyruvate forms acetyl CoA, one NADH
•• The most likely sites for regulation in TCA are the is produced (= 2.5 ATPs). Then acetyl CoA gives 10
non-equilibrium reactions catalysed by Pyruvate ATPs from TCA cycle. So, total is 12.5 ATPs from one
Dehydrogenase, Citrate Synthase, Isocitrate Dehyd- pyruvate.
rogenase and Alpha-Ketoglutarate Dehyd-rogenase.
•• There is allosteric inhibition of Citrate Synthase by 44. Ans. (d) Iodoacetate inhibits Glycerol-3-phosphate
ATP and long chain fatty Acyl CoA and allosteric Dehydrogenase


activation of Isocitrate Dehydrogenase by ADP.

[Ref: Harper 30th/e pg. 163]
•• Succinate Dehydrogenase is inhibited by Oxaloace-
tate. •• Iodoacetate inhibits Glyceraldehyde-3-phosphate
Dehydrogenase, (not Glycerol-3-phosphate Dehy-
38. Ans. (b) Conversion of Pyruvate to Oxaloacetate drogenase). Other options given in question are true.
[Ref: Harper 30th/e pg. 162] 45. Ans. (c) Mitochondria
•• Anaplerotic reactions are those reactions which
[Ref: Harper 30th/e pg. 119]
synthesize TCA intermediates. Conversion of pyruv-
ate to Oxaloacetate is the most important anaplerotic •• Complete oxidation of glucose to CO2 and H2O occurs
reaction. in mitochondria. (when TCA and ETC takes place).

39. Ans. (b) It is both endothermic and exothermic 46. Ans. (b) Oxaloacetate

[Ref: Harper 30th/e pg. 162] [Ref: Harper 30th/e pg. 163]
•• TCA is Amphibolic means it is both Catabolic (i.e. •• The two molecules of CO2 which are lost in TCA, are
Exothermic) and Anabolic (i.e. Endothermic). from Oxaloacetate. These are not from that acetyl
CoA, which has immediately entered the cycle.
40. Ans. (d) Malonate •• Acetyl CoA in one turn of the cycle will give its
A carbons to oxaloacetate, not CO2. In next turn of the
[Ref: Harper 30th/e pg. 161]
N cycle, these two carbons from Oxaloacetate (given by
S •• Malonate (3C) is the competitive inhibitor of Acetyl CoA in the previous turn of cycle) will be lost
Succinate Dehydrogenase. in the form of CO2.
W
•• If oxaloacetate not given in option, then you should
E 41. Ans. (a) H2O mark Acetyl CoA.
R
S [Ref: Harper 30th/e pg. 163]
•• Water is liberated when citrate converted to cis-
WITH
aconitate.
E
X
P
L
A
N
A
T
I
O
N
S
47. Ans. (b) NADH 54. Ans. (c) Competitive

[Ref: Harper 30th/e pg. 165] [Ref: Harper 30th/e pg. 164] 107
•• Main source of energy in TCA is from NADH (enters •• Malonate competes with succinate for enzyme
ETC to give ATPs) (1NADH = 2.5 ATPs). Succinate Dehydrogenase as malonate resembles
in structure with succinate (Competitive inhibitor

CHAPTER 3  CARBOHYDRATE METABOLISM

48. Ans. (b) Pyruvate Dehydrogenase always resembles in structure with the substrate of
[Ref: Harper 30th/e pg. 163] the reaction).

•• Pyruvate Dehydrogenase is involved in link reaction 55. Ans. (b); (c); (d)
(conversion of Pruvate to acetyl CoA). •• Lactate dehydrogenase is working in anaerobic
•• There are 4 Dehydrogenases in TCA - Isocitrate conditions. It converts pyruvate to lactate. This
Dehydrogenase, Alpha-Ketoglutarate Dehydro- enzyme is normally required during exercise. So,
genase, Succinate Dehydrogenase and Malate Dehy- deficiency will lead to exercise intolerance and
drogenase.
muscle cramps (Cori’s cycle cannot occur to provide
49. Ans. (d) All glucose to exercising muscles).

[Ref: Lehninger 7th/e pg. 641] 56. Ans. (b); (c); (d)
There is not one rate limiting enzyme of TCA cycle. [Ref: Harper 30th/e pg. 163,164]
3 enzymes can be rate limiting depending on various
•• NAD acts as a cofactor for Isocitrate Dehydrogenase,
complex conditions. They are Citrate Synthase,
a-Ketoglutarate Dehydrogenase and Malate Dehy-
Isocitrate Dehydrogenase and Alpha-Ketoglutarate
drogenase.
Dehydrogenase.
•• Coenzyme A acts as cofactor for Citrate Synthase.
50. Ans. (d) Succinate Dehydrogenase Succinyl Thiokinase uses GDP or ADP.

[Ref: Harper 30th/e pg. 164] 57. Ans. (b); (d)

All enzymes of TCA are in mitochondrial matrix. But [Ref: Harper 30th/e pg. 163,164]
Succinate Dehydrogenase is present in IMM.It is also part
of complex II of ETC. It is inhibited by Malonate. FAD is the •• There are two oxidative decarboxylation reactions in
hydrogen acceptor. TCA i.e CO2 is released in two steps of TCA
1. Conversion of Isocitrate (6C) to Alpha-Keto-
51. Ans. (b) Availability of Oxaloacetate glutarate (5C) by enzyme Isocitrate Dehyd-
rogenase
[Ref: Lehninger 7th/e pg. 641] 2. Conversion of Alpha-Ketoglutarate (5C) to Succi-
A
TCA has no hormonal control. It is a vital cycle for the nyl CoA (4C) by enzyme Alpha-keto-gluta-rate
N
cells. So, it does not depends on insulin or glucagon. Dehydrogenase.
S
TCA depends on the availability of oxaloacetate (not
58. Ans. (d) Acetyl CoA W
acetyl CoA) and energy status of cells.
E
[Ref: Harper 30th/e pg. 162]
52. Ans. (b) Irreversible R
•• First Substrate of TCA cycle
S
[Ref: Harper 30th/e pg. 162] ƒƒ Best answer—Oxaloacetate
ƒƒ Second best—pyruvate (because it can form WITH
The initial reaction of TCA between acetyl CoA and
Oxaloacetate by enzyme pyruvate Carboxylase)
oxaloacetate to form citrate is catalysed by Citrate E
ƒƒ If these not given, then go for acetyl CoA.
Synthase. This reaction is irreversible and exothermic. X
Two reactions in TCA are irreversible: Citrate Synthase 59. Ans. (b) Oxaloacetate P
and Alpha-Ketoglutarate Dehydrogenase steps. L
[Ref: Harper 30th/e pg. 164]
A
53. Ans. (a) Carbohydrates •• Oxaloacetate has a catalytic role in TCA. It is unaltered N
during the cycle. A
[Ref: Harper 30th/e pg. 164]
•• Oxaloacetate is regarded as: T
•• Thiamine is a coenzyme mainly for pyruvate Dehy- ƒƒ Carrier of TCA cycle I
drogenase (Link reaction), Alpha Ketoglutarate ƒƒ 1st substrate of TCA O
Dehydrogenase (TCA)and Transketolase (HMP). ƒƒ Has a catalytic role in TCA. N
•• All these enzymes are important in carbohydrate S
metabolism.
 60. Ans. (a) TCA cycle 67. Ans. (d) 0.5

108 (Ref: Harper’s illustrated biochemistry, 30th ed., Pg. 162) [Ref: Harper 30th/e pg. 131]
•• ATPs given by complex I and III are 1 ATP each. But
•• Fluoroacetate is the non-competitive inhibitor of
Aconitase (TCA Cycle). Flourocitrate is the com- ATP given by complex IV is 0.5 ATPs. As complex I
petitive inhibitor of same enzyme. and III transfers 4 protons. Complex IV transfers 2
CRO BIOCHEMISTRY

protons.
61. Ans. (a) Glycolysis
68. Ans. (a) Inner Mitochondrial Membrane
[Ref: Lehninger 7th/e pg. 687]
•• Malate shuttle and Glycerol phosphate shuttle [Ref: Harper 30th/e pg. 131]
transports NADH from cytoplasm to mitochondria. •• All components of ETC are located in Inner mito-
So, any pathway which occurs in cytoplasm requires chondrial Membrane (IMM). Other options given in
these shuttles. But any pathway which occurs in question are true.
mitochondria does not require shuttles.
•• Glycolysis occurs in cytoplasm. Pyruvate Dehydr- 69. Ans. (b) Occurs in mitochondrial matrix
ogenase complex i.e. link reaction and TCA cycle


[Ref: Harper 30th/e pg. 131]

occurs in mitochondria.
ETC occurs in inner mitochondrial membrane, not in
62. Ans. (b) 5 ATP mitochondrial matrix.
[Ref: Harper 30th/e pg. 134]
70. Ans. (d) ATP
•• In Glycerol-3-phosphate shuttle, NADH is taken
from cytoplasm but FADH2 is delivered into the [Ref: Harper 30th/e pg. 131]
mitochondria. So, now 2 FADH2 gives 3 ATPs. Mitochondrial membrane has ADP –ATP Translocase,
•• 2 ATPs used at Hexokinase and PFK-I (Phospho- which transports ADP in and ATP out.
fructokinase-I). 4 ATPs are produced at Substrate
Level Phosphorylation steps. 71. Ans. (d) NAD+/NADH
•• So, 3 + 4 – 2 = 5 ATPs.
[Ref: Harper 30th/e pg. 120, table 12-1]
63. Ans. (d ) Zero
•• NADH has minimum redox potential
[Ref: Harper 30th/e pg. 134] •• Oxygen has maximum R.P
•• NADPH does not take part in ATP formation. It takes (Refer Fig. 3.26)
part in reductive biosynthesis.
72. Ans. (a) Oxidative Phosphorylation
64. Ans. (c) After going in brain in ETC, it gives high
energy [Ref: Harper 30th/e pg. 131]
A Most of the ATPs in body are derived from ETC or
N [Ref: Harper 30th/e pg. 134]
oxidative phosphorylation. But only few ATPs are
S •• There are two shuttles for transport of NADH from
derived from substrate level phosphorylation.
W cytoplasm to mitochondria. In brain, Glycerol-P-
E shuttle is present which gives 1.5 ATPs. This shuttle is 73. Ans. (a) Inhibition of ATP synthesis only not ETC
R a shorter shuttle, so, it is a quick source of ATP.
[Ref: Harper 30th/e pg. 132]
S 65. Ans. (b) Coenzyme Q Coupling in ETC means oxidation (electron flow)
WITH [Ref: Harper 30th/e pg. 131] and phosphorylation (ATP synthesis) occur together.
•• All members of ETC (Complex I to V) and Cyto- Uncoupling means electron flow is occurring but ATP
E synthesis is not occurring. So, electron transport is
X chrome c are proteins but Coenzyme Q is not a
protein. It is hydrophobic, a lipophilic compound, occuring in chain without ATP formations.
P
L made up of isoprene units, present within the inner
mitochondrial membrane. 74. Ans. (a) Complex I to Coenzyme Q
A
N [Ref: Harper 30th/e pg. 132]
66. Ans. (a) Cu
A •• Barbiturates such as Amobarbital inhibit electron
T [Ref: Harper 30th/e pg. 131] transport via Complex I (by blocking the transfer
I •• All Oxidases requires copper. Complex IV (Cyto- from Complex I to Coenzyme Q).
O chrome C oxidase) of ETC also requires copper. Two
N oxidases donot require Cu i.e. Xanthine Oxidase,
S Sulfite Oxidase.
75. Ans. (d) Complex IV 83. Ans. (b) Prevents ATP synthesis and electron trans-
port chain is increased
[Ref: Harper 30th/e pg. 132] 109
[Ref: Harper 30th/e pg. 131]
•• Carbon Monoxide (Co2), Hydrogen Sulphide (H2S )
and Cyanide (CN–) are inhibitors of Complex IV of •• Here best answer is that Dinitrophenol prevents ATP
ETC. synthesis and electron transport chain is increased.

CHAPTER 3  CARBOHYDRATE METABOLISM

•• Dinitrophenol is an uncoupler. Uncoupler means
76. Ans. (c) Fo-F1 ATPase electron flow (oxidation) is occurring but phosho-
rylation is not occurring. So, this will lead to reduced
[Ref: Harper 30th/e pg. 132]
ATP formation as phosphorylation is not occurring.
•• F0-F1 ATPase is Complex V of ETC, which is So, it will stimulate the electron transport in order to
responsible for generation of ATP. try to produce more ATPs.

77. Ans. (c) NADH Dehydrogenase 84. Ans. (a); (d)

[Ref: Harper 30th/e pg. 128; Fig.13–3] [Ref: Harper 30th/e pg. 132]
•• NADH Dehydrogenase / Ubiquinone is the compo- •• In normal mitochondria, the rate of electron transfer
nent in ETC, also known as Coenzyme Q. Or is tightly coupled with ATP formation (phosphor-
sometimes Complex I is called NADH Dehydrogenase. ylation).
•• Succinyl CoA Thiokinase or Succinate Thiokinase is •• Uncouplers are substances which disrupt the
involved in TCA. coupling of oxidation and phosphorylation in ETC.
•• Pyruvate Kinase is involved in glycolysis. •• 2,4 DNP and Thermogenin are uncouplers.
Thermogenin, present in brown fat is responsible for
78. Ans. (d) Cyt aa3 non-shivering thermogenesis. It is a natural/ physi-
ological uncoupler. (Refer Fig. 3.28)
[Ref: Harper 30th/e pg. 129; Fig.13–5]
•• H2S and Cyanide are inhibitors of Complex IV of ETC.
•• Cyt aa3 is complex IV of ETC, which directly reacts •• Carboxin is a drug which act as Fungicide, inhibits
with oxygen. complex II of ETC

79. Ans. (a) Oxygen 85. Ans. (b) Low to high potential

[Ref: Harper 30th/e pg. 3] [Ref: Harper 30th/e pg. 129]

•• Electron transfer in ETC starts from NADH and last Electrons in electron transport chain travel from low
electron acceptor in the chain is oxygen. redox potential to high redox potential. Redox potential
is the tendency to gain electrons.
80. Ans. (b) Inhibitor of oxidative phosphorylation A
86. Ans. (c) Nicotinic acid N
[Ref: Harper 30th/e pg. 132]
[Ref: Harper 30th/e pg. 131] S
•• Atractyloside is inhibitor of ADP-ATP translocase, W
which transfers ADP and ATP across the inner mito- Both vit B2 (Riboflavin) and B3 (Niacin) are used ETC as E
chondrial membrane. So, it is inhibitor of oxidative ETC starts from NADH or FADH2. R
phosphorylation i.e. ETC.
87. Ans. (b) Riboflavin S
81. Ans. (d) Creatine phosphate WITH
[Ref: Harper 30th/e pg. 133]
[Ref: Harper 30th/e pg. 116] •• In ETC, complex I (FMN, Fe-S ) and complex II (FAD, E
•• Creatinine is the breakdown product of Creatine Fe-S) are involved, which contains the component of X
phosphate. This reaction occurs in muscles and does riboflavin (FAD, FMN). P
not involve any enzyme. •• Nicotinic Acid or NAD is not a component of ETC, it L
acts as Substrate or starting material in ETC. A
82. Ans. (d) Non-competitive irreversible inhibition •• Riboflavin is component as well as starting material N
in ETC. A
[Ref: Harper 30th/e pg. 132] T
CN, CO and H2S are inhibitors of Cytochrome c Oxidase 88. Ans. (d) Thermogenin I
(Complex IV of ETC). They inhibit by non-competitive [Ref: Harper 30th/e pg. 131] O
inhibition and this type of inhibition is mostly irrever- Oxidative phosphorylation means ETC. Inhibitors N
sible. of ETC are Malonate (Complex II inhibitor), CO S
 (Complex IV inhibitor) & Antimycin A (Complex III 96. Ans. (c) Leucine
inhibitor). But Thermogenin is uncoupler i.e. it inhibits
110 Phosphorylation but not Oxidation. [Ref: Harper 30th/e pg. 171]
•• 18 amino acids are glucogenic (out of 20). These
89. Ans. (b) II
all can form glucose. Only 2 amino acids are purely
[Ref: Harper 30th/e pg. 135] ketogenic which cannot form glucose i.e. Leucine,
CRO BIOCHEMISTRY

MELAS inhibits complex I, III & IV. MELAS is Lysine.

mitochondrial Encephalopathy, Lactic Acidosis and •• Now in this question both are given. Leucine is
Stroke like episodes. This is a mutation in mitochondrial the best answer to be marked because Lysine is
DNA, which affects brain and causes Lactic acidosis due in controversy. (Some books say that it is both
to anaerobic metabolism. glucogenic and ketogenic).

90. Ans. (a); (e) 97. Ans. (c) Acetoacetate

DNP is uncoupler of ETC. Oligomycin inhibits Complex
[Ref: Harper 30th/e pg. 171]
V of ETC. Hence, oxidative phosphorylation is inhibited
by 2, 4 DNP and oligomycin. Flouride inhibits glycolysis •• Glucose can be synthesized from amino acids. Most


(Enolase). Carboxin is a fungicide, inhibits complex II of glucogenic amino acid is Alanine.

ETC. Ouabain is a cardiac glycoside like Digitoxin. •• From Glycerol, glucose can be formed.
•• Lactic acid can form pyruvate by LDH, which further
91. Ans. (a); (b); (e) forms glucose.
[Ref: Harper 30th/e pg. 135] •• Ketone bodies, fats, or acetyl CoA can never be
NADH Co-Q Reductase is Complex I of ETC. converted into carbohydrates. Acetoacetate is a
Cytochrome C Oxidase is Complex IV of ETC. Co-Q ketone body.
Cytochrome C Reductase is Complex III of ETC.
98. Ans. (a) Acetyl CoA
Glutathione Reductase is involved in reduction of
glutathione. Isocitrate Dehydrogenase is involved in [Ref: Harper 30th/e pg. 171]
TCA.
•• Acetyl CoA is also not considered the intermediate of
92. Ans. (b); (c) TCA cycle.
•• Acetyl CoA is never glucogenic i.e. It can never form
[Ref: Harper 30th/e pg. 120]
glucose because it cannot be converted to pyruvate
High energy compounds in biological systems are : ATP,
GTP, UTP, Creatine phosphate, Phosphoenol pyruvate, 99. Ans. (c) Palmitate
Coenzyme A, 1,3-bisphospho glycerate and Carbamoyl
phosphate. [Ref: Harper 30th/e pg. 171]
A •• Palmitate, a fatty acid (16C), is not a substrate of
93. Ans. (d) 2.6 ATP
N gluconeogenesis.
S [Ref: Harper 30th/e pg. 132] •• Odd chain fatty acid do act as substrates of
W Number of ATPs produced from NADH in ETC is 2.5. gluconeogenesis, since Propionyl CoA- the product
E Here best answer is 2.6 of their oxidation can enter TCA cycle through
R formation of Succinyl CoA, hence can contribute
94. Ans. (a); (c)
S towards glucose production.
Complex I (NADH-Q Oxidoreductase) and III (CoQ •• Alanine, pyruvate and Glutamate are glucogenic.
WITH Cytochrome c Reductase) of ETC transfers 4 protons.
Complex II transfers 2 protons. 100. Ans. (d) Decreased uptake of glucose by peripheral
E cells
X 95. Ans. (b); (d) ; (e)
P [Ref: Harper 30th/e pg. 131] [Ref: Harper 30th/e pg. 171]
L
Breaking of high energy bond to low energy bond •• In diabetes, lack of insulin occurs. Normally insulin
A
releases energy. Electrical gradient across inner and recruits GLUT-4 which helps in entry of blood
N
outer side of mitochondrial membrane generates ATP glucose into the peripheral tissues (skeletal muscles,
A
using ETC. Passage of electron through FADH2 in ETC cardiac muscles, adipose tissues). In diabetes, there
T
also gives ATP using Complex II. is decreased uptake of glucose by peripheral cells,
I
due to lack of insulin and thus GLUT-4.
O
N
S
 •• Increased ketone body production can occur in a 106. Ans. (b) Gluconeogenesis
diabetic patient (Option a), but that is not the reason
for the patient's inability to maintain a normal blood [Ref: Harper 30th/e pg. 171] 111
glucose level. Odd chain fatty acid can act as substrates of Gluconeo-
genesis, since Propionyl CoA, the product of their
101. Ans. (a) Mitochondria → Cytoplasm → ER

CHAPTER 3  CARBOHYDRATE METABOLISM

oxidation can enter TCA cycle through formation of
[Ref: Harper 30th/e pg. 171] Succinyl CoA, hence can contribute towards glucose
production.
First step of gluconeogenesis (Pyruvate to oxaloacetate)
occurs in mitochondria, then next many steps occur in 107. Ans. (a) Glycerol
cytoplasm and the last step (glucose-6-Phosphatase)
occurs in endoplasmic reticulum (ER). [Ref: Harper 30th/e pg. 171]
Fats can never be converted to carbohydrates. But
102. Ans. (b) Acetyl CoA activates Pyruvate Carboxylase
Glycerol and Propionic acid (breakdown products of fat)
[Ref: Harper 30th/e pg. 171] can be converted to carbohydrates. Adenine is a purine
nitrogenous base. Guanosine is a purine nucleoside.
•• Acetyl CoA can never be converted to glucose but
acetyl CoA activates the first step of gluconeogenesis 108. Ans. (d) Leucine
(pyruvate Carboxylase). Option (a) is also correct but
here the Question is asking which is most effective. [Ref: Harper 30th/e pg. 171]
Most effective is Option (b) •• Only 2 amino acids are purely ketogenic which cannot
•• Option (c) is also correct but that occurs in FA form glucose i.e. Leucine, Lysine. They cannot form
synthesis. glucose, So, they cannot form glycogen.
•• Alanine is glucogenic. Phenylalanine and threonine
103. Ans. (c) glucose-6-Phosphatase
belongs to both Ketogenic and Glucogenic category.
[Ref: Harper 30th/e pg. 171]
109. Ans. (c) Liver
Glucose-6-Phosphatase enzyme is common to both
pathways; Gluconeogenesis and Glycogenolysis. So, [Ref: Harper 30th/e pg. 171]
Glucose-6-phosphatase is affected. •• Gluconeogenesis occurs in liver and kidney. If both
are given, then best answer is liver as it mainly occurs
104. Ans. (c) Alanine, (a) Glycerol
in liver.
[Ref: Lehninger 7th/ed pg 559 ]
110. Ans. (b) PFK-I
•• Substrates of Gluconeogenesis: Pyruvate, Lactate,
Glycerol, Propionic acid, glucogenic amino [Ref: Harper 30th/e pg. 173]
A
acids, Both Glucogenic and Ketogenic amino •• PFK-I is an enzyme involved in glycolysis not gluco- N
acids, intermediates of TCA (acetyl CoA is not an neogenesis. Pyruvate carboxylase and Glucose-6- S
intermediate of TCA) phosphotase involved in gluconeo-genesis. Enolase W
•• Generally fats cannot be converted to carbohydrates is involved in both glycolysis and gluconeogenesis. E
but there are two EXCEPTIONS: Glycerol and
Propionic acid can form glucose R
111. Ans. (b) Aldolase B
•• Among 13 purely glucogenic amino acids, Alanine is S
most glucogenic. [Ref: Harper 30th/e pg. 171]
WITH
•• Lysine and leucine are purely ketogenic and can •• Aldolase B is an enzyme involved in fructose
never form glucose. metabolism, not in gluconeogenesis. E
•• In gluconeogenesis, three enzymes are rate limiting X
105. Ans. (a) Phosphoenol pyruvate depending on various complex conditions in the P
[Ref: Harper 30th/e pg. 171] body. → These are pyruvate carboxylase, PEP carbo- L
xykinase and fructose 1,6-bisphosphatase. A
Phosphoenol pyruvate to pyruvate conversion is a step N
in glycolysis catalysed by enzyme pyruvate Kinase. This 112. Ans. (c) Phosphoglucomutase A
step is irreversible. So, Pyruvate cannot be converted T
directly into phosphoenol pyruvate. Pyruvate forms [Ref: Harper 30th/e pg. 171]
I
Alanine by SGPT enzyme. Pyruvate forms acetyl CoA by •• Phoshoglycero mutase is involved in Gluco- O
PDH enzyme. Pyruvate forms lactate by LDH enzyme. neogenesis (not phosphogluco mutase). There N
S
 are four enzymes involved in gluconeogenesis, 117. Ans. (a) Increased alanine levels in liver
which are not in glycolysis. They are Pyruvate
112 Carboxylase, PEPCK, fructose 1,6 bisphosphatase [Ref: Harper 30th/e pg. 171]
and glucose-6-Phosphatase. Enolase, Phoshoglycero •• During fasting/ starvation, pyruvate in muscles gets
Mutase, phospho glycerate Kinase, glyceraldehyde- converted to alanine, which enters liver and forms
3-phosphate Dehydrogenase, Phospho Triose glucose via gluconeogenesis (Fig. 5.15 Cahill cycle).
CRO BIOCHEMISTRY

isomerase and Aldolase A are enzymes common to

glycolysis and gluconeogenesis. Phoshogluco mutase 118. Ans. (c) Tyrosine
is involved in glycogen metabolism. [Ref: Lehninger 7th/e pg. 563]
113. Ans. (d) Synthesis of glycogen from non- •• Six amino acids form pyruvate during catabolism.
carbohydrate sources They are Glycine, Alanine, Serine, Threonine,
Cysteine and Hydroxy-Proline. Then pyruvate forms
[Ref: Harper 30th/e pg. 171] acetyl CoA via pyruvate Dehydrogenase.
•• Glyconeogenesis is synthesis of glycogen from non-
119. Ans. (b) Gluconeogenesis
carbohydrate precursors, without using glucose


or other carbohydrates. Glycogenesis – synthesis [Ref: Harper 30th/e pg. 188, 189]
of glycogen from glucose. This pathway is same
•• Gluconeogenesis occurs in mitochondria &
like gluconeogenesis, just last step is different i.e.
cytoplasm. Glycogenesis and glycogenolysis occur
in glyconeogenesis, glucose-6-phosphate directly
in the cytoplasm. Glycolysis also occurs in the
enters glycogen synthesis. So, glucose-6-Phosphatase cytoplasm. (Three pathways which occur both in
step is not required. mitochondria & cytoplasm are gluconeogenesis, urea
cycle and haem synthesis).
114. Ans. (a) Level of fructose 1,6 bisphosphate is higher
than normal 120. Ans. (a) Fructose 6 phosphate to glucose 6 phosphate
•• Normally fructose 2, 6 bisphosphate decreases the
activity of fructose1,6 bisphosphatase (enzyme of [Ref: Harper 30th/e pg. 188, 189]
gluconeogenesis). Now due to mutation, activity is •• Glucose 6 phosphate to fructose 6 phosphate
not decreased. So, Fructose 1, 6-bisphosphatase will occurs in glycolysis but opposite of this i.e. Fructose
increase. So, its substrate (fructose 1, 6 bisphosphate) 6 phosphate to glucose 6 phosphate occurs in
is decreased. This is committed molecule of gluconeogenesis. Conversion of pyruvate to lactate
glycolysis. So, glycolysis decreases. Therefore, ATP is in anaerobic glycolysis. Opposite of Phosphoenol
and pyruvate decreases. So, option (a) is wrong. pyruvate to oxaloacetate occurs in gluconeogenesis.
Pyruvate to acetyl CoA is in link reaction.
115. Ans. (a) Malate
121. Ans. (b) Glucose – 6 – phosphate
A
[Ref: Lehninger 7th/e pg. 558]
N [Ref: Harper 30th/e pg. 203]
S •• During gluconeogenesis, pyruvate to oxaloacetate
conversion occurs in mitochondria But next step, Glucose–6–phosphate is the common metabolite
W
i.e. oxaloacetate to phosphoenol pyruvate formation formed in glycogenesis, glycogenolysis, glycolysis and
E gluconeogenesis.
R occurs in cytoplasm. So, there is a need for the
transport of oxaloacetate from mitochondria to 122. Ans. (c) Pyruvic acid (pyruvate)
S
cytoplasm but oxaloacetate cannot cross inner
WITH mitochondrial membrane. So, malate is used here. [Ref: Harper 30th/e pg. 186]
(but be careful, this is not malate shuttle). •• Pyruvic acid or pyruvate (3C) is the most important
E
substrate of gluconeogenesis. It forms oxaloacetate in
X 116. Ans. (a) Glycolysis
the first step of gluconeogenesis by enzyme pyruvate
P
[Ref: Harper 30th/e pg. 186; Fig.19-1] carboxylase.
L
•• Fatty acids or Palmitic acid (16 C fatty acid) and acetyl
A •• Malate shuttle is for the transport of NADH from CoA are not the substrates of gluconeogenesis.
N cytoplasm to mitochondria, so, that it can take part
A in ETC. Glycolysis occurs in cytoplasm. So, glycolysis 123. Ans. (c) Thyroxine
T needs this malate shuttle.
I •• There is no malate shuttle in gluconeogenesis. Its just [Ref: Harper 30th/e pg. 192]
O that malate is used in gluconeogenesis to transport •• Thyroid hormones increase cellular metabolism.
N oxaloacetate from mitochondria to cytoplasm. Initially, they stimulate transcription and protein
S
 synthesis in various tissues, resulting in positive 128. Ans. (c) Insulin
nitrogen balance. Glucose absorption from the
intestine and its utilization in various tissues is
[Ref: Harper 30th/e pg. 150] 113
•• Insulin increases the activity of muscle glycogen
enhanced. It increases gluconeogenesis & lipolysis
Synthase after the intake of carbohydrates and
leading to increased concentration of glucose and
increases glycogen reserves in liver and muscles.
free fatty acids in plasma. Epinephrine and nor-

CHAPTER 3  CARBOHYDRATE METABOLISM

Insulin is anabolic hormone, So, increases the rate of
epinephrine leads to glycogenolysis.
anabolic pathway – glycogenesis.
•• Glucagon leads to both glycogenolysis and glucone-
ogenesis glucocorticoids increase gluconeogenesis 129. Ans. (a) Glucose-6-Phosphatase
and inhibit utilization of glucose in extra-hepatic
tissues. 130. Ans. (b) Glycogen Phosphorylase
•• So, thyroid hormones and glucocorticoids don’t
affect glycogen metabolism. [Ref: Lehninger 7th/e pg. 604]
•• But catecholamines and glucagon affects glycogen These are 2 different questions. Please read carefully
metabolism. Q129 is asking why muscles cannot maintain blood
glucose levels. This is because muscles lack the enzyme
124. Ans. (a); (c); (d); (e) Glucose-6-Phosphatase. Liver cells have this enzyme
[Ref: Harper 30th/e pg. 188, 189] and therefore liver can only maintain blood glucose.
•• Increased fatty acid breakdown release acetyl CoA However Q130 is asking why muscle cannot make
which activates the first enzyme of gluconeogenesis, use of glycogen. It is because of deficiency of Glycogen
i.e. Pyruvate Carboxylase(Option a and c). Phosphorylase. Muscles normally lack enzyme glucose-
•• PFK-II (Phopho fructo kinase-II) is inhibited in 6-phosphatase, but they can make use of their glycogen
fasting state (Option d). reserves for muscle activity. Glucose-6-phosphatase
•• Alanine is most glucogenic amino acid which is is required for maintainence of blood glucose. This
mainly released from muscles (Option e) enzyme is present in Liver.
•• Increased conversion of Phosphoenol Pyruvate to 131. Ans. (a) Hepatic Glycogen
pyruvate by activation of Pyruvate Kinase occurs in
glycolysis not Gluconeogenesis. [Ref: Harper 30th/e pg. 150]
Hepatic glycogen is the major storage form of
125. Ans. (b) PLP (Pyridoxal phosphate) carbohydrates in animals. Starch is the major storage
form of carbohydrates in plants.
[Ref: Harper 30th/e pg. 178]
•• Rate limiting enzyme of glycogenolysis is glycogen 132. Ans. (b) Glycogen Phosphorylase
Phosphorylase which requires PLP i.e. vitamin B6.
[Ref: Harper 30th/e pg. 178]
This vitamin is required as donor of phosphate as
Glycogen phosphorylase transfers phosphate to Glucagon (Catabolic hormone) activates the rate
A
glucose, releasing glucose-1-phosphate. limiting enzyme of glycogenolysis (catabolic pathway)
i.e. Glycogen Phosphorylase. N
S
126. Ans. (d) Glycogenin
133. Ans. (b) Polypeptide W
[Ref: Harper 30th/e pg. 153] [Ref: Harper 30th/e pg. 177] E
Protein Kinase, Calmodulin and cAMP increase the R
Glycogenin is a protein which acts as primer in glycogen
activity of Glycogen Phosphorylase. But Glycogenin is a synthesis, on which glucose residues are added. S
protein, which acts as primer in glycogen synthesis, not
WITH
in glycogen breakdown. 134. Ans. (c) 3
E
127. Ans. (b) Glucose-6-phosphate 135. Ans. (c) Glycogen X
[Ref: Harper 30th/e pg. 178] [Ref: Lehninger 7th/e pg. 604] P
Q134, 135 Normally in anaerobic glycolysis, energetics L
•• Both Insulin & glucose-6-P are activators of Glycogen A
is 4-2 = 2 ATPs i.e. 4 ATPs from substrate level
Synthase. But answer is glucose-6-P because insulin N
phosphorylation and two ATPs used by Hexokinase and
will do covalent modification (by dephosphorylation) A
PFK-I.
and activation. Allosteric Modulator/ Activator is T
•• But the end product of muscle glycogenolysis is
glucose-6-P. I
glucose-6-phosphate. This Glucose-6-Phosphate
•• Glucagon (option c) is inhibitor of this enzyme. O
directly enters glycolysis, without the need for
Fructose 1,6 bisphosphate (Option d) is intermediate N
Hexokinase step. So, now in anaerobic glycolysis; 4-1
in glycolysis. S
= 3 ATPs.
 136. Ans. (c) Muscle Glycogen 142. Ans. (b) More glucose-1-P than glucose

114 [Ref: Lehninger 7th/e pg. 604] [Ref: Harper 30th/e pg. 178]
Liver glycogen can maintain blood glucose levels. •• In glycogenolysis, 90% glucose is in the form of Glu-
But muscle glycogen cannot provide glucose to 1-P, released by Glycogen Phosphorylase and 10%
blood, because of deficiency of enzyme glucose- glucose is in the form of free glucose, released by
CRO BIOCHEMISTRY

6-phosphatase. Alanine is most glucogenic amino debranching enzyme.

acid. Gluconeogenesis synthesize glucose from non-
143. Ans. (d) UTP
carbohydrate sources.
[Ref: Harper 30th/e pg. 178]
137. Ans. (a) Liver
•• Glycogenesis requires UTP, making UDP-glucose
[Ref: Lehninger 7th/e pg. 604] which is the activated form of glucose required for
In liver, glycogen is stored. The function of this liver Glycogen synthesis.
glycogen is to maintain blood glucose levels.
144. Ans. (a) HMP


138. Ans. (b) Glucose-1-P [Ref: Harper 30th/e pg. 178]

[Ref: Harper 30th/e pg. 178] •• UDP-glucose is used in many pathways → Glycogen
•• In glycogenolysis, 90% of glucose is released by synthesis, Galactose metabolism and Uronic acid
Glycogen Phosphorylase in the form of glucose- pathway.
1-P. Glycogen Phosphorylase breaks alpha (1 → 4)
145. Ans. (c) Amylo- (1 → 4), (1 → 6) Transglycosylase
bonds. 10% glucose is released as free glucose by
Debranching enzyme, which breaks alpha (1 → 6) [Ref: Harper 30th/e pg. 176]
bonds. So major end product is Glucose-1-P. •• Branches are made by Glycogen branching enzyme
(also known as amylo-a(1:4) → a(1:6) trans
139. Ans. (c) Requires a bifunctional enzyme
glycosylase), which transfers the end of the chain
(debranching and transferase)
onto an earlier part via a-1:6 glycosidic bond,
[Ref: Harper 30th/e pg. 178] forming branches, which further grow by addition of
Debranching and Glucan Transferase are bifunctional more a-1:4 glycosidic units.
enzymes i.e. two enzymatic activities present on a single
protein. 146. Ans. (b) 18
•• Glycogenolysis involves enzymes cleaving alpha [Ref: Harper 30th/e pg. 149]
(1-4) glycosidic linkage (not beta).
•• Glycogen reserves can be used for 12-18 hours.
•• Glycogenolysis requires activation of Glycogen
A
phosphorylase (not Glycogen synthase). 147. Ans. (c) Epinephrine
N •• It requires activation of Phosphorylase Kinase which
S phosphorylates Glycogen Phosphorylase and makes [Ref: Harper 30th/e pg. 178]
W it active. •• Glycogenolysis is activated by glucagon and epine-
E
140. Ans. (d) Glucose residues in a-1,6 glycosidic linkages phrine. Glucagon receptors are not found in muscles.
R
S During the breakdown of glycogen, free glucose is 148. Ans. (d) Limit Dextrins, Maltose, and Maltotriose
formed from glucose residues in a-1,6 glycosidic
WITH
linkages (by debranching enzyme) and glucose-1- [Ref: Harper 30th/e pg. 178]
E phosphate is formed from glucose residues in a-1,4 •• The hydrolysis of starch is catalyzed by salivary
X glycosidic linkages, by enzyme Glycogen Phosphorylase. and pancreatic amylases, which catalyze random
P hydrolysis of alpha (1–4) glycosidic bonds, yielding
L 141. Ans. (c) It is not a major pathway in brain limit dextrins, and further hydrolysis yields a
A [Ref: Harper 30th/e pg. 178] mixture of glucose, Maltose, Isomaltose (from the
N branch points in Amylopectin) and Maltotriose
•• Glycogen metabolism occurs mainly in liver and
A (a trisaccharide made up of 3 glucose units).
kidney and to a lesser extent in brain. Phosphorylase
T •• Sucrose, galactose and fructose are not the compo-
cleaves glucose residues from the non reducing end
I nents of starch.
O of glycogen.
N
S
149. Ans. (b) Glucose-6-Phosphatase is present in endo- •• Normally lactate levels are increased in blood after
plasmic reticulum while Glycogen is in the cytoplasm exercise.
•• Glucose-6-phosphate in cytoplasm of Liver cells is •• But lactate normal after exercise is a characteristic 115
not acted upon by glucose-6-phosphatase enzyme as feature of McArdles’s disease. Because the first
it is present in ER. enzyme to breakdown glycogen in muscles is absent
i.e. muscle glycogen phosphorylase. So, glycogen

CHAPTER 3  CARBOHYDRATE METABOLISM

150. Ans. (b) Phosphoglucomutase cannot be broken down to give glucose, which gets
•• Enzyme common between glycogenesis and glyco- converted to lactate during exercise.
genolysis is Phosphoglucomutase. Whereas enzyme
156. Ans. (a) Von gierke’s disease
in glycolysis is phosphoglyceromutase.
[Ref: Harper 30th/e pg. 183]
151. Ans. (c) More commonly seen in fasting state than in
fed state •• If beta oxidation of FA cannot occur, then ketone
bodies cannot be formed. Carnitine deficiency and
•• The enzyme in this question is rate-limiting enzyme of
MCAD are defects in beta oxidation, So, they both
glycogenolysis, which is active in the phosphorylated
lead to non-ketotic hypoglycemia. Von gierke’s
state. Phosphorylation always activates this enzyme.
disease leads to ketotic hypoglycemia. In insulinoma,
Catecholamines stimulate this enzyme through
non ketotic hypoglycemia occurs.
cAMP, not directly. Glycogenolysis is occurring
in fasting state, not in fed state. So, this enzyme is 157. Ans. (a) Type I
also active during the fasting state. cAMP activates
this enzyme but Calcium and Calmodulin directly [Ref: Harper 30th/e pg. 183]
activates this enzyme without phosphorylation and •• In Von Gierke’s disease, - hyperuricemia, (i.e.
without cAMP. increased uric acid level) occurs both due to increased
production and decreased excretion of uric acid.
152. Ans. (a) Von Gierke’s disease
158. Ans. (c) Liver Phosphorylase
[Ref: Harper 30th/e pg. 183]
•• In Von Gierke’s disease (most common glycogen [Ref: Harper 30th/e pg. 179]
storage disease), patient has severe hypoglycemia, Her’s disease is Type VI glycogen storage disease in
ketosis, enlarged liver and kidneys, hypertri- which enzyme deficient is liver glycogen phosphorylase.
glyceridemia, lactic acidosis and hyperuricemia.
Spleen is not enlarged in this case. 159. Ans. (b) Pompe’s disease

153. Ans. (d) Phosphorylase enzyme [Ref: Harper 30th/e pg. 179 table, 18-2]
Pompe’s disease is Type II glycogen storage disease,
[Ref: Harper’s illustrated biochemistry, 30th ed., Pg 178] A
which is due to deficiency of Acid Maltase enzyme. This
•• This is McArdle’s disease and the enzyme deficient is enzyme is present in lysosomes, catalyzing the minor N
muscle phosphorylase. If GSD affects muscles, then pathway of glycogen breakdown. Only this pathway S
patient presents with muscle cramps and exercise occurs in lysosomes. All other pathways of glycogen W
intolerance because function of muscle glycogen is metabolism occurs in cytoplasm. So, Pompe’s disease is E
muscle contraction. the only GSD which is a lysomal storage disease. R
S
154. Ans. (b) Branching enzyme 160. Ans. (a) No Gluconoegenesis in type I disease
WITH
[Ref: Harper 30th/e pg. 176] [Ref: Harper 30th/e pg. 179 table, 18-2]
•• The enzyme deficient is branching enzyme. Type I glycogen storage disease is known as Von Gierke’s E
•• Branching enzyme deficiency, also known as Amylo- disease. Enzyme deficient is glucose-6-Phosphatase. X
pectinosis or Anderson’s disease. This enzyme is common for two pathways – P
glycogenolysis and gluconoegenesis. Therefore in Type L
155. Ans. (a) Mc Ardle's disease I disease, both glycogenolysis and gluconoegenesis A
cannot occur leading to very severe hypoglycemia. N
[Ref: Harper 30th/e pg. 176] A
In Type VI, enzyme deficient is liver Glycogen
•• Blood glucose is normal so, liver is not affected in this Phosphorylase. So, in this disease only Glycogenolysis T
glycogen storage disease. If liver glycogen storage cannot occur but gluconoegenesis can occur which will I
disease, then hypoglycemia occurs. Glycogen storage give some glucose in blood. O
disease type I & VI affects liver. N
S
 161. Ans. (d) Debranching enzyme •• NADP gets converted to NADPH. NAD is an active
form of vitamin B3 i.e. Niacin
116 [Ref: Harper 30th/e pg. 179 table, 18-2]
•• If FAD/FADH2 is used then vitamin used is B2 i.e.
•• Debranching enzyme deficiency, also known as Riboflavin
Limit Dextrinosis. Normally this enzyme releases •• Vitamin B1/Thiamine is used in oxidative decar-
just 10% glucose. So, patient has only early morning boxylation and Transketolase in HMP.
CRO BIOCHEMISTRY

hypoglycemia in its deficiency. (No hypoglycemia •• Biotin/ B7 is used in carboxylation reactions.

during day time, when gap b/w two meals is just 6-8
hours). 167. Ans. (d) All
•• Limit Dextrins are accumulated in this patient’s liver
[Ref: Harper 30th/e pg. 197]
during fasting. So, if glucagon given during fasting,
Phosphorylase will be activated which cannot break NADPH is mainly derived from HMP but it has two other
limit dextrins. So, this will not raise blood glucose. minor sources also; i.e. Malic enzyme and cytoplasmic
•• In fed state glycogen is formed. So, if glucagon given Isocitrate Dehydrogenase.
during fed state, Phosphorylase will be activated
which can break glycogen and can raise blood 168. Ans. (b) HMP


glucose. [Ref: Harper 30th/e pg. 196]

HMP is not involved in production or generation of ATP.
162. Ans. (d) OAA is required for Gluconoegenesis
[Ref: Harper 30th/e pg. 179 table, 18-2] 169. Ans. (b) Decreased RBC transketolase activity
•• Type I glycogen storage disease is known as Von [Ref: Harper 30th/e pg. 174-176]
Gierke’s disease. Enzyme deficient is glucose-6- Marker for vitamin B1 (thiamine) deficiency is dec-
Phosphatase. This enzyme is common for two reased activity of transketolase in RBCs.
pathways – Glycogneolysis and Gluconeogenesis.
Therefore in type I disease, both Glycogneolysis and 170. Ans. (c) HMP
Gluconeogenesis cannot occur leading to very severe [Ref: Harper 30th/e pg. 174-176]
hypoglycemia and ketosis occurs because of severe
HMP is the only source for the synthesis of Ribose-5-P,
hypoglycemia.
which is required for the synthesis of nucleic acids.
•• In this Q, both (c) and (d) options are wrong. Question
is asking the wrong statement. Better answer is d. 171. Ans. (b) Non lactating mammary glands
163. Ans. (d); (e) [Ref: Harper 30th/e pg. 174-176]
[Ref: Harper 30th/e pg. 179] •• Skin and non lactating mammary glands are never
the site of HMP. Also, HMP activity is low in skeletal
Pompe’s disease and Mc Ardles disease are glycogen muscles.
A storage diseases. Niemann-Pick disease, Gaucher •• HMP shunt occurs in liver, adipose tissue, lactating
N disease and Tay-Sachs disease are sphingolipidoses or mammary glands, adrenal cortex, gonads and RBCs.
S lipid storage diseases.
W 172. Ans. (c) Tripeptide
164. Ans. (b) Glycerol-3- P
E [Ref: Harper 30th/e pg. 39]
R [Ref: Harper 30th/e pg. 196] Glutathione is a tripeptide made up of three amino
S Products of HMP are Glyceraldehyde-3-P, 6 NADPH, 3 acids – Glutamate, Cysteine and Glycine. It is a reducing
CO2 and two molecules of glucose-6-phosphate. agent. Reducing property is due to –SH (sulfhydryl)
WITH group in cysteine.
165. Ans. (c) Ribose-5-P
E
[Ref: Harper 30th/e pg. 196] 173. Ans. (b) Hexose Monophosphate Pathway (HMP
X
shunt)
P HMP synthesizes NADPH and Ribose-5-phosphate.
L The only source for Ribose-5-phosphate is HMP but [Ref: Harper 30th/e pg. 197]
A NADPH has two other sources also; i.e. Malic enzyme G6PD is glucose-6-phosphate dehydrogenase is rate
N and cytoplasmic Isocitrate Dehydrogenase. limiting step in HMP. This enzyme converts NADP to
A NADPH.
T 166. Ans. (c) Niacin
I 174. Ans. (b) NADP+
O [Ref: Harper 30th/e pg. 196]
[Ref: Harper 30th/e pg. 198]
N •• Glucose-6-phosphate Dehydrogenase is a rate
Dehydrogenases of HMP shunt uses NADP and
S limiting enzyme of HMP pathway and forms NADPH.
produces NADPH.
175. Ans. (b) G6PD
[Ref: Harper 30th/e pg. 234] 117
G6PD is glucose-6-phosphate dehydrogenase is rate
limiting step in HMP. This enzyme converts NADP to
NADPH.

CHAPTER 3  CARBOHYDRATE METABOLISM

176. Ans. (b) Steroids
[Ref: Harper 30th/e pg. 503]
NADPH is formed mainly by HMP in cytoplasm, where
it takes part in biosynthesis reactions like synthesis of
steroids, fatty acids, cholesterol and destruction of H2O2
in RBCs.

177. Ans. (a) Vitamin C

[Ref: Harper 30th/e pg. 202, 203]
•• Humans cannot synthesize vitamin C due to
deficiency of enzyme, L-Gulonolactone Oxidase.
178. Ans. (b) Oxidation of terminal alcohol
[Ref: Harper 30th/e pg. 201]
Oxidation at C-6 gives uronic acid and oxidation at C-1
gives aldonic acid. Oxidation at both C1 and C6 gives
saccharic acid.
183. Ans. (a) Galactosemia
179. Ans. (c) L-Gulonolactone oxidase
[Ref: Harper’s illustrated biochemistry, 30th ed., Pg. 200] [Ref: Harper 30th/e pg. 203]
L-Gulonolactone oxidase produces vitamin C via uronic •• The clinical manifestations are typical of classical
acid pathway, but only in plants and most animals. galactosemia. Bilateral cataract rules out the
Humans cannot make vitamin C due to absence of this possibility of Von Gierke's disease and hereditary
enzyme. fructose intolerance although other symptoms are
180. Ans. (c) Xylulose Reductase there in both these diseases.
•• In juvenile diabetes mellitus, jaundice and hepato-
[Ref: Harper 30th/e pg. 202] megaly are not observed.
Essential pentosuria occurs due to deficiency of enzyme
Xylulose Reductase. 184. Ans. (a) Galactose-1-phosphate Uridyl Transferase/ A
GALT N
181. Ans. (d) Produces Ribose 5-phosphate in oxidative
phase of pathway S
[Ref: Harper 30th/e pg. 203]
W
[Ref: Harper 30th/e pg. 174] •• In classical type galactosemia, enzyme deficient is E
HMP produces Ribose–5–Phosphate in non-oxidative Galactose 1- phosphate Uridyl Transferase (GALT).
R
phase (not oxidative) other options given in question •• Deficiency of GALT: This is known as classical type
are true. Galactosemia. Excess Galactose-1-P accumulates S
182. Ans. (a); (b); (c); (d); (e) in Liver and brain leading to jaundice and mental WITH
retardation. Excess Galactose-1-P also inhibits
UDP-glucose used is a key intermediate in carbohydrate
enzyme Galactokinase by product inhibition. If this E
metabolism: Galactose metabolism, Disaccharides,
patient still continue to take Galactose in diet then X
Glycogen metabolism, GAGs synthesis, Glycolipids/
Glycosphingolipids formation, Bilirubin metabolism. excess Galactose is converted to Galactitol leading to P
oil drop cataract. L
A
N
A
T
I
O
N
S
 185. Ans. (b) Disease manifest only at adolescence diet), known as post-prandial hypoglycemia i.e.
hypoglycemia after ingestion of fruit juices or fruits.
118 [Ref: Harper 30th/e pg. 203]
This occurs because fructose-1-P inhibits Glycogen
Galactosemias manifest within one week or one month
phosphorylase.
of life as the diet given to newborn is milk, which
•• This is also known as fructose induced Hypoglycemia
contains galactose.
despite the presence of high Glycogen reserves.
CRO BIOCHEMISTRY

186. Ans. (a) Galactosemia

193. Ans. (a) Aldolase B
[Ref: Harper 30th/e pg. 203]
[Ref: Harper 30th/e pg. 202]
In galactosemia, galactose is excreted in urine, which is
Aldolase–B is involved in fructose metabolism.
a monosaccharide. All monosaccharides are reducing.
Aldolase–A is involved in glycolysis. Deficiency of
187. Ans. (a) Aldose Reductase Aldolase–B is hereditary fructose intolerance.

[Ref: Harper 30th/e pg. 203] 194. Ans. (c) Sucrose

Aldose reductase enzyme reduces galactose to [Ref: Harper 30th/e pg. 202]


galactitol/ dulcitol, which is responsible for oil drop

cataract. •• Sucrose can be digested but not metabolized as
enzyme sucrose is only present in intestine.
188. Ans. (c) Glucokinase •• Rest all – glucose, fructose and Sorbitol can be meta-
bolized in the body.
[Ref: Harper 30th/e pg. 203]
This is a case of galactosemia in which liver is affected 195. Ans. (b) Glyceraldehyde-3-P Dehydrogenase
and oil drop cataract occurs. Glucokinase is not the •• Enzymes involved in phase II of glycolysis are also
enzyme of galactose metabolism. Rest all are enzymes used in fructose metabolism.
of galactose metabolism, which can be deficient in
galactosemias. 196. Ans. (c) Fructokinase
[Ref: Harper 30th/e pg. 202]
189. Ans. (c) Galactosemia
Deficiency of fructokinase is a rare & benign condition
[Ref: Harper 30th/e pg. 203] known as essential fructosuria. It is asymptomatic and
•• Galactosemia patients have increased risk for E.coli is detected accidentally on urinalysis. Benedicts test is
neonatal sepsis. also positive in this case.

190. Ans. (b) Galactokinase 197. Ans. (c) Galactosemia

[Ref: Harper 30th/e pg. 203] [Ref: Harper 30th/e pg. 202]
A •• Galactokinase is enzyme of galactose metabolism, Benedict’s test positive means reducing sugar is
N which can be deficient in Galactosemia. present in urine. Both fructose and galactose are
S
reducing. But if blood glucose is normal, then it cannot
W 191. Ans. (a) Hexokinase can phosphorylate fructose
be fructosemia because fructose accumulation causes
E [Ref: Harper 30th/e pg. 202] hypoglycemia as fructose-1-P inhibits Glycogen
R
•• Fructokinase is the first enzyme for fructose phosphorylase. In galactosemia, blood glucose is
S metabolism. It converts fructose to fructose-1-P. In normal. In both fructosemia and galactosemia liveris
fructokinase deficiency, hexokinase (a non specific affected but hypoglycaemia occurs only in fructosemia.
WITH
enzyme) can phosphorylate fructose as well as other
E sugars but it has high km (low affinity) for fructose. 198. Ans. (a) Aldose Reductase
X Glucose is the true substrate for this enzyme. Fructose [Ref: Harper 30th/e pg. 202]
P 6-phosphate-the end product of hexokinase reaction Glucose is reduced by Aldose Reductase enzyme to
L can enter glycolytic pathway to be utilized further. So, produce sorbitol/ polyol, which is responsible for snow
A it does not accumulate to produce toxic effects. flake cataract in a diabetic patient.
N
A 192. Ans. (a) Fructose 199. Ans. (b) Cataract
T [Ref: Harper 30th/e pg. 202] [Ref: Harper 30th/e pg. 202]
I •• Hereditary fructose Intolerance (HFI) - Deficiency
O Glucose is reduced by Aldose Reductase enzyme to
of Aldolase B – is asymptomatic but fructose produces sorbitol/ polyol, which is responsible for snow
N ingestion causes symptoms (typically child presents
S flake cataract in a diabetic patient. Drugs which are
after weaning when fruit juices are started in the inhibitors of Aldose Reductase can prevent this cataract.
UNIT
III

ENZYMES, AMINO
ACIDS AND PROTEINS
Unit Outline
Chapter 4 E nzymes
Chapter 5 C  hemistry and Metabolism of Amino
Acids
Chapter 6 Urea Cycle
Chapter 7 Proteins
Enzymes 4
Overview of Chapter Active Site (AS)
•• General Enzymology zz Active site = Binding site + Catalytic site

•• Isoenzymes
•• Serine Proteases
•• Classification and Nomenclature of Enzymes
•• Enzyme Kinetics
•• Enzyme Inhibitors
•• Enzyme uses
•• Regulation of Enzyme Activity

GENERAL ENZYMOLOGY
Enzymes make life on earth possible. All enzymes are proteins Active site can be visualized as being similar to the mouth of
except Ribozyme . Pac-Man doing cutting work.
Definition: Enzymes are high molecular weight, highly
efficient and specialized proteins, which catalyze the bio-
chemical reactions. They catalyze the conversion of one or
more substrates into one or more products. They increase the
rate of reaction, without being changed in the overall process.

Properties of Enzymes
zz Highly efficient catalyst (increase the rate of reaction by a
factor of 106 or even more)
zz Group or substrate specificity (Enzymes are specific not
only for the type of reaction, but also for substrate or a
small set of closely related substrates)
zz Stereospecificity—They are specific for only one stereoi-
somer of a given compound e.g. only D-Glucose and not
L-Glucose
zz They are not used up in the reaction
zz Thermolabile (affected by temperature changes)
122
CRO BIOCHEMISTRY


Fig. 4.1: Formation of ES complex and release of Product. S is complementary to the active site. So ES complex is formed. Then reaction
occurs and EP complex is formed and then P is released. (Note: Enzyme is unchanged during the conversion of substrate to the product)
Part of enzyme that binds to Substrate. It is a 3D space complementary to the Substrate (Fig. 4.1)

Fig. 4.2: Amino acids in Active Site

How Enzymes Work?
For any thermodynamically favorable reaction, the free energy of the product is lower than that of substrate.

T
H
E
O
R
Y
Fig. 4.3: Gibb’s Free Energy graph for enzyme catalyzed and enzyme uncatalyzed reaction. On X-axis, reaction progress taken (i.e. S getting
converted to P). For any spontaneous reaction, S has more free energy as compared to P. For S to be converted to P, first S has to acquire
Activation energy (ΔG uncatalyzed) and reach high energy state (Transition state). Only then it can be converted to P
zz Any spontaneous reaction can occur like this even in the
absence of enzyme. But time taken is in years (very slow)
zz In the presence of enzyme, the activation energy required 123
is less (ΔG catalyzed). So, enzymes increase the rate of
reaction.

F undamental B ox

CHAPTER 4  ENZYMES
Amount of ∆G indicates nothing about rate of reaction. It tells
that the reaction is thermodynamically favorable or not.
•• Negative ∆G (∆G < 0) Thermodynamically stable reaction
•• Positive ∆G (∆G > 0) Thermodynamically unfavorable
reaction (i.e. energy required)
Fig. 4.5: Induced Fit Theory
•• ∆G = 0 Reaction is at equilibrium
(i.e. freely reversible)
Cofactor and Prosthetic Group
All enzymes are proteins. But most of the enzymes require a
non-protein portion to function. In such cases, the protein
portion is known as Apoenzyme. The complete enzyme here
is known as Holoenzyme.

Fig. 4.4: Suppose in this figure there is a mountain. A person

has to go to other side of the mountain. This takes lot of time as
the person has to go to the peak first. But the time duration will
be decreased if a tunnel is created in the mountain. Enzymes
decrease the time required by the reaction by doing this kind of
action (decrease activation energy).
Enzyme is doing Enzyme is not doing
Decreases Activation Do not affect the position and
energy direction of equilibrium of the reaction
Decrease time required Not changing free energy of reactants
by the reaction or products
Increases rate/velocity Not used up in the reaction
of reaction

Multiple Mechanisms for Catalysis

zz Lock and Key Hypothesis was the first model which
proposed that the active site has a rigid/fixed 3-D
structure that is complementary to the substrate to zz One third of all human enzymes are Metallo Enzymes
facilitate complementary binding, but this theory is now zz Apoenzyme alone is inactive
discarded.
zz Coenzyme may be covalently or non-covalently linked
zz Induced Fit Theory: It postulates that the catalytic site
zz Coenzyme Q and Glutathione also acts as Coenzyme T
of enzymes is not rigid. It is flexible. Initially active site
zz Some Metallo enzymes contains more than one metal H
is not complementary to substrate. But when substrate
approaches the active site, then active site is induced
e.g. Cytochrome Oxidase contains both iron and copper E
to fit the substrate. Substrate’s presence induces zz Role of coenzyme → acts as acceptor for intermediate O
conformational changes in the active site (Fig. 4.5). products of substrate. e.g. R
Y
 1. Act as hydrogen carrier LDH—Lactate Dehydrogenase Isoenzymes
2. Acts as carrier of moieties other than Hydrogen
124 A tetramer, made up of 2 types of subunits – H and M
Protein portion Non-protein portion zz Subunit H – Heart
High Molecular Weight Low Molecular Weight zz Subunit M – Muscle
Thermolabile Relatively Thermostable
CRO BIOCHEMISTRY

Not consumed in Not consumed in Reaction. But are LDH—Lactate Dehydrogenase Isoenzymes
Reaction chemically changed during the reaction
Table 4.1: Isoenzymes of LDH

Isoenzyme Subunit Location

LDH-1 HHHH Heart
LDH-2 HHHM Blood (WBC) → Functional Plasma
Enzyme
LDH-3 HHMM Brain, Kidney, Lungs and Pancreas


LDH-4 HMMM Muscle and Liver

LDH-5 MMMM Muscle and Liver

zz Normal person LDH-2 > LDH-1 (in blood)

zz Myocardial infarction patient LDH-1 > LDH-2 (in
blood)

H igh R eturn This is known as Flipped ratio of LDH in MI (Myocardial

infarction)
Cu – for all Oxidases
� Cytochrome c Oxidase � Tyrosinase
zz Ascorbic acid Oxidase � Amino acid Oxidase
zz Lysyl Oxidase
zz Cytoplasmic SOD (Super Oxide Dismutase)

2 Oxidases do not require Cu

1. Xanthine Oxidase
2. Sulfite Oxidase
They require Molybdenum

zz All Kinases need Mg2+ (But Pyruvate Kinase need K+>>

Mg2+)
zz All Carboxylases and synthetases also need Mg2+
zz Cytoplasmic SOD requires copper but mitochondrial Fig. 4.6: Electrophoretic Pattern of LDH
SOD requires Manganese (Mn)
CK (Creatine Kinase) Isoenzymes
ISOENZYMES
Made up of two types of subunits B and M
Definition: Physically distinct form of enzyme which catalyse
zz Subunit B–Brain
same biochemical reaction
zz Subunit M–Muscle
Table 4.2: CK (Creatine Kinase): 3 Isoenzymes

Isoenzyme Subunit Location

CK-1 BB Brain
T
H CK-2 MB Heart
E CK-3 MM Skeletal muscle
O
R
Y
 Hexokinase Glucokinase
• Phosphorylates all hexoses • Phospharylates only Glucose 125
• Present in all cells • Present in liver and pancreas
(GLUT-2)
• High affinity, less [S] • Low affinity, more [S]

CHAPTER 4  ENZYMES
required, active in fasting required, active in fed state,
state, low km, low Vmax high km, high Vmax
• Feedback inhibition by • Induced by Insulin
Glucose-6-Phosphate

SERINE PROTEASES
Fig. 4.7: Electrophoretic Pattern of CK
zz Three amino acids present at AS of all serine proteases.
Table 4.3: Enzymes and Proteins raised in Myocardial This is known as catalytic triad
Infarction
Catalytic Triad
Enzymes Raised in MI
Serine proteases are proteolytic enzymes with serine at
CK-2 / CK-MB 4–6 hours the catalytic site, i.e. responsible for catalysis, so known as
AST/SGOT 6–8 hours serine proteases. This is a big class of enzymes which have
LDH-1 8–10 hours serine present at the catalytic site, helping in catalysis. All
serine proteases have same 3 amino acids at the active site
Proteins Raised in MI
(Histidine and Aspartate at the binding sites and Serine at the
Myoglobin 2–6 hours catalytic site). This is known as Catalytic Triad.
Trop T and Trop I 3–6 hours
Proteases–Examples of Serine
Troponin type Property zz Chymotrypsin, Trypsin, Elastase
• Troponin C Calcium binding zz Thrombin, Plasmin
zz Complements
• Troponin I Inhibits actomyosin ATPase
zz Clotting factors X and XI
• Troponin T Tropomyosin binding element zz PSA (Prostate Specific Antigen)
Trop T In cardiac and skeletal muscle
Trop I Only cardiac muscle Serine Proteases has a role in tumor cell metastasis
• Cardiac troponin is an early, accurate and stable biomarker
Serine Proteases Differ in Substrate Specificity
zz Earliest marker for MI → Myoglobin but it is non specific
zz Last marker to rise in MI → LDH zz Chymotrypsin: Cleaves at carboxy terminal of bulky
hydrophobic amino acids
zz Most specific marker for MI → Troponin I >Troponin T
zz Trypsin: Cleaves at carboxy terminal of basic amino
zz First marker to fall in MI → Myoglobin
acids
zz Second marker to fall in MI → CK-MB
zz Elastase: Cleaves at carboxy terminal of small neutral
zz Last marker to fall in MI → LDH
amino acids e.g. Glycine, Alanine
A New Biomarker for Cardiac Ischemia → IMA
Bi-Bi reaction
IMA- Ischemia Modified Albumin→ very useful for Acute
Myocardial Ischemia zz Two substrate, two product reaction, e.g. when one
Albumin in blood gets modified on Cardiac Ischemia. substrate is oxidized, other is reduced. So, there are two
IMA offers an early test (along with ECG changes and cardiac substrates and two products in such a reaction.
Troponin) for the exclusion of acute coronary syndrome as
IMA gives a good discrimination between ischemic and non-
ischemic patients. T
H
Hexokinases – Isoenzymes are of 4 types → • E
Type I, Type II, Type III, Type IV O
R
zz Hexokinase II → most abundant, overexpressed in Y
cancerous cells
zz Hexokinase IV → known as Glucokinase
 Each enzyme is given a systematic name and a unique 4-digit
identification number for identification by the Enzyme
126 Commission (EC) of IUBMB (since 1964).
CRO BIOCHEMISTRY

Fig. 4.8: Types of Bi-Bi Reaction

Site Specific/Directed Mutagenesis

In the figure 4.9 (A), Chymotrypsin gene is shown, which As per IUBMB (International Union of Biochemistry and
forms its mRNA (having a codon for serine) and then it forms Molecular Biology), enzymes are divided into six major
the protein/ enzyme having serine at the catalytic site. classes with several sub classes:


To increase the rate of enzyme chymotrypsin, a mutation

is created in chymotrypsin gene at a very specific site of serine
i.e. U → C (Fig. 4.9 B). With this mutation, serine is changed
to proline. This technique is known as Site specific/Directed
mutagenesis. (Mostly in such experiments, enzymatic
activity cannot be increased as nature has already provided
us with the best thing).

Fig. 4.9: Site Specific/Directed Mutagenesis

H igh R eturn
zz Oligonucleotide with single base change is used in – Site
directed mutagenesis (not RFLP)
zz Single mutation detected by – RFLP
zz Multiple mutations detected by – Micro Array

CLASSIFICATION AND NOMENCLATURE OF

ENZYMES
T
H F undamental B ox
E zz Mostly we add suffix ‘ase’ to the name of the substrate e.g.
O Sucrase, Urease
R zz Sometimes description of action performed e.g. Lactate
Y Dehydrogenase
zz Systematic name: It’s a unique 4 digit identification number
given to each enzyme
 Table 4.4: Enzyme Commission Numbers
Enzyme Commission Number Name of Enzyme 127
EC No. 1 Oxidoreductase
EC No. 2 Transferase
EC No. 3 Hydrolase

CHAPTER 4  ENZYMES
EC No. 4 Lyase
EC No. 5 Isomerase
EC No. 6 Ligase

Table 4.5: Classification of Enzymes

Classification Distinguishing feature

1. Oxidoreductases Transfer of electron (Hydride ions or H atom)
ƒƒ Oxidases • Use Oxygen as an electron acceptor e.g. Cytochrome c Oxidase, Tyrosinase
ƒƒ Dehydrogenases • Use molecules other than oxygen (NAD, FAD, NADP) as an electron acceptor
ƒƒ Peroxidases • Use H2O2 as an electron acceptor e.g. Glutathione Peroxidase, Catalase
ƒƒ Oxygenases • Directly incorporate O2 into the substrate
ƒƒ Reductases • Cause reduction (add Hydrogen or removes Oxygen)
2. Transferases Carry group transfer reactions
ƒƒ Methyl Transferase • Transfer one carbon units between Substrates
ƒƒ Amino Transferase • Transfer NH2 from amino acids to keto acids
ƒƒ Kinase • Transfer phosphate from ATP to a substrate
ƒƒ Phosphorylase • Transfer phosphate from inorganic phosphate to a substrate
ƒƒ Glycogen Synthase • Transfer glucose from UDP-Glucose to glycogenin
3. Hydrolases Hydrolysis of substrate using water
ƒƒ All digestive enzymes • Enzymes which break down macro-molecules
ƒƒ Phosphatase • Remove phosphate from a substrate
4. Lyases Addition of groups to double bonds or formation of double bonds by removal of groups
ƒƒ Decarboxylases • Produce CO2 via elimination reactions
ƒƒ Aldolases • Produce aldehydes via elimination reactions
ƒƒ Synthase • Link two molecules without involvement of ATP
ƒƒ Hydratase/Hydrase • Removes water to make double bond or add water across a double bond, without breaking bond e.g
Enolase, Aconitase, Fumarase, PEPCK
ƒƒ Desulfhydrase • Remove H2S from substrate
5. Isomerases Transfer of group within same molecule to form isomers
ƒƒ Racemase • Interconvert L and D stereo isomers
ƒƒ Mutase • Transfer groups between atoms within a molecule
ƒƒ Epimerase • Interconvert Epimers e.g. Glucose and Galactose
6. Ligase Formation of C-C, C-N, C-O, C-S bonds by condensation reaction using ATP
ƒƒ Carboxylase • Use CO2 as a substrate
ƒƒ Synthetase • Link two molecules using ATP

Important Points No. is 5 (Isomerase) because Molecular formula is not

changed when Mutase work.
zz Xanthine Oxidase produce H2O2 but it is not a peroxidase zz All Synthases are Lyases but
zz Oxygenation means O2 is incorporated. In Hemoglobin, 1. Nitric Oxide synthase - is an oxidoreductase (Mono-
there is oxygenation. If oxidation of Hb occurs, then met oxygenase) {EC No- 1} T
Hb is formed. 2. Glycogen Synthase- is a Transferase {EC No.- 2} H
zz Whenever Transferases work, molecular formula is 3. Citrate Synthase is a Tranferase {EC No. -2} E
changed.
4. ATP Synthase – Hydrolase {EC No.- 3} O
zz Whenever Isomerases work, molecular formula is not
zz Phosphatase (for removal of Phosphate) - is a Hydrolase R
changed.
zz Oxygenases: Directly incorporate O2 into the substrate. Y
zz Mutase enzyme is involved in intra-molecular transfer
They are of two types:
of groups. But EC No. is not 2 (Transferase) instead EC
 1. Mono-oxygenases/Hydroxylases/Mixed Function Ox- 2. Dioxygenases- incorporate two atoms of molecular
idases: incorporate one atom of molecular oxygen. e.g. oxygen. e.g. Homogentisate Dioxygenase, Tryptophan
128 Phenyl alanine Hydroxylase, Cytochrome P450, Nitric Pyrrolase
Oxide Synthase
 Most Hydroxylases are monooxygenases but Prolyl
Hydroxylase and Lysyl Hydroxylase are Dioxygenase.
H igh R eturn
CRO BIOCHEMISTRY

 Monooxygenases are known as Mixed Function zz EC Number of Hydroxylase is 1

Oxidases because they incorporate one [O] into the zz EC Number of Hydrolase is 3
substrate and other into H2 to make H2O. zz EC Number of Hydratase/Hydrase is 4


ENZYME KINETICS AND ENZYME USES Effect of Enzyme Concentration

In Enzyme Kinetics, we study the effect of various variables
on the Rate (Velocity) of Reaction.

Fig. 4.10: Effect of Enzyme Concentration

Each time we make a graph, in which velocity is always taken This graph between velocity and enzyme Concentration is a
on y-axis and the particular variable is taken on x-axis. The straight line graph (linear graph). This indicates that velocity
various variables we study are : of reaction is directly proportional to the substrate.
zz Enzyme concentration
zz Substrate concentration
zz Temperature
zz pH
zz Presence or absence of inhibitors
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Effect of Substrate Concentration [S]
129

CHAPTER 4  ENZYMES
Fig. 4.11: Graph showing relationship between [S] and velocity
in the presence of constant enzyme. Initially velocity is directly
proportional to the substrate concentration (First order kinetics)
but later on (increasing more substrate) velocity is independent of
substrate concentration (Zero order kinetics). This graph is known
as Michaelis Menten graph. Fig. 4.12: Shape of graph for simple and allosteric enzymes

Table 4.6: Difference between Simple and Allosteric

F ormula B ox Enzymes
Michaelis-Menton Equation Simple enzymes Allosteric/Regulatory enymes
Vmax × S (most enzymes)
Vi =
Km + S • Most of the enzymes are • Few enzymes in our body are
Where Vi  Initial Velocity simple enzymes regulatory enzymes
Vmax  Maximal Velocity • Follow Michaelis • Do not follow Michaelis Menton
Menton kinetics Kinetics (follow Hill’s equation)
S  Substrate Concentration
Km  Michaelis Menten constant • Rectangular hyperbola • Sigmoidal/S-shape graph with
graph with [S] [S]

H igh R eturn
Km →Michaelis Menten constant.
zz Km is that [S] at which velocity of reaction is half of Vmax.
zz Km is characteristic of an enzyme and its particular substrate
and reflects the affinity of enzyme for that substrate.
zz Km is inversely proportional to the affinity between enzyme
and substrate Simple enzymes → have just one site, i.e. active site. If
substrate is present, then reaction will occur. If substrate is
 Small Km→ means a high affinity i.e. a low concentration
not present, then reaction will not occur.
of substrate is needed to attain half of Vmax.
 Large Km→ means low affinity i.e. high concentration of Regulatory/Allosteric enzymes→ have active site where
substrate is needed to attain half of Vmax. substrate binds. But they also have regulatory/allosteric site,
zz Km – is signature of the enzyme. Means Km is: where the regulator (activator/inhibitor) binds. Just presence
 Constant value for each enzyme of substrate does not causes the reaction to occur. Infact,
 Each enzyme has its own Km value. (Even Isoenzymes have activator should be present and inhibitor should be removed,
different Km values) only then reaction can occur.
zz Km is expressed in the same units as [S] i.e. mole/L
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 Table 4.7: Oxygen Dissociation Curves

130 Hemoglobin Myoglobin

• 4 chains • Only one chain
• Shows cooperativity • No cooperativity
CRO BIOCHEMISTRY

• Sigmoidal/S-shape • Rectangular hyperbola

Modification of Michaelis Menton Graph →

Lineweaver Burk Plot or Double Reciprocal Curve
A straight line graph is of much use as it is very easy to
calculate so many things from a straight line graph by using
mathematical formulas. So, rectangular hyperbola Michaelis
Menton graph is converted to a more useful straight line
graph by doing some modifications.


Plotting the reciprocals of Velocity and [S] yields a

“Double-Reciprocal” or Lineweaver-Burk plot. This provides
a more precise way to determine Vmax and Km.

Fig. 4.13: Allosteric regulation of Enzyme

H igh R eturn
zz Regulator (Activator/ Inhibitor) is always more important than
substrate
Allosteric enzymes are usually multi-subunit enzymes (so
have multiple active sites). They show a very important
property of cooperativity that when one substrate binds at
active site, this binding induces conformational changes in
the other active sites so that next substrates binds with a very Fig. 4.14: Lineweaver Burk Plot
high affinity. Its like family members, they cooperate with
each other. EFFECT OF TEMPERATURE AND pH
The graph between temperature and velocity is bell shaped
graph. See extremes of temperature, enzyme is not active
because enzymes are proteins, They get denatured at very
high or low temperature. For pH also, graph is bell shaped as
enzymes get denatured at extremes of pH.

The vertically straight line in the graph is because of

T cooperativity. Here binding of more substrates has increased
H the affinity of enzyme and substrate to a greater extent that
minor changes in substrate leads to an exponential increase
E
in the rate of reaction.
O Fig. 4.15: Effect of Temperature
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Table 4.8: Shapes of Enzyme Kinetics Graphs ENZYME INHIBITORS
Enzyme concentration [E] Linear/ Straight line Types 131
Substrate concentration [S] for Rectangular Hyperbola
simple or most enzymes 1. Competitive inhibitors
Substrate concentration [S] for Sigmoidal or S shape 2. Non-Competitive inhibitors
Allosteric or Regulatory enzymes 3. Uncompetitive inhibitors

CHAPTER 4  ENZYMES
Temperature Bell shaped 4. Suicidal inhibitors
pH Bell shaped 5. Feedback inhibitors
6. Allosteric inhibitors

Competitive Inhibitor
Inhibitor binds with free enzyme, reversibly, to form enzyme-inhibitor complex that is catalytically inactive and cannot bind
substrate.

Fig. 4.16: Lineweaver Burk Plot for Competitive and Non-competitive inhibitors
So a higher concentration of substrate is required in order to offset the binding of inhibitor with the enzyme. Thus Km is
increased. Examples- Table 4.10 (a)
T
Non-Competitive Inhibitor H
Inhibitor binds at regulatory site of enzyme and induces changes in the active site so that substrate cannot bind and thus E
decreases the velocity. But there is no change in Km or Substrate concentration. O
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 Table 4.9: Difference between Competitive and Non-Competitive Inhibitors

132 Competitive Inhibitors Non-Competitive Inhibitors

CRO BIOCHEMISTRY


Inhibitor and substrate resemble each other Does not resemble in structure
Inhibitor binds active site Inhibitor does not bind the active site
Vmax is same Vmax is lowered

Km increased Km unaltered
Inhibitor cannot bind with ES complex Inhibitor can bind with ES complex
Lowers the substrate affinity to enzyme Does not change substrate affinity for the enzyme
This inhibition is irreversible This inhibition is mostly irreversible
Table 4.10: Examples of (a) Competitive and (b) Non-Competitive Inhibitors

Competitive Inhibitors Non-Competitive Inhibitors

Inhibitor Enzyme inhibited Inhibitor Enzyme inhibited
Arsenate Glyceraldehyde3-P Dehydrogenase Iodoacetate Glyceraldehyde3-P DH
Oxamate Lactate dehydrogenase NaF Enolase
Malonate Succinate dehydrogenase Fluoroacetate Aconitase
Fluorocitrate Aconitase Arsenate Alpha-Ketoglutarate DH
Sulfonamide (Antibiotic) Dihydropteroate synthase (DHPS) (Inhibits Cyanide Complex IV (ETC)
synthesis of folic acid in bacteria)
Methotrexate Dihydrofolate reductase (DHFR) Heavy Metals & Dimercaprol Inhibits SH group
Dicumarol (Anticoagulant) Quinone reductase Di-iso-propyl fluorophosphate Serine protease

Uncompetitive Inhibitor Suicidal Inhibition (also Known As Mechanism

In this case, both Km and Vmax are decreased. This is a rare
Based Inhibition)
type of inhibition. A special class of irreversible inhibitors in which enzyme
E.g. Acetyl Choline inhibits placental ALP (Alkaline Phos- converts the inhibitor into a reactive form in its active site. e.g.
phatase) zz Allopurinol inhibits Xanthine Oxidase
T
Table 4.11: Km and Vmax for different type of Inhibitors zz Aspirin inhibits cycloxygenase
H
E Type of inhibitor Km Vmax Feedback inhibition/ End-Product inhibition
O
Competitive Increased Same
R Feedback inhibition controls the amount of product the cell
Y Non-Competitive Same Decreased needs.
Uncompetitive Decreased Decreased
 133

CHAPTER 4  ENZYMES
FUNCTIONAL AND NON-FUNCTIONAL PLASMA ENZYMES

Fig. 4.17: Functional and Non-Functional Plasma Enzymes

ENZYMES USES Lactate DH (LDH) Myocardial Infarction, Liver diseases,

Leukemia
Enzymes can be used in clinical diagnosis, as markers of
organelles, used in lab tests and they can also have therapeutic Transaminases Myocardial infarction (SGOT), Liver
diseases (SGPT)
roles

Enzymes Used In Clinical Diagnosis Enzymes used in Lab Tests

Table 4.12: Enzymes of Diagnostic Importance Table 4.13: Analyte estimated by different enzymes

Enzyme Diagnostic use Enzyme Analyte which is to be

estimated or analyzed T
Amylase and Lipase Pancreatitis
Uricase Uric acid H
Acid Phosphatase Prostate cancer E
Urease Urea
Alkaline Bone, Liver disease and Hyperthyroidism O
Phosphatase (ALP) GOD-POD and Hexokinase Glucose
R
Creatine Kinase (CK) Recent Myocardial Infarction Cholesterol Oxidase Cholesterol Y
Lipase Triglycerides (TGs)
Contd…
 Therapeutic Role REGULATION OF ENZYMATIC ACTIVITY
134 Table 4.14: Therapeutic Role of Enzymes Enzymes can be regulated in following five ways:
1. Allosteric Regulation
Enzymes Therapeutic use
2. Covalent Regulation
• Streptokinase/ Lysis of intravascular clots e.g. In
CRO BIOCHEMISTRY

3. Compartmentalization of cell
Urokinase myocardial infarction (Fig. 4.18)
4. Control of rate of synthesis/degradation
• Asparaginase/ ALL (Acute Lymphoblastic Leukemia) 5. Synthesis of inactive zymogens
Glutaminase (Fig. 4.19)
• Pepsin Chronic indigestion and pancreatic Allosteric Regulation
insufficiency
• Alpha-1-antitrypsin Emphysema (caused by deficiency Key regulatory reactions or committed steps of metabolic
of alpha-1-antitrypsin), pathways are often subjected to allosteric regulation.
Hyaluronidase traumatic or post- Allosteric regulators/effectors binds to a site other than active
operative edema site (i.e. allosteric/regulatory site) e.g. Rate limiting enzyme


• Uricase Gout of glycolysis is PFK-1 (Phosphofructokinase-I) is allosterically

inhibited by ATP, Citrate. It is allosterically activated by
• Lactase Lactose intolerance
Fructose 2,6 Bisphosphate (Fig. 4.20)
• Lactase Penicillin Allergy
• Trypsin and Inflammation
Chymotrypsin
• Collagenase Skin ulcers

Fig. 4.18: Streptokinase and Urokinase used for lysis of

intravascular clots

Fig. 4.20: Effect of an Allosteric activator/inhibitor on Enzyme

activity

Covalent Regulation
zz Most common covalent modification is phosphorylation
and dephosphorylation.
zz Phosphate group is added most commonly at the OH
group of Serine (less common at Threonine)
zz Other covalent modifications → Adenylation, Uridy-
Fig. 4.19: ALL (Acute Lymphoblastic Leukemia) cell has high lation, ADP Ribosylation, Methylation, Acetylation
demand of asparagine and glutamine amino acids, which has to (Fig. 4.21)
be taken from blood. So for the treatment of ALL, the enzymes
Asparaginase and Glutaminase are given, which can destroy the
T asparagine and glutamine present in blood depriving cancer cells
H of their important nutrient. So ALL cell cannot survive.
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 135

CHAPTER 4  ENZYMES
Fig. 4.21: Enzyme Regulation by Covalent Modification

Enzymes Active in Phosphorylated State

zz Glycogen Phosphorylase
zz Phosphorylase Kinase
zz Key enzymes of Gluconeogenesis
zz HMG CoA Reductase Kinase
zz Citrate Lyase

Enzymes Active in Dephosphorylated State

zz Glycogen Synthase
zz Acetyl CoA Carboxylase
T
zz Pyruvate Dehydrogenase
H
zz HMG CoA Reductase
E
zz PFK-II (Phosphofructokinase-II)
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 Compartmentalization of the Cells 2. Inducible Genes→ Genes that can be activated
whenever required e.g. enzymes of Gluconeogenesis
136 Compartments of cells are specified for various processes.
This gives a regulation to control the enzymatic activity. Degradation of Enzymes/Proteins
For protein degradation, there are two enzyme systems:
1. ATP dependent - Ubiquitin Proteasome Pathway (UPP):
CRO BIOCHEMISTRY

zz Highly regulated
zz Uses ATP
zz Occurs in cytoplasm and nucleus
zz For damaged or short lived or misfolded proteins
zz Ubiquitin protein is highly conserved, small, glob-
ular, non enzymic protein and is recycled
zz ε – amino of Lysine in protein is attached with covalent

Fig: 4.22: Transfer of Acetyl CoA from mitochondria to cytoplasm. bond to C-terminal Glycine of ubiquitin. This bond is
For example Fatty Acid Synthesis occurs in Cytoplasm. The starting isopeptide bond.
material is Acetyl CoA, which is formed in Mitochondria via


Pyruvate Dehydrogenase enzyme. So this is a point of control that

if Fatty Acid Synthesis has to occur, only then this Acetyl CoA is
allowed to reach Cytoplasm from Mitochondria.

Control of Rate of Synthesis/

Degradation of Enzyme
In body, there is a control of rate of synthesis of enzyme/
protein by controlling the rate of transcription of that gene.
Enzyme synthesis can be induced or repressed at the level of
genes. zz Ubiquitin molecules are attached to each other also by
isopeptide bond.
2 Types of Genes zz Occurs in two steps:
1. Tagging of the protein by the covalent attach-
1. House Keeping/Constitutive Genes→ Always active ment of multiple ubiquitin molecules (Conju-
e.g. genes of TCA cycle enzyme as TCA is a vital cycle. gation)
Levels of these enzymes are not controlled and remains 2. Degradation of the tagged protein by the protea-
almost constant. some

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Fig. 4.23: The Ubiquitin-Proteasome pathway

2. ATP Independent proteolytic system of lysosomes – zz Proteins which are rich in following amino acids are also
enzyme used is Acid Hydrolase e.g. Cathepsins (Mostly rapidly degraded: Proline (P), Glutamate (E), Serine (S)
Cysteine Proteases) and Threonine (T) —known as PEST sequence. Ubiquitin is 137
Table 4.15: Two pathways of protein degradation not covalently attached to these amino acids. (Ubiquitin is
covalently attached to Lysine in protein)
ATP Dependent Protein ATP Independent Protein

CHAPTER 4  ENZYMES
Degradation Degradation zz Mono ubiquitylation of histone proteins is associated mainly
• Ubiquitin Proteasome • Proteolytic enzyme system of with gene repression and heterochromatic gene silencing
Pathway (UPP) lysosomes zz Sumoylation of histones (SUMO stands for Small Ubiquitin like
• Proteolytic complex • Acid hydrolase enzyme involved Modifiers) is associated with transcription repression.
called proteasome used zz Ubiquitination – conjugated with Ubiquitin for degradation
• Degrade damaged or • Degrade extracellular proteins zz Sumoylation – addition of SUMO (no conjugation with
short lived proteins e.g. e.g. Plasma Proteins (taken into ubiquitin). It is not involved in protein degradation.
Intracellular Protein, cells by endocytosis) i.e. long
Regulatory Protein lived structural proteins. Synthesis of Inactive Zymogens

A dditional E dge Many enzymes are first synthesized as inactive precursors,

known as Zymogens or Proenzymes, mainly digestive
Destabilizing N-terminal amino acids (i.e. those proteins having enzymes. This is done to prevent the digestive enzymes from
these amino acids at N-terminal end are rapidly degraded or have digesting the cells that produce them. In a zymogen, part of
short half-lives):
the protein blocks the active site of the enzyme. Cleaving off
1. Arginine
2. Acetylated alanine this peptide activates the enzyme, e.g. Trypsinogen
3. Aspartate
Contd…

Summary Table of Important EC Numbers

Enzyme Enzyme Commission Number
Any Kinase 2
Thiokinase 6
Any Phosphorylase 2
Any Phosphatase 3
Any Mutase 5
Any Carboxylase 6
Any Dehydrogenase (Oxidation) 1
Any Reductase (Reduction) 1
Aconitase (TCA) 4
Fumarase (TCA) 4
Hydratase
Enolase (Glycolysis) 4
PEPCK (Gluconeogenesis) 4
Most Synthases 4 T
EXCEPTIONS H
Glycogen Synthase 2
E
Citrate Synthase 2
ATP Synthase 3
O
Nitric Oxide Synthase (NOS) 1 R
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 Pearls of the Chapter
138 zz All Enzymes are proteins except Ribozyme zz Mostly Rate Limiting enzymes show cooperative Kinetics
zz Enzymes are substrate specific, stereospecific and thermolabile zz Sigmoidal graph is because of a special property of Allosteric
zz Active site is flexible, not rigid enzymes i.e. Cooperativity
CRO BIOCHEMISTRY

zz The active site of an enzyme is formed of amino acids which zz Allosteric enzymes are usually multi-subunit enzymes
may be placed at long distances on the primary structure, but zz Km → Michaelis Menton constant, signature of enzyme and is
are brought nearer to each other by the 3-D conformation of inversely proportional to affinity between enzyme and substrate
the enzyme. zz Hb oxygen dissociation curve → Sigmoidal/S-shape
zz Enzyme does not affect free energy or equilibrium of reaction zz Mb oxygen dissociation curve → Rectangular Hyperbola
zz Enzymes which require metal ion as a cofactor are known as zz LineweaverBurk plot or Double Reciprocal Curve is a modification
metal activated Enzymes. of Michaelis Menton graph (to convert Michaelis Menton graph
zz Enzymes which require metal ion as a prosthetic group are into a straight line graph)
known as Metallo Enzymes. zz Functional plasma enzymes are those enzymes which function
zz All Carboxylases need Mg in plasma e.g. Blood coagulation enzymes


zz IMA- Ischemia Modified Albumin → very useful for Acute zz Cell damage leads to increase concentration of non-functional
Myocardial Ischemia plasma enzymes in plasma
zz All Serine Proteases have catalytic triad of histidine, serine and zz Km does not changes with change in [E] or [S]. It is a constant
aspartate value.
zz Serine proteases and aminotransferases have ping pong zz Key regulatory reactions or committed steps of metabolic
mechanism of Bi-Bi reaction pathways are often subjected to Allosteric Regulation
zz Covalent catalysis often has Ping-Pong mechanism (first zz Most common covalent modification is phosphorylation and
Substrate binds and Product released and then the second dephosphorylation
Substrate will bind) zz House Keeping or Constitutive genes are those genes which are
zz Oxidoreductases carries oxidation-reduction reactions always active
zz Transferases transfer groups zz Zymogens are inactive precursors of enzymes
zz Isomerases interconvert isomers into each other

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 Multiple Choice Questions 139

Basic Concepts about Enzymes 10. Coenzymes are ….. organic compounds-
1. Enzyme DOES NOT act by – (PGMEE 2014)  (PGMEE 1984)
a. Forming non-covalent interactions a. Lipoprotein b. Proteinaceous
b. Catalyzing the reaction c. Non-protein d. Any of the above
c. Increasing activation energy 11. Apoenzyme is – (PGMEE 2013-12)
d. Increasing the rate of reaction a. Cofactor b. Conezyme
2. Mechanism of action of enzymes is all EXCEPT: c. Protein moiety d. None
 (PGMEE 2013) 12. Aldehyde Dehydrogenase requires NAD+ to act. Here
NAD+ is termed as (PGMEE 2013)
a. Acid – Base catalysis
a. Cofactor b. Coenzyme
b. Catalysis by proximity
c. Hypoenzyme d. Abenzyme
c. Catalysis by denaturation
13. FAD linked Dehydrogenase is - (PGMEE 2014)
d. Catalysis by strain
a. Enoyl Reductase
3. Specific activity of enzyme is- (PGMEE 2015,12)
b. Pyruvate Dehydrogenase
a. µ mol of Enzyme per gram of Substrate
c. Succinate Dehydrogenase
b. Enzyme units per mg of protein d. Isocitrate Dehydrogenase
c. Concentration of Substrate transformed per minute 14. Kinases require: (PGMEE 2008)
d. None a. Mn+2 b. Inorganic phosphate
4. Abzyme is a/an (PGMEE 2013) c. Cu+2 d. Mg+2
a. Isoenzyme 15. Zinc is cofactor for (PGMEE 2013)
b. Abnormal enzyme a. Carbonic Anhydrase b. Phosphofructokinase
c. Antibody with a catalytic activity c. Hexokinase d. Aldolase B
d. Allosteric enzyme 16. Cofactor for Xanthine Oxidase: 
5. Definition of Ribozyme: (PGMEE 2013)  (FMGE June 2018)
a. t-RNA a. Zinc b. Copper M
b. RNA molecule that acts catalytically to change it self c. Molybdenum d. Selenium C
or another RNA molecule 17. Carboxylases require (PGMEE 2013-12) Qs
c. Ribonucleoprotein a. Vitamin B7 b. Vitamin B2 Ans.
d. Ribosome c. Vitamin B12 d. Vitamin B1
18. Cofactor for Glutathione Peroxidase- 1. c
Serine Proteases  (PGMEE 2012-13) 2. c
6. Serine of Chymotrypsin is changed with Proline. a. Ca+2 b. Se 3. b
Which of the following will happen ? c. Mn+2 d. Mg+2 4. c
a. Chymotrypsin can catalyze the protein but cannot 5. b
bind Isoenzymes 6. b
b. Chymotrypsin can bind the protein but cannot 19. LDH has how many isoenzymes (PGMEE 2015) 7. b
catalyze a. 3, based on B and M polypeptide subunits 8. a
c. Chymotrypsin can bind the protein as well as can b. 5, based on B and M polypeptide subunits 9. c
catalyze c. 7, based on H and M polypeptide subunits 10. c
d. Cannot decide from given information d. 5, based on H and M polypeptide subunits 11. c
7. Trypsin cleaves carboxy terminal of: 20. Which of the following is true for MI (Myocardial 12. b
a. Glutamate b. Arginine Infarction): 13. c
c. Glycine d. Proline a. LDH-1>LDH-2 b. LDH-1=LDH-2 14. d
8. Which of the following is a Serine Protease ? c. LDH-3>LDH-4 d. LDH-2>LDH-1 15. a
 (Recent Question June 2018) 21. True regarding isozymes is:  (FMGE June 2018) 16. c
a. Chymotrypsin b. Pepsin a. Forms of the same enzymes that catalyze different 17. a
c. Carboxypeptidase d. Caspases reaction 18. b
b. Forms of the same enzymes that catalyze same 19. d
Co-factors and Coenzymes reaction 20. a
9. Non vitamin coenzyme is: c. Forms of the different enzymes that catalyze 21. b
a. Niacin b. Coenzyme A different reaction
c. Lipoic acid d. SAM d. Forms of the different enzymes that catalyze same
reaction
 22. The predominant isoenzyme of LDH occurring in Enzyme Kinetics and Enzyme Uses
liver injury is- (PGMEE 2011) 33. Which among the following is a feature of non
140 a. LDH – 1 b. LDH – 2 competitive inhibition ? (PGMEE 2013-14)
c. LDH - 4 d. LDH-5
a. Increased Km
23. Which isoform of LDH is raised in hemolytic anemia:
b. Decreased Vmax
 (PGMEE 2013, 2012)
c. Decreased Km
a. LDH- 5 b. LDH-3
d. Increased Vmax
c. LDH -4 d. LDH-2
34. Km of an enzyme is:
24. Which of the following enzyme shows liver
 (PGMEE 2008) (PGI-Nov 2009)
obstruction?  (Recent Pattern Jan 2018)
a. Numerically identical for all isoenzymes that
a. ALP b. AST
catalyze a given reaction
c. ALT d. Aminotransferase
25. True about isoenzymes is: b. Dissociation constant
a. Catalyse the same reaction c. The substrate concentration at half maximum
b. Same quaternary structure velocity
c. Same distribution in different organs d. The normal physiological substrate concentration
d. Same enzyme classification with same number and 35. Which of the following is a functional plasma
name enzyme ? (PGMEE 2015, 2008)
a. LDH b. Acid phosphatase
Classification of Enzymes c. Prothrombin d. Amylase
26. Hydrolase belongs to enzyme category number: 36. Km increases, but Vmax remains same. This is….
 (Recent Question 2017, 2012) Inhibition: (FMGE June 2018)(PGI Nov 2011)
a. 1 b. 2 a. Competitive
c. 3 d. 4 b. Non-competitive
27. Which of the following is a Lyase-  c. Irreversible
 (PGMEE 2012, 2013, 2015) d. Uncompetitive
a. Aldolase 37. Which of the following is a suicide enzyme?
b. Fumarase  (PGMEE 2015, 2011)
c. Decarboxylase a. Thromboxane synthase
d. All of the above b. 5’ Nucleotidase
M 28. Carbon Monoxide (CO) is released in reaction c. Lipoxygenase
C catalyzed by- (PGMEE 2015) d. Cyclooxygenase
Qs a. Decarboxylases 38. Which of the following estimates blood creatinine
Ans. b. Carboxylases level most accurately?
22. d c. Heme oxygenase a. Jaffe method b. Kinetic Jaffe method
23. d d. Pyruvate Dehydrogenase c. Technicon method d. Enzyme assay
24. a 29. The difference in MW between Phenylalanine and 39. An enzyme-catalyzed reaction was carried out with
25. a Tyrosine is by: (Recent Question 2016) the initial substrate concentration 1,000 times greater
26. c a. 17 b. 16 than the Km for that substrate. After 9 minutes, 1% of
27. d c. 64 d. 32 the substrate had been converted to the product, and
28. c 30. Enzyme which cleaves C-C bond:  the amount of product was 12 mmol. If, in a separate
29. b  (Recent Question 2016) experiment, one-third as much enzyme and twice as
30. a a. Lyase b. Ligase much of the substrate is combined, how long it would
31. d c. Transferase d. Isomerase to take for the same amount (12 mmol) of product to
32. c 31. All are true about oxygenase EXCEPT: be formed?  (AIIMS May 2018)
33. b  (Recent Question 2015) a. 13.5 mins b. 27 mins
34. c a. Incorporate one atom of O2 c. 8 mins d. 9 mins
35. c b. Incorporate both atoms of O2
36. a c. Hydroxylation of steroids Enzyme Regulation
37. d d. Help in carboxylation of drugs 40. Which statement is FALSE about covalent modi-
38. d 32. Which of the following enzymes DOES NOT parti- fication?  (Recent Question 2018)
39. b cipate in oxidation-reduction reactions?  a. It is reversible
40. c  (AIIMS May 2013) b. It is slower than allosteric regulation
a. Oxygenases b. Peroxidases c. It uses the same enzyme for activation and
c. Hydrolases d. Dehydrogenases inactivation
d. Phosphorylation is a common covalent modification
41. Allosteric modulators seldom resemble the substrate 44. Mechanism of conversion of trypsinogen to trypsin-
or product of the enzyme. What does this observation  (PGMEE 2015)
show: (Recent Question 2018) a. Hydrolysis 141
a. Modulators likely bind at a site other than the active b. Removal of Carboxyl group
site c. Phosphorylation
b. Modulators always act as activators d. Removal of part of protein
c. Modulators bind and inhibit the enzyme 45. All of the following are covalent modifications of
d. The enzyme catalyzes more than one reaction enzyme regulation EXCEPT:
42. Which statement is FALSE about allosteric regu- a. Phosphorylation b. ADP Ribosylation
lation? c. Acetylation d. Glycosylation
a. It is usually the mode of regulation for the last step in 46. Mechanisms for regulating enzyme activity is/are:
reaction pathways  (PGI May 2017)
b. Cellular response is faster with allosteric control a. Covalent modification
than by controlling enzyme concentration in the cell b. Allosteric activation
c. The regulation usually is important to the c. Competitive inhibition
conservation of energy and materials in cells d. Induction of genes for enzyme synthesis
d. Allosteric modulators bind non-covalently at sites e. Repression of gene for inhibition of enzyme syn-
other than the active site and induce conformational thesis
changes in the enzyme 47. Which of the following is/are Not raised in liver
43. Defective proteins are degraded after attaching disorder:  (PGI May 2013)
a. Lipase b. Urease
covalently to- 
c. ALP d. AST
 (PGMEE 2018, 2015, 2013, 2010)
e. ALT
a. Pepsin b. Laminin
c. Clathrin d. Ubiquitin

M
C
Qs
Ans.
41. a
42. a
43. d
44. d
45. d
46. a,b
d,e
47. a,b
 Answers with Explanations
142
1. Ans. (c) Increasing activation energy 7. Ans. (b) Arginine
CRO BIOCHEMISTRY

[Ref: Harper 30th/e pg. 92] [Ref: Lehninger 7th/e pg. 215]
•• Enzyme decreases the activation energy and thus •• Trypsin cleaves at the carboxy terminal of basic
they increase the rate of reaction. Mostly in Enzyme- amino acid (Arginine, Lysine).
Substrate interactions, non-covalent interactions are
formed (Hydrogen, Hydrophobic and Ionic bonds). 8. Ans. (a) Chymotrypsin

[Ref: Harper 30th/e pg. 687]

2. Ans. (c) Catalysis by denaturation
•• Serine protease is a category of enzyme having
[Ref: Harper 30th/e pg. 92] Serine at its active site. Examples include: Trypsin,
•• There are four general mechanisms to facilitate Chymotrypsin, Elastase, Thrombin, Plasmin, Clotting


catalysis: Acid – Base catalysis, Catalysis by proximity, factor X and XI, Complement protein, PSA (Prostate
Catalysis by strain and Covalent Catalysis Specific Antigen)
•• Denaturation destroys enzymatic activity, so this is •• Cysteine proteases are also known as Thiol proteases.
not a mechanism of enzyme action. Examples are: Papain and Caspases
•• Carboxy proteases hydrolyze peptide bond at
3. Ans. (b) Enzyme units per mg of protein carboxy terminal Examples are pepsin and lysosomal
cathepsins.
[Ref: Harper 30th/e pg. 63]
•• Metalloproteinases any protease enzyme where
•• Specific activity is the activity of an enzyme per catalytic mechanism involves a metal. Examples are
milligram of total protein (expressed in μmol carboxypeptidase A & B.
min−1mg−1). It is a measure of enzyme processivity,
at a particular specific substrate concentration and is 9. Ans. (c) Lipoic acid
usually constant for a pure enzyme. Specific activity [Ref: Lehninger 7th/e pg. 621]
gives a measurement of enzyme purity in the mixture.
•• Lipoic acid is a coenzyme (required for Pyruvate
4. Ans. (c) Antibody with a catalytic activity Dehydrogenase and Alpha-Ketoglutarate
Dehydrogenase) but it is not a Vitamin (Option c).
[Ref: Harper 30th/e pg. 62] Niacin is Vitamin B3 (Option a). Coenzyme A is a
•• Abzyme is antibody acting as enzyme. It is a derivative of Vitamin B5 – Pantothenic acid (Option
monoclonal antibody and is also called catalytic b). SAM – is S-Adenosyl Methionine, which is a
A methyl donor. It is neither a Coenzyme, nor a Vitamin
N antibody or catmab.
(Option d).
S 5. Ans. (b) RNA molecule that acts catalytically to
W change it self or another RNA molecule 10. Ans. (c) Non-protein
E
[Ref: Harper 30th/e pg. 62] [Ref: Lehninger 7th/e pg. 188; Harper 30th/e pg. 64]
R
S •• Ribozyme is when RNA is acting as enzyme. Mostly •• Coenzymes are organic but they are not proteins
they cleave phosphodiester bonds and they catalyze e.g. Lipoic acid, water soluble vitamins, NAD, NADP,
WITH FMN, FAD.
themselves or other RNA molecules.
E 11. Ans. (c) Protein moiety
X 6. Ans. (b) Chymotrypsin can bind the protein but
P cannot catalyze [Ref: Harper 30th/e pg. 62]
L [Ref: Lehninger 7th/e pg. 215] •• The protein portion of holoenzyme is known as
A Apo enzyme. The non protein portion is known as
N •• Serine is the catalytic amino acid in Chymotrypsin.
Whenever catalytic amino acid is changed, then the cofactor, coenzyme or prosthetic group.
A
T enzymatic activity decreases. But binding amino
12. Ans. (b) Coenzyme
I acids in chymotrypsin are Histidine and Aspartate.
O They are not changed so the enzyme is able to bind [Ref: Harper 30th/e pg. 62]
N but not catalyze. •• NAD+ is organic non protein portion which is required
S for the enzyme, so it is a Coenzyme.
13. Ans. (c) Succinate Dehydrogenase 21. Ans. (b) Forms of the same enzymes that catalyze
same reaction
[Ref: Harper 30th/e pg. 62] 143
[Ref: Lippincott’s 4th/e pg.65]
•• Succinate Dehydrogenase is FAD linked and it is an
enzyme involved in both TCA and ETC. •• Isozymes/Isoenzymes are different forms of same
enzyme, catalyzing same reaction in same species.

CHAPTER 4  ENZYMES
14. Ans. (d) Mg2+ Example: There are five isozymes of Lactate
Dehydrogenase.
[Ref: Harper 30th/e pg. 62]
•• Isoenzymes have different structure, electrophoretic
•• All Kinases require Mg, but Pyruvate Kinase requires mobility, Km and Vmax. Isoenzymes are present in
K+>> Mg2+. different cells of the body.
15. Ans. (a) Carbonic Anhydrase 22. Ans. (d) LDH-5
[Ref: Harper 30th/e pg. 62] [Ref: Harper 30th/e pg. 66]
•• Carbonic Anhydrase requires zinc. Zinc is a cofactor •• In liver, both LDH-4 and 5 are present but LDH-5
for over 300 enzymes e.g. Carbonic Anhydrase, isoenzyme is predominant.
Alcohol Dehydrogenase, Alkaline Phosphatase, DNA
Polymerase which are essential for growth, wound 23. Ans. (d) LDH-2
healing, reproductive function and protection from
[Ref: Harper 30th/e pg. 66]
free radical damage.
•• LDH-2 is raised in haemolytic anemia. LDH-1 is
16. Ans. (c) Molybdenum raised in Myocardial Infarction.
[Ref: Harper 30th/e pg. 120] 24. Ans. (a) ALP
•• All oxidases require copper EXCEPT: Xanthine •• ALP – Alkaline Phosphatase, active at alkaline pH
Oxidase and Sulfite Oxidase has the physiological role of dephosphorylating
•• They both require Molybdenum. compounds.
•• There are 4 Isozymes:
17. Ans. (a) Vitamin B7 ƒƒ ALP-I – Intestinal
[Ref: Harper 30th/e pg. 62] ƒƒ ALP-L – Tissue-non specific (expressed mainly in
Liver/Bone/Kidney)
•• All Carboxylases require Vitamin B7 i.e. Biotin. ƒƒ ALP-P – Placental (Regan isozyme)
ƒƒ GC-ALP – Germ cell
18. Ans. (b) Se
•• Elevated levels of ALP are seen in:
[Ref: Harper 30th/e pg. 62] ƒƒ Liver diseases: Biliary obstruction, Hepatitis, A
Cirrhosis N
•• Selenium is required Glutathione Peroxidase.
ƒƒ Bone Diseases: Osteoblastic bone tumors, S
19. Ans. (d) 5, based on H and M polypeptide subunits Osteomalacia, Osteoporosis, Paget’s disease W
ƒƒ Other conditions: Myelofibrosis, Leukemoid re- E
[Ref: Harper 30th/e pg. 66] action, Lymphoma, Sarcoidosis, Hyperthyroid- R
ism, Hyperparathyroidism, Myocardial infarction,
•• Lactate Dehydrogenase (LDH) to catalyze the S
Pregnancy.
reversible conversion of pyruvate and lactate. There
•• Decreased levels of ALP are seen in: Hypophospha- WITH
are 5 isoenzymes. This enzyme is a tetramer, made up
tasia, Oral contraceptives, Postmenopausal women
of two subunits H and M. (H stands for Heart and M E
receiving Estrogen therapy, Hypothyroidism.
stands for Muscle). X
25. Ans. (a) Catalyse the same reaction P
20. Ans. (a) LDH-1>LDH-2
L
•• LDH-1 (HHHH) → Present in Heart [Ref: Harper 30/e p63] A
•• LDH-2 (HHHM) → Present in RBCs (Blood) •• Isoenzymes catalyse the same reaction. For example, N
•• In normal person, LDH-2 is more than LDH-1 in LDH-1 to LDH-5 all convert Pyruvate to Lactate. A
serum. But in myocardial infarction, LDH-1>>LDH-2. •• They have different quaternary structure. For T
This is known as Flipped ratio of LDH in Myocardial example, the subunits in LDH-1 is different from LDH- I
Infarction. 2. Tissue distribution of each isoform is different. O
Enzyme name and number can be different. N
S
 26. Ans. (c) 3 one atom of O2 and Dioxygenase will incorporate two
atoms of molecular oxygen. All monoxygenases are
144 [Ref: Harper 30th/e pg. 61]
known as hydroxylases.
•• Hydrolase is EC No. 3. These enzymes use water •• Oxygenases have no role in carboxylation (option d).
to break the bond. E.g. Phosphatase, all digestive
enzymes 32. Ans. (c) Hydrolases
CRO BIOCHEMISTRY

•• Hydroxylase is also known as Mono-Oxygenase/ •• Enzyme of oxidoreductase category are Oxidases,

Mixed function Oxidase. Mono-Oxygenases incorpo- Reductases, Peroxidases, Oxygenases (Mono or Di)
rate one atom of molecular Oxygen into the Substrate. and Dehydrogenases.
They belong to EC No. 1 i.e. Oxidoreductase.
33. Ans. (b) Decreased Vmax
27. Ans. (d) All of the above
[Ref: Harper 30th/e pg. 92]
[Ref: Harper 30th/e pg. 61]
•• In Competitive inhibition, Km increases and Vmax
•• Lyases are enzymes which can either break a bond or remains same. But in Non competitive inhibition, Km
make a bond but they do not require water or ATP. The is same and Vmax decreases.
EC Number is 4. Fumarase is a Hydratase (enzyme


which can add or remove water but bond is not 34. Ans. (c) The substrate concentration at half maxi-
broken). Other examples of Hydratase are Enolase, mum velocity
PEPCK and Aconitase. Aldolase A and B both are
[Ref: Harper 30th/e pg. 79]
Lyases. Simple Decarboxylases are also Lyases.
•• Km is Michaelis Menton constant. Km is that substrate
28. Ans. (c) Hemeoxygenase concentration at which velocity of reaction is half
of Vmax. Km is inversely proportional to affinity.
[Ref: Harper 30th/e pg. 81] Km is signature of enzyme. It is not association or
•• Heme Oxygenase catalyze degradation of Heme. dissociation constant.

35. Ans. (c) Prothrombin

[Ref: Medical biochemistry by Sheriff p. 94]
•• Prothrombin >> LDH
•• Functional plasma enzymes are those enzymes
which are present in plasma as they have function in
plasma e.g. Lipoprotein Lipase, clotting factors e.g.
Prothrombin.
•• Non–functional plasma enzymes do not have
A function in plasma, they are present inside cells. E.g.
N SGOT is present in liver and heart.
29. Ans. (b) 16
S •• Only LDG-2 is a functional plasma enzyme.
W [Ref: Lehninger 7th/e pg. 697]
36. Ans. (a) Competitive
E •• Usually students tend to mark 17 as the answer.
R •• Phenylalanine gets converted to Tyrosine by enzyme [Ref: Lippincott’s 4th/e pg.29]
Phenylalanine Hydroxylase. Most Hydroxylases are
S •• In competitve inhibition, Km increases and Vmax is
Mono-oxygenases i.e. EC No. 1. In hydroxylation
WITH reactions, only one atom of molecular oxygen is same
added. So, change in MW is by 16. •• In non-competitve inhibition, Km is same and Vmax
E decreases
X 30. Ans. (a) Lyase •• In uncompetitive inhibition, both Km and Vmax
P decreases.
L [Ref: Harper 30th/e pg. 61]
A •• Lyase breaks C-C bond. E.g. Aldolase, Synthase. (In 37. Ans. (d) Cyclooxygenase
N this case, ATP is not used). [Ref: Harper 30th/e pg. 200]
A
31. Ans. (d) Help in carboxylation of drugs •• Suicide inhibition is also known as mechanism
T
based inhibition. E.g. Allopurinol inhibits Xanthine
I [Ref: Harper 30th/e pg. 61] Oxidase, Aspirin inhibits Cycloxygenase.
O
N •• There are two types of oxygenases i.e. Monoxygenase
S and Dioxygenase. Monoxygenase will incorporate
 38. Ans. (d) Enzyme assay 40. Ans. (c) It uses the same enzyme for activation and
inactivation
[Ref: Varley’s Practical Clinical Biochemistry 6/e p352] 145
[Ref: Harper 30th/e pg. 90-91]
There are two methods for estimation of creatinine in
blood: •• Covalent modification is a long term regulation (by
•• Jaffe’s test: In alkaline medium, creatinine forms a hormones). It is slower than allosteric regulation. It

CHAPTER 4  ENZYMES
red coloured tautomer of Creatinine Picrate which is reversible.
is measured colorimetrically. This method can be •• Most common covalent modification is
automated in auto-analysers and kinetic method can phosphorylation and dephosphorylation. It uses
be used. Kinetic jaffe is more accurate than Jaffe’s different enzymes for activation and inactivation.
method. The enzyme which adds phosphate is Protein Kinase
•• Enzymatic method: By employing enzymes, and the enzyme which removes phosphate is Protein
Creatininase or Creatinine Deaminase. It is more Phosphatase.
specific and accurate. There is no interference
by ketones, bilirubin or glucose. Hence measure 41. Ans. (a) Modulators likely bind at a site other than
creatinine accurately. the active site
[Ref: Harper 30th/e pg. 90-91]
39. Ans. (b) 27 mins
•• Allosteric modulators can be activators or inhibitors.
[Ref: Lehninger principles of biochemistry, 6th ed., pg,204] They bind non-covalently to the allosteric/ regulatory
Let’s say: site and induce changes in the active site, (where
At time t1= [S] = 1000 x Km, [E] = z substrate binds). Therefore, modulate the binding of
At time t2= [S] = 2000 x Km, [E]=z/3 substrate.

42. Ans. (a) It is usually the mode of regulation for the

Formula is: Vmax = K catalysis × [E]t
last step in reaction pathways
Vmax × [S] [Ref: Harper 30th/e pg. 90-91]
V0 = (Michaelis Menton equation)
Km +[S] •• Allosteric regulation is usually the mode of regulation
for the first step in reaction pathways
Put formula of Vmax
Refer Q.41 for explanation.
K × [E]t × [S]
Vo = cat
Km +[S] 43. Ans. (d) Ubiquitin
Kcat × z × 1000 Km
At t1, V1 = [Ref: Harper 30th/e pg. 289]
Km +1000 Km
•• The Ubiquitin Proteosome Pathway (UPP) is the main
and highly regulated mechanism for intracellular A
K × z/3 × 2000 Km
At t2, V2 = cat protein catabolism. It occurs in cytoplasm and N
Km +2000 Km S
nucleus. Proteins which are bound to ubiquitin are
Kcat × z × 1000 Km Km + 2000 Km W
V1/V2 = × degraded in proteosomes.
Km + 1000 Km Kcat × z / 3 × 2000 Km E
44. Ans. (d) Removal of part of protein R
For same enzyme K catalysis is same and Km <<<<< [S] so
we can neglect Km [Ref: Dinesh Puri 3rd/e pg. 118] S

kcat × z × 1000 Km Km + 2000 Km •• Many enzymes are first synthesized as inactive WITH
V1/V2 = × precursors known as Zymogens or Proenzymes,
Km + 1000 Km Kcat × z / 3 × 2000 Km mainly digestive enzymes. In a zymogen, part of the E
protein blocks the active site of the enzyme. Cleaving X
1
V1/V2 = 1/3 = 3 off this part of protein activates the enzyme. E.g. P
Trypsinogen to Trypsin. L
V2 = V1/3 A
45. Ans. (d) Glycosylation N
Rate of reaction is decreasing three times so time taken A
will increase three times i.e. more time taken now Various ways of covalent modifications of enzymes are: T
for same reaction as velocity or speed of reaction is •• Most common is Phosphorylation and Dephos- I
decreased. phorylation. O
So time = 9 × 3 = 27 minutes. N
S
 •• Other covalent modifications: Adenylation, Uridy- 47. Ans. (a); (b)
lation, ADP Ribosylation, Methylation and Acety- •• Lipase and Amylase are used in diagnosis of
146 lation. Pancreatitis (Option a)
•• Urease is used to estimate urea in lab. (Option b)
46. Ans. (a); (b); (d); (e)
•• ALP (Alkaline Phosphatase), AST (Aspartate Transa-
•• Covalent modification (Option a) and allosteric
CRO BIOCHEMISTRY

minase), ALT (Alanine Transaminase) are specific for

activation (Option b) are methods of enzyme liver. (Option c, d, e).
regulation. Enzyme’s quantity i.e. amount of enzyme
synthesized can be controlled at the level of gene
expression, by inducing or repressing the gene
(Option d, e).


A
N
S
W
E
R
S
WITH

E
X
P
L
A
N
A
T
I
O
N
S
 5
Chemistry and
Metabolism of Amino
Acids
Overview of Chapter The carboxy and amino groups of all amino acids are joined
by peptide bonds in proteins and these groups are therefore
•• Amino acid basics not free for any chemical reactions (they can just make
•• Buffers and Titration curve hydrogen bonds). So the side chain of amino acid decides the
role of that amino acid in protein.
•• Essential amino acids
The central carbon of any amino acid is asymmetric. So
•• Classification of amino acids with diseases amino acids show both optical and structural isomerism.
•• Polyamine Pathway (Any compound having asymmetric carbon shows both
•• Fish odour syndrome optical and structural isomerism)
•• Color reaction of proteins
H igh R eturn
AMINO ACIDS BASICS “All amino acids have one assymetric C”
Exceptions:
There are around 300 amino acids in nature, out of which zz 0 → Glycine
22 amino acids are found in mammalian proteins. 21st and zz 2 → Isoleucine, Threonine
22nd are Selenocysteine and Pyrrolysine respectively. There
is nothing like 1st or 2nd or 5th amino acid (number does not
specify any sequence but just total of these are 22 ). 21st and Zwitterion
22nd were discovered later that’s why they are given a specific Amino acid ionizes to give negative charge on carboxy group
number. These 22 amino acids are encoded by DNA i.e. they and positive charge on amino group.
are not formed by post translational modifications. They have zz Net charge is zero
codons in DNA. But derived amino acids are those which do zz This is known as Zwitterion (also known as ampholyte)
not have codons. zz This zwitterion is insoluble (solubility is because of
DERIVED AMINO ACIDS charges). Hence precipitates

Found in proteins Not in proteins

• OH-Proline • Ornithine
• OH-Lysine }
• Citrulline in Urea cycle
• ϒ-carboxy Glutamate (in • Homocysteine- in
clotting proteins II, VII, IX, X) methionine metabolism

Why the name ‘‘Amino Acid” ?

Amino group (–NH2) is always on left side and acid group Fig. 5.2: Zwitterion
(–COOH) is always on right side. That’s why the name– Amino
acid (Fig. 5.1) pI – Isoelectric pH – that pH at which zwitterion exist. So at pI, a
protein precipitates

Isoelectric Focussing (IEF)

zz This is a special type of electrophoresis in which
isoelectric pH is used for separation of proteins. Different
pH gradient is taken in electrophoretic plate. When
proteins move then they get focussed or precipitated
in the region of their pI as they are insoluble at this pH.
(Fig. 5.3)
zz pH gradient (different pH in different region) is taken in an
electrophoretic gel. Protein solution applied (all proteins
are negatively charged). When electric field switches on,
Fig. 5.1: Common structure of all amino acids. Each amino acid then negative charge proteins will start moving towards
has a carboxyl group, a primary amino group and a side chain.
 the positive end of the electrode. The region at which pH zz Form of amino acid present in protein → always L
of protein matches pI protein becomes insoluble and zz But Free amino acid (i.e. amino acid which is not in
148 thus precipitates. protein) → can be -L or- D e.g. D-Serine and D-Aspartate
are present in brain. Also, D-amino acids are found in
some antibiotics and in bacterial cell walls
CRO BIOCHEMISTRY

H igh R eturn
Q. Which form of amino acid is abundant in protein?
A. L
Q. Which form of amino acid is abundant in body?
A. L
Q. Which form of amino acid is present in body?
A. → Both –L and -D

Meaning of Alpha, Beta and Gamma



zz a means that alpha carbon (C-2) contains the amino

group.

Fig. 5.3: Isoelectric focussing (also known as Electro Focussing)

D and L Amino Acids

zz Amino group is a special thing in amino acids (amino
Fig. 5.5: In case of fatty acids, the carbon containing functional
group is not found as main constituent in any other
group (–COOH) is given number 1 (C1). And the carbon adjacent
macromolecule). So, everything is in relation to amino to functional carbon is always α i.e. C-2 is always a, C-3 is always β
group (–NH2), in case of amino acids. and C-4 is always γ
zz L means that amino group is on left side. D means that
amino group is on right side.

Fig. 5.6: Now consider amino acid is a kind of acid having two
carbons. Out of these two carbons, alpha carbon (–C2) has amino
group (–NH2). That’s why all amino acids are called ‘alpha’ amino
acids

F undamental B ox

T
H
E
O
R
Y
Fig. 5.4: L and D isomers (Enantiomers or Mirror Images)
of amino acids
 BUFFERS AND TITRATION CURVE
zz A buffer is a solution whose pH does not changes when a 149
small amount of strong acid or base is added to it. These
are usually solutions of weak acid with its conjugate base.
zz A buffer is most effective when pH = pKa
(pKa = Dissociation constant)

CHAPTER 5  CHEMISTRY AND METABOLISM OF AMINO ACIDS

zz Buffering range =pKa ± 1 (means maximum buffering
capacity)
zz Handerson Hasselbalch equation
[Base]
pH = pKa + log
[Acid]
zz When pH = pI (isoelectric point), then solution has
minimum buffering capacity and precipitation occurs
due to insolubility.
zz Amino acids have buffering action
zz Histidine has maximum buffering capacity because pKa
of imidazole group of histidine (6.5-7.4) is very near to
physiological pH (7.4).
H igh R eturn zz
zz
Hb has buffering action due to Histidine
Major extra-cellular buffer – Bicarbonate buffer
zz Name of any amino acid written without mentioning about zz Major intra cellular buffer – Phosphate buffer
–L/–D or Alpha/beta/gamma → then obviously it is –L Alpha
zz Major urinary buffer – Phosphate buffer
amino acid (there is no need to mention about that)
zz Bicarbonate buffer (H2CO3/HCO3-) is ideal buffer as
zz But if we have to write the name of some –D amino acid then
both the constitutes of this buffer can be altered by body
it is written like this → D-Serine
(HCO3- can be altered by renal system and H2CO3 can be
zz Similarly, if we have to write some beta or gamma amino acid
maintained by respiratory system)
then it is written like this → beta-Alanine, gamma amino
butyric acid
zz NOTE: e.g. of non α-amino acids β-alanine, β amino iso- Titration Curve
butyrate, ϒ-amino iso butyrate
It is a graphical plot of pH against the amount of alkali (OH-)
added. pH is usually taken on y-axis.

T
H
E
O
R
Y
 Titration Curve of a Weak Acid

150
CRO BIOCHEMISTRY


Titration Curve of Glycine

T
H
E
O
R
Y
Titration Curve of Aspartic Acid (Three Ionizable Groups) ESSENTIAL AMINO ACIDS
Amino acids which cannot be synthesized in body are known 151
as essential amino acids.
Q. How many amino acids are essential in diet?
A. 8.

CHAPTER 5  CHEMISTRY AND METABOLISM OF AMINO ACIDS

These are Valine, Leucine, Isoleucine, Phenylalanine, Try-
ptophan, Lysine, Threonine and Methionine
Q. How many amino acids are essential in body or in
proteins?
A. 20. Because all 20 amino acids are required in body for
synthesizing proteins. (The eight amino acids are essential
only in diet).
Q. Which are the two semi essential amino acids?
A. Arginine and Histidine.
Q. Which is semi essential?
  a. Arginine
b. Histidine }
If both given, then mark Arginine
A. Arginine. (Histidine can be synthesized in adults so arginine
Amino Acids in Alkaline & Acidic Medium is more essential as compared to histidine)
Q. Which amino acid can be formed in adults but cannot be
zz Amino acids are negatively charged in alkaline medium formed in children?
(pH > pI) due to carboxy group A. Histidine
zz Amino acids are positively charged in acidic medium
(pH < pI) due to amino group
zz When pH = pI, then zwitterion exists i.e. net charge is F undamental B ox
zero. zz Essential amino acids: Those which cannot be synthesized in
body and are required in diet.
zz Non-essential amino acids: Those which can be synthesized in
body and are not required in diet.
zz Semi-essential amino acids: Those which can be synthesized
in body but to some extent.

CLASSIFICATION OF AMINO ACIDS

Aliphatic Amino Acids
1. Glycine 4. Leucine
2. Alanine 5. Isoleucine
Q. Why albumin is negatively charged? 3. Valine
A. Isoelectric point (pI) of albumin is 4.7
T   pH of blood is 7.4 i.e. (pH > pI) , means albumin is Glycine
H present in alkaline medium. So, it is negatively
I
charged.
N
K

F undamental B ox
zz pI of acidic amino acids is 2-3. pH of blood is 7.4 i.e. (pH > pI),
means acidic amino acids are present in alkaline medium. So,
they are negatively charged.
zz pI of basic amino acids is > 8. pH of blood is 7.4 i.e. (pH < pI), T
Fig. 5.7: Glycine: simplest, smallest amino acid H
means basic amino acids are present in acidic medium. So,
they are positively charged. E
Uses of Glycine
O
zz Formation of Glutathione R
zz Formation of Creatine Y
zz Haem synthesis
 Q. Which amino acid is responsible for flexibility of pro- Glycine is not found in alpha helix. Because alpha helix is a very
teins? symmetrical, helical structure (like cylindrical structure). There is
152 A. Glycine. Because Glycine has smallest side chain. So it can no bend in the structure of alpha helix.
fit in a small space and therefore it creates bends (a sharp
bend has very less space) in the structure of protein. Ability
to bend means flexibility. So Glycine is responsible for Glycine Metabolism
CRO BIOCHEMISTRY

flexibility of proteins.
  Glycine is mostly found in beta turns/beta bends (also Glycine metabolism is very much linked with THF and PLP.
Proline). Beta turns allow the polypeptide to turn and get glycine can be synthesized from:
folded, but there is very less space present at the bend.
1. CO2 and NH4+ by Glycine Synthase (N5, N10 methylene
THF involved)
2. Glyoxylate by Glycine Amino Transferase or Glycine
Transaminase (PLP required)
3. Serine by Serine Hydroxyl Methyl Transferase (reversible)
– PLP and Folic acid required –N5, N10 methylene THF
involved


4. Threonine – by Threonine Aldolase glycine cleavage

Glycine Cleavage
(1) Glycine cleavage system: Occurs in liver mitochondria.
It is a major pathway for Glycine degradation. Glycine
(a) (b) is converted to CO2 and THF bound one carbon unit.
Enzyme is also known as Glycine Synthase for the reverse
reaction.
(2) A minor pathway is conversion to glyoxylate by glycine
Figs. 5.8: (a & b): (a) Shows beta turn (less space in a bend/turn);
oxidase. Glyoxylate can then be either converted to
(b) Shows very less space in a bend or turn
formate or oxalate.
(3) Also glycine can be converted to serine, which further
forms pyruvate. Glycine is the precursor for many
compounds. (Fig. 5.9)

T
H
E
O
R
Y

Fig. 5.9: Glycine metabolism

Summary Box
Glycine 153
zz Simplest, smallest, no asymmetric carbon
zz Gives flexibility to proteins
zz Not found in alpha helix, but found in beta turns/bends
zz Non-essential, non-polar

CHAPTER 5  CHEMISTRY AND METABOLISM OF AMINO ACIDS

zz Glucogenic (as catabolic end product is Pyruvate)

Glycine as Precursor of Many Compounds

Fig. 5.10: Glycine is the precursor for many important compounds.

Fig. 5.11: Synthesis of Choline & Betaine from Glycine (Choline and Betaine Metabolism is linked with Folate, SAM & PLP)

T
H
E
O
R
Y
154
CRO BIOCHEMISTRY


Fig. 5.12: Formation of creatine from glycine: Creatine synthesis starts from glycine and arginine in kidney making guanidino acetic acid.
Then creatine formation occurs in liver after addition of methyl group from SAM. Creatine reaches muscles and gets converted to creatine
phosphate

Diseases in Glycine Metabolism 3. Glycinuria: Defect in transporter for Glycine and Proline
which results in:
1. Oxaluria  Glycine and Proline in urine
 Defect in Glycine Transaminase is associated with  Serum Glycine is normal
impaired Glyoxylate to Formate conversion which  Increased risk for oxalate stones but urine oxalate is
diverts excess Glyoxylate to form Oxalate, which can normal
form Oxalate stones in kidneys. This is known as
4. Non-Ketotic Hyper-Glycinemia:
Primary Hyperoxaluria. In treatment, restriction of
oxalate rich foods done e.g. green leafy vegetables,  Defect in glycine cleavage system which is the major
beet root, tea. (Fig. 5.9) pathway of glycine catabolism.
 Also, Oxalate in urine can appear in B6 deficiency.  Increased glycine in blood, CSF, urine
Treatment is B6 supplementation  Brain affected as glycine is a neurotransmitter, leads
2. Secondary Hyperoxaluria: Occurs due to: to mental retardation, seizures, lethargy, apnea.
T This is known as Glycine Encephalopathy.
 Vitamin B6 deficiency (Glycine Transaminase
H affected leading to Glyoxylate accumulation, which  No effective treatment.
E forms excess Oxalate)
O  Vitamin C toxicity (as Dehydro Ascorbic acid is Alanine
R converted to Oxalic acid) zz Alanine (3C) is the most glucogenic amino acid
Y  Ethylene glycol poisoning (as Ethylene glycol gets zz The side chain of alanine has simple methyl group which
converted to Glyoxylate) can be easily synthesized in the body. So, alanine is non-
essential. (Fig. 5.13)
Summary Box
155

CHAPTER 5  CHEMISTRY AND METABOLISM OF AMINO ACIDS

Fig. 5.13: Structure of Alanine

Q. Why in exercising muscle Lactate is produced while

in fasting muscle Alanine is formed?
T A. During exercise, anaerobic glycolysis occurs which
H produces Lactate. (Fig. 3.3)
I
During fasting, transamination reaction occurs
N
K which produces alanine. (5.14)

Fig. 5.16: Catabolism of Leucine, Valine and Isoleucine. It shows

that Leucine is purely ketogenic, Valine is purely glucogenic,
Isoleucine is both ketogenic and glucogenic

Summary Box
Branched Chain Amino Acids
zz All are essential
Fig. 5.14: Cahill Cycle (Glucose-Alanine Cycle) zz All are non-polar (Isoleucine is most non-polar)
zz Leucine is Ketogenic
zz Valine is Glucogenic
Summary Box zz Isoleucine- both Ketogenic and Glucogenic (Fig. 5.16)
Alanine zz Amino acid that can be a fuel for brain is Isoleucine
zz Non-essential
zz Non-polar
Maple Syrup Urine Disease (MSUD)
zz Most glucogenic (catabolic end product is Pyruavte)
This AR (autosomal recessive) disease is a defect in the
catabolism of branched chain amino acids. During their
Valine, Leucine and Isoleucine catabolism, there is a step of oxidative decarboxylation,
which is defective in MSUD.
These are branched chain amino acids. Branching is a difficult
Defective enzyme → Branched chain alpha-keto acid
thing to be done (just an idea to learn). So, all these branched
dehydrogenase, also known as, branched chain alpha-keto
chain amino acids are essential. (required in diet)
acid decarboxlyase. This enzyme is a multienzyme complex.

Clinical Features
1. Burnt sugar like odour (because of isoleucine)
2. Ketosis, vomiting, feeding problems
T
3. Mental retardation (because of increased leucine)
H
4. Abnormal muscle tone
E
5. If not treated, then coma (because of increased leucine)
O
6. Death can occur (High Mortality rate)
R
Diagnosis: DNPH test
Y
Fig. 5.15: Structure of branched chain amino acids
 Treatment

156 zz Diet given which is restricted in branched chain amino

acids, but some amount given to allow normal growth.
zz Patients with a rare thiamine dependent variant of MSUD
gives response when large doses of vitamin B1 is given.
CRO BIOCHEMISTRY

Isovaleric Academia – (Rare Autosomal Disorder)

Defect in Dehydrogenase (FAD dependent) which specifically
degrades isovaleric acid (involved in metabolism of leucine). (a) (b)
It is characterized by a sweaty feet odour or rancid cheese
odour caused by isovaleric acid in body fluids and urine, Fig. 5.17 (a and b): Structure of (a) Phenyl Alanine and (b) Tyrosine
especially during acute illness. Patient has neurological
symptoms, metabolic acidosis (usually with an elevated
anion gap) and ketosis.


Non Polarity of All 5 Aliphatic Amino Acids

zz All are non-polar
zz Most non-polar is Isoleucine > Valine
zz Least non polar is Glycine

H igh R eturn
Controversy for the Polarity of Glycine
Q1. Which is Polar?
a. Glycine    b. Alanine    c. Valine
A. (a) Glycine
All are non-polar. Glycine is least non-polar. So here we
can mark Glycine as polar.
Q2. Which is Polar?
a. Glycine    b. Alanine    c. Aspartate
A. (c) Aspartate Fig 5.18: Conversion of Phenylalanine to Tyrosine
Aspartate is negatively charged. So it is very clear in this
question that aspartate is polar. So here consider Glycine H R
igh eturn
as non-polar.
All three aromatic amino acid hydroxylases are similar
1. Phenylalanine hydroxylase
Aromatic Amino Acids
1. Phenylalanine
2. Tyrosine hydroxylase
3. Tryptophan hydroxylase }
Require NADPH and THB

THB is mainly required for hydroxylases [Nitric Oxide Synthase

2. Tyrosine (NOS) also requires THB].
3. Tryptophan
4. Histidine
F undamental B ox
Phenylalanine (Fig. 5.17a) Phenylalanine add OH Tyrosine
Structure: Alanine + a phenyl ring added to alanine Alanine add OH Serine
So this amino acid has a big side chain (use this idea to
learn), which cannot be synthesized in body. So it is essential
in diet. And it is a non-polar amino acid as rings are mostly
H igh R eturn
T
non-polar. Controversy for the polarity of Tyrosine
H
zz Because of –OH group, it is polar and because of phenyl ring,
E Tyrosine (Fig. 5.17b) it is non-polar. So according to question, you have to decide
O Tyrosine is polar or non-polar (depends what are other options
R Phenylalanine is converted to tyrosine by enzyme given in the question)
Y Phenylalanine Hydroxylase. So therefore tyrosine is non-
essential as it can be synthesized in body from phenylalanine.
Summary Box H igh R eturn
Phenyl-Alanine zz Mousy/musty odour → due to Phenyl Acetate 157
zz Essential zz Mental Retardation → due to Phenylalanine
zz Non polar zz FeCl3 Urine test → due to Phenyl Pyruvate
zz Both glucogenic and ketogenic

CHAPTER 5  CHEMISTRY AND METABOLISM OF AMINO ACIDS

Tyrosine # Maternal Phenylketonuria
zz Non-essential If mother has PKU and levels of Phenylalanine are raised
zz Controversy for polarity during pregnancy then infant has mental retardation,
zz Both glucogenic and ketogenic microcephaly, low birth weight, growth retardation and
congenital heart defects.
Phenylketonuria (PKU)
zz Most common metabolic disorders of amino acid Q. Why brain is affected in PKU?
PKU is of 2 types: T 1. Excess Phenylalanine from blood enters brain
zz Classical – Deficiency of Phenylalanine Hydroxylase H through L- aromatic amino acid transporter in brain
zz Non- classical – Deficiency of DHB. Reductase or GTP I capillaries. Other aromatic amino acids (Tyrosine,
Cyclohydrolase. N Tryptophan) cannot enter brain due to competitive
K inhibition. Tyrosine and Tryptophan synthesize
many neurotransmitters which are required in brain.
2. Deficiency of Thyroxine as it is synthesized from
Tyrosine.
3. Cerebral serotonin deficiency (as it is synthesized
from tryptophan)

Diagnosis
1. FeCl3 test is less sensitive and detects Phenyl Pyruvate
Fig. 5.19: Phenylketonuria (Defect in Phenyl Alanine Hydroxylase) in urine (Pyruvate is a keto acid). Positive test gives blue
green color.
Why this name of disease: Phenyl ketone in urine i.e. Phenyl
Pyruvate found in urine of these patients (Pyruvate is a Keto- 2. Guthrie’s test or Bacterial inhibition test is more reliable
than FeCl3 urine test. Bacteria used is Bacillus subtilis
acid).
(detects serum Phenylalanine levels). But Guthrie’s
Child is normal at birth as no problem in fetal development test should be done 1-2 weeks after birth when the
because maternal enzyme breaks down phenylalanine.
body has ingested some protein containing amino
acid phenylalanine in diet to ensure accurate results.
Clinical features: (Phenylalanine is present in both human and cow’s milk.)
zz Body odour – mousy or musty because of phenylacetate 3. Confirmatory diagnosis is by measuring amino acid
zz Severe mental retardation due to excess Phenylalanine. levels in blood by Tandem Mass Spectrometry (can
zz Tyrosine becomes essential detect smaller increase of phenylalanine present at the
zz Deficiency of pigment Melanin (formed from Tyrosine), time of birth.)
leads to fair skin, blue eyes and light hair color.
Prenatal diagnosis also possible

Treatment of Phenyl Ketonuria

zz Give tyrosine
zz Restrict Phenylalanine in diet (lifelong restriction)
zz Aspartame contraindicated (because it is made up of
aspartate and phenyl-alanine)
zz THB given for Non Classic PKU. T
H
Uses of Tyrosine E
zz OTHER FEATURES: Microcephaly, rash, hypertoma, Synthesis of catecholamines, thyroid hormones and the O
seizures, hyperactivity, exaggerated tendon reflexes, pigment melanin. R
wide spaced teeth, enamel hypoplasia. Y
 Catecholamine Synthesis
158 zz Catecholamines are epinephrine, norepinephrine and
dopamine
zz Precursor of all three catecholamines is L-DOPA
First catecholamine to be synthesized is Dopamine
CRO BIOCHEMISTRY

zz
zz Dopamine is deficient in Parkinsonism
zz NE is synthesized in sympathetic ganglia and nerve endings.
zz Catecholamine with methyl group: Epinephrine
zz Conversion of NE to Epinephrine occurs in periphery, not in
CNS. 80 % of it occurs in adrenal medulla.


Fig. 5.20: Synthesis of Catecholamines (Dopamine, NE and E) from Tyrosine

zz Patients also have vision defects and photophobia. This

H igh R eturn is also called oculo-cutaneous albinism.
VMA (Vanillyl Mandelic Acid) zz Vitiligo: lack of melanoblast in some regional areas, but
zz Catabolic end product of epinephrine and nor-epinephrine enzyme Tyrosinase is normal.
zz It is increased in pheochromocytoma and neuroblastoma
zz Normal levels = 2.6 mg/ 24 hour urine sample

Synthesis of Melanin Pigment

T zz It occurs in melanosomes present in melanocytes of skin
H and hair
E zz Albinism – absence of pigment from skin, hair and eyes.
O Defect is in enzyme Tyrosinase.
R Fig. 5.21: Melanin synthesis
Y
 159

CHAPTER 5  CHEMISTRY AND METABOLISM OF AMINO ACIDS

T
Fig. 5.22: Catabolism of Phenylalanine and Tyrosine: From diet, Phenylalanine is derived which gets converted to Tyrosine in body. H
Phenylalanine has just one role in body i.e. formation of Tyrosine. All the very important functions in body are done by this important E
amino acid – Tyrosine. So therefore we can conclude that catabolism of Phenylalanine and Tyrosine is same. O
R
Y
160
CRO BIOCHEMISTRY

Fig. 5.23: Tyrosine gets converted to same product DOPA by two

different enzymes.

NOTE: These enzymes are present in different cells of the body.

Tyrosinemia


zz Tyrosinemia Type I – Defect in Fumaryl Acetoacetate Fig. 5.24: Deposition of black higments
Hydrolase. Also known as tyrosinosis or hereditary zz Treatment: Dietary restriction of PhenylAlanine and
tyrosinemia or hepato renal tyrosinemia Tyrosine. Also vitamin C is found to be helpful in few
zz Tyrosinemia Type II – Defect in Tyrosine Transaminase. patients as it prevents the oxidation of homogentisic
Also known as Richner/Hanhart syndrome or Oculo acid.
cutaneous Tyrosinemia. Oculo is due to corneal ulcer
and photophobia, cutaneous is due to hyperkeratotic Tryptophan
plaques.
zz Tyrosinemia Type III – Defect in PHPP hydroxylase.
Also known as neonatal Tyrosinemia.

Alkaptonuria
zz Defect in catabolism of Tyrosine and Phenylalanine
zz Enzyme deficient is Homogentisate Dioxygenase.
Urine turns black on exposure to air due to oxidation of
homogentisic acid. Homogentisic acid is an intermediate
in the catabolism of Tyrosine.
zz Dark staining of diapers
zz Deposition of black pigments (polymerization of Catabolic end product of Tryptophan is Alanine. (Alanine further
homogentisic acid occurring in body over years) in joints, forms pyruvate by transamination) so Tryptophan is glucogenic.
cartilage and collagenous tissue, known as ochronosis. Also Tryptophan gets converted to acetoacetyl CoA (Ketone
(Fig. 5.24) Body). So, Tryptophan is also ketogenic.
zz First pigment gets deposited in sclera, ear and nose
cartilage, intervertebral discs, joints (ochronotic arthritis)
zz Later knee, shoulders, hips are also affected (small joints Summary Box
of hands and feet are spared) Tryptophan
zz Essential
zz Non-polar
zz Both ketogenic and glucogenic

T
H
E
O
R
Y
Uses of Tryptophan “Usually vitamins cannot be synthesized in body. So, Atypical
1. Tryptophan forms 5’OH-Tryptophan which is converted vitamin is that vitamin which can be synthesized in body E.g.
to serotonin (5-hydroxytryptamine) and then melatonin Vitamin B3 and Vitamin D’’. Vitamins which can be synthesized 161
by intestinal bacteria are not atypical as they are synthesized by
bacterial enzymes, not human enzymes.
# For Niacin formation, vitamin B2 and B6 and iron are

CHAPTER 5  CHEMISTRY AND METABOLISM OF AMINO ACIDS

required.

Hartnup’s Disease
zz AR (autosomal recessive), rare disorder, named after the
first affected family.
zz Defect in neutral amino acid transporter also called
mono-amino mono carboxylic amino acid transporter
which transports neutral amino acid including
tryptophan, encoded by gene SLC 6A19 on chromosome
5
zz Failure to absorb tryptophan from intestine and also
reabsorb it from kidneys
zz C/F → Aminoaciduria i.e. Tryptophan in urine without a
corresponding increase in plasma tryptophan levels
zz Pellagra (niacin deficiency) like symptoms (neurological
and dermatologic), despite an adequate intake of both
Tryptophan and niacin.
zz It is a part of Garrod’s tetrad
zz Age of onset is 30-40 years, usually with lower backache
Fig. 5.25: Synthesis of serotonin and melatonin zz No mental retardation
zz Fresh urine of these patients- normal color
2. Tryptophan also forms Niacin (Vit B3). So Niacin is zz Benedict’s test is positive as Homogentisic acid is a
considered an atypical vitamin. reducing substance
zz FeCl3 test is positive.
zz Bluish urine stained diapers
zz Treatment:
1. Niacin 2. High protein diet

Carcinoid Syndrome
Carcinoid syndrome occurs in 5% cases of carcinoid
tumor which produces serotonin. In this case, much of the
Tryptophan is diverted to the formation of serotonin so
that Tryptophan cannot synthesize niacin. So pellagra like
symptoms occur. (Fig. 5.27)
zz C/F – sweating, cutaneous flushing, GI motility affected,
abdominal cramps, diarrhoea.
zz Urine has increased 5 HIAA (5-Hydroxy Indole Acetic
Acid)

Fig. 5.26: Formation of Niacin: Kynurenine Anthranilate Pathway

3-hydroxy kynurenine is one of the intermediate in Tryptophan T
catabolism, which gets converted to next intermediate and
this conversion requires vitamin B6 (Pyridoxine). In vitamin B6 H
deficiency, this reaction does not occur and 3-hydroxy kynurenine E
is diverted to form alternate metabolite, xanthurenic acid, which is O
excreted in urine. R
Y
(60 mg tryptophan gets converted to 1mg of Niacin) Fig. 5.27: Carcinoid syndrome
 Basic Amino Acids
162 Basic amino acids are also known as positively charged
amino acids e.g. Arginine, Lysine and Histidine. All these are
essential. All are polar but arginine is most polar, then lysine
and then histidine.
CRO BIOCHEMISTRY

H igh R eturn
zz Arranged in decreasing order of polarity:
Arginine > Lysine > Histidine


Fig. 5.30: Basic amino acids

Catabolic end product of arginine is alpha-keto glutarate, so it is
also glucogenic.
Fig. 5.28: Normal and Abnormal routes of Tryptophan.

Histidine
zz Positive charge but less positive charges
zz Polar but less polar
zz Essential but semi-essential
zz Maximum buffering capacity because histidine is the Summary Box
only amino acid with a side chain which can ionize
within the physiological pH range Basic Amino Acids Arginine Lysine
zz Catabolic end product – glutamate formed via FIGLU. So • Semi-essential • Essential
it is glucogenic. • Polar • Polar
• Glucogenic • Ketogenic

Acidic amino acids and their amides

Acidic amino acids are also known as negatively charged
amino acids e.g. Aspartate and Glutamate. All these are
non-essential because they can be synthesized from TCA
intermediates. All are polar.
Fig. 5.29: Histidine structure
Their amides are Asparagine and Glutamine. Asparagine
contains amide (–CONH) linkage at beta carboxy carbon
T Summary Box and glutamine contains amide linkage at gamma carboxyl
H carbon. They are uncharged but polar, due to –CONH (amide
E Histidine group)
O zz Semi-essential
zz Polar but less polar
R
zz Glucogenic
Y
 163

Fig. 5.34: Conversion of asparagine to aspartate and then

CHAPTER 5  CHEMISTRY AND METABOLISM OF AMINO ACIDS

oxaloacetate

F undamental B ox
zz Cation + charge � Cathode – charge
zz Anion – charge � Anode + charge

–OH containing Amino Acids

All these amino acids are polar because –OH group can
participate in hydrogen bond formation. E.g. Serine,
Threonine and Tyrosine. So Tyrosine is aromatic also and OH
containing also. OH containing amino acid are mostly the site
of enzyme regulation.
Tyrosine is less common than serine for phosphorylation
and is the site for Iodination in Thyroglobulin and site of
glycosylation in glycogenin.

Fig. 5.31: Glutamine synthesis requires ATP

Summary Box
Acidic amino acids and their amides
zz All are non-essential
zz All are polar
zz All are glucogenic
zz Catabolic product of all these amino acids are TCA inter-
mediates, so all are glucogenic.

Fig. 5.35: Metabolism of Serine, Glycine and Cysteine and also

selenoCysteine is interlinked
Fig. 5.32: Asparaginase synthetase requires ATP. Glutamine is the
NH2 down

T
H
E
Fig. 5.33: Conversion of glutamine to glutamate and then alpha-
ketoglutarate Fig. 5.36: Acetyl choline synthesis from serine O
R
Y
 Summary Box Sulfur containing Amino Acids
164 Serine They are Cysteine and Methionine
zz Non-essential zz Cysteine has a special group- SH (sulfhydryl group).
zz Polar SH group is present at the active site of many enzymes.
zz Glucogenic Cysteine imparts reducing nature to glutathione.
CRO BIOCHEMISTRY

Because of SH cysteine is polar.

Threonine zz Methionine is required in diet but in body it can be
zz Essential
converted to Cysteine. So methionine is essential but
zz Polar
Cysteine is non essential.
zz Both glucogenic and ketogenic

Trick For Q Solving

Whenever these three questions asked, then answer is
T OH-containing amino acid
H Q. Which amino acid has maximum tendency to bind Cysteine is oxidized to cystine (2 sulfhydryl groups of cysteine
I


phosphate? join to form a covalent cross link known as disulphide bond).

N
K A. OH-group has maximum tendency to bind phosphate
Q. Which amino acid is responsible for covalent
modifications?
A. OH-Containing amino acid most common covalent
modifications are phosphorylation and dephospho-
rylation
Q. Which amino acid is the site for O-glycosidic Catabolic product of Cysteine is pyruvate. And sulphate
bonds?
released is used to form PAPS (3’ phospho adenosine 5’
A. Oxygen is taken from –OH group of OH-Containing
phospho sulphate), which acts as sulphate donor.
amino acid
Q. Which amino acid is the site for N-glycosidic Methionine forms SAM (S-Adenosyl Methionine), which
bonds? acts as a methyl donor. Methionine also forms HomoCysteine.
A. Asparagine

F undamental B ox
2 types of bonds in case of Glycoproteins:
1. O- glycosidic bond → Carbohydrate is joined with protein with
the help of oxygen (O)
2. N- glycosidic bond → Carbohydrate is joined with protein with
the help of nitrogen (N)

Fig. 5.38: All three phosphate bonds of ATP are used in the
formation of SAM
MAT 1 and 3 - in Liver
MAT 2 - Extra-hepatic tissues
zz SAM and Coenzyme A are high energy compounds but they
don’t have any phosphate
T zz Cysteine is not an essential amino acid as long as methionine
H is available in diet (as cysteine is synthesized from methionine)
E zz Tyrosine is not an essential amino acid as long as
O Phenylalanine is available in diet (as Tyrosine is synthesized
R from Phenylalanine)
Y Fig. 5.37: O- and N-glycosidic bonds in glycoproteins
 165

CHAPTER 5  CHEMISTRY AND METABOLISM OF AMINO ACIDS

Fig. 5.39: Metabolism of methionine

Homocystinuria [HCU]
zz ↑ Homocysteine (SH) in blood
zz ↑ Homocystine (S-S) in urine
zz Homocystine is made up of 2 homocysteines
zz Types → Acquired, Genetic
Acquired → occurs due to vitamin B6, B9 or B12 deficiency
Genetic (AR) – further of 2 types: Type I and Type II (see table below)

Type I HCU or Classic HCU/ Typical Type II or Non- Classic

More common Less common
Defect: Enzyme cystathionine β- synthase Defect: is in the formation of active form of folate or vitamin B12
or HomoCysteine N-methyl transferase.

Methionine↑ Methionine ↓

Cysteine ↓ Cysteine normal or ↑

Treatment: Treatment:
Give Cysteine Increase methionine by giving Betaine (Betaine can convert
Decrease methionine in diet. Homocysteine to methionine by giving methyl group)

C/F of HCU
zz Vascular Diseases: Stroke, Atherosclerosis, Myocardial Infarction, Pulmonary Embolism.
zz Ectopia lentis, elongated limbs, Pectus carinatum T
zz Seizures, Mental Retardation H
zz Osteoporosis E
Lab diagnosis (HCU): positive Cyanide Nitroprusside Test (CNT test) O
R
Y
166
CRO BIOCHEMISTRY


Cystinuria – Autosomal Recessive Summary Box

zz Most common inborn error of amino acid transport
zz Part of Garrod’s tetrad Methionine Cysteine
zz Defect in dibasic amino acid transporter • Essential • Non-essential
zz In urine: Cysteine, Ornithine, Lysine, Arginine • Non-polar • Polar
(mnemonic COLA) • Glucogenic • Glucogenic
zz Cystine stones
zz Treatment: chelating agent Penicillamine (forms comp-
lex with cysteine) IMINO ACID – Proline
zz All amino acids have a primary amino group except
Cystinosis Proline which has a secondary amino group i.e. N is
zz A generalized LSD (Lysosomal Storage Disease) attached to 2 carbons.
zz Defect in Cystine transporter in lysosomes known as zz In proline, the side chain and the alpha amino nitrogen
Cystinosis joins to form a rigid pentacyclic ring
zz Cystine deposits in tissues (Cornea, Liver, Kidney, Bone zz This unique structure of proline helps in the fibrous
marrow) structure of collagen and proline is found in beta turns/
zz Treatment: Cysteamine (forms complex with Cystine) bends. But proline cannot be accommodated in alpha
Diseases of S- containing Amino Acids helix.
Disease Cyanide Defect Proline is not found in alpha helix, but it may be present in the
Nitroprusside first turn of the helix.
test (CNT) Because:
zz Proline has unusual shape as the side chain & alpha amino
Homocystinuria + Defect in the conversion
of HomoCysteine to nitrogen of proline joins to form a rigid pentacyclic ring.
Cystathionine or Methionine zz It is unable to complete the H-bonds in the chain of the helix
(Alpha helixes are held together by hydrogen bonds between
Cystinuria + Defect in Dibasic Amino Acid the alpha amino- and alpha carboxy- groups of the backbone).
(Part of Garrod’s Transporter (for Cystine & So, Prolines in alpha helices after the first turn (4th residue) cause
Tetrad) Basic Amino Acids) a kink in the helix.
Cystinosis + Defect in Cystine
(a Lysosomal Transporter in Lysosomes zz Catabolic end product: Proline is oxidized to glutamate.
Storage Disease) So it is glucogenic as glutamate can form alpha-keto
T glutarate (intermediate of TCA cycle).
Cystathioninuria - Defect in Cystathionase
H enzyme. (In treatment, give
E vitamin B6)
O
Note: In blood, Cysteine if accumulated, then in urine it gets
R converted to Cystine. So it is always Cysteinemia & Cystinuria.
Y
 POLYAMINE PATHWAY
Polyamines are polycations which interact with negatively 167
charged DNA, RNA and proteins. Polyamines are synthesized
from non-protein amino acid – ornithine. Major polyamines
are: putescine (diamine), spermidine (triamine) and
spermine (tetramine).

CHAPTER 5  CHEMISTRY AND METABOLISM OF AMINO ACIDS

Fig. 5.40: Structure of Proline Rate limiting enzyme of polyamine biosynthesis is Orn-
ithine decarboxylase, for which substrate is ornithine.
Summary Box
Proline
zz Non-essential Roles of polyamines are:
zz Non-polar zz Cell growth, survival, regulation and proliferation
zz Glucogenic zz DNA stability & chromatin modulation
zz Signal transduction & cell migration
zz Used as a growth factor in cell cultures
zz Changes in polyamine levels are associated with aging
and diseases.
zz Therapeutic use of polyamine biosynthesis inhibitor
– DFMO (Di Fluoro Methyl Ornithine) is in sleeping
sickness & hirsuitism.

H igh R eturn
zz Most polar amino acid – Arginine
zz Most non-polar amino acid – Isoleucine
zz Polar amino acids: They can form ionic or hydrogen bonds
zz Non-polar amino acids: These amino acids cannot gain or
Fig. 5.41: Polyamine pathway
loose protons and therefore cannot participate in ionic or
hydrogen bonds. These can form hydrophobic interactions.
FISH ODOUR SYNDROME/ TRI METHYL
H igh R eturn AMINURIA
zz 21 protein forming amino acid → SelenoCysteine → UGA
st zz Fish odour syndrome (autosomal recessive) is a rare
zz Selenocysteine is found in approximately 25 human proteins. metabolic disorder in which enzyme defect is Flavin
Enzymes which use selenocysteine at their active sites are containing mono-oxygenase -3 (FMO3). This enzyme
known as Selenoproteins e.g. uses riboflavin as a co-factor.
1. Glutathione peroxidase zz In this syndrome, trimethyl amine is not broken down,
2. Thioredoxin reductase which gives a strong fishy odour as it is excreted in body
3. Iodothyronine deiodinase fluids (sweat, urine & breath).
To synthesize selenocysteine, serine is attached to the tRNA.
So precursor amino acid for selenocysteine is serine. T
zz 22nd protein forming amino acid → Pyrrolysine → UAG H
 Not present in humans. E
 Precursor- Lysine O
 Co-translational modification. R
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  Glutamine and Asparagine give brown color (due to
amide group).
168  Mostly used to detect finger prints.
2. Biuret reaction to detect peptide linkages (general test
for proteins). Minimum two peptide linkages must be
present so free amino acid and dipeptides do not give
CRO BIOCHEMISTRY

positive Benedict’s test.

 # Free amino acids will give Ninhydrin positive but
Biuret negative.
3. Xanthoproteic reaction to know the presence of
aromatic amino acid. Positive test gives yellow color.
 Phenylalanine do not give positive test.
4. Modified Millon’s reaction (Cole’s test) to know the
presence of Tyrosine (only amino acid with phenol
group)
5. Aldehyde test/Hopkins-Cole’s Test- to detect the


presence of tryptophan ( indole ring). Positive test gives

purple color.
6. Obermeyer test
 Test for Indican or Indoxyl compound
 Positive test is Blue or Violet color.
 Positive in Hartnup disease.
7. Pauly’s Test for Imidazole Group (Histidine) and
Phenolic Group (Tyrosine).
8. Sakaguchi Test for Guanidine Group (Present in
Arginine). Positive test is red color.
9. Test for Sulphur containing amino acids (Cysteine and
Cystine gives test positive).
Fig. 5.42: Fish Odour Syndrome  Methionine does not give this test because sulfur is
zz Affected individuals have a reduced capacity to blocked in between 2 carbons with strong bond.
metabolize trimethylamine into trimethylamine N-oxide 10. Cyanide Nitroprusside Test (CNT)
(odourless).  Cyanide breaks disulfide bond into sulfhydryl group.
zz Trimethylamine is volatile, which gives off a strong fishy  Sodium Nitroprusside reacts with sulfhydryl group
smell in urine, sweat, and expired air. to give magenta color.
 Cysteine, Cystine, Homocysteine and Homocystine
Treatment gives this test positive. So this test is positive in
Homocystinuria (HCU), Cystinuria and cystinosis.
zz Restriction of foods rich in trimethyl amine (fish, egg,
11. Ferric chloride test
liver, nuts, grains)
 Detects Phenylpyruvate and Branched chain amino
zz Riboflavin supplement
acid in urine.
 Gives various colors depending on the substance.
COLOR REACTIONS OF PROTEINS  Positive in Phenylketonuria (PKU), Maple Syrup
Proteins react with a variety of reagents to give colored Urine Disease
compounds. These reactions are known as color reactions 12. DNPH test – (Dinitro Phenyl Hydrazine)
of proteins. These can be used for qualitative & quantitative  Positive test in PKU, MSUD
detection of proteins. Some are based on peptide bonds and  Detects α –Keto acids in urine.
others are based on type of amino acid residues These are: 13. Guthrie’s Test- or Bacterial Inhibition Assay
1. Ninhydrin reaction – for α- amino acids (i.e. free amino  Detects Phenyl-pyruvate, Phenyl-lactate and
group) and gives purple color. Phenyl-acetate in serum
T
 Proline and Hydroxy-Proline react in a different way  Positive in PKU
H
due to 2º amino group (not 1º) and they give yellow 14. VMA spot test – for diagnosis of Pheochromocytoma
E
color. 15. 5-HIAA test (Hydroxy Indole Acetic Acid)
O
 Used for diagnosis of Carcinoid syndrome.
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 169

CHAPTER 5  CHEMISTRY AND METABOLISM OF AMINO ACIDS

Table 5.1: All amino acid classes

Aliphatic AA Aromatic AA
Valine Tyrosine
Isoleucine Tryptophan
Leucine Phenylalanine
Alanine Histidine
Glycine
Sulphur containing Imino acid
Cysteine Proline
Methionine
Acidic side chain Basic side chain
Aspartic acid Histidine
Glutamic acid Arginine
Lysine
Essential Amino Acids (OH group)
• Methionine • Valine Tyrosine T
• Arginine • Isoleucine Threonine H
• Threonine • Leucine Serine E
• Tryptophan • Lysine O
• Phenyl Alanine R
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 Summary
170
CRO BIOCHEMISTRY


Fig. 5.43: Showing the end products of amino acids. Amino acids which form Acetyl CoA or Acetoacetyle CoA, are ketogenic. Amino
acids which form pyruvate or any TCA intermediate, they are glucogenic. Some amino acids are both glucogenic and ketogenic

Polar amino acids further divided into charged polar and uncharged polar. Uncharged polar amino acids have no charge but
they are polar because of some other group e.g. SH in Cysteine, OH- group in serine, Threonine, Tyrosine and amide group
and carbonyl group in asparagine and glutamine. These special groups can form hydrogen bonds therefore they contributes
to polarity.

POLAR AND NON-POLAR AMINO ACIDS

POLAR AMINO ACIDS NON-POLAR AMINO ACIDS
CHARGED POLAR UNCHARGED POLAR
• Basic amino acids • Cysteine (Sulhydryl group) • Aliphatic amino acids
e.g. Arginine, Lysine, Histidine • OH-containing amino acids e.g. e.g. Glycine, Alanine, Valine, Leucine, Isoleucine
• Acidic amino acids Serine, Threonine, Tyrosine • Aromatic amino acids:
e.g. Glutamate, Aspartate • Amides of Acidic amino acid e.g. PhenylAlanine, Tyrosine, Tryptophan
e.g. Glutamine, Asparagine • Proline
• Methionine
# Note: There is a controversy for the polarity of Tyrosine and Glycine. According to the question, one has to analyse what should be
marked. Once you know it is a controversy, you will be able to solve the question.
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Peculiar Odours in Different Amino Acidurias
Inborn Error of Metabolism Urine Odour
171
Phenylketonuria (PKU) Mousy/ Musty
Maple Syrup Urine Disease (MSUD) Maple Syrup/ Burnt Sugar

CHAPTER 5  CHEMISTRY AND METABOLISM OF AMINO ACIDS

Isovaleric Acidemia Sweaty feet/ Cheesy
Hawkinsinuria Swimming Pool
Glutaric Acidemia Sweaty Feet
3-Hydroxy-3-Methyl Glutaric Aciduria Cat Urine
Multiple Carboxylase Deficiency Tomcat Urine
Hypermethioninemia Boiled Cabbage
Tyrosinemia Boiled Cabbage, Rancid Butter
Trimethylaminuria Rotten fish
Diabetic Ketoacidosis Fruity
Urinary tract infections Foul smell

Ketogenic and Glucogenic Amino Acids

Ketogenic Amino Acids Glucogenic And Ketogenic Amino Acids Glucogenic Amino Acid
• Leucine • Tyrosine • Rest all are Glucogenic
• Lysine • Tryptophan (NO NEED TO CRAM)
• Threonine
• Isoleucine
• PhenylAlanine
Note: There is controversy for Lysine that it is purely ketogenic or in both categories. So, according to the question, one has to analyse
what to mark. Once you know it is a controversy, you will be able to solve.

Amino Acids and their Special Groups

Amino acid Special groups

PhenylAlanine Benzene/ Phenyl
Tyrosine Phenol
Tryptophan Indole
Histidine Imidazole
Arginine Guanidino
Proline Pyrrolidine/imino group
• Guanidino group is a special group in Arginine, but Guanidine is a chemical which act as Denaturing Agent for Proteins
• Guanidine is neither a Purine, nor a Pyrimidine

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 Amino Acid Derivatives
172
CRO BIOCHEMISTRY


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 Pearls of the Chapter
zz Aromatic amino acid with basic properties – Histidine
173
zz Aromatic amino acid with –OH group – Tyrosine
zz 21st and 22nd are Selenocysteine and Pyrrolysine respectively
zz 22 amino acids are encoded by DNA i.e. they are not formed by post translational modifications

CHAPTER 5  CHEMISTRY AND METABOLISM OF AMINO ACIDS

zz All amino acids have 1 assymetric carbon. Except – Glycine, Threonine and Isoleucine
zz pI – Isoelectric pH – that pH at which zwitterion exist
zz Amino acid abundant in protein - L - form
zz Amino acid abundant in body → L-form
zz Amino acid present in body→ Both –L and -D forms
zz Arginine and Histidine are semiessential amino acids
zz Precursor amino acid for SelenoCysteine is Serine.
zz Amino acid responsible for flexibility of proteins is Glycine
zz Alanine is the most glucogenic amino acid (3C)
zz Wheat lacks lysine
zz Pulses lack methionine
zz Conservative or Homologous substitution of amino acids is that when one amino acid is replaced by another amino acid with similar
characteristics. E.g. Glutamate replaced by aspartate (both are acidic amino acids).
zz Non-Conservative or Non-Homologous substitution: of amino acids is that when one amino acid is replaced by another amino acid with
different characteristics. E.g. Glutamate replaced by valine.

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174 Multiple Choice Questions
Chemistry of Amino Acids 13. Polyamine like putrescine is derived from:
1. Indole ring is present in: (PGMEE 2013, 12) a. Arginine b. Ornithine
a. Tryptophan b. Phenylalanine c. Yohimibine d. Arginosuccinate
c. Tyrosine d. Threonine 14. Non-essential amino acid group is? (PGMEE 2015)
2. Which of following is polar: (PGMEE 2013,12) a. Acidic amino acid
a. Tryptophan b. Branched chain amino acid
b. Methionine c. Basic amino acid
c. Glutamic acid d. Aromatic amino acid
d. Isoleucine 15. Which amino acid has maximum tendency to bind
3. Strength and rigidity in keratin is due to: phosphate?
 (PGMEE 1995) a. Serine b. Alanine
a. Leucine b. Cysteine c. Phenylalanine d. Tryptophan
c. Lithium d. None of the above 16. After a point mutation, glutamic acid replaced by
4. Creatine is made up of all, EXCEPT: (PGMEE 2013) valine, which leads to formation of sickle cell Hb. The
a. Arginine b. Alanine mobility of HbS as compared to normal Hb on gel
c. Methionine d. Glycine electrophoresis will be :
5. Amino acid with double chiral carbon:  a. Decreased
 (PGMEE 2013) b. Increased
a. Tyrosine b. Threonine c. Dependent on HbS concentration
c. Tryptophan d. Phenylalanine d. Unchanged
6. All of the following amino acids forms acetyl CoA via 17. Xanthurenic acid is the metabolite in the metabolism
Pyruvate Dehydrogenase EXCEPT: of: (PGMEE 2009)
a. Glycine b. Hydroxyproline a. Uric acid b. Xanthine
c. Tyrosine d. Alanine c. Tryptophan d. Uronic acid
M 7. Amino acid which is not stable in (incompatible with) 18. Glycine is used in the synthesis of all EXCEPT:
C alpha-helix is:  (PGMEE 2007)  (PGMEE 2015)
Qs a. Proline b. Glutamine a. Purines b. Creatine
Ans. c. Alanine d. Tryptophan c. Heme d. Pyrimidines
8. Sulphur containing amino acids metabolism needs: 19. Both glucogenic and ketogenic amino-acids are all
1. a  (PGMEE 2014) EXCEPT: (PGMEE 2015)
2. c a. Pyridoxine b. Folic acid a. Leucine b. Tryptophan
3. b c. Vitamin B12 d. All of the above c. Phenylanine d. Tyrosine
4. b 9. Coenzyme for Phenylalanine hydroxylase is: 20. Substitution of which one of the following amino acids
5. b  (PGMEE 2015) in place of alanine would increase the absorbance of
6. c
protein at 280 nm? (PGMEE 2005)
a. S-Adenosyl Methionine
7. a
a. Leucine b. Arginine
b. Tetra Hydro Biopterin
8. d
c. Tryptophan d. Proline
c. Tetra Hydro Folate 21. What is isoelectric point?
9. b d. Pyridoxal phosphate
10. b
a. When pH = pI
10. N Methyl Glycine is known as: (PGMEE 2015) b. When zwitterion exists
11. c a. Betaine b. Sarcosine
12. c
c. Protein precipitation occurs
c. Carnosine d. Ergothionine d. All
13. b 11. Taurine is synthesized from which amino acid?
14. a
22. Which of the following amino acid is extracted
 (PGMEE 2011) predominantly by muscle, having been spared by the
15. a a. Tryptophan b. Phenyl Alanine
16. a
liver in post prandial state?
c. Cysteine d. Alanine a. Valine
17. c 12. Which of the following does not contain β-alanine?
18. d
b. Glutamine
 (PGMEE 2014,13) c. Glutamate
19. a a. Carnosine b. Anserine
20. c
d. Alanine
c. Homocarnosine d. Pantothenic acid
21. d
22. a
23. tRNAsec during the co-translational synthesis of 35. Cereals are deficient in which amino acid:
selenocysteine is initially charged by which amino  (Recent Question June 2018)
acid ? a. Leucine b. Lysine 175
a. Selenocysteine b. Methionine c. Phenylalanine d. Tryptophan
c. Serine d. Cysteine 36. Amino Acid having role in sleep wake cycle:
24. The formation of glycine takes place by transamina- (Recent Question June 2018)
tion of :
a. Alanine b. Glyoxylate a. Tryptophan b. Phenylalanine
c. Aspartate d. Glutamate c. Tyrosine d. Glutamine
25. Aspartame is composed of: 37. Glycine is present in all EXCEPT:
a. Aspartic acid and Methionine  (Recent Question June 2018)
b. Aspartate and Phenylalanine a. Glutathione b. Creatine
c. Asparagine and Methionine c. Glutamine d. Purine nucleotide
d. Asparagine and Phenylalanine 38. Melanin is synthesized from :  (FMGE Nov 2018)
26. Out of the following, which is substrate for the rate a. Tyrosine b. Tryptophan
limiting enzyme of polyamine biosynthesis? c. Phenyl-alanine d. Threonine
a. Anandamide b. Cadaverine 39. Adenosine receptor stability is because of extensive
c. Ornithine d. Histidine disulfide bonds formed between: (FMGE Nov 2018)
27. 3-phosphoglycerate is the precursor for the synthesis a. Cysteine b. Methionine
of which amino acid? c. Arginine d. Alanine
a. Threonine b. Proline 40. Atherosclerosis is associated with:
c. Serine d. Aspartate  (FMGE June 2018 )
28. Out of the following statement which is NOT TRUE a. Lysine b. Histidine
about phenylalanine hydroxylase: c. Homocysteine d. Leucine
a. NADPH provides the reducing power 41. Fibrinopeptide A and B are highly negatively charged
b. Tetrahydrobiopterin (THB) is a cofactor proteins made up of:  (Recent Question Jan 2018)
c. Mixed function oxidase a. Serine and Threonine
d. Vitamin C is a cofactor b. Lysine and Histidine
29. Amino acids which contribute to biosynthesis of c. Aspartate and Glutamate
purine nucleotides are all EXCEPT: d. Leucine and Lysine
a. Aspartate b. Histidine M
c. Glutamine d. Glycine Diseases of Amino Acid metabolism C
30. Essential amino acids are named so: 42. In Maple syrup urine disease, the amino acids Qs
a. Because they are produced in thebody excreted in urine are all EXCEPT: (PGMEE 2007) Ans.
b. Because they are not produced in thebody a. Leucine 23. c
c. They are the only amino acids important forlife b. Phenylalanine 24. b
d. Every food stuff essentially contains them c. Isoleucine 25. b
31. Aspartate is similar to glutamate in the same way d. Valine 26. c
that: 43. A child with pellagra like symptoms, amino acids in 27. c
a. Valine is similar to threonine urine, family history of two siblings affected and two 28. d
b. Asparagine is similar to glutamine normal. Parents are normal. What is the diagnosis? 29. b
c. Phenylalanine is similar to tryptophan 30. b
d. Phenylalanine is similar to tyrosine a. Phenylketonuria 31. b
32. Essential amino acids are all EXCEPT:  b. Alkaptonuria 32. d
 (Recent Question June 2018) c. Maple syrup urine disease 33. b
a. Leucine b. Lysine d. Hartnup’s disease 34. a
c. Methionine d. Proline 44. Guthrie’s bacterial inhibition test detects: 35. b
33. Which of the following is a feature of Phenylketonuria? a. Phenyl pyruvate 36. a
 (Recent Question Jan 2018) b. Phenyl alanine 37. c
a. Loss of deep tendon reflexes c. Phenyl lactate 38. a
b. Mental retardation d. All of the above 39. a
c. Macrocephaly 45. First line therapy in Phenylketonuria is? 40. c
d. All a. Limiting the substrate for deficient enzyme. 41. c
34. Serotonin is also known as?  b. Replacement of the deficient product. 42. b
 (Recent Question Jan 2018) c. Replacement of the defective enzyme. 43. d
a. 5-hydroxytryptamine (5-HT) d. Giving the missing amino acid by diet. 44. d
b. N-methyl phenylamine 45. a
c. 3-Methoxytyramine
d. Phenethylamine
 46. True regarding phenylketonuria is?  51. In Cystinuria, all of the following amino acids are
 (PGI May 2018) excreted in urine, EXCEPT: 
176 a. Musty odour is due to phenylalanine in sweat  (Recent Question Jan 2018)
b. Deficient enzyme is phenylalanine hydroxylase a. Cystine b. Ornithine
c. Autosomal dominant c. Leucine d. Arginine
d. May be associated with impaired mental devel- 52. VMA is excreted in urine in : 
opment  (Recent Question Jan 2018)
e. Infants are normal at birth a. Alkaptonuria
47. True regarding PKU is all EXCEPT:  b. Phenyl ketonuria
 (Recent Question Jan 2018) c. Diabetic ketoacidosis
a. Due to deficiency of phenyl alanine hydroxylase d. Pheochromocytoma
b. Neurological symptoms are due to excess phenyl 53. HIAA is excreted in urine in : 
alanine  (Recent Question Jan 2018)
c. Blood phenyl alanine level > 20 mg/dl causes severe a. Alkaptonuria
disease b. Carcinoid syndrome
d. Method of choice for screening is urinary phenyl c. Albinism
alanine by Guthrie’s test d. Phenyl ketonuria
48. Tyrosinosis is caused due to deficiency of which 54. Mutation in Hartnup disease:  (JIMPER May 2018)
enzyme?  (Recent Question Jan 2018) a. SLC5A 18
a. Fumaryl acetoacetate hydrolase b. SLC6A 19
b. p-hydroxy phenyl pyruvate dehydrogenase c. SLC7A 9
c. Tyrosine transaminase d. SLC3A 1
d. Tyrosine ligase 55. In homocystinuria, what should be given to the
49. Fish odour syndrome is caused due to deficiency of patient?
which enzyme :  (Recent Question Jan 2018) a. Pyridoxine
a. Fumaryl acetoacetate hydrolase b. Folic acid
b. Methane monoxygenase c. Vitamin B12
c. Mono oxygenase 3 d. All
d. D-amino acid oxidase 56. In cystinuria, following amino acids are reabsorbed
50. All are true regarding this disease EXCEPT:  EXCEPT?
M  (AIIMS May 2018) a. Lysine
C a. Due to PAH enzyme defect b. Arginine
Qs b. White patch of hair due to tryptophan deficiency c. Citruline
Ans. c. Phenyl acetate positive in urine d. Ornithine
46. b,d,e d. Mental retardation is present
47. d
48. a
49. c
50. b
51. c
52. d
53. b
54. b
55. d
56. c
 Answers with Explanations
177
1. Ans. (a) Tryptophan •• 6 amino acids (Glycine, Alanine, Serine, Threonine,
Cysteine and Hydroxy-Proline) forms Pyruvate and
[Ref: Harper 30th/e pg. 17] then it forms Acetyl CoA via Pyruvate Dehydrogenase

CHAPTER 5  CHEMISTRY AND METABOLISM OF AMINO ACIDS

•• Indole ring is present in Tryptophan. complex. Alanine is the most glucogenic. Tyrosine is
•• Benzene ring is present in Phenylalanine, Tyrosine. not in this list.
•• Imidazole ring is present in Histidine.
•• Guanidinium group is associated with Arginine. 7. Ans. (a) Proline

[Ref: Harper 30th/e pg. 38]

2. Ans. (c) Glutamic acid
•• Amino acids not found in alpha helix are: Proline,
[Ref: Harper 30th/e pg. 17] Glycine, Tryptophan, Aspartate, Glutamate and
•• Glutamate/Glutamic acid is a negatively charged Valine
polor acidic amino acid. Other polar amino acids are
Aspartic acid, Histidine, Arginine, Lysine, Cysteine. 8. Ans. (d) All of the above
Tyrosine, Threonine, Serine, Asparagine and [Ref: Harper 30th/e pg. 269]
Glutamine. (There is a controversy for the polarity of
Glycine and Tyrosine. So, according to the question •• Sulphur containing amino acid metabolism requires
you have to mark answer) three vitamins i.e. Vitamin B6 (Pyridoxine), Vitamin
B9 (Folate) and Vitamin B12. Folate and vitamin B12
3. Ans. (b) Cysteine are required for one carbon transfer and Pyridoxine is
required in Cystathionine Synthase, which converts
[Ref: Harper 30th/e pg. 666] Homocysteine to Cystathionine.
•• Keratin is a protein present in the hair, nails and outer
layer of skin. Strength and rigidity in keratin is due 9. Ans. (b) Tetrahydrobiopterin
to Cysteine because of its ability to form disulphide [Ref: Harper 30th/e pg. 304]
bonds.
•• Number of disulphide bonds is directly proportional •• Tetrahydrobiopterin is a cofactor for:
to rigidity of keratin i.e. the more the disulphide ƒƒ Tryptophan Hydroxylase
bond, harder the keratin. ƒƒ Phenylalanine Hydroxylase
ƒƒ Tyrosine Hydroxylase
4. Ans. (b) Alanine ƒƒ Nitric Oxide Synthase (NOS)

[Ref: Harper 30th/e pg. 269] 10. Ans. (b) Sarcosine A

•• Creatine = Arginine + Methionine + Glycine. Creatine •• Sarcosine/ N-Methyl Glycine, is an intermediate and N
does not contain alanine by product in Glycine synthesis and degradation S
•• Glutathione = Glutamate + Cysteine + Glycine •• Ergothioneine is a thiourea derivative of histidine, W
•• Carnitine = Lysine + Methionine containing a sulfur atom on the imidazole ring E
•• Methionine (SAM - S-Adenosyl Methionine) is •• Betaine is trimethyl glycine, present in sugar beet R
used actually as methyl donor in both Creatine and •• Carnosine (beta-alanyl-L-histidine), has imidazole- S
Carnitine ring like Histidine, is a dipeptide molecule, made up
of beta-alanine and histidine. It is present in muscle WITH
5. Ans (b) Threonine and brain tissues.
E
[Ref: Harper 30th/e pg. 22] 11. Ans. (c) Cysteine X
•• All amino acids have 1 assymetric C/chiral C. •• Taurine, major constituent of bile is synthesized from
P
Exceptions: cysteine.
L
0 - Glycine A
2 - Isoleucine, Threonine N
A
6. Ans. (c) Tyrosine T
I
[Ref: Harper 30th/e pg. 165 figure,16-4] O
•• Glucogenic amino acids are those that forms Pyruvate N
or any TCA intermediate. S
 12. Ans. (c) Homocarnosine 15. Ans. (a) Serine
178 [Ref: Harper 30th/e pg. 318] •• OH containing amino acid has maximum tendency
to bind phosphate. OH containing amino acids are
Homocarnosine donot contain b-Alanine Tyrosine, Threonine and Serine.
CRO BIOCHEMISTRY

16. Ans. (a) Decreased

[Ref: Lehninger 7th/e pg. 172-173]
•• HbA (normal) and HbS (Sickle cell Hb) differ only in
one amino acid. In HbA, there is Glutamate, which
has one negative charge. HbS has Valine, which is
non polar, so no charge. So we can say that HbS has
one charge less as compared to HbA.
•• Movement in electrophoresis is mainly because of
charge. Here charge is decreased, so movement also


decreases.

17. Ans. (c) Tryptophan

[Ref: Harper 30th/e pg. 308]
•• 3-Hydroxy Kynurenine is one of the intermediate in
tryptophan catabolism, which gets converted to next
intermediate and this conversion requires vitamin
B6/ PLP. So, in B6 deficiency, this reaction does not
occur and 3-Hydroxy Kynurenine is diverted to form
alternate metabolite, Xanthurenic acid, which is
excreted in urine
   Refer Fig. 5.24

18. Ans (d) Pyrimidines

[Ref: Harper 30th/e pg. 52]

•• Glycine is required for synthesis of purines, heme
and creatine.
•• Three amino acids are required for purine synthesis.
13. Ans. (b) Ornithine They are Aspartate, Glutamine and Glycine.
A
N [Ref: Harper 30th/e pg. 314] •• For pyrimidines, two amino acids are required i.e.
S Aspartate and Glutamine.
•• The enzyme Ornithine Decarboxylase (ODC) •• Creatine synthesis requires three amino acids-
W catalyzes the decarboxylation of ornithine (an Arginine + Methionine + Glycine
E intermediate in urea cycle) to form putrescine. •• Haem synthesis requires one amino acid i.e. Glycine
R This reaction is known to be the committed step in
S polyamine synthesis. 19. Ans. (a) Leucine
•• Yohimbine is an indole alkaloid derived from the bark
WITH
of yohimbe tree. Uses are reversal of sedation in dogs [Ref: Harper 30th/e pg. 17]
E and deer, fat burning drug & treatment for erectile •• There are only two ketogenic amino acids i.e.
X dysfunction but there are multiple side effects. Leucine and Lysine. There are five amino acids
P in both glucogenic and ketogenic category i.e.
14. Ans. (a) Acidic Amino Acid Tyrosine, Tryptophan, Threonine, Isoleucine and
L
A [Ref: Harper 30th/e pg. 282] Phenylalanine. Rest 13 amino acids are in glucogenic
N category
A •• All the acidic amino acids (Aspartate, Glutamate) are
T non-essential i.e. they can be synthesized from TCA 20. Ans (c) Tryptophan
I intermediates.
•• But all the basic amino acids (Arginine, Lysine, [Ref: Harper 30th/e pg. 27 and 28]
O
N Histidine) are essential. Arginine and Histidine are •• Basically, this absorption of UV-light is because of
S semi-essential. conjugated double bonds present in the ring.
 •• Proteins absorb UV Light at 280 nm due to aromatic
amino acids. This absorption is maximum for
tryptophan. 179
•• Nucleic acids (DNA) also absorb UV Light at 260 nm
due to nitrogenous-base. This absorption is more for
Purines as compared to pyrimidines.

CHAPTER 5  CHEMISTRY AND METABOLISM OF AMINO ACIDS

•• NAD/NADP absorb at 340 nm
•• Porphyrins absorb at 400 nm (this absorption band of
porphyrins is known as Soret Land)

21. Ans. (d) All

[Ref: Harper 30th/e pg. 20-21]
•• At Isoelectric pH (pI), protein’s net charge is zero,
24. Ans. (b) Glyoxylate
known as Zwitter ion. This structure is insoluble, so
precipitates. [Ref: Harper’s 30thed pg. 283]
•• At acidic pH, protein has positive charge.
•• The formation of glycine from glyoxylate takes place
•• At alkaline pH, protein has negative charge.
by enzymes glycine transaminase which requires
22. Ans. (a) Valine vit B6/PLP as coenzyme.

[Ref: Harper’s 30thed pg. 289, 291]

•• After a protein rich meal, splanchnic tissues release
branched chain amino acids (Valine, Leucine
and Isoleucine) and they are mainly extracted by
peripheral tissues like skeletal muscles.
•• These branched chain amino acids provide energy to 25. Ans. (b) Aspartate and Phenylalanine
brain during fasting state. •• Aspartame is used as artificial sweetner. It is a
•• Therefore, branched chain amino acids have a special dipeptide made up of aspartate & phenyl alanine.
role in nitrogen metabolism. •• It is contraindicated in case of PKU (Phenylketonuria).
•• It is 200 times sweeter than sucrose.

A
N
S
W
E
R
S
WITH

E
X
P
23. Ans. (c) Serine L
26. Ans. (c) Ornithine A
[Ref: Harper’s 30thed pg. 286] N
[Ref: Harper’s 30thed pg. 314] A
•• Selenocysteine is the 21st amino acid, synthesized
co-translationally by modification of a stop codon – •• Rate limiting enzyme of polyamine biosynthesis T
UGA. The precursor amino acid of selenocysteine is is Ornithine decarboxylase, for which substrate is I
Serine, not cysteine. ornithine. O
•• Biosynthesis of selenocysteine requires ATP & the •• Arginine can also be marked as arginine is the source N
carbon skeleton is provided by Serine. of ornithine, by enzyme Arginase. S
 27. Ans. (c) Serine •• Aspartate is similar to glutamate in the same way
that asparagine is similar to glutamine (polar amino
180 [Ref: Harper’s 30thed pg. 284] acids)
•• The first step in the biosynthesis of serine begins with the
oxidation of the hydroxyl group of 3-phosphoglycerate
by NAD+ to produce 3-phosphohydroxy pyruvate.
CRO BIOCHEMISTRY

This reaction is followed by a transamination

reaction in which glutamate transfers an amine
group, becoming –ketoglutarate in the process, to
3-phosphohydroxypyruvate. Finally, hydrolysis and
removal of the phosphate group yields serine.


•• Glycine is similar to alanine in the same way that

valine is similar to leucine

28. Ans. (d) Vitamin C is a cofactor

[Ref: Harper’s 30thed pg. 285; Fig. 27-12]

•• Phenylalanine hydroxylase requires NADPH & THB.
•• All hydroxylases are known as mono-oxygenases or
mixed function oxidases.
A •• All hydroxylases do not require Vitamin C.
N 29. Ans. (b) Histidine
S
•• In purines, N1 is derived from Aspartate, N3 and N9
W
from Glutamine, C4, C5, N7 from Glycine, C6 from
E
CO2 , C2 and C8 from THF (Tetra Hydro Folate).
R •• Alanine is similar to serine in the same way, phenyl-
(Refer Fig. 11.1)
S alanine is similar to tyrosine.
30. Ans. (b) Because they are not produced in the body
WITH 32. Ans. (d) Proline
[Ref: Harper’s 30thed pg. 282]
E [Ref: Harper 30th/e pg. 285]
X •• Essential amino acids are those which cannot be
P produced in the body and are required in the diet. •• Leucine is a branched chain amino acid and all
L These are Methionine, Threonine, Tryptophan, branched chain amino acids are essential amino
A Valine, Isoleucine, Leucine, Phenylalanine, Lysine acids.
N •• Semi-essential amino acids are Histidine and •• Lysine is a basic amino acid and all the basic amino
A Arginine. acids are essential
T •• Rest all are non-essential amino acids •• Methionine is S containing amino acid where S is
I surrounded by two carbons. This kind of amino acid
O 31. Ans. (b) Asparagine is similar to glutamine is difficult to synthesize in body so is essential.
N •• Proline is non essential amino acid and can be
[Ref: Harper’s 30thed pg. 17]
S formed from derived amino acid i.e. ornithine.
 Ornithine forms proline by transamination in the •• Tryptophan synthesizes Melatonin, Niacin, Sero-
presence of enzyme ornithine amino transferase tonin.
(OAT) and vitamin B6. Deficiency of OAT is known to •• Phenylalanine synthesize Tyrosine. 181
cause gyrate atropy of choroid and retina.
39. Ans. (a) Cysteine
33. Ans. (b) Mental retardation [Ref: Harper 30th/e pg. 666]

CHAPTER 5  CHEMISTRY AND METABOLISM OF AMINO ACIDS

[Ref: Harper 30th/e pg. 304] •• Disulfide bonds can only be formed between one
•• Severe mental retardation occurs due to excess amino acid i.e. cysteine, as it has sulfhydryl group
phenylalanine (-SH). Two cysteine gets joined to form cystine
•• Microcephaly (NOT macrocephaly) is a fracture of (having disulfide bond)
PKU.
40. Ans. (c) Homocysteine
•• Tendon reflexes are exagerrated in these patients.
[Ref: Lehninger 7th/e pg. Fig. 821]
34. Ans. (a) 5-Hydroxy Tryptamine (5-HT)
•• Accumulation of derived amino acid homocysteine,
[Ref: Lehninger 7th/e pg. 79] results due to deficiency of Cystathionine Synthase.
•• 5-hydroxytryptamine (5-HT) is also known as This enzyme synthesize cysteine and requires
Serotonin. It is formed by hydroxylation and decar- vitamin B6.
boxylation of tryptophan. •• Homocysteine accumulated causes dangerous
•• Serotonin is a mono-amine neurotransmitter. vascular diseases like atherosclerosis, stroke, myo-
•• 5-hydroxytryptophan is also known as Oxitriptan. cardial infarction and pulmonary embolism.
Refer Fig. 5.24
41. Ans. (c) Aspartate and Glutamate
35. Ans. (b) Lysine
[Ref: Harper 30th/e pg. 286]
[Ref: Harper 30th/e pg. 306] •• Fibrinopeptides (A & B) are formed from fibrinogen
•• Cereals lack lysine and pulses lack methionine on cleavage by thrombin, during blood coagulation.
These are negatively charged due to the presence of
36. Ans. (a) Tryptophan acidic amino acids present in them i.e. aspartate &
glutamate.
[Ref: Harper 30th/e pg 308]
•• When fibrinopeptides are removed from fibrinogen,
•• Tryptophan is responsible for the formation of this forms fibrin monomers. The removal of
melatonin, which regulates the sleep-wake cycle. fibrinopeptides allows ‘knob-hole’ interactions
•• Chemically melatonin is acetyl methyl serotonin. between fibrin monomers, causing the monomers to
polymerize and form protofibrils and fibers. A
37. Ans. (c) Glutamine
N
[Ref: Harper 30th/e pg 283] S
W
•• Glutamine is an amide of glutamate
•• Glutathione is a tripeptide composed of glutamate, E
cysteine and glycine. R
•• Creatine is composed of glycine and arginine. This S
reaction takes place in kidneys.
WITH
•• Purine nucleotides are formed from aspartate,
glutamine and glycine E
•• Pyrimidines are formed form aspartate and gluta- X
mine. P
L
38. Ans. (a) Tyrosine A
[Ref: Harper 30th/e pg. 269] N
A
•• Melanin is synthesized from Tyrosine by enzyme 42. Ans. (b) Phenylalanine T
Tyrosinase which converts tyrosine to melanin. I
•• Maple syrup urine disease is a defect in the
Deficiency of the enzyme causes albinism. O
catabolism of branched chain amino acids i.e. Valine,
•• Tyrosine synthesize melanin, catecholamines, N
Leucine and Isoleucine. As a result these amino acids
Thyroid hormone. S
are excreted in urine. This defect occurs due to the
 inherited deficiency of branched Chain Keto-acid •• Phenylacetate formed is responsible for mousy/
Decarboxylase. musty odour and phenyl pyruvate is responsible for
182 positive ferric chloride test in urine.
43. Ans. (d) Hartnup’s disease •• At birth the infant is normal, as during pregnancy
maternal enzyme converts phenylalanine to tyrosine.
[Ref: Harper 30th/e pg. 308, 557] So screening is done usually 7 days after birth.
CRO BIOCHEMISTRY

•• Hartnup’s disease is autosomal recessive, rare dis-

order. There is failure to reabsorb Tryptophan from 47. Ans. (d) Method of choice for screening is urinary
kidneys. As a result, massive aminoaciduria occurs phenylalanine by Guthrie’s test
(Tryptophan in urine). Tryptophan forms Niacin [Ref: Harper 30th/e pg. 304]
(Vitamin B3) in the body, so deficiency of Tryptophan
will lead to the deficiency of Niacin in the body which •• Guthrie’s test (was the first method used to detect
will further lead to pellagra like symptoms. PKU) is replaced by tandem mass spectrometry
•• Guthrie test detects serum phenyl alanine levels (not
44. Ans. (d) All of the above urinary)
•• FeCl3 detects phenyl alanine levels in urine.


[Ref: Harper 30th/e pg. 304]

•• Guthrie’s bacterial inhibition test is a screening 48. Ans. (a) Fumaryl acetoacetate hydrolase
test for phenylketonuria (PKU) used to detect the [Ref: Harper 30th/e pg. 304]
abnormal presence of phenylalanine metabolites in
blood i.e. Phenyl pyruvate, Phenyl alanine & Phenyl •• Most common tyrosinemia is known as Tyrosinosis
lactate. or tyrosinemia type I. It is caused due to deficiency
•• A small amount of blood sample taken from heel of Fumaryl acetoacetate hydrolase. This enzyme
of the infant and placed in a medium with a strain catalyze the final step in the catabolism of tyrosine.
of Bacillus subtilis, a bacteria that cannot grow
without phenylalanine. β-2-Thienylalanine, an 49. Ans. (c) Mono oxygenase 3
amino acid, is added which inhibits the growth of [Ref: Harper 30th/e pg. 304]
bacteria Bacillus subtilis.
•• If phenylalanine metabolites are present, then these •• Fish odour syndrome/Tri methyl aminuria (auto-
bacterias reproduce, and the test result is positive, somal recessive) is a rare metabolic disorder in which
indicating that patient has PKU. enzyme defect is Flavin containing mono-oxygenase
-3 (FMO3) and malnutrition. Patient has blue eyes,
45. Ans. (a) Limiting the substrate for deficient enzyme. blonde hair and fair skin. On investigation, Guthrie
•• First line therapy in Phenylketonuria & in most test was found to be positive.
metabolic disorders is reducing the substrate. In Refer Fig. 5.41.
A PKU, this is phenyl alanine. The target is to keep
50. Ans. (b) White patch of hair due to tryptophan
N phenyl alanine levels low enough to prevent mental
deficiency
S retardation but should be enough for optimal growth.
W [Ref: Devlin’s textbook of Biochemistry, 7th ed., pg. 769]
E 46. Ans. (b); (d); (e)
•• This is Phenyl ketonuria which is due to PAH enzyme
R [Ref: Harper 30th/e pg. 304] defect
S •• Guthrie test detects serum Phenyl alanine levels
•• Most inborn errors of metabolism are Autosomal
Recessive (AR). Phenyl ketonuria (PKU) is also an •• FeCl3 detects Phenyl alanine levels in urine.
WITH
autosomal recessive disorder. •• Severe mental retardation occurs due to excess
E •• In phenylketonuria, there is absence of enzyme phenylalanine.
X Phenylalanine Hydroxylase. This enzyme converts
P 51. Ans. (c) Leucine
phenyl alanine to tyrosine. As a result, there is
L accumulation of phenylalanine in blood and tyrosine [Ref: Harper 30th/e pg. 269]
A becomes essential. Pheny-lalanine gets converted to
N •• In Cystinuria, there is a mutation in gene which is
phenyl acetate, phenyl lactate and phenyl pyruvate.
A responsible for the reabsorption of basic amino acids
•• Accumulation of phenylalanine causes mental
T & cysteine from kidneys. So amino acids found in
retardation, as excess phenylalanine enters the brain
I high concentration in urine are cystine, ornithine,
and competitively inhibits the uptake of tyrosine
O arginine and lysine. High cysteine in blood will lead
and tryptophan which are important for synthesis of
N to the formation of cystine in urine (made up of two
neurotransmitters.
S cysteines).
 •• These patients are prone to have cystine stones in 54. Ans. (b) SLC6A 19
kidneys. [Ref: Harper 30th/e pg. 308, 557] 183
52. Ans. (d) Pheochromocytoma Hartnup’s disease is due to defect in transport of nuetral
amino acid transporter. This transporter is a protein
[Ref: www.ncbi.nlm.nih.gov/pubmed/17635948] encoded by gene SLC6A19 (located on chromosome 5).

CHAPTER 5  CHEMISTRY AND METABOLISM OF AMINO ACIDS

•• VMA is Vanillyl Mandelic Acid, which is the catabolic There is mutation in this gene in Hartnup's disease.
end product of Catecholamines. It is increased in
Pheochromocytoma & Neuroblastoma. 55. Ans. (d) All
•• All these three vitamins, i.e. Pyridoxine, Folic
53. Ans. (b) Carcinoid syndrome acid, Vitamin B12 are involved in metabolism of
homocysteine.
[Ref: Harper 30th/e pg. 316]
•• Carcinoid syndrome occurs in tumor of argentaffin 56. Ans. (c) Citrulline
cells, which secrete lots of serotonin.
[Ref: Harper 30th/e pg. 269]
•• Develops in some patients of carcinoid tumor and
is characterized by: abdominal cramps, cutaneous •• Amino acids in urine in Cystinuria are Cystine, Orni-
flushing & sweating and diarrhoea. thine, Lysine and Arginine. Citrulline is a derived amino
•• In this syndrome, Tryptophan is excessively used acid, involved in urea cycle.
for the formation of serotonin, which forms excess
HIAA (5-hydroxy indole acetic acid), which is the end
product of serotonin. Refer Fig. 5.26.

A
N
S
W
E
R
S
WITH

E
X
P
L
A
N
A
T
I
O
N
S
 6 Urea Cycle
Overview of Chapter First step in catabolism of amino acids: Transfer of NH3
group from amino acid to alpha keto glutarate forming
•• Transamination glutamate- known as Transamination.
•• Why Ammonia is toxic?
•• Urea cycle and regulation
•• Urea cycle disorders

Fig. 6.2: Amino acid forming its corresponding keto acid by

removal of amino group. On the other hand this amino group is
taken by alpha-keto glutarate forming glutamate
Transamination reactions are reversible. During protein
synthesis, amino acid is formed (reaction goes in left direction).
But during protein catabolism, amino acid lose amino group
to form glutamate (reaction goes in right direction). EXCEPT
TRANSAMINATION for enzyme AST which forms aspartate from glutamate during
During catabolism of amino acids, the first group to be protein catabolism, because aspartate is used in urea cycle as
removed is NH2 group, which is highly toxic for body, it contributes one of the nitrogen of urea.
especially for brain. So there is a special mechanism to throw
this NH2 group out of body, i.e. in the form of urea. Urea cycle
only occurs in liver.
zz Ammonia in aqueous solution is → NH4+ (ionized form)
zz Form that crosses cell membrane → NH3 ( unionized form)

Fig. 6.3: On addition of amino group, alpha keto glutarate forms

glutamate and further it forms glutamine. Similary oxaloacetate
forms aspartate and then it forms asparagine.
All the amino acids undergo transamination to form a
common amino acid, i.e. glutamate. So glutamate is also
regarded as collector of amino groups. Enzymes are
transaminases or aminotransferases, e.g. SGOT and SGPT
zz SGOT: Serum Glutamate Oxaloacetate Transaminase
– also known as AST i.e. Aspartate transaminase –
aspartate loses amino group and forms oxaloacetate. On
Fig. 6.1: Aspartate is a 4 C amino acid and oxaloacetate is a 4C the other hand, alpha keto glutarate takes amino group
keto acid. Glutamate is a 5C amino acid and alpha keto glutarate and forms glutamate.
is a 5C keto acid. Whenever amino group is removed, then keto
group is formed. So on removal of amino group aspartate gets
converted to oxaloacetate and glutamate gets converted to alpha
keto glutarate.
zz SGPT: Serum Glutamate Pyruvate Transaminase – also zz OAT is responsible for the formation of non-essential amino
known as ALT i.e. Alanine transaminase. Alanine loses acid- Proline, from ornithine.
amino group and forms pyruvate. On the other hand zz OAT is present in liver, kidney, retina, brain
185
alpha keto glutarate takes amino group and forms zz Deficiency of OAT is known as Gyrate atrophy of choroid
glutamate. and retina: Ornithine accumulates. This is a rare, autosomal
recessive disease. Clinical features are Vision loss and Cataract

CHAPTER 6  UREA CYCLE

zz Treatment of OAT deficiency → Restrict Arginine as it is a
source of ornithine

zz Transfer of amino group occurs (not loss of NH2 group)

zz Reversible
zz Require PLP (derivative of vitamin B6) as coenzyme

Fig. 6.5: NH2 group of lysine at active site of enzyme is covalently

linked to PLP.

zz Common amino acid – Glutamate

zz Covalent catalysis
zz Involves ping pong mechanism of Bi-Bi reaction
zz Play role in synthesis of TCA intermediates like
oxaloacetate, alpha keto glutarate
zz Also play role in synthesis of non-essential amino acids
e.g. Aspartate, Alanine
zz These enzymes are in mitochondria and cytoplasm of all
cells of body

Synthesis of Glutamine from Glutamate

Enzyme is glutamine synthetase. ATP is used at this step.

Fig. 6.4 Transamination is first reaction in the

catabolism of amino acid

Transamination Reactions
zz Usually named after amino acid that serve as amino
Glutamine is a non-toxic storage and transport form of
group donor, e.g. AST–Aspartate Transaminase (Aspar-
ammonia in blood because it is neutral in nature and can
tate donates amino group to α keto glutarate)
readily move across cell membrane by facilitative diffusion.
zz Only α- amino group can take part in transamination.
It is transported to liver. In liver, glutamine is again converted
A dditional E dge to glutamate by enzyme glutaminase, which is a hydrolase.

zz One exception δ-amino group (non α) of ornithine can take

part in transamination, where enzyme is OAT (Ornithine T
Amino Transferase) H
E
O
R
Y
Contd…
 Then in liver glutamate undergo oxidative deamination is Glutamate dehydrogenase (GDH). This enzyme is very
to form alpha keto glutarate, releasing ammonia, which special as it can use NAD or NADP both. Alpha keto glutarate
186 can enter urea cycle. The enzyme of oxidative deamination formed is the carbon skeleton left which can be degraded via
TCA cycle.
CRO BIOCHEMISTRY


Fig. 6.6: Transamination in peripheral tissues to form glutamate and then glutamine formed by glutamine synthetase. Glutamine is the
transport form in blood from tissues to liver. In liver glutamine converted to glutamate by glutaminase and then glutamate is oxidatively
deaminated to alpha-keto glutarate, releasing NH3, which enters urea cycle.

Glutamate is the only amino acid that can undergo rapid

F undamental B ox
zz
oxidative deamination.
zz 17 amino acids can undergo transamination to form glutamate
zz But rest 3 amino acids cannot undergo transamination
reactions and hence does not form glutamate. They are:
POLYTHENE (Mnemonic)
zz PO — Proline (and hydroxyl-proline also)
zz LY — Lysine
zz THENE — Threonine

Q. Why Glutamine Synthetase is first line NH3 trapping,

why not transamination?
A. Because Glutamine Synthetase uses free NH3.
Transamination is using NH2 group from amino acid.

Transdeamination = Transamination (All organs) + Oxidative H igh R eturn

deamination (Liver) zz Major transporter of NH3 from:
T
BODY → Glutamine
H
BRAIN → Glutamine
E MUSCLE → Alanine (Fig. 6.14)
O
R
Y
 A dditional E dge C linical B ox
Why hyperammonemia occurs in kidney failure? 187
Asparagine synthetase Glutamine synthetase
Normally once urea is formed in liver, then it is transported
Source of amino group is Source of amino group is Free from liver to kidneys. But some of it enters intestine where it is
Glutamine NH3 converted to ammonia by bacteria. This ammonia released in
+NH2 +NH2 faeces but also some of it re-enters circulation. In case of kidney

CHAPTER 6  UREA CYCLE

Asparatate Asparagine Glutamate Glutamine
failure, increased urea will form increased intestinal NH3, which
No role in transport of NH3 Very important role in transport is reabsorbed into blood. So hyperammonemia in kidney failure.
of NH3 from peripheral tissue Oral Antibiotics reduces these bacteria, which produce ammonia.
to liver

UREA CYCLE
WHY AMMONIA IS TOXIC?
H igh R eturn
zz Organ – Liver
zz Organelle – both mitochondria and cytoplasm (Any pathway
which occurs both in mitochondria and cytoplasm – starts
from mitochondria, so urea cycle starts from mitochondria)
zz Rate limiting enzyme – CPS-I

Sources of Urea
Excess ammonia (or NH2 group) present will increase the 1. α amino group of amino acid by transamination (most
formation of glutamate from alpha keto glutarate and further abundant source)
increased glutamine formation from glutamate. So glutamine 2. Porphyrins (PBG deaminase reaction)
increases and alpha keto glutarate and glutamate decreases. 3. Purines and pyrimidines
So effects are:
1. Depletion of alpha-keto glutarate – This is intermediate
of TCA cycle, so TCA cycle stops and brain is affected as
brain is highly dependent on this aerobic pathway. As a
result, there is reduced availability of ATPs in brain cells.
2. Glutamate decreased- normally glutamate forms GABA,
which is inhibitory neurotransmitter. So if glutamate
is decreased then GABA also decreases. This leads to
excitation in brain presenting as fine tremors in patients
of hyperammonemia. Fig. 6.8: Source of urea Carbon and Nitrogen.
3. Increased glutamine is osmotically active, which leads
to cell swelling, especially in brain cells. In patient,
cerebral oedema and vomiting occurs. This is known as Steps of Urea Cycle (Fig. 6.9)
Ammonia Encephalopathy.
First 2 steps occur in mitochondria and rest in cytoplasm
1. CO2 and NH3 combines to form carbamoyl phosphate.
Two ATPs are used at this step. Enzyme is CPS-I
(Carbamoyl phosphate synthetase-I). This is the rate
limiting enzyme (CPS-II is in pyrimidine synthesis in
cytoplasm).
2. Carbamoyl phosphate combines with ornithine to
form citrulline and phosphate released as inorganic
phosphate. Enzyme is Ornithine Transcarbamoylase
(OTC). This enzyme is absent in brain (therefore brain T
cannot synthesize urea) and is the most common urea H
cycle enzyme defect. E
Fig. 6.7: Clinical features of Hyperammonemia O
R
Y
 zz Citrulline is then transported out from mitochondria into 4. Arginosuccinate is cleaved to fumarate and arginine by
cytoplasm. Ornithine and Citrulline antiport present in IMM. enzyme Argininosuccinate Lyase.
188 Both these are basic amino acids. They are not present in 5. Enzyme Arginase breaks arginine into urea and
proteins as there are no codons for them. ornithine. Urea is transported to kidneys from liver via
blood and then excreted in urine.
3. Citrulline gets combined with aspartate to form
CRO BIOCHEMISTRY

arginosuccinate. In this step ATP gets converted to AMP. zz Ornithine is transported from cytoplasm into mitochondria.
This is equivalent to 2 molecules of ATP used. Name of
enzyme is Argininosuccinate synthetase.


Fig. 6.9: Urea Cycle

H igh R eturn
Q. How many ATPs used to synthesize one molecule of Urea?
zz It is a controversial thing. So understand carefully.
zz Actually 3 ATPs (2 ATPs at CPS-I step and one ATP at arginino succinate synthetase step) but 4 high energy phosphates (2 high energy
phosphates at CPS-I step and two at arginino succinate synthetase step because here ATP gets converted to AMP) are used to make
one urea.
zz So now depending upon the question, you have to mark the answer. E.g.
Q. Which of the following used to make one molecule of urea:
a. 3 ATP b. 4 ATP
T c. 3 ATP & 4 high energy phosphates d. 2 ATP
H A. (c) (3 ATP & 4 high energy phosphates). This is the best answer to be marked in these options
E Q. How many ATPs are used to make one molecule of urea:
O a. 3 ATP b. 4 ATP c. 1 ATP d. 2 ATP
R A. (b) 4 ATP.
Y Because if you count total, then it is 4 ATP. ATP to AMP conversion is counted as 2 ATPs.
 A dditional E dge
zz No (nitric oxide) or Endothelium derived relaxing factor (EDRF) 189
is synthesized from arginine.

CHAPTER 6  UREA CYCLE

H igh R eturn
zz Enzyme deficient in brain: OTC (Ornithine Trans Carbamoylase)
zz Enzyme deficient in Kidneys: Arginase
But only last enzyme is deficient in kidneys. Therefore:
zz End product in kidneys → Arginine
zz Major source of arginine (a semi-essential amino acid) → Kidneys

Regulation of Urea Cycle

NAG is an allosteric activator of CPS-I (Rate Limiting Enzyme).
NAG is synthesized from N-acetyl glutamate synthase or NAG
synthase, for which arginine is the activator. NAGS is regarded
as 6th enzyme of urea cycle. UREA CYCLE DISORDERS
Main clinical features:
zz Hyperammonemia
zz Encephalopathy
zz Respiratory alkalosis
zz No mental retardation

Table 6.1: Urea Cycle Disorders (UCDs)

Disease name Enzyme deficiency Substrate accumulated

} Hyperammonemia Type I CPS-I (Carbamoyl Phosphate NH3

Most synthetase-I)
severe
Hyperammonemia Type II OTC (Ornithine Transcarbamoylase) NH3
(Most common UCD) OMP
UMP (Fig. 6.10)
Orotic acid
Citrullinemia Type I Argino-Succinate synthetase NH3 (UCDs)
Citrulline

Least
Hyper
} Arginosuccinic aciduria Argino-Succinate Lyase NH3
Arginosuccinic acid

}
ammo Hyperargininemia Arginase NH3
nemia T
Arginine
H
Citrullinemia Type II Citrin (Asparate-Glutamate Transporter) Citrulline E
Urea cycle O
HHH Syndrome Ornithine transporter Hyperammonemia transporter
R
(Autosomal Recessive) Hyperornithinemia defect
Y
Homocitrullinuria
Hyperammonemia Type I and II are most severe as ammonia is present in inorganic form. Ammonia is made organic in
citrulline.
 OTC Deficiency
190 zz Most common urea cycle disorder
zz Only this is X-linked partial dominant inheritance (rest all are AR), predominantly affecting males
N-Acetyl Glutamate Synthase Deficiency Same like Hyperammonemia Type I
CRO BIOCHEMISTRY

Urea Cycle Transporter Defects

1. HHH syndrome or Ornithine Transporter Defect → Autosomal Recessive
2. Citrullinemia Type II → Defect of Citrin transporter.
 Autosomal Recessive
 Citrin transports aspartate from mitochondria to cytoplasm, to be used in urea cycle
zz Citrullinemia Type I — Defect of AS Synthetase
zz Citrullinemia Type II — Defect of Citrin Transporter


Abbreviations: OMP-Orotidine monophosphate; UMP-Uridine monophosphate

Fig. 6.10: OTC deficiency (Excess carbamoyl phosphate from mitochondria enters cytoplasm and takes part in pyrimidine synthesis)

Treatment of Urea Cycle Disorders

1. Give Arginine (1st line Treatment )
 An essential amino acid
 Provides ornithine (required in 2nd step of urea
cycle)
 Activator of NAGS
T  But it is contraindicated in Arginase deficiency
H 2. Acylation therapy or NH3 scavenging agents (N2 excretion
E in form other than urea) (Fig. 6.11)
O 3. Restrict Protein intake. Restrict to 50% as some amount is
R required for optimal growth Fig. 6.11: With Phenyl acetyl glutamine two NH2 groups are
Y excreted. With Hippuric acid only one NH2
is excreted. So Phenyl Butyrate is more effective
 Pearls of the Chapter
zz Glutamate is the only amino acid that can undergo oxidative deamination
191
zz Arginine is the immediate precursor of urea
zz Urea cycle only occurs in liver
zz Transamination reactions are Reversible and require PLP (derivative of vitamin B6) as coenzyme

CHAPTER 6  UREA CYCLE

zz Glutamine synthetase converts glutamate to glutamine (ATP used) and glutaminase converts glutamine to glutamate (no ATP used)
zz Glutamine is a non-toxic storage and transport form of ammonia in blood
zz GDH (glutamate dehydrogenase) helps in oxidative deamination of glutamate to release ammonia in liver for entry into urea cycle. This
enzyme can use either NAD or NADP.
zz Amino acids which cannot form glutamate and cannot undergo transamination are Proline, hydroxyl-proline, Lysine and Threonine
zz Major transporter of NH3 in body is glutamine, in brain → glutamine and in muscle → alanine
zz Urea cycle occurs in both Mitochondria and Cytoplasm
zz Rate limiting enzyme of Urea cycle – CPS-I
zz Ornithine and Citrulline antiport is present in IMM, which transports citrulline from mitochondria to cytoplasm and ornithine is
transported into the mitochondrial matrix.
zz NAG synthase is regarded as 6th enzyme of urea cycle, which synthesize NAG (N-Acetyl glutamate). NAG is the allosteric activator of
CPS-I (Rate limiting enzyme of Urea cycle)
zz 4 ATPs are used to synthesize one molecule of Urea
zz Ornithine has a catalytic role in urea cycle
zz Carbon and oxygen of urea is derived from CO2 or bicarbonate (HCO3–). Two nitrogens are derived from aspartate and ammonia
zz Urea cycle enzyme deficient in brain: OTC (Ornithine Trans Carbamoylase)
zz Urea cycle enzyme deficient in kidneys: Arginase
zz End product of urea cycle in kidneys → arginine
zz Major source of arginine (a semi-essential amino acid) → kidneys
zz Main clinical features of urea cycle disorders are hyperammonemia, encephalopathy and respiratory alkalosis
zz OTC (Ornithine Transcarbamoxylase) Deficiency is the most common urea cycle disorder
zz Phenyl Butyrate is more effective than sodium benzoate in treating urea cycle disorders
zz Increased gluconeogenesis from amino acids increase urea formation

T
H
E
O
R
Y
 192 Multiple Choice Questions
Transamination and NH3 Transport 10. Which of the following enzyme(s) is/are NOT
1. The process in which amino group of amino acid is involved in urea cycle: (PGI May 2013)
transferred to keto acid and keto group of keto acid is a. Glutamate dehydrogenase
transferred to amino acid is called b. Argininosucinate synthetase
 (Recent Question 2018) c. α- KG dehydrogenase
a. Phosphorylation b. Transamination d. Isocitrate dehydrogenase
c. Deamination d. Decarboxylation e. Fumarase
2. Glutamate is formed from which amino acid? 11. Compound that enters into urea cycle and regene-
 (Recent Question 2017) rated is
a. Threonine b. Alanine a. Ornithine b. Citrulline
c. Proline d. Lysine c. Arginine d. Aspartate
3. Ammonia from brain is trapped by: 12. Urea is formed from which substrate:
 (Recent Question 2017) a. Arginine b. Ornithine
a. Urea b. Glutamate c. Citrulline d. Alanine
c. Glutamine d. Glycine 13. How many ATPs are used by urea cycle ?
4. In transaminases, PLP is covalently attached to a. 1 b. 3
which amino acid? c. 2 d. 4
a. Glutamate b. Lysine 14. Carbamoyl group in ureotelic animals is transferred
c. Alanine d. Threonine to:
5. Co-enzyme required for the synthesis of storage and a. Urea b. Uric acid
transport form of ammonia in blood and brain is: c. Creatine d. Ornithine
a. ATP 15. Creatinine, NO & Urea are synthesized from which
b. GTP amino acid ? (Recent Question Jan 2019)
c. Thiamine pyrophosphate a. Arginine b. Citrulline
M d. NADPH c. Aspartate d. Glycine
C 16. Nitric oxide is synthesized from which amino acid:
Qs Urea Cycle  (Recent Question Jan 2018)
Ans. 6. In urea cycle, hydrolysis occurs during a. Arginine b. Serine
 (PGMEE 2013) c. Threonine d. Lysine
1. b a. Cleavage of arginine 17. Nitric oxide acts by increasing: 
2. b b. Formation of ornithine  (Recent Question Jan 2018)
3. c c. Formation of argininosuccinate a. BRCA 1 b. BRCA 2
4. b d. Formation of citrulline c. Interleukin d. cGMP
5. a 7. Carbamoyl Phosphate Synthetase I (CPS-I) is which 18. Amino acid linking Kreb’s cycle & urea cycle: 
6. a one of the following? (PGMEE 2006) (Recent Question Jan 2019 Q)
7. b a. Cytosolic enzyme a. Aspartate b. Fumarate
8. b b. Hepatic mitochondrial enzyme c. Alanine d. Arginine
9. c,d,e c. Lysosomal enzyme 19. Ammonia causes depletion of which of the following
10. a,c d. All of the above in TCA cycle?  (Recent Question Jan 2019 Q)
d,e 8. Not a metabolic product of urea cycle-(PGMEE 2015) a. Oxaloacetate
11. a a. Ornithine b. Alanine b. Alpha-keto glutarate
12. a c. Citrulline d. Arginine c. Fumarate
13. d 9. True about urea cycle: (PGI May 2015) d. Malate
14. d a. Nitrogen of the urea comes from alanine and 20. Glutamine is increased in CSF, blood & urine in which
15. a ammonia defect:  (Recent Question Jan 2019 Q)
16. a b. Uses ATP during conversion of Argininosuccinate to a. Argininosuccinate synthetase
17. d arginine b. OTC
18. a c. On consumption of high amount of protein, excess c. CPS-I
19. b of urea formed d. Arginase
20. c d. Occur mainly in cytoplasm 21. Urea cycle occurs in:  (FMGE Nov 2018)
21. c e. Synthesis of Argininosuccinate consumes energy a. Mitochondria b. Cytoplasm
c. Both a and b d. ER
Urea Cycle Disorders 24. Phenylbutyrate is used in urea cycle disorders
22. Which intermediate of citric acid cycle is used in because it (Recent Question 2016)
detoxification of ammonia in brain?  a. Scavenges nitrogen 193
 (Recent Question 2018) b. Activates enzymes
a. Citrate c. Maintains renal output
b. Succinate d. Maintains energy production
c. Alpha-keto glutarate 25. In all of the following enzyme deficiencies, hyper-
d. Oxalo-acetate ammonemia is a common feature, EXCEPT:
23. A patient presented to casualty with nausea, vomiting. a. Argininosuccinate synthetase
Intravenous glucose was given and the patient b. Carbamoyl Phosphate synthetase (CPS-I)
recovered. After few months, patient presented with c. Ornithine transcarbamoylase (OTC)
same complaints. Blood glutamine was found to be d. Ornithine amino transferase
increased. Also uracil levels were raised. What is the 26. Cause of HHH syndrome (Hyperornithinemia-Hyp-
diagnosis? (May AIIMS 2015) erammonemia-Homocitrullinuria) is:
a. CPS-I deficiency a. Deficiency of ornithine transcarbamoylase
b. Arginino succinate synthetase deficiency b. Defect in ornithine transporter
c. CPS-II deficiency c. Deficiency of ornithine aminotransferase
d. Ornithine trans carbamoylase deficiency d. All of the above

M
C
Qs
Ans.
22. c
23. d
24. a
25. d
26. b
 Answers with Explanations
194
1. Ans. (b) Transamination •• Arginase, the final enzyme of urea cycle is a hydrolase
i.e. it uses water to break the bond in arginine and
CRO BIOCHEMISTRY

[Ref: Harper 30th/e pg. 679-680] release urea.

•• Transaminase or amino-transferase transfers alpha
amino nitrogen to alpha keto glutarate, forming 7. Ans. (b) Hepatic mitochondrial enzyme
glutamate. [Ref: Harper 30th/e pg. 293]
•• Alanine amino-transferase (ALT) and Aspartate
amino-transferase (AST) catalyze the transfer of •• CPS-I (Carbamoyl Phosphate Synthetase I) is the
amino group forming glutamate. These reactions are rate limiting enzyme of urea cycle. Urea cycle
reversible. occurs in liver. The first step i.e. CPS-I step occurs in
mitochondria. So CPS-I is a hepatic mitochondrial
2. Ans. (b) Alanine enzyme.


[Ref: Harper 30th/e pg. 679-680] 8. Ans. (b) Alanine

•• Amino acids undergo transamination to form [Ref: Harper 30th/e pg. 291]
Glutamate. But 4 amino acids cannot undergo
transamination to form Glutamate. These are Proline, •• Metabolic products of urea cycle are: Ornithine, Ar-
Lysine Threonine and hydroxy proline. ginine, Argininosuccinate, Citrulline and Carbamoyl
Phosphate.
3. Ans. (c) Glutamine
9. Ans. (c); (d); (e)
[Ref: Harper 30th/e pg. 291]
[Ref: Harper 30th/e pg. 291]
•• The transport form of ammonia in body and from
brain is Glutamine. From muscles, it is Alanine. •• Consumption of high amount of protein, produces
more amino groups, which leads to the formation of
4. Ans. (b) Lysine excess urea.
•• Urea cycle occurs both in mitochondria and cyto-
[Ref: Harper’s 30thed pg. 291] plasm (only two reactions occur in mitochondria).
•• Transaminases transfer amino groups in reactions. •• Synthesis of argininosuccinate converts ATP to AMP.
Coenzyme required is Vitamin B6 i.e. active form •• Carbon of urea is derived from CO2 or bicarbonate.
PLP (Pyridoxal phosphate). This PLP (carboxyl Nitrogen is derived from ammonia and aspartate.
A group) binds to ɛ – amino group of lysine residue in
the enzyme with imine bond known as Schiff base. 10. Ans. (a); (c); (d); (e)
N
S [Ref: Harper 30th/e pg. 291]
5. Ans. (a) ATP
W •• Enzymes of urea cycle are: CPS-I (Carbamoyl Phos-
E [Ref: Harper 30th/e pg. 679-680] phate Synthetase-I), Ornithine Transcarbamoylase
R •• Storage and transport form of ammonia in blood and (OTC), Argininosuccinate Synthetase, Argininosuc-
S brain is glutamine. cinate Lyase and Arginase

WITH 11. Ans. (a) Ornithine

E [Ref: Harper 30th/e pg. 293]

X
•• Ornithine has a catalytic role in urea cycle (same like
P
•• For the synthesis of glutamine, glutamine synthetase oxaloacetate in TCA cycle)
L
A binds with coenzyme ATP and then reacts with
glutamate. (All synthetases require ATP) 12. Ans. (a) Arginine
N
A 6. Ans. (a) Cleavage of Arginine
[Ref: Harper 30th/e pg. 293]
T •• Arginine as substrate is broken down by enzyme
I [Ref: Harper 30th/e pg. 290-91] Arginase to release Ornithine & urea. On the other
O hand, nitrogens of urea are derived from ammonia
N and aspartate.
S
13. Ans. (d) 4 17. Ans. (d) cGMP

[Ref: Harper 30th/e pg. 289] 195

Energetics of urea cycle:
•• 2 ATPs are used in CPS-I step
•• At argininosuccinate synthetase step, ATP gets

CHAPTER 6  UREA CYCLE

converted to AMP (equivalent to 2 ATPs used)
•• So in total, 4 ATPs are used to make one molecule of
urea
•• But to be very precise, best answer is 3 ATP & 4 high
energy phosphates (if given in option).

14. Ans. (d) Ornithine

[Ref: Harper’s 30thed pg. 291, 293]

•• In urea cycle, Carbamoyl phosphate combines with
Ornithine to form Citrulline. This reaction occurs
in mitochondria & is catalyzed by enzyme OTC
(Ornithine Transcarbamoylase).

15. Ans. (a) Arginine

[Ref: Harper 30th/e pg. 289]

•• Creatinine formation uses Arginine, Glycine & SAM. •• NO (Nitric oxide) activates enzyme Guanyl cyclase,
Urea is formed from Arginine by enzyme Arginase. which converts GTP to cyclic GMP, which causes
NO is synthesized from Arginine by enzyme NOS relaxation of smooth muscle cells.
(Nitric Oxide Synthase).
18. Ans. (a) Aspartate
16. Ans. (a) Arginine [Ref: Harper 30th/e pg. 290]
[Ref: Harper 30th/e pg. 289] •• Aspartate and fumarate makes a link between Kreb’s
•• NO is also called as Endothelium Derived Relaxing cycle & urea cycle. But fumarate is not an amino acid.
Factor (EDRF).
•• NO (nitric oxide) is synthesized from arginine
by enzyme NOS (Nitric oxide synthase) in the
endothelial cells. A
•• The vasodilator – nitroglycerin also enters smooth N
muscle cells, where its metabolism also leads to the S
formation of NO. W
•• There are three isoforms of NOS (Nitric oxide E
synthase) R
a. nNOS - neuronal S
b. iNOS- inducible
c. eNOS- endothelial WITH

Kreb’s Bicycle E
X
P
L
A
N
A
T
I
O
N
S
 19. Ans. (b) Alpha-keto glutarate 25. Ans. (d) Ornithine amino transferase

196 [Ref: Lehninger 7th/e pg. 686, 687] [Ref: Harper 30th/e pg. 291]
•• In hyperammonemia (i.e. excess amino groups in •• Excess ornithine from urea cycle is taken by enzyme
cells), alpha keto glutarate (intermediate of TCA) tries OAT (Ornithine amino transferase).
•• OAT deficiency leads to gyrate atrophy. In this
CRO BIOCHEMISTRY

to use excess amino group in cells to form glutamate.

This leads to depletion of Alpha-KG. deficiency, plasma, urine, spinal fluid and aqueous
humour concentration of ornithine levels are 10-20
20. Ans. (c) CPS-I times higher than normal.
•• Hyperammonemia is absent because urea cycle is
[Ref: Harper 30th/e pg. 291]
not disrupted in this deficiency. (Hyperammonemia
•• In urea cycle defect, hyperammonemia occurs. So, occurs in enzyme deficiency of Urea cycle).
alpha keto glutarate gets converted to glutamate, All the other options given in the question are enzyme
which further forms glutamine. So glutamine is deficiencies of urea cycle.
increased in all urea cycle defects. Maximum increase
occurs in CPS-I deficiency.


21. Ans. (c) Both

[Ref: Harper 30th/e pg. 49]

•• Urea cycle occurs both in cytoplasm and mitochondria.
•• Other pathways which occur both in mitochondira
and cytoplasm are Haem synthesis, Gluconeogenesis

22. Ans. (c) Alpha-keto glutarate

[Ref: Lehninger 7th/e pg. 686, 687]

Glutamine is an amide of Glutamic acid which is formed
from alpha-keto glutarate. 26. Ans. (b) Defect in ornithine transporter
•• Glutamine is a safe form of ammonia storage and
transport. [Ref: Harper 30th/e pg. 291]
•• Defect in ornithine transporter leads to HHH
23. Ans. (d) Ornithine trans carbamoylase deficiency
syndrome (Hyperornithinemia-Hyperammonemia-
[Ref: Lehninger 7th/e pg. 688, 690] Homocitrullinuria).
•• This prevents ornithine transport into mitochondria
A •• Increase in blood glutamine indicates hyper-
and hence if in the cytoplasm (Hyperornithinemia)
N ammonemia, which occurs in all enzyme defects of
and its reduced ability to clear Carbamoyl phosphate
S urea cycle. Increased uracil indicates Ornithine Trans
and Ammonia (Hyperammonemia).
W Carbamoylase deficiency. If Citrulline is increased
•• Mitochondrial Carbamoyl Phosphate carbamoylates
E then it is argininosuccinate synthetase deficiency.
Lysine to Homocitrulline leading to Homocitrulli-
R •• CPS-II (Carbamoyl Phosphate Synthetase-II) is
nuria.
enzyme involved in pyrimidine synthesis, not in urea
S cycle.
WITH
24. Ans. (a) Scavenges nitrogen
E
X [Ref: Lehninger 7th/e pg. 689, 689f ]
P •• One of the treatment of urea cycle disorder is the use
L of NH3 scavenging agents.
A •• These are Phenylbutyrate and Sodium benzoate.
N
A
T
I
O
N
S
Proteins
7
Overview of Chapter AMIDE/PEPTIDE BOND
•• Amide/Peptide bond Peptide bond is always formed between the alpha carboxy
group of one amino acid and the alpha amino group of
•• Protein structures
another.
•• Protein sequencing zz If amide bond is present in proteins → known as peptide
•• Chaperones bond
•• Glycoproteins zz Peptide bond has partial double bond character
•• Haem zz Rigid and planar
•• Fibrous Proteins zz Uncharged (neither gain nor loose protons) but polar
•• Plasma Proteins (involved in hydrogen bonds).
•• Chromatography zz The double bond is in trans configuration. (Fatty acids
double bond is in cis configuration).
•• Electrophoresis
zz If < 50 amino acids in a chain → known as a peptide
•• Protein Precipitation Reactions
zz If > 50 amino acids then it is known as polypeptide or
protein.
zz Proteins are polymers of amino acids.
zz Amino acids are joined by strong covalent amide/pep-
tide bond.

Fig. 7.1: Formation of amide/peptide bond by joining two amino acids. This amide/peptide bond is a strong covalent bond. Sequence of
amino acid is read from N- to C- terminal

Fig. 7.2: Towards left is N-terminal and towards right is C-terminal of this tripeptide
(Sequence is always read from –N to –C terminal).
 Peptide bond or Eupeptide bond – between α NH2 group PROTEIN DIGESTION
and α- carboxy group
198 Isopeptide bond – An amide bond which is formed with non Digestion begins in stomach. HCl from parietal cells just
α-amino group or non α-carboxy group. denature the proteins and activates pepsinogen to pepsin.
zz Occurs post translationally Pepsin is an endopeptidase.
Makes proteins resistant as proteases cannot break
CRO BIOCHEMISTRY

zz
isopeptide bond Pancreatic Enzymes
zz Can be formed spontaneously or enzymatically Endopeptidases: Cut at the carboxy terminal of specific
zz Usually seen with those amino acids which have NH2 or amino acids.
COOH group in their side chain e.g. Lysine, Glutamate, zz Trypsin: Cleaves at carboxy terminal of basic amino
Aspartate acids e.g. Arginine and Lysine
zz Chymotrypsin cleaves at carboxy terminal of bulky
hydrophobic amino acids e.g. Phenylalanine, Tyrosine,
Tryptophan, Leucine and Methionine
zz Elastase: Cleaves at carboxy terminal of small neutral


aliphatic amino acids e.g. Glycine, Alanine, Serine

Exopeptidases: Cut peptide bond at the C-terminal of protein
Carboxypeptidases A: Preferably acts on alanine and
branched Chain Amino Acids (BCAA)
Carboxypeptidases B: Preferably acts on basic amino acids
e.g. Arginine and Lysine

Examples: Small Intestinal Enzymes

1. Glutathione – (ϒ-carboxy of Glutamate binds with α NH2
of Cysteine) zz Aminopeptidase: An exopeptidase, cut peptide bond at
2. Ubiquitin (Glycine of ubiquitin) binds with Lysine of N-terminal of protein
target protein for protein degradation.) zz Dipeptidases and Tripeptidases: Cleaves di- and tri-
3. Ubiquitin with each other. peptides into their individual amino acids, which are
absorbed.

T
H
E
O
R
Y
Fig. 7.3: Digestion of proteins
ABSORPTION OF AMINO ACIDS STRUCTURE OF PROTEINS
1. Na dependent secondary active transport at the apical zz Primary: The sequence of amino acids joined by peptide 199
side of membrane (same like glucose transport) bond is known as primary structure
2. At basolateral membrane – Sodium Independent trans- zz Secondary: Folding of primary structure is secondary
porter – transports free amino acids into portal vein. Then structure e.g. alpha helic and beta sheet, beta turn
either they are metabolized in liver or in general circulation

CHAPTER 7  PROTEINS
zz Tertiary: Single fully folded functional polypeptide chain
(specially BCAA are metabolized in general circulation).
zz Quarternary: More than one fully folded functional
polypeptide chains

Myoglobin has only single chain. So Myoglobin (or any

monomeric protein) does not contain quaternary structure.
Insulin has 2 chains but it does not has quaternary structure
(as these chains are joined by disulfide bonds).

Transporters in Small Intestine for the Absorption

of Amino Acids
These transporters are common in small intestine and PCT
of kidneys so that any defect in these will lead to problem in
both intestine and kidneys. They are:
1. For Cysteine and basic amino acids e.g. Ornithine,
Arginine, Lysine (represented as COAL)
2. Neutral Amino Acid Transporter (including Tryptophan)
3. For Proline and Glycine
4. Acidic amino acids
5. Beta amino acids e.g. Beta-Alanine Fig. 7.4: Protein structures

Amino Acid Transporter Defect Secondary Structures

Disorder Defect in transporter Amino acid in urine zz Alpha helix
zz Beta pleated sheet
CYSTINURIA Common transporter Cysteine,
(Most for Basic Amino Acids Ornithine, zz Beta bend or turn
Common and Cysteine Arginine, zz Alpha chain in collagen (not alpha helix)
Transporter Lysine
Defect) Mnemonic: COAL Alpha Helix
HARTNUP’S Common transporter Tryptophan (mainly) zz Most common helix found in nature
DISEASE for Tryptophan and and Neutral Amino T
zz Right handed, rigid H
neutral amino acid Acids
(mono-amino-mono- zz Side chains extend outward from central axis E
carboxy amino acid zz Each turn contains 3.6 amino acids O
transporter) R
GLYCINURIA Common transporter Glycine & Proline Y
for Glycine and Proline
 H igh R eturn Beta Turn/Bend
200 Amino Acids not Found in Alpha Helix are: zz They reverse the direction of a polypeptide chain.
zz Proline: Because of secondary amino group, it introduce kink zz Usually found on the surface of a protein
in helix zz Mostly contains 4 residues
zz Glycine: Because of small side chain it creates a sharp bend in zz Hydrogen bonds between 1st and last residue stabilize
CRO BIOCHEMISTRY

the structure (Glycine imparts flexibility to proteins) beta bends

zz Amino acids with bulky R groups: e.g. Tryptophan zz Glycine and Proline are frequently found (More abun-
zz Charged amino acids like Aspartate, Glutamate dant is Proline)
zz Amino acid with branch chain at beta carbon – Valine

Alpha Helix
• One polypeptide chain coiled in spiral
structure
• Hydrogen bonds are parallel to the
backbone


• Only intrachain Hydrogen bonds

• Chain is not fully extended
• Most abundant amino acid is Methionine

Beta Sheet
• 2 or more polypeptide chains arranged in
sheet like structure
• Hydrogen bonds between the strands are
perpendicular to the backbone
• Both intrachain and interchain hydrogen
bonds
• Almost fully extended chain. They may be Tertiary Structure
parallel or anti-parallel
• Most abundant amino acid is Valine. Hydrophobic amino acids lie in the interior and hydrophilic
residues lie on the surface. Amino acids which come together
in the tertiary structure may lie far apart in the primary
structure. But they are brought closer during folding of
Fig. 7.5: Difference between alpha helix and beta sheet protein to make tertiary structure.

Differences between Various Protein Structures

Features 1° 2° 3° 4°
Bond Covalent/Peptide/Amide Hydrogen-bond (H-bond) S~ S Hydrophobic
Hydrophobic HHI H-bond HHI
Hydrogen (Mnemonic)
(Mnemonic) Ionic
Ionic
Functional activity Absent Absent Present Present
Denaturation Retained because peptide Lost Lost Lost
bond is very strong.
T Detection Mass spectrometry, X-ray crystallography
H Edman’s Technique NMR spectrometry
E • Ionic bond is also known as electrostatic bond.
O • S-S stands for disulphide bond
R zz Mass Spectrometry is the best technique to determine the primary structure of proteins as this technique is qualitative
Y as well as quantitative. It can also detect post translational modifications (like hydroxylation) or covalent modifications of
enzymes (like phosphorylation and dephosphorylation). It can also tell the exact molecular weight of the protein (as the
name says - MASS).
zz X-ray Crystallography is the best technique to detect the secondary, tertiary or quaternary structure of proteins
zz NMR-Spectrometry is the best technique to detect the secondary, tertiary or quaternary structure of non-crystallizable
proteins or membrane proteins 201
F undamental B ox
zz Denaturation of proteins: Sometimes reversible.

CHAPTER 7  PROTEINS
zz Denaturing agents are → Heat, Urea, Guanidine, UV rays, Infra red rays, strong acids or alkali. Denaturation leads to loss of activity
and opening of structure.
zz Coagulation: Clumping of denatured protein. Always irreversible.

m nemonic
Enzyme substrate interaction
zz Hydrophobic In enzyme substrate interactions, usually weak bonds are used i.e. Hydrogen, Hydrophobic and Ionic/
zz Hydrogen
Electrostatic. Sometimes it can be covalent bond. Vander waals forces are never present.
zz Ionic
HHI Mnemonic
Protein DNA interaction
zz Hydrogen
zz Ionic In protein-DNA interactions, bonds used are hydrogen, ionic or vander waals forces and never by covalent
zz Vander waals
bond
HIV Mnemonic

PROTEIN SEQUENCING • Chaperonins - They help Chaperones, and act late in the
Direct Method process of protein folding
ƒƒ HSP - 70 class is Chaperones
1. Using Ninhydrin ƒƒ HSP - 60 class is Chaperonins
2. Edman’s degradation (Edman’s reagent is Pheny-liso-
thiocyanate) – Determine sequence from N-terminal end Note- Calbindin is not a chaperone. It stands for a group of
and can sequence 30-60 amino acid residues. But it can- proteins which bind calcium.
not work if N terminal amino acid is chemically modified. • Enzymes which assist in folding are:
3. Use of various chemical or enzymes to cleave peptides – ƒƒ PDI (Protein DisulfideIsomerase)
method used for large polypeptides ƒƒ PPI (PeptidylProlylCis-Trans Isomerase)
4. Mass spectrometry (3 parts are ionization source, mass
analyzer and ion detector)
Indirect Method
GLYCOPROTEINS
By DNA sequencing— First nucleotide sequence of coding Glycoproteins Functions
region is determined and then amino acid sequence can be
Collagen Structural
easily determined. DNA sequencing can be done by Sanger's
technique. Sanger's reagent is DNFB (Di Nitro Fluoro Mucins Lubricative
Benzene). This method can sequence a short peptide only. Transferrin Fe transport
Ceruloplasmin Cu transport
CHAPERONES
Immunoglobulin, Immunity
• They are proteins which assist in Protein Folding
Histocompatibilty antigen
• Located in RER ( Rough Endoplasmic Reticulum)
Examples: TSH, hCG Hormones
• BIP (Immunogloulin Heavy Chain Binding Protein) ALP (Alkaline Phosphatase) Enzyme
• GRP - 78 (Glucose Regulated Protein) Lectins, Selectins Carbohydrate binding for
• GRP - 94
recognition and attachment T
• GRP -170
• Calnexin Calnexin, Calreticulin Chaperones (for Protein H
• Calreticulin Folding) E
• HSP - 40 (also known as Cochaperone) Glycoproteins on platelet surface Homeostasis O
• HSP - 47 (Heat Shock Protein) R
• HSP - 70
• HSP - 90 HAEM Y
• ERp - 29 (Endoplasmic Reticulum protein) Haem is present in haemoglobin, myoglobin and cyto-
• ERp - 57 chromes. Cytochromes are components of ETC, which are
Contd…
 required in all cells of the body. So Haem is required in all Structure
cells of the body.
202 zz Haem is a metalloporphyrin (Iron + Protoporphyrin).
Heme as a Component of Important Structures zz Pyrrole rings are derived from Porphobilinogen (PBG)
zz 4 Pyrrole rings = Porphyrin (cyclic structure)
zz Hemoglobin (transport oxygen) zz Porphyrin with attachment to 4 Methyl, 2 Propionyl and
CRO BIOCHEMISTRY

zz Myoglobin (stores oxygen) 2 Vinyl groups = Protoporphyrin

zz Cytochromes (in ETC and hydroxylation of xenobiotics) zz Protoporphyrin ring + Iron (Fe) in center = Heme
zz Enzymes:
The side chain groups in porphyrins may be arranged in four
 Catalase (degradation of Hydrogen Peroxide) different structural configurations (known as Porphyrin Type
 Peroxidase (degradation of Hydrogen Peroxide) I to IV). Heme is Type III Porphyrin.
 Guanylate Cyclase (stimulated by Nitric Oxide)
 Tryptophan Pyrrolase (oxidation of Tryptophan)


Haem Synthesis
zz Occurs in all cells of body. But Haem synthesis does not
occur in mature RBCs as they lack mitochondria. 85 % of
Haem synthesis occurs in liver and erythroid tissues
zz Compartment → both mitochondria and cytoplasm (It
starts from mitochondria)
zz Starting material – Succinyl CoA and Glycine
zz RLE – ALA Synthase. It requires Vitamin B6 (Pyridoxal
Phosphate) as a cofactor
zz It is repressed by haem.
T
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 ALA-Synthase I ALA-Synthase II
• Expressed throughout the body • Expressed in erythroid 203
tissues
• Rate limiting step in Liver –
• Haem is the negative regulator • No feedback inhibition

CHAPTER 7  PROTEINS
Porphyrias
zz Porphyria is deficiency of one of the haem synthesis
enzymes, other than ALA-synthase
zz ALA-Synthase I deficiency is lethal
zz ALA-Synthase II deficiency is known as X-linked Side- Lead poisoning is the most common cause of acquired
roblastic anemia (It is not a porphyria) porphyria. Although iron deficiency is more common. But
iron deficiency leading to porphyria is a rare thing. But lead
A dditional E dge poisoning will almost always lead to porphyria.
Isoniazid-induced Pyridoxine deficiency causes decreased • Erythropoietin activates ALA synthase, So erythropoietin
heme formation in RBCs, leading to Sideroblastic Anemia with deficiency which occurs in chronic renal failure, leads to
Isoniazid Therapy (ALA Synthase requires Vit. B6) anemia.
• Haem synthesis continues in liver cells according to metabolic
needs. But in RBCs, it is a one time event, so no rate limiting
enzyme here.

Fig. 7.6: Lead Poisoning

T
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204
CRO BIOCHEMISTRY


Fig. 7.7: Haem synthesis steps and porphyrias. Lead poisoning (plumbo porphyria) is an acquired porphyria. Genetic Porphyrias are
deficiency of other enzymes in Haem synthesis pathways. (NOTE: Deficiency of ALA Synthase is not a Porphyria)

 Motor: Muscle weakness

H igh R eturn  Excitation of visceral pain fibers leading to Acute pain
Hydroxymethylbilane is the 1st porphyrin of the pathway which Crisis, Abdominal pain, Vomiting, Constipation,
is able to absorb light and cause photosensitivity. Hence, defects Hypertension and Tachycardia
above HMB will have“No Photosensitivity” and defects below
will have “Photosensitivity“.

Porphyrins are colored compounds e.g. Hydroxymethylbil-

ane, Uroporphyrin, Coproporphyrin, Protoporphyrin

A dditional E dge
zz Uroporphyrin is the most water soluble porphyrin.
zz Protoporphyrin is least water soluble

Fig. 7.8: Acute pain crisis

Clinical Features of Porphyria
T zz Neurological symptoms Dark red/brown colored urine: ALA and PBG in urine
H zz Photosensitivity
E Photosensitivity
O Neurological Symptoms
zz Block later in the pathway leads to accumulation of Por-
R zz Occurs due to block early in the pathway phyrinogens
Y zz Due to accumulation of ALA and PBG zz Porphyrinogens absorbs light and auto-oxidizes to co-
 Block the action of GABA leading to neuropsychiatric rresponding porphyrin derivatives, so accumulation of
symptoms Porphyrin occurs
 Drugs like Barbiturates, Phenytoin, Griseofulvin are contra-
indicated in Porphyria as they induce cytochrome P450 and
precipitate Acute Porphyric episode 205
Fasting-induced Porphyria and role of glucose infusion:
Fasting increases production of ALAS-1, which leads to a
build-up of precursor heme molecules, which causes acute

CHAPTER 7  PROTEINS
attacks of porphyria
So Glucose infusion given to relieve acute attacks of
porphyria as high glucose will prevent induction of ALAS-1.

Haem Catabolism
Site: Macrophages of reticuloendothelial system (Spleen,
Bone Marrow)

Steps of Haem Degradation

Formation of Bilirubin: Haem first gets broken down
1. 
and forms Biliverdin and then Bilirubin. Haemoxygenase
catalyze cleavage of Haem ring by NADPH- cytochrome
P450 reductase dependent oxidation of Haem to Biliverdin,
CO and free iron (a complex oxidation-reduction
reaction). This enzyme is also known as NADPH: Ferri-
hemoprotein oxidoreductase.

2. Uptake of bilirubin by liver: Bilirubin is transported

to liver via blood. In blood, bilirubin is complexed with
albumin.
3. Conjugation of bilirubin: Bilirubin is conjugated with
H igh R eturn two molecules of glucuronic acid, making conjugated
zz Most common Porphyria → Porphyria Cutanea Tarda (PCT) bilirubin (bilirubin diglucuronide).
zz Most common Acute Porphyria → Acute Intermittent 4. Fate of bilirubin: Bile is secreted from liver into the
Porphyria (AIP) intestine. In intestine, glucuronic acid is removed
zz Most common porphyria in children → Erythropoietic by bacteria. The resultant bilirubin is converted to
urobilinogen. Some of the urobilinogen is reabsorbed T
Protoporphyria
from gut and enters portal circulation. Some of this H
zz Most common symptom – Abdominal pain
zz Most common physical sign – Tachycardia urobilinogen enters enterohepatic recirculation and E
All are AD Except: remaining enters kidneys via blood where it is converted O
zz Plumbo Porphyria to urobilin, which is responsible for giving yellow color R
zz Congenital Erythropoietic Porphyria to urine. Remaining urobilinogen in intestine is oxidized Y
zz Erythropoietic Protoporphyria
 by intestinal bacteria to form stercobilinogen, which gets zz So, Every 3rd amino acid in Collagen is Glycine
converted to stercobilin, giving stools its characteristic zz Collagen is made up of 3 alpha- chains (not alpha helix)
206 color.

FIBROUS PROTEINS
CRO BIOCHEMISTRY

Collagen and elastin are two well characterized fibrous

proteins, that serve structural function in body. Other Fibrous
proteins are Keratin, Fibrillin, Laminin. zz These 3 chains together make a superhelix → Tropocollagen
(a rope like triple helix)


zz Intrachain H-bonds are not favoured

zz Interchain H-bonds are favoured
Tropocollagen is Resistant to unwinding because:
zz Connective tissue – present in bones, tendon, cartilage, zz Individual alpha chains are left handed → This is
ligament, skin, blood vessels → gives structure and favoured by Proline residues. Proline facilitates the
support formation of this helix conformation of each alpha-
zz Proteoglycans embedded in water and minerals mixture chain because of its ring structure which causes kink in
zz Ground substance acts as a cushion or lubricant the peptide chain.
zz Superhelix is right handed → This is favoured by
3 main molecular components of connective tissue:
Glycine residues. Glycine has the smallest side chain so
zz Collagen it fits into the restricted space where three chains of helix
zz Elastin come together.
zz Proteoglycan

Collagen Elastin
Cannot be stretched Can be stretched
Has hydroxy-Lysine No hydroxy Lysine
A Glycoprotein Only protein
(no carbohydrate attached)
Has many types Only of one type
(28 approximately)
Intramolecular cross links are Intramolecular cross links
known as Aldol condensation are known as Desmosine
condensation

Collagen
Types of Collagen Location
Type I (most abundant) Skin
Type II Connective tissue, Cartilage
T Type III Arteries and CVS
H
E Type IV Basement membrane
O Type VII Dermis and Epidermis junction
R
Y zz Primary structure of collagen → (Gly – X – Y)n
Where X and Y can be Proline, OH-Proline, Lysine or OH-
Lysine. Most of Proline and Lysine are hydroxylated.
zz Tropocollagen is resistant to digestion by proteases zz Epidermolysis bullosa
because all peptide bonds are internal
207
Post translational Modifications (PTMs) of Collagen
PTMs of collagen are done in order to make more bonds in
the structure of collagen, so that this protein is very strong.

CHAPTER 7  PROTEINS
1. Hydroxylation → This makes Hydrogen bonds. Indivi-
dually H-bonds are weak bonds. But when these are
more in number then they provide a good strength.
Hydroxylation is done on Proline and Lysine residues.

Fig. 7.10: Epidermolysisbullosa

zz Osteogenesis Imperfecta
2. Glycosylation → i.e. carbohydrate (Glucose or Gala-  Characterized by Fragile bones, Pathological frac-
ctose) is added on some of the hydroxy-lysine residues ture. So known as Brittle bone syndrome.
of collagen. Enzyme Galactosyl Transferase or Glycosyl  Biosynthesis of type I collagen Defective
Transferase is required.
 Extra skeletal manifestations - Blue discoloration of
sclera, hearing loss, poor teeth development
Formation of Covalent Cross Links
Enzyme Lysyl oxidase (requires Cu), oxidatively deaminates
some of the lysyl and hydroxy lysyl residues, resulting in the
formation of allysine.

Aldol condensation is formed between two Allysine residues.

Disorders of Collagen
zz Ehlers-Danlos Syndrome (EDS) – Heterogenous group Fig. 7.11: Osteogenesis Imperfecta
of disorders characterized by stretchy skin and loose zz Alport Syndrome
joints  X-linked disorder affecting kidneys
 Type IV collagen affected, which is present in
basement membrane of glomerulus
 C/F → Hematuria, ESRD
 Deafness and visual abnormalities
 Lenticonus together with Hematuria is a character-
istic feature
zz Scurvy – Vit C deficiency
 Hydroxylation reactions are affected
 Patient has Petechiae and Haemorrhages

Fig. 7.9: Ehlers-Danlos Syndrome T

H
 Type IV EDS – is most serious – this is vascular type.
E
There is defect in Type III Collagen
O
 Type VI EDS – Lysyl Hydroxylase defect
R
Y

Fig. 7.12: Scurvy

 zz Menke’s Syndrome
 Dietary deficiency of Copper
208  Defect in production of Cu binding P-type ATPase in
intestinal mucosa
 X-linked
Lysyl Oxidase reaction is affected
CRO BIOCHEMISTRY


 C/F – Kinky hair, greying hair, growth retardation
 Can be severe leading to mental retardation

Marfan syndrome

Keratin
zz Present in hair, nails, skin
zz Rich in non-polar amino acids.
zz Cysteine also present which forms disulfide bond.


Fig. 7.13: Menke’s syndrome

PLASMA PROTEINS
Elastin
Plasma contains about 200 different proteins which are
Elastin is rich in Proline, Glycine, Lysine and Valine. Its structurally and functionally different. They are divided into
primary structure has alternating hydrophilic (Lysine) and three categories:
hydrophobic (Valine) amino acids. Every seventh amino zz Albumin
acid is Valine. The intramolecular cross links are named zz Globulins – a1, a2, β and γ
Desmosine condensation. zz Fibrinogen
zz Desmosine: Crosslinks are formed by the condensation All these proteins except γ-globulins (Immunoglobulins) are
of two Allysine residues on one Tropoelastin and one synthesized by liver. γ-globulins are antibodies released by
Allysine residue and one Lysine residue on another plasma cells. Immunoglobulins constitute the second most
molecule. abundant category of serum proteins after albumin.
Emphysema
It is a disorder of Elastin. Alpha-1 Antitrypsin (α1-AT) is
produced by hepatocytes and macrophages. It inhibits
Elastase, which degrades Elastin of alveolar walls.
Deficiency of α1-AT causes elastin degradation in alveolar
walls, which leads to Emphysema (destruction of connective
tissue of alveolar walls).

Fig. 7.14: Serum Protein Electrophoresis

Albumin/Globulin Ratio (A/G Ratio)

zz Normal serum albumin concentration is 3.6 – 5.4 mg/dl
zz Normal serum globulin concentration is 1.8 - 3.6 mg/dl
zz So normal A/G ratio is 1.2 -1.6
It is decreased if serum albumin is low or globin is increased.
T Fibrillin E.g. conditions like liver disease, malnutrition, nephrotic
H zz A glycoprotein synthesized by Fibroblasts. syndrome, chronic infections, multiple myeloma.
E zz A major type – Fibrillin I. Albumin is the most abundant protein in plasma. It is
O exclusively produced by liver. So albumin levels are measured
R Marfan Syndrome to assess the function of liver. It has low MW (molecular
Y zz Mutation in gene for Fibrillin I. weight) so it is the first protein to appear in urine after
zz Clinical Features: Tall structure, long arms and legs, renal damage. All plasma proteins are glycoproteins except
loose joints, lens dislocation, rupture aorta. albumin. Functions of albumin are:
zz To maintain oncotic pressure. This is because of CHROMATOGRAPHY
high concentration and low MW of albumin. So Chromatography – ‘chroma’ means color; ‘graphy’ means
hypoalbuminemia decreases oncotic pressure and thus separation. So this name given because earlier this technique 209
leads to oedema. was used to separate colored compounds but now colorless
zz Transport role because albumin is polar e.g. transport of substances can also be separated. So Chromatography is
calcium, bilirubin, free fatty acids, steroid hormones and separation of closely related compounds from a mixture.

CHAPTER 7  PROTEINS
many non polar drugs
There are two phases in chromatography :
zz Buffering action: Albumin has maximum buffering
zz Mobile phase: The phase which is moving e.g. mixture
capacity out of all plasma proteins
which is to be separated constitutes the mobile phase. It
zz Nutritive function is dissolved in liquid or gas.
zz Stationary phase: Various things can be used as
Prealbumin/Transthyretin stationary phase in different chromatographies e.g.
column, paper. This is the porous solid medium through
zz ‘Thy’ stands for thyroxine and ‘retin’ stands for retinol
which the sample mixture percolates.
binding protein.
zz This is a plasma protein which is responsible for Type of chromatography
binding of thyroxine and retinol binding protein. Stationary phase
Earlier it was called ‘prealbumin’ or thyroxine binding Column chromatography Column
prealbumin because it moves slightly ahead of albumin Paper chromatography Paper
in electrophoresis.
Thin layer chromatography Thin layer of silica
zz It is present in both blood and CSF.
zz Liver secretes retinol binding protein in circulation.
In circulation, retinol-RBP makes a ternary complex Column Chromatography
with transthyretin. Due to the formation of this ternary
complex, retinol-RBP is not lost in urine. It uses column as Stationary phase. It can be based on size,
charge or affinity.
Table 7.1: Plasma Proteins and functions

Plasma Protein Function A dditional E dge

I. Albumin Maintain oncotic pressure and HPLC is high performance liquid chromatography in which high
transport of many compounds pressure is applied to column chromatography in order to increase
the speed of chromatography and decrease the time required.
II. P
 realbumin/ Transport thyroxine and retinol binding
transthyretin protein
III. a1 Globulins
ƒƒ a1Anti-trypsin Inhibitor of Serine protease
ƒƒ Orosomucoid Binds progesterone
ƒƒ Thyroxine Major thyroid binding protein (TBG)
binding globulin.
ƒƒ Transcortin CorticoSteroid binding protein (CBG)
IV. a2 Globulins
ƒƒ a2macroglobulin Non-specific protease inhibitor
ƒƒ Ceruloplamin Copper transport and ferroxidase
activity
ƒƒ Haptoglobin Scavenger of free intravascular Hb
ƒƒ Prothrombin clotting
V. β- globulins
ƒƒ Haemopexin Scavenger of Hb (this is used if
haptoglobin is saturated)
Fig. 7.15: Common principle of column chromatography
ƒƒ Transferrin Iron transport
ƒƒ Plasminogen Retraction of fibrin clot Column Chromatography is of three types:
VI. γ-globulins Immunoglobulins (Ig G type) 1. Size exclusion Chromatography T
2. Ion exchange Chromatography H
VII. Fibrinogen Precursor of fibrin clot E
3. Affinity Chromatography (most specific)
NOTE: Aldosterone (The most potent neutral mineralocorticoid, 1. Gel filtration/Size Exclusion Chromatography (Fig. 7.16) O
does not have a specific Plasma Transport Protein). R
 Also known as molecular sieve or molecular
exclusion chromatography Y
 Separation is based on size/molecular weight
  Procedure: A mixture of two differently sized
proteins is taken and poured on a column (filled
210 with gel molecules). The size of the gel is such that
the smaller protein can enter the gel but the bigger
protein cannot enter the gel. Now both proteins
move down with gravity but smaller protein takes
CRO BIOCHEMISTRY

time to come down as it has to enter the gel. So,

smaller protein is given hinderance from the gel
molecules. But the bigger sized protein comes down
from in between spaces and is eluted first. This way
separation can be done.
 The commercial name of gel which is used in gel
filtration chromatography is ‘Sephadex’.

Fig. 7.16: Size Exclusion Chromatography

2. Ion exchange chromatography


Further of two types:

 Cation exchange chromatography positively charged compound is retained in column
 Anion exchange chromatography negatively charged compound is retained in column
In this, a particular charge is taken on the column e.g. CMC (Carboxy Methyl Cellulose) is taken on the column. It is having
negative charge. Now mixture which contains both negative and positive charges is poured on the column. Now column is
negatively charged. So positive charged particles are attracted to CMC and therefore they stay in column. But the negative
charged particles are repelled out and they are eluted first. This is known as Cation Exchange Chromatography.

Fig. 7.17: Cation Exchange Chromatography

If DEAE (Di Ethyl Amino Ethyl) Cellulose is taken on the column, which is positively charged due to amino group (NH3+).So
positive charged particle will be eluted first. This is known as Anion exchange chromatography.

T
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Figs 7.18: (A) Anion Exchange Chromatography (B) Cation Exchange Chromatography
Affinity Chromatography
In this chromatography, affinity (by specific but non-covalent interactions) between two compounds is used for separation e.g. 211
enzyme-coenzyme, antigen –antibody, hormone-receptor.

CHAPTER 7  PROTEINS
Figs 7.19 A and B: Affinity Chromatography (A) Separation of Enzyme (B) Separation of Antigen

Enzymes, antigens, hormones, drugs, vitamins, nucleic acids antibodies can be separated by Affinity chromatography.

Paper Chromatography/TLC (Thin Layer Chromatography)

Both these have same principle. Paper Chromatography is the older but cheaper version, used only for teaching purposes. But
TLC is a better and rapid but costly version used in research.
Mobile phase: mixture of amino acid and solvent
Stationary phase: paper or thin layer of silica
Use: separation of amino acids, sugars, sugar derivatives or peptides

Common Principle (Fig. 7.21)

A paper or thin layer of silica is taken and some amino acids are applied at the lower end of the paper. Then dip this paper in
the solvent (methyl cellulose media, which is a non polar solvent). The solvent will move upwards by capillary action and this
solvent will also take along the amino acids. So mixture of solvent and amino acids are moving upwards (this is mobile phase).
When the solvent has reached the upper end of paper, then paper is taken out and stained. Ninhydrin is used for staining the
T
α- amino acids, which forms a purple complex.
H
Analysis of result: Any polar compound is soluble in polar solvent and a non polar compound is soluble in non polar
E
solvent. So if a particular amino acid is soluble in this solvent then this amino acid will reach the farthest point on the paper
O
(this amino acid will give a late stain on the paper), as solvent has moved till the last point. But if the amino acid is not soluble
then it will give a very late stain on the paper. So in case of a non polar solvent, a late stain means this amino acid is non polar
R
and an early stain means this amino acid is polar. Y
212
CRO BIOCHEMISTRY

Fig. 7.20: Steps of thin layer chromatography

Gas –Liquid Chromatography – for Separation of Gels used in Electrophoresis

Volatile Substances zz Agar
zz Agarose


Mobile phase – an unreactive gas zz Polyacrylamide gel: very efficient, high molecular sieving
Stationary phase – inert solid material effect
zz Mixture of volatile liquid is injected into the column
along with mobile phase (gas) Types of Electrophoresis
zz This method is sensitive, rapid and reliable for lipids, zz Isolelectric focussing
drugs and vitamins. zz SDS-PAGE
zz Immuno electrophoresis (Antigen-Antibody
interactions)
SDS-PAGE → only depends on size.
Full form: Sodium Dodesyl Sulfate - Poly Acrylamide Gel
Electrophoresis
zz SDS is a salt derivative of Lauric Acid (12 C). This salt
is applied to the mixture of proteins, which are to be
separated by SDS-PAGE.

Properties of SDS
zz It denatures the proteins, so secondary, tertiary or
quaternary structure of proteins is lost. So shape of
proteins is lost. So difference of movement in this
electrophoresis does not depend on the shape.
zz SDS is an anionic detergent. So it will coat all the proteins
with a negative charge. So difference of movement in this
Fig. 7.21: Gas liquid chromatography electrophoresis does not depend on their charge because
all proteins are negatively charged here.
ELECTROPHORESIS zz 1.4 gm SDS binds to 1 gm Protein. So the size difference is
Electrophoresis is movement of charged particles in electric not lost as SDS binds depending on the size of the protein.
field. Movement in electrophoresis depends on charge, size SDS cannot break disulphide bonds. So to break disulphide
and shape, out of which charge is the main factor. bond, either performic acid is used or mercapto ethanol is
used. Performic acid oxidize the disulphide bond. (Mercapto
ethanol reduces the disulphide bond).

PROTEIN PRECIPITATION REACTIONS

These are various methods which are used for precipitation
of proteins.
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Use of Protein Precipitation zz Organic solvents (ether and alcohol) Causes
zz Neutral salts (e.g. Ammonium sulfate) dehydration
Purification of enzymes/proteins
zz
zz Preparation of PFF (plasma free filtrate) for various
213
Precipitation of Albumin and Globulin by Ammonium
biochemical tests
Sulfate Salt
Various Methods of Protein Precipitation zz Albumin precipitated by full saturation and Globulins

CHAPTER 7  PROTEINS
zz Head are precipitated by half saturation (See Figure Below)
Causes zz This is known as salting out i.e. salt used to precipitate
zz Strong mineral acids (H2SO4, HNO3, HCl) denaturation the protein. (See Figure Below)
zz Heavy metal salts
zz Alkaloidal reagents (Trichloroacetic Causes
acid, Phospho-tungstic acid, neutralization
Sulfosalicylic acid) of charges

Pearls of the Chapter

zz Peptide bond → partial double bond character → “trans” configuration
zz Unsaturated fatty acid double bond → “cis”
zz Monomeric protein) does not contain quaternary structure.
zz Proline is never found in alpha-helix
zz Proline and Glycine are found in beta turns
zz Mass spectrometry is the best technique to determine the primary structure of proteins
zz X-ray crystallography is the best technique to detect the secondary, tertiary or quaternary structure of proteins
zz Enzyme decreased in lead poisoning → ALA-Dehydratase
zz Enzyme increased in lead poisoning → ALA-Synthase
zz In lead poisoning, acid excreted in urine→δ -ALA (δ- amino levulinic acid)
zz ALA-synthase I deficiency is LETHAL
zz ALA-synthase II deficiency is X-linked Sideroblastic anemia
zz Most common porphyria is Porphyria CutaneaTarda (PCT)
zz Every 3rd amino acid in collagen is GLYCINE
zz Every seventh amino acid in elastin is valine
zz 3 enzymes required for collagen formation are prolyl hydroxylase, lysyl hydroxylase and lysyl oxidase.
zz Desmosine cross links in elastin
zz Aldol condensation in collagen
zz Menke’s disease is dietary deficiency of Copper
zz The most sensitive acute phase reactant is CRP (C-reactive protein).
zz Albumin is the most abundant protein in plasma T
zz Normal A/G ratio is 1.2 -1.6 H
zz Ceruloplasminis a Cu containing enzyme, which also has ferroxidase activity E
zz Most rapid method of Protein separation- Capillary Electrophoresis
O
zz Zwitter Ion → Net charge is zero, so Insoluble, so Precipitation occurs
R
zz pI – Isoelectric pH – that pH at which zwitterion exist
Y
zz Heat and Strong mineral acids causes denaturation
zz Heavy metal salts and Alkaloidal reagents causes neutralization of charges
zz Organic solvents and Neutral salts causes Dehydration
 214 Multiple Choice Questions
Proteins 11. Alpha helix of primary polypeptide structure is
1. Which of the following groups of proteins assist in the stabilized by: (PGMEE 2013)
folding of other proteins? a. Disulfide linkage
 (PGMEE 2018, FMGE JUNE 2018) b. Covalent bonding
a. Proteases b. Proteosomes c. Hydrophobic interactions
c. Templates d. Chaperones d. Hydrogen bonds
2. Examples of chaperone includes all EXCEPT:  12. All are consequences of protein denaturation
 (PGI May 2018) EXCEPT- (PGMEE 2013)
a. Calreticulin b. Calnexin a. Loss of solubility
c. Calbindin d. BiP b. Change in viscosity
e. Ubiquitin c. Loss of primary structure
3. Which of the following is a powerful qualitative and d. Loss of function
quantitative technique to assess molecular mass 13. Which among the following is the weakest
and primary amino acid sequence of peptides and interaction? (PGMEE 2013-14)
proteins? : (Recent Question 2017) a. Hydrophobic b. Electrostatic
a. Colorimetry c. Hydrogen d. Vanderwall’s
b. Spectrophotometry 14. Xanthoproteic test is used for - (PGMEE 2014)
c. X-Ray Crystallography a. Ketone bodies b. Vitamin B6
deficiency
d. Mass spectrometry c. Amino acids d. Reducing sugars
4. A peptide bond: (Recent Question 2017) 15. Spectroscopy is used for the interaction of-
a. Has partial double bond character  (PGMEE 2015)
b. Ionized at physiological pH a. Electromagnetic radiation
c. Occurs in cis configuration b. Alpha particles
d. All c. Positrons
M 5. Myoglobin has which structure? d. Protons
C a. Primary, secondary, tertiary 16. In forming 3D structure of protein following
Qs b. Primary and secondary components help: (PGI May 2014)
Ans. c. Primary, secondary, tertiary, quarternary a. Hydrogen bonds
d. Primary and secondary b. Amino acid sequence
1. d
6. Structure of protein is best detected by: c. Interaction between amino acid side chains
2. c d. Chaperones
 (Recent Question 2017)
3. d e. Ubiquitin
a. MS (Mass Spectrometry)
4. a 17. Which group of amino acid is responsible for peptide
b. NMR spectrometry
5. a bond: (PGI May 2013)
c. X-ray crystallography
6. c a. Amino group b. Carboxyl group
d. Edman’s technique
7. d c. Side chain d. Aldehyde group
7. Which structure of Hb can do function?
8. a e. Amide group
a. Primary, secondary, tertiary
9. c 18. Which one of the following statements about protein
b. Primary and secondary
10. d structure is correct: (PGI May 2015)
c. Primary, secondary, tertiary, quaternary
11. d a. Proteins consisting of one polypeptide can have
d. Tertiary and quaternary
12. c quaternary structure
8. Denaturation is resisted by which of the following
13. d b. The formation of a disulfide bond in a protein
bond? (PGMEE 2015)
14. c requires that the two participating cysteine residues
a. Peptide bond b. Electrostatic bond
15. a be adjacent to each other in the primary sequence of
c. Hydrogen bond d. Hydrophobic bond
16. a,b the protein
9. Amide group is present in which part of protein-
c,d c. The stability of quaternary structure in proteins
 (PGMEE 2015)
17. a,b is mainly a result of covalent bonds among the
a. Amino-terminal b. Disulfide bond
18. e subunits
c. Peptide bond d. Carboxy-terminal
10. Shortest peptide is:  (PGMEE 2007, 2001) d. The denaturation of proteins always leads to
a. Angiotensin-I b. Vasopressin irreversible loss of secondary and tertiary structure
c. Angiotensin-II d. Angiotensin-III e. The information required for the correct folding of
a protein is contained in the specific sequence of
amino acids along the polypeptide chain.
19. Which of the following is/are storage protein: 28. Which of the following is a negative acute phase
 (PGI Nov 2011) proteins:  (Recent Question June 2018)
a. Myoglobin b. Ovalbumin a. Transthyretin 215
c. Ricin d. Ferritin b. C-Reactive protein
e. Glutelin c. Ferritin
20. True about isopeptide bond: (PGI Nov 2011) d. Ceruloplasmin
a. It makes protein resistant
b. Bond is formed between carboxyl terminus of one Heme Metabolism
protein and the amino group of a lysine residue on 29. In first step of heme synthesis, which amino acid is
another required :- (PGMEE 2012,13)
c. Involves in post-transcriptional modification of a. Histidine b. Folate
protein c. Glycine d. Fe
d. Enzyme acts as catalyst for the bond formation 30. Heme is which porphyrin- (PGMEE 2013)
e. Exists in ubiquitin a. Type I
21. Defect in Amyloid protein folding occurs in: b. Type II
 (Recent Question June 2018) c. Type III
a. Alzheimer disease d. Type IV
b. Creutzfeldt-Jakob disease 31. Number of pyrrole rings in Porphyrins:
c. Scrapie disease  (PGMEE 2015)
d. Bovine spongiform encephalopathy (BSE) a. 2 b. 3
22. Collagen most commonly contains which of the c. 4 d. 5
following amino acid:  (Recent Question June 2018) 32. Heme oxygenase produces all, EXCEPT:
a. Proline and hydroxyproline a. CO b. CO2
b. Cysteine and Cystine
c. Glycine and Cysteine c. Fe d. H2O
d. Methionine and Proline 33. Haem is not synthesized in :
23. Which of the following is the best method for measur- a. Osteocyte
ement of HbA1C?  (AIIMS May 2018) b. RBCs
a. Ion exchange chromatography c. Liver parenchyma
b. Affinity chromatography d. Erythropoietic precursor cells in Bone Marrow
34. δ-aminolevulinic acid is a metabolic product in M
c. Immunoassay C
d. Electrophoresis synthesis of- (PGMEE 2015)
a. Glycosaminoglycans Qs
24. Best investigation for metabolic disorders is: 
b. Collagen Ans.
 (AIIMS May 2018)
a. Western blot c. Tryptophan 19. b,d,e
b. Tandem mass spectrometry d. Heme 20. a,b
c. PCR 35. Which is an inhibitor of ferrochelatase? d,e
d. Gel electrophoresis  (PGMEE 2015) 21. a
25. Methods that can be used to see protein – protein a. Arsenic b. Mercury 22. a
interaction include: (PGI May 2018) c. Lead d. Iron 23. a
a. Fluorescence life imaging 36. A boy with staining of teeth and raised Coproporphy- 24. b
b. Fluorescence resonance energy transfer rin-I levels and increased risk of photosensitivity, the 25. e
c. Fluorescence polarization enzyme deficient is: (PGMEE 2015) 26. b
d. Fluorescence complementation 27. b
a. Coprophorphyrinogen Oxidase
e. All of the above 28. a
b. Uroporphyrinogen Synthase
26. Protein not synthesized in liver :  29. c
c. Uroporphyrinogen Decarboxylase
 (Recent Question Jan 2019 Q) 30. c
d. Uroporphyrinogen III Synthase
a. Albumin 31. c
37. Pruritis (itching) is caused by deficiency of-
b. Immunoglobulins 32. b
 (PGMEE 2015)
c. Plasma enzyme 33. b
a. Uroporphyrinogen – III Synthase
d. Acute phase proteins 34. d
b. Uroporphyrinogen – I Synthase 35. c
27. Which is/are not a transport protein: c. HMB Synthase
 (PGI May 2018, 2013 36. d
d. δ– ALA Dehydratase 37. a
a. Transferrin 38. Variegate porphyria enzyme defect is: (PGMEE 2015)
b. Collagen 38. b
a. Coproporphrinogen Oxidase
c. Ceruloplasmin b. Protophorphyrin Oxidase
d. Hemoglobin c. Uroporphyrinogen Decarboxylase
e. Albumin d. Uroporphyrinogen Synthase
 39. Enzyme deficient in acute intermittent porphyria- 50. True about collagen: (PGI May 2015)
 (PGMEE 2015 2013) a. Double helix structure
216 a. ALA synthase b. Single helix structure
b. Uroporphyrinogen – III synthase c. Triple helix structure
c. Porphobilinogen Deaminase d. β-pleated structure
d. δ-ALA dehydratase e. Component of connective tissue of body
40. Paroxysmal nocturnal haemoglobinuria is seen in: 51. Which type of collagen is present in Basement
 (PGMEE 2011) membrane? (Recent Question 2016)
a. Deficiency in synthesis of G-6 phospahte a. Type I b. Type II
dehydrogenase c. Type III d. Type IV
b. Deficiency in synthesis of glycosylphosphatidyl 52. Collagen is composed of all EXCEPT:
inositol in haemopoietic cells a. Proline b. Glycine
c. Deficiency in synthesis of glucose -6-phosphatase in c. Hydroxy Lysine d. Desmosine
haemopoietic cells 53. The fold in collagen is because of presence of –
d. Deficiency in synthesis of prostaglandins (PGMEE 2012,13)
41. A 10 year old boy presented with muscle weakness, a. Histidine b. Glycine
fatigue, increased lead in blood. Which enzyme is c. Arginine d. Glutamate
increased? (Recent Question 2016) 54. Type of collagen found in space of Disse in liver is-
a. ALA Synthase (PGMEE 2013)
b. Porphobilinogen Deaminase a. Collagen II and V b. Collagen III and IV
c. Ferrochelatase c. Collagen II and III d. Collagen I and III
d. ALA Dehydratase 55. Most abundant in bone – (PGMEE 2012,13)
42. A girl with habit of eating paint from the wall a. Arginine b. Hydroxyproline
complaints of acute abdominal pain on and off and c. Lysine d. Alanine
tingling sensation of limbs and at times weakness of 56. Which of the following type of collagen is present in
limbs. She had a history of eating paint from the wall healing and granulation tissue? (AIIMS May 2018)
of newly built house. Which of the following enzyme a. Type I
deficiency will be the cause of her condition? b. Type II
 (AIIMS May 2017) c. Type III
a. ALA Synthase b. ALA Dehydratase d. Type IV
M c. Heme Synthase d. Coproporphyrinogen
C 43. In a patient with lead poisoning, which heme pathway Chromatography and Electrophoresis
Qs intermediate will increase in urine? 57. Which of the following technique does not depend on
Ans.  (AIIMS Nov 2015) molecular weight of protein: (Recent Question 2018)
39. c a. ALA b. Prophobilinogen a. Thin layered Chromatography
40. b c. Protoporphyrin d. Coproporphyrinogen III b. Sucrose density gradient centrifuge
41. a 44. Which is not involved in iron metabolism? c. Gel filtration
42. a a. Transthyretin d. SDS-PAGE
43. a b. Ceruloplasmin 58. All of the following are true EXCEPT:
44. a c. Hepcidin a. Electrophoretic mobility is function of length of
45. b d. Ferritin polypeptide chain, higher order protein folding and
46. b 45. Plasma Ceruloplasmin is a – (PGMEE 2009) post translational modification
47. a,b a. β-2 globulin b. Alpha-2 globulin b. SDS is anionic detergent
c,e c. β-1 globulin d. α -1 globulin c. SDS break H bond and S-S bond
48. c 46. Plasma protein which binds and transports free fatty d. SDS binds in a ratio of appx. 1.4 gm SDS per one gm
49. a acids is (PGMEE 2009) of protein.
50. c,e a. Prealbumin b. Albumin 59. Which of the following fatty acid derivative is used in
51. d c. Transthyretin d. Ceruloplasmin purification of proteins: (Recent Question 2017)
52. d 47. Hemoproteins are: (PGI Nov 2011)
a. Myristic acid
53. b a. Cytochrome C b. Cytochrome 450
b. lauric acid
54. d c. Myoglobin d. Hemoglobin
c. Oleic acid
55. b e. Catalase
d. Caproic acid
56. c 48. Iron in haemoglobin binds with:
60. Farthest moving amino acid on paper chromato-
57. a a. Alanine b. Serine
graphy on methylcellulose medium:
58. c c. Histidine d. Glycine
a. Glycine  (Recent Question 2017)
59. b 49. Type of collagen maximum in skin: 
b. Aspartic acid
60. c  (Recent Question Jan 2019)
c. Valine
a. Type I b. Type II
d. lysine
c. Type III d. Type IV
61. Which is not a method of protein precipitation? 66. Proteins are separated on the basis of size by:
a. Salting out with heavy metals  (PGMEE 2003)
b. Acetone and Alcohol a. SDS- PAGE 217
c. Changing pH other than isoelectric pH b. Affinity chromatography
d. Trichloroacetic acid c. HPLC
62. Which of the following method of protein separation d. Ion-exchange chromatography
is not dependent on molecular size – (PGMEE 2013) 67. Two same charged proteins can be separated by all
a. SDS-PAGE EXCEPT – (PGMEE 2012,13)
b. Ultracentrifugation a. Agarose b. DEAE cellulose
c. Ion-exchange chromatography c. Sephadex d. None of these
d. Gel filtration chromatography 68. Plasma proteins can be separated by:
63. Separation of proteins by their mass is by- a. Fluorometry b. Salting out
 (PGMEE 2013) c. Electrophoresis d. Both b and c
a. Gel filtration chromatography 69. Haemoglobin electrophoresis is based on:
b. Ion exchange chromatography a. Size b. Charge
c. Isoelectric focusing c. Solubility d. Calorimetric property
d. Affinity chromatography 70. Which method is used to separate a mixture of
64. Molecular separation of two proteins with same lipids- (PGMEE 2015)
charge can be done by - (PGMEE 1997) a. Isoelectric focusing
a. Electrophoresis b. PAGE
b. Dialysis c. Chromatography
c. Gel filtration chromatography d. Electrophoresis
d. Ion exchange chromatography 71. What is forward scatter in flow cytometry used to
65. The following separation technique depends on the assess? (AIIMS May 2015)
molecular size of the protein- (PGMEE 2003) a. Cell death b. Cell size
a. Chromatography on diethyl amino ethyl (DEAE) c. Cell granules d. Cell fluorescence
cellulose column 72. Molecular weight of protein can be determined by:
b. Iso- electric focusing a. SDS-PAGE
c. Gel Filtration chromatography b. Gel filtration chromatography
d. Chromatography on a carboxymethyl (CM) cellulose c. Agarose gel eletrophoresis
column d. Ultra Centrifugation M
e. FRET microscopy C
Qs
Ans.
61. c
62. c
63. a
64. c
65. c
66. a
67. b
68. d
69. b
70. c
71. b
72. a,b,d
 Answers with Explanations
218

1. Ans. (d) Chaperones structure. In this case, consider tertiary structure

CRO BIOCHEMISTRY

(the 3D structure present in vivo). So, mark X-ray

[Ref: Harper 30th/e pg.45] crystallography (which is best for tertiary).
•• Chaperones are proteins which assist in protein •• Mass spectrometry and Edman’s technique can
folding. They are located in endoplasmic reticulum. determine the primary structure of proteins. NMR-
They mediate post translational assembly of protein spectrometry is the best technique to detect the
complexes. (Refer: Chaperones in Theory) secondary, tertiary or quaternary structure of non
crystallizable proteins or membrane proteins
2. Ans. (c) Calbindin
7. Ans. (d) Tertiary and quaternary
[Ref: Harper 30th/e pg.45]
[Ref: Lehninger 7th/e pg. 131-133]


•• Calbindin is not a chaperone. It stands for a group of

proteins which bind calcium. Other options given in •• In case of any protein, primary and secondary
Question are examples of Chaperones. structures cannot do function. Function starts from
tertiary structure.
3. Ans. (d) Mass Spectrometry
8. Ans. (a) Peptide bond
[Ref: Lehninger 7th/e pg. 100]
[Ref: Harper 30th/e pg. 38]
•• Best method to detect the primary structure
(sequence of amino acids) of protein is Mass •• Peptide/ amide bond is a strong bond which is not
Spectrometry. This technique is qualitative as well broken down on denaturation.
as quantitative. It detects the molecular mass of
9. Ans. (c) Peptide bond
each particle in the protein. In this technique, after
ionization, sorting of ions is done based on mass to [Ref: Harper 30th/e pg. 38]
charge ratio.
•• Peptide/ amide bond is a strong covalent bond which
4. Ans. (a) Has partial double bond character is present in primary structure of proteins.

[Ref: Lehninger 7th/e pg. 85-86] 10. Ans. (d) Angiotensin-III

•• Peptide bond is always formed between the alpha [Ref: Harper 30th/e pg. 513]
carboxy group of one amino acid and the alpha amino
A •• Human angiotensinogen has 452 amino acids.
group of another amino acid. It has partial double
N Angiotensin I (decapeptide -10 amino acids) is formed
bond character and exists in ‘trans’ configuration. It
S from angiotensin by the action of renin. Angiotensin
is rigid and planar.
W •• Note: The unsaturated fatty acid double bond is in I is converted to Angiotensin –II (8 amino acids) by
E ‘cis’ configuration. removal of 2 carboxy terminal residues by enzyme
R ACE (Angiotensin converting enzyme).
S 5. Ans. (a) Primary, secondary, tertiary •• Angiotensin- III (heptapeptide -7 amino acids) is
formed from Angiotensin-II by removal of an amino
WITH [Ref: Lehninger 7th/e pg. 131-133] acid from N-terminus by enzyme aminopeptidase A.
•• Myoglobin has only one chain. So it does not has •• Posterior pituitary has two hormones i.e. Vasopressin
E
quaternary structure. Monomeric proteins do not (ADH- Anti- diuretic hormone) and Oxytocin,
X
contain quarternary structure. [Quarternary structure containing 9 amino acids (option –b).
P
L is more than one functional polypeptide chain]
11. Ans. (d) Hydrogen bonds
A
6. Ans. (c) X-ray crystallography
N [Ref: Harper 30th/e pg.47]
A [Ref: Lehninger 7th/e pg. 134b-135b] •• Secondary structures are stabilized by hydrogen
T bonds.
•• In the question, they have not mentioned that
I
it is primary, secondary, tertiary or quaternary
O
N
S
 •• Bond in primary structure is covalent/amide/ 18. Ans. (e) The information required for the correct
peptide bond. In Tertiary, there are many bonds i.e. folding of a protein is contained in the specific
disulphide, hydrogen, hydrophobic and ionic bond. sequence of amino acids along the polypeptide chain. 219
•• Quaternary structure has hydrogen, hydrophobic
and ionic bonds. [Ref: Harper 30th/e pg.47]
•• More than one polypeptide chains are required to

CHAPTER 7  PROTEINS
12. Ans. (c) Loss of primary structure make quaternary structure (Option-a). Amino acids
[Ref: Harper 30th/e pg. 38] which may be placed at long distances on the primary
structure, are brought nearer to each other by the
•• Primary structure is not lost on denaturation. Only 3-D conformation of the enzyme (Option-b). Bonds
secondary, tertiary and quaternary structures are lost in quaternary structure are hydrogen, hydrophobic
on denaturation. and ionic bonds (Option-c). Denaturation can be
sometimes reversible (Option d).
13. Ans. (d) Vander waal’s
19. Ans. (a); (b); (d); (e)
[Ref: Harper 30th/e pg. 39]
•• Weakest bond – vander waal’s interactions. They [Ref: Harper 30th/e pg. 567]
arise from the rapid movement of electrons of all •• Ovalbumin is the main protein in egg albumin.
neutral atoms Glutelin is present in wheat. Ferritin is a storage
•• Strongest covalent bond is peptide bond. protein for iron.
•• Myoglobin carries and stores O2 in muscle cells. Ricin
14. Ans. (c) Amino acids
(a lectin produced in seeds of castor oil plant) is
[Ref: Harper 30th/e pg.22] inhibitor of protein synthesis.
•• Xanthoproteic test is a color reaction of proteins. This 20. Ans. (a); (b); (d); (e)
is positive because of aromatic amino acids. It gives
yellow color. [Ref: Harper 30th/e pg.47]
•• Note: Xanthurenic acid is the metabolite in the Isopeptide bond is an amide bond formed between non
metabolism of Tryptophan, which is excreted in α-NH2 or non-α-carboxy groups. Option a, b, d, e are
urine in vitamin B6 deficiency. correct.
•• It occurs post translationally (not transcriptionally)
15. Ans. (a) Electromagnetic radiation
•• It makes protein resistant as proteases cannot
[Ref: Harper 30th/e pg. 43] hydrolyse this bond.
•• Glycine of ubiquitin binds with lysine of target protein,
•• Spectroscopy is the study of interaction between with isopeptide bond, for protein degradation.
matter and electromagnetic radiations (‘scopy’ A
means looking at). Spectrometry (‘metry’ means 21. Ans. (a) Alzheimer disease N
measurement of ). These two terms are used S
interchangeably. [Ref: Harper 30th/e pg 46]
W
•• Alzheimer’s disease is a disease characterized by the E
16. Ans. (a); (b); (c); (d)
aggregation of amyloid β proteins to form flexible R
[Ref: Harper 30th/e pg.47] soluble oligomers. These oligomers are toxic to nerve S
cells.
•• Chaperones assist in folding of protein. Bonds •• Amyloid β are composed of peptides of 30-40 amino WITH
in tertiary structure are disulphide, hydrogen, acids and formed from the proteolytic cleavage of
hydrophobic and ionic bonds. Sequence of amino precursor, APP (Amyloid Precursor Protein). Amyloid E
acid is folded due to interaction of side chains of plaques get deposited in this disease. X
amino acids to make 3-D structure of protein. P
•• Ubiquitous is used in protein degradation 22. Ans. (a) Proline and Hydroxyproline L
A
17. Ans. (a); (b) [Ref: Harper 30th/e pg. 627] N
•• Primary structure of collagen is (Gly-X-Y) repeats. A
[Ref: Harper 30th/e pg. 38]
•• Every third amino acid in collagen is Glycine T
•• Peptide bond is always formed between the alpha •• X and Y are Proline, Hydroxy Proline, Lysine or I
carboxy group of one amino acid and the alpha Hydroxylysine. This hydroxylation confers strength O
amino group of another. to collagen. N
S
 23. Ans. (a) Ion exchange chromatography 28. Ans. (a) Transthyretin

220 [Ref: Tietz Textbook of clinical chemistry and molecular [Ref: Harper 30th/e pg 672]
biology, 5th ed., Pg. 1443-1444] •• Acute phase reactant proteins are those plasma-
•• Best method for measurement of HbA1C is enzymatic proteins whose concentration changes during infla-
mmation.
CRO BIOCHEMISTRY

assay, but has high cost.

•• So, commonly used method for measurement of •• The proteins whose concentration is decreased are
HbA1C is Ion exchange HPLC. called negative acute phase reactant proteins. The
proteins whose concentration is increased are called
24. Ans. (b) Tandem mass spectrometry positive acute phase reactant proteins.
•• Negative acute phase reactant proteins are Albumin,
(Ref: Tietz, Textbook of clinical chemistry and molecular Transferrin, RBP, Transthyretin.
biology, 5th ed., pg. 2052) •• Positive acute phase reactant proteins are: CRP, D
•• Most powerful technique of screening for inborn error dimer protein, Mannose binding protein, α 1 an-
of metabolism is TMS (Tandem mass spectrometry). titrypsin, α 1 anti-chymotrypsin, α 2 macroglobu-
lin, Fibrinogen, Prothrombin, Clotting factor VII,


25. Ans. (e) All of the above Von-Willibrand Factor, Plasminogen, Complement
factor, Ferritin, SAP complement [Serum Amyloid P
[Ref: Harper 30th/e pg 43-46]
complement], SAA [Serum Amyloid A], Ceruloplas-
•• Methods that can be used to see protein – protein min, Hapto-globin, Hepcidin, orosomucoid. The
interaction include: most sensitive is C-Reactive Protein (CRP) which is
ƒƒ Fluorescence Life Imaging Microscopy, Fluores- raised within few hours after an inflammation event
cence Resonance Energy Transfer (FRET), Fluo- but it is non-specific.
rescence Polarization, Fluorescence Complemen-
tation, Im-munoprecipitation, Affinity Electropho- 29. Ans. (c) Glycine
resis, Step Protein Interaction Experiment (SPINE),
[Ref: Harper 30th/e pg. 52]
Proximity Ligation Assay, MALDI- MS, Biomolecu-
lar Fluo-rescence Complementation, Interferom- •• Glycine and Succinyl CoA are the two starting
etry, Static And Dynamic Light Scattering, Surface materials used in Haem synthesis.
Plasmon Resonance, Flow Induced Dispersion
30. Ans. (c) Type III
Analysis, Fluorescence Correlation Spectroscopy,
Microscale Thermophoresis, Rotating Cell Based [Ref: Dinesh Puri 3rd/e pg. 335]
Ligand Based Assay, Single Color Reflectometry
(SCORE). •• The side chain groups in porphyrins may be arranged
A in four different structural configurations (I to IV).
N 26. Ans. (b) Immunoglobulins Only Type I and Type III series are found in nature
S and the porphyrins of type III series are not only more
[Ref: Harper 30th/e pg 570] abundant but are also physiologically important in
W
E •• All these are plasma proteins. All plasma proteins are humans. Heme is type III porphyrin.
R synthesized by liver except Immunoglobulins, which
31. Ans. (c) 4
are produced & released from plasma cells (in RER).
S
[Ref: Harper 30th/e pg 322]
WITH 27. Ans. (b) Collagen
•• There are four pyrrole rings in Porphyrins e.g. Haem
E [Ref: Harper 30th/e pg 673]
X 32. Ans. (b) CO2
•• Transferrin is for transport of Fe. Ferritin is for
P storage of iron. Ceruloplasmin is for transport of Cu. [Ref: Harper 30th/e pg. 330]
L Haemoglobin is for transport of oxygen and carbon
A dioxide. Albumin is for transport of steroids, fatty •• Heme Oxygenase uses Oxygen & it produces CO, Fe3+
N and water. But it does not produces CO2.
acids, thyroid hormones, bilirubin, cations and many
A
drugs. Collagen is not a transport protein, rather it is a
T
fibrous protein present in skin and connective tissue.
I
O
N
S
33. Ans. (b) RBCs 40. Ans. (b) Deficiency in synthesis of glycosyl-phos-
phatidyl inositol in haemopoietic cells
[Ref: Harper 30th/e pg. 328] 221
[Ref: Harper 30th/e pg. 580]
•• Haem synthesis occurs in all cells of the body, and it
occurs both in cytoplasm and mitochondria. But in •• Paroxysmal Nocturnal Haemoglobinuria (PNH)
RBCs, no mitochondria. So haem synthesis cannot is an acquired hematopoietic stem cell disorder in

CHAPTER 7  PROTEINS
occur in RBCs. which PIG-A gene is mutated. This gene synthesizes
GPI anchor (Glycosyl-Phosphatidyl Inositol). Intra-
34. Ans. (d) Heme vascular haemolysis occurs due to absence of GPI-
[Ref: Harper 30th/e pg. 328]
linked proteins from the surface of RBC’s.

•• δ-aminolevulinic acid or δ-ALA is formed in haem 41. Ans. (a) ALA Synthase
synthesis from Succinyl CoA and Glycine.
[Ref: Harper 30th/e pg. 325]
35. Ans. (c) Lead •• In lead poisoning, the enzyme affected is ALA
Dehydratase/ PBG Synthase. This is an enzyme in
[Ref: Harper 30th/e pg. 326]
Haem synthesis pathway, so Haem not formed. So
Lead inhibits two important enzymes in the heme the rate limiting enzyme of Haem synthesis – ALA
synthetic pathway: Synthase is induced due to lack of product inhibition.
•• ALA dehydratase (Enzyme contains Zinc)
•• Ferrochelatase or Heme Synthase (this enzyme 42. Ans. (a) ALA Synthase
incorporates Fe in protoporphyrin)
[Ref: Harper 30th/e pg. 325]
36. Ans. (d) Uroporphyrinogen III Synthase •• Lead is found in both exterior and interior wall
paints. Many countries have restricted the use of lead
[Ref: Harper 30th/e pg. 328]
in paints (only upto 90 ppm allowed), whereas some
•• Uroporphyrinogen – III Synthase deficiency is countries have > 10,000 ppm (parts per million) lead
a Congenital Porphyria, known as Congentinal concentration in paints. Leads inhibits two enzymes
Erythropoietic Porphyria (CEP) substrate HMB of haem synthesis pathway (ALA Dehydratase and
(Hydroxymethylbilane) accumulated and gets Ferrochelatase), leading to Porphyria.
converted to Uroporphyrinogen I (non enzymatically),
which further forms Coproporphyrinogen I 43. Ans. (a) ALA
•• These patients have red or brown discoloured teeth
[Ref: Harper 30th/e pg. 325]
due to deposition of porphyrin pigment.
•• In lead poisoning, ALA Dehydratase affected, so
37. Ans. (a) Uroporphyrinogen – III synthase substrate ALA accumulates and excreted in urine. A
N
[Ref: Harrison 19th/e pg. 328] 44. Ans. (a) Transthyretin S
•• In CEP, itching occurs due to excess porphyrins W
[Ref: Harper 30th/e pg 672]
in skin and light radiation causes photo-oxidative E
damage. Other options are enzyme deficiencies early •• Iron storage: Ferritin and Haemosiderin R
in the pathway, where porphyrins are not formed. •• Haemosiderin has higher iron content than Ferritin
S
•• Iron transport: Transferrin
38. Ans. (b) Protophorphyrin Oxidase •• Hepcidin: regulate iron transport in circulation WITH
•• Ceruloplasmin: a Cu containing enzyme in ferro-
[Ref: Harper 30th/e pg. 328] E
xidase activity
•• Variegate Porphyria is due to defect in enzyme •• Transthyretin: transports Thyroxine and Retinol X
Protophorphyrin Oxidase. binding protein P
L
39. Ans. (c) Porphobilinogen Deaminase 45. Ans. (b) Alpha-2 Globulin A
N
[Ref: Harper 30th/e pg. 328 [Ref: Harper 30th/e pg. 675] A
•• Enzyme deficient in Acute Intermittent Porphyria is •• Plasma Ceruloplasmin is α-2 Globulin. Other α-2 T
Porphobilinogen Deaminase Globulins are α-2 Macroglobulin, Haptoglobin, I
Prothrombin. O
N
S
 46. Ans. (b) Albumin 51. Ans. (d) Type - IV

222 [Ref: Harper 30th/e pg. 223] [Ref: Harper 30th/e pg 631]
•• Albumin is the most abundant protein in plasma. It Type IV collagen is present in basement membrane
binds and transports free fatty acids. (specially of Glomerulus).
•• Type I is most abundant, present in skin
CRO BIOCHEMISTRY

47. Ans. (a); (b); (c); (d); (e) •• Type II is in Connective tissues
•• Type III is in CVS and Arteries
[Ref: Harper 30th/e pg 328]
•• Hemoproteins have Haem in their structure eg. Hb, 52. Ans. (d) Desmosine
Mb, Cytochromes, Catalase, Peroxidase, Guany- [Ref: Harper 30th/e pg 627]
latecyclase, Tryptophan Pyrrolase. Desmosine is not present in Collagen. It is present in
48. Ans. (c) Histidine Elastin.

[Ref: Harper 30th/e pg 59] 53. Ans. (b) Glycine

•• There are 38 histidine residues in haemoglobin


[Ref: Harper 30th/e pg. 578]

molecule. The imidazole rings in histidine residues •• Folds or turns in collagen are because of Glycine &
of haemoglobin can be protonated or de-protonated Proline
to absorb or release H+ ions. This helps haemoglobin
to act as buffer. 54. Ans. (d) Collagen I and III

[Ref: Harper 30th/e pg. 631]

•• Space of Disse or Peri-sinusoidal space in liver is
between hepatocytes and sinusoid, which contains
blood plasma. Microvilli of hepatocytes extend into
this space, allowing absorption of proteins and other
plasma components from sinusoids into liver cells.
Here Collagen Type I & III are found.

55. Ans. (b) Hydroxyproline

[Ref: Harper 30th/e pg. 579]

•• Hydroxyproline is found in collagen, which is present
in bones.

A 56. Ans. (c) Type III

N
[Ref: Harper 30th/e pg.631]
S
W During wound healing, first Type III Collagen is
E deposited. Later it is replaced by a matrix mainly
R containing Type I Collagen.
S 57. Ans. (a) Thin layered Chromatography
WITH 49. Ans. (a) Type I
[Ref: Harper 30th/e pg.748]
th
E [Ref: Harper 30 /e pg.631]
•• Thin layered Chromatography depends on the affinity
X •• Type I collagen is present in skin, which is most of the amino acid towards the stationary phase (thin
P abundant collagen present in body. layer of silica) or mobile phase (solvent). It does not
L depend on the molecular weight of the amino acid.
A 50. Ans. (c); (e) •• Sucrose density gradient centrifuge, Gel filtration
N and SDS-PAGE – all these techniques depend on
[Ref: Harper 30th/e pg 579]
A molecular weight.
T •• Collagen is triple helix, made up of 3 alpha chains
I (not α-helix) It is a component of connective tissue
O of body.
N
S
58. Ans. (c) SDS break H bond and S-S bond chromatography can be done for the separation of
differently charged proteins. And dialysis cannot be
[Ref: Harper 30th/e pg.695] done at molecular level. 223
•• SDS (Sodium Dodecyl Sulfate) can break H-bonds
but cannot break disulfide bonds. So option c is 65. Ans. (c) Gel Filtration Chromatography
wrong. Other options given in question are true. [Ref: Harper 30th/e pg. 42]

CHAPTER 7  PROTEINS
59. Ans. (b) Lauric acid •• DEAE Cellulose is used in Anion Exchange
Chromatography. Isoelectric Focusing is a kind of
[Ref: Harper 30th/e pg.695] electrophoresis in which isoelectric pH is used for
•• Lauric acid synthesizes SDS (Sodium Dodecyl protein separation. Gel filtration chromatography
Sulfate), which is used in separation of proteins on is based on size. CMC Cellulose is used in Cation
the basis of size and also purification of proteins. Exchange Chromatography.

60. Ans. (c) Valine 66. Ans. (a) SDS- PAGE

[Ref: Harper 30th/e pg.23] [Ref: Harper 30th/e pg. 42]

•• In this Paper Chromatography, a non polar medium •• SDS –PAGE is a special kind of electrophoresis
(Methyl Cellulose medium) is used. So, the amino which separates molecules on the basis of size. But a
acid which is most non polar out of these four, will be normal electrophoresis depends on many factors like
moving farthest on the paper. charge, size and shape. In affinity chromatography,
•• Aspartate (negatively charged) and Lysine (positively interaction (or affinity) between two compounds is
charged) are polar. Glycine is least non polar out of all used for separation. HPLC is high performance liquid
aliphatic amino acids. chromatography in which high pressure is applied
to column chromatography in order to increase the
61. Ans. (c) Changing pH other than isoelectric pH speed of chromatography and decrease the time
[Ref: Harper 30th/e pg.20] required.

Only at Isoelectric pH, protein’s net charge is zero, 67. Ans. (b) DEAE cellulose
known as Zwitterion. This structure is insoluble, so
precipitates. [Ref: Harper 30th/e pg. 41]
•• At acidic pH, protein has positive charge. •• DEAE Cellulose is used in Anion Exchange Chroma-
•• At alkaline pH, protein has negative charge. tography.

62. Ans. (c) Ion Exchange Chromatography 68. Ans. (d) Both b and c
[Ref: Harper 30th/e pg. 43] [Ref: Wilson and Walker 7th/e pg 407] A
N
•• Ion Exchange Chromatography is based on charge. •• Plasma Protein Electrophoresis is the most commonly
S
Rest all depend on size or molecular weight. used method. Salting out i.e. precipitation of proteins
W
by ammonium sulphate salt can also be used for
63. Ans. (a) Gel Filtration Chromatography E
plasma protein separation.
R
[Ref: Harper 30th/e pg. 44]
69. Ans. (b) Charge S
•• Gel filtration chromatography is based on mass.
Ion exchange chromatography is based on charge. [Ref: Wilson and Walker 7th/e pg 399] WITH
In affinity chromatography, interaction (or affinity) •• Electrophoresis depends on three factors (charge, E
between two compounds is used for separation. size and shape). Main factor is charge. X
P
64. Ans. (c) Gel Filtration Chromatography 70. Ans. (c) Chromatography
L
[Ref: Harper 30th/e pg. 44] [Ref: Harper 30th/e pg. 213] A
N
•• Separation at molecular level of two proteins •• Gas Liquid Chromatography is used to seperate A
with same charge can be done by gel filtration mixture of lipids T
chromatography. Electrophoresis and ion exchange
I
O
N
S
 71. Ans. (b) Cell size 72. Ans. (a); (b); (d)
•• Flow cytometry is a laser based, biophysical
224 technique used in cell counting, cell sorting,
[Ref: Harper 30th/e pg. 42]
biomarker detection and protein engineering, by •• SDS-PAGE → Sodium Dodecyl Sulphate - Poly Acryla-
suspending cells in a stream of fluid and passing mide gel Electrophoresis → Used for molecular
CRO BIOCHEMISTRY

them through an electronic detection apparatus. A weight determination of proteins

flow cytometer allows simultaneous multiparametric •• Gel Filtration chromatography→ for separation of
analysis of the physical and chemical characteristics proteins, purification of proteins and molecular
of up to thousands particles per second. weight determination.
•• Ultra Centrifugation→ for isolation of sub cellular
organelles, proteins and nucleic acids. It also
determines molecular weight of macromolecules.
•• FRET → Fluorescence Resonance energy transfer
→ used to measure distances between domains in
a single protein. Therefore helps in determining
protein conformations.


A
N
S
W
E
R
S
WITH

E
X
P
L
A
N
A
T
I
O
N
S
UNIT
IV

LIPIDS
Unit Outline
Chapter 8 Chemistry of Lipids
Chapter 9 Lipoproteins
Chapter 10 Metabolism of Lipids
Chemistry of Lipids
8
Overview of Chapter
•• Fatty acids
•• Classification of Fatty acids
•• Classification of lipids
•• Sphingolipidoses
•• Bile Acids
Fig. 8.1: Synthesis of fat. Whenever a macromolecule is
Lipids Definition synthesized, water molecule is removed. So here non-polar fat/
It is a simple definition just based on physical property that lipid/ester is formed by joining fatty acid (polar) and alcohol
any compound which is insoluble in water and soluble in (polar).
non-polar organic solvents (like ether, chloroform) is a lipid.

FATTY ACIDS
Fatty Acid is polar. Fat is non-polar.
The hydrocarbon chain of fatty acid is non-polar
(hydrophobic) but the carboxy group ionize to give a negative
charge, so polar. So we can say that fatty acid is amphipathic.

Fig. 8.2: Difference between ‘ol’ and ‘thiol’ & ‘ester' and
‘thioester’ linkage

Numbering of Fatty Acids

The functional group (carboxy) is given number 1. The carbon
adjacent to functional C is alpha. So C2 is always alpha, C3 is
beta, and C4 is gamma. The terminal methyl group is called
The short name of fatty acid is “ACYL” For e.g. Acyl CoA omega carbon.
means a fatty acid is joined to Coenzyme A.
Numbering of Double Bond
zz Delta system—when counting starts from carboxy end
zz Omega system—when counting starts from the methyl
group (omega C)

How polar fatty acid gets converted to non-polar fat?

Fatty Acid Alcohol Fat
(Polar) (Polar) (Non-Polar)

Fig. 8.3: This double bond can be delta-9 or omega -7, depending
on end of fatty acid from which the numbering is taken
 CLASSIFICATION OF FATTY ACIDS zz Oleic acid (18 C,one double bond) is the most common
fatty acid in natural fats.
228 Depending on the Length of the Carbon Chain
zz Elaidic acid is present in hydrogenated oils (e.g. vanaspati
Fatty Acid Length of Chain ghee) and ruminant fats.
Short chain 2-4 C
CRO BIOCHEMISTRY

Medium chain 6-12 C Polyunsaturated Fatty Acids (PUFAs)

Long chain 14-20 C
They are also known as Essential Fatty Acids (EFA) as they
Very Long chain >20 C
are essential in diet. We cannot synthesize them in the body.
zz Very long chain Fatty Acids (VLFAs) are usually found in Enzymes that can introduce double bonds beyond 9th carbon
brain. are absent in humans.
Coconut and Coconut Oil Essential fatty acids (EFA) are divided into 2 categories :
zz Has highest content of medium chain fatty acids
1. Omega-3
zz They are absorbed directly into blood.
2. Omega -6


zz They can cross inner mitochondrial membrane without

carnitine shuttle system zz Two fatty acids which are essential in diet are → Linoleic
zz Has no effect on atherosclerosis and α-Linolenic acid.
zz Most Essential Fatty Acid– Linoleic acid (because it can
Depending On Saturation synthesize Arachidonic acid)
zz Saturated fatty acids: No double bond is present. zz But all six are essential in body. (Fig. 8.4)
Number of hydrogens in case of a saturated fatty acids
are double the number of carbons
Omega 3 Category
Saturated Fatty Acid No. of Carbons No. of Hydrogens
zz Cervonic acid: also known as DHA (Docosa Hexaenoic
Acetic acid 2C 4H
Acid)– This is having 22 C and 6 double bonds (Docosa
Propionic acid 3C 6H means 22, Hexaenoic means 6 double bonds).
Butyric acid 4C 8H  These days health drinks (bournvita, horlicks) are
Valeric acid 5C 10 H for-tified with DHA, as it is important for the brain
Caproic acid 6C 12 H and retina development in infants and children.
Lauric acid 12 C 24 H Decreased amounts lead to increase risk of Retinitis
Myristic 14 C 28 H Pigmentosa.
Palmitic 16 C 32 H  DHA is present in high concentration in sperms,
Stearic 18 C 36 H retina, cerebral cortex.
 Constant source of DHA is breast milk.
zz Unsaturated fatty acids: They have double bonds. This
double bond is in ‘cis’ configuration. But on prolonged zz Alpha-Linolenic Acid: 18 C, 3 double bonds. This Fatty
heating, it gets converted to ‘trans’ configuration. Also Acid is the precursor of omega-3 category. So if this is
trans fats are found in dairy products and partially available in diet, then the other two fatty acids of omega
hydrogenated oils. Trans fats are dangerous as they
increase TG, LDL and decrease HDL. So trans fats 3 category can be synthesized from this.
increase the risk of cardiovascular diseases. They also zz Timnodonic acid– 20 C, 5 double bonds.
increase body’s inflammatory response.
1. Monounsaturated fatty acids Omega-6 Category
2. Polyunsaturated fatty acids
zz Gamma-Linolenic acid– 18 C, 3 double bonds

T Monounsaturated Fatty Acids zz Linoleic acid– 18 C, 2 double bonds. This fatty acid is the
H One double bond is present precursor of omega-6 category. So if this is available in
E Fatty Acid Number of Carbons Number of Double Bonds
diet, then the other two fatty acids of omega 6 category
O can be synthesized from this.
R Palmitoleic 16 C 1
zz Arachidonic acid– 20 C, 4 double bonds. This is very
Y Oleic 18 C 1 (cis)
important for the synthesis of prostaglandins and
Elaidic 18C 1 (trans)
leukotrienes.
 229

CHAPTER 8  CHEMISTRY OF LIPIDS

Fig. 8.4: Classification of PUFAs

Uses of Essential Fatty Acids

A dditional E dge
zz Important constituents of phospholipids. They are Lipotropic factors
mostly present at position 2 of glycerol in phospholipids.
Factors which are required for the synthesis of phospholipids are
zz Decrease the risk of atherosclerosis, cardiovascular
known as lipotropic factors. Their deficiency leads to fatty liver.
disease, fatty liver
Examples of Lipotropic factors:
zz Deficiency of essential fatty acids leads to fatty liver, � Essential fatty acids � Choline
hyperkeratosis, acanthosis of skin and reduced efficiency � Methionine � Betaine
of oxidative phosphorylation.

CLASSIFICATION OF LIPIDS

Fig. 8.5: Classification of Lipids

Mono-acyl Glycerol Di-acyl Glycerol Tri-acyl Glycerol

Simple Lipids (MAG) (DAG) (TGs)
The alcohol here is Glycerol (3C). One Fatty Acid + Two Fatty Acid + Three Fatty Acid +
Glycerol Glycerol Glycerol

T
H
E
O
R
Y
Where R1, R2, R3 are fatty acids
 Triacylglycerol: Non-polar (also known as Neutral Fat/
A dditional E dge
zz
Triglycerides/TGs). TGs are the main lipids present in
230 diet and also the main storage form of lipids in the body. Lecithin
They are present in the adipose tissues. They typically zz Largest body store of Choline
have saturated fatty acid at C1, unsaturated fatty acid at zz Most abundant phospholipid of cell membrane
C2 and fatty acid at C3 can be saturated or unsaturated. Present in high amount in HDL
CRO BIOCHEMISTRY

zz
The enzyme which breaks down TG in adipose tissue zz Dipalmitoyl lecithin is used as a lung surfactant
is Hormone Sensitive Lipase (HSL) present in adipose zz L/S ratio (Lecithin: Sphingomyelin ratio) > 2 indicate fetal lung
tissues. But HSL cannot break fatty acid at position 2. maturity, which occurs at 32 weeks of gestation
Complex Lipids
1. Phospholipids = Alcohol + FA + Phosphate Phospholipases
Phospholipids are complex lipids made up of 3 components
Enzymes which break down phospholipids. They are 4 in
i.e Alcohol, FA and Phosphate. They are also known as phos-
number – A1, A2, C and D
phoglycerides.
Phospholipids are of two types:


1. Glycero-phospholipids— Parent alcohol is Glycerol (3C)

2. Sphingo-phospholipids— Parent alcohol is Sphingosine
(20C)
(a) Glycero-Phospholipids
The parent alcohol is Glycerol here, which is a 3C polyhydroxy
alcohol. It is synthesized from glucose.

Fig. 8.7: Phospholipases. Phospholipase A1 breaks the first fatty

acid, Phospholipase A2 breaks the fatty acid from C2, Phospholipase
C breaks phosphoryl base from rest of the molecule and
Phospholipase D acts between Phosphate and the base

Cardiolipin
zz Cardiolipin was earlier found in heart that’s why named
so.
Phosphatidic acid (PA) = Glycerol + 2 FA + Phosphate. zz A phospholipid present in IMM (Inner Mitochondrial
Phosphatidic acid is the simplest phospholipid and is also the Membrane)
precursor of all phospholipids. zz Has a very complex structure
To complete the valency of phosphate, a base is attached zz It maintains the structure and function of ETC complexes
to phosphate. If this base is choline, then the phospholipid zz Can be antigenic (Usually phospholipids are not anti-
is known as Lecithin. If this base is ethanolamine, then the genic)
phospholipid is known as Cephalin. zz The structure contains one glycerol attached to two
phosphatidic acids. So also known as diphosphatidyl
H igh R eturn glycerol (Fig. 8.8).
zz Lecithin = Glycerol + 2 FA + Phosphate + Choline zz Role of Cardiolipin:
zz Cephalin = Glycerol + 2 FA + Phosphate + Ethanolamine  Make ETC more effective by acting as Proton Trap
zz Phosphatidyl Serine = Serine + Phosphatidic Acid (PA)  In apoptosis signaling
zz Phosphatidyl Inositol = Inositol + Phosphatidic Acid (PA)
zz Phosphatidyl Glycerol = Glycerol + PA
zz Lecithin and Cephalin are the most abundant phospholipids in
T most eukaryotic cells.
H
E
O
R Fig. 8.8: Structure of Cardiolipin
Y

Fig. 8.6 SAM (S-Adenosyl Methionine) used in the synthesis of

lecithin from cephalin
 2. Glycosphingolipids/Glycolipids = Alcohol + FA +
A dditional E dge Carbohydrate
zz Barth Syndrome – a rare X-linked disorder, occurs due to defect 231
in cardiolipin remodelling. Patients have muscle weakness,
neutropenia and cardiomyopathy.
F undamental B ox
zz Antiphospholipid Syndrome – a thrombotic condition which zz Alcohol glycerol is never present

CHAPTER 8  CHEMISTRY OF LIPIDS

leads to abortion. Anti-cardiolipin antibodies are found. zz Phosphate and base is never present
So always in glycolipids, sphingosine alcohol is taken
(therefore they are also known as Glycosphingolipids), one fatty
Platelet Activating Factor (PAF)
acid attached and carbohydrate is either glucose or galactose.
It is a glycerophospholipid in which C1 of glycerol is attached They are also known as cerebrosides.
to saturated alkyl group with an ether linkage and C2 is having
an acetyl residue (not fatty acid). PAF activates inflammation, zz If Glucose added, then it is known as Gluco-cerebroside
thrombosis and aggregate platelets. It lowers the blood or Glucosyl ceramide.
pressure. It is one of the most potent bioactive molecules zz If Galactose added, then it is known as Galacto-
known, effective at very low concentrations. cerebroside or Galactosyl ceramide.
(b) Sphingo-phospholipid = Alcohol + FA + Phosphate zz Gluco-cerebrosides are never found in CNS and Galacto-
zz The parent alcohol is Sphingosine, which is a 20 C amino cerebrosides are always found in CNS.
alcohol. It is synthesized from serine and palmitate. zz In Galacto-cerebrosides, fatty acid mostly present is
Cerebronic Acid (C-24)
zz Sphingosine + Fatty Acid + Monosaccharide (Glu/Gal) =
Cerebroside
zz Sphingosine + Fatty Acid + Oligosaccharide = Globoside
zz Sphingosine + Fatty Acid + Oligosaccharide + NANA
= Ganglioside or
Globoside + NANA = Ganglioside
zz Ceramide: Sphingosine + FA (mostly long chain FA)
Ceramide has amide bond. (whenever amino and
carboxy groups combine, amide(-CONH) bond is
formed)
zz Sphingomyelin: Sphingosine + FA + Phosphate + Cho-
line
 Phosphoryl choline is added to C1 of sphingosine
attached to fatty acid. So sphingomyelin is Ceramide
+ Phosphoryl Choline. It is present in myelin sheath of
nerve cells.
Fig. 8.10: Gangliosides representation (More than 60 gangliosides
are known which have different number and position of NANA)

Fig. 8.9: Structure of sphingomyelin

F undamental B ox Fig. 8.11: Chromatographic separation of Gangliosides. Mono-

Sialo Gangliosides are GM1, GM2, GM3
Mostly lipids have ester bond. But ceramide is a lipid having
amide bond. Gangliosides T
Note: With alcohol glycerol, two fatty acids are attached. But
zz They are never found in liver. H
with alcohol Sphingosine, only one fatty acid is attached.
zz Name is because these were first isolated from ganglion E
cells of brain O
H igh R eturn zz Predominantly found in nervous system (specially in cell R
zz For sugar activation, UDP (Uridine Diphosphate) is used in membranes) Y
carbohydrate synthesis zz Oligosaccharide chain is towards extracellular side and
zz Similarly, in phospholipid synthesis, CDP (Cytidine Diphosphate) has role in signal transduction.
is used
 SPHINGOLIPIDS AND DISEASES i.e. SPHINGOLIPIDOSES (SLP)
232 zz The common non-polar component in all sphingolipids is ceramide
zz Sphingolipidoses are a group of Lysosomal Storage Disorders.
CRO BIOCHEMISTRY


Fig. 8.12: Sphingolipids and their diseases

zz Niemann Pick Disease
Disease Enzyme Deficiency Substrate
accumulated  Mental retardation
 Hepatosplenomegaly
Niemann Pick Sphingomyelinase Sphingomyelin present
disease  Cherry red spots present
Farber’s disease Ceraminidase Ceramide  Lymphadenopathy
zz Farber’s Disease:
Gaucher’s β-Glucosidose or Glucosyl Ceramide ƒƒ Resemble rheumatoid arthritis.
disease β-Glucocerebrosidase ƒƒ Patient has painful joint swelling due to accumu-
lation of ceramide in joints.
Krabbe’s disease β-Galactosidase for Galactosyl Ceramide
Galactosyl Ceramidase
zz Gaucher’s Disease
 Enzyme deficient – Gluco-cerebrosidase
Fabry’s disease α- Galactosidase CeramideTrihexoside  Most common Lysosomal Storage Disease
or Globotriosyl  ERT (Enzyme Replacement Therapy) is available.
Ceramide Recombinant acid β-Glucosidase (Velaglucerase,
Sandhoff’s Hexosaminidase A & B Globoside Taliglucerase, Imiglucerase).
disease  Substrate reduction therapy with oral Miglustat as it
inhibits synthesis of glucosyl ceramide.
Tay Sach’s Ηexosaminidase A GM2 Ganglioside  No cherry red spots
disease  Bony pain, pathological fractures
GM1 β – Galactosidase GM1 Gangliosides  Hepatosplenomegaly
gangliosidosis  No Mental Retardation
(or generalized  Macrophages affected: Their nucleus becomes
gangliosidosis) eccentric due to accumulation of glucocerebrosides
and cytoplasm shows a typical crumpled tissue
Metachromatic Αrylsulfatase A or Cerebroside Sulfate
T paper appearance (due to large elongated lysosomes
Leukodystrophy Cerebroside Sulfatase or Sulfatide
filled with glucocerebrosides).
H
E Fucosidosis α- Fucosidase H- iso antigen  Only extra neurological manifestations: Pancyto-
(Fucose present in penia, Thrombocytopenia and bleeding, pain and
O
globoside) pathological fractures of long bones, osteoporosis,
R avascular necrosis of femur, Erlenmeyer flask
Y deformity in X-ray femur, Gaucher cell in bone
marrow biopsy.
 zz Metachromatic Leukodystrophy:

233

CHAPTER 8  CHEMISTRY OF LIPIDS

H igh R eturn
zz All Sphingolipidoses are autosomal recessive except Fabry’s
Fig. 8.13: Crumpled tissue paper appearance of Gaucher’s cells which is X- linked recessive
shown zz No hepatosplenomegaly in Tay Sach’s & Krabbe’s disease.
zz Krabbe’s Disease or Globoid cell Leukodystrophy: zz Sphingolipidosis with no cherry red spot – Fabry’s, Krabbe’s &
 Increased Galactocerebroside in white matter of Gaucher’s disease
brain. zz Sphingolipidosis with no mental retardation in Gaucher’s &
 Severe neurological deficit (no extra neurological Fabry’s disease
manifestations) zz Sphingolipidosis with angiokeratoma – GM1 gangliosidosis &
 No hepatosplenomegaly Fabry’s disease
zz 2 Classic Leukodystrophies which are Sphingolipidosis:
 Inclusion bodies known as multi nucleated globoid
1. Krabbe’s (Globoid cell Leukodystrophies)
cell inclusion bodies Globoid cells 2. Metachromatic Leukodystrophies
are found
zz Other 2 Leukodystrophies are Adreno Leukodystrophy and
 Unmetabolized
Alexander disease
substances
zz Diseases of myelin are leukodystrophies (congenital) and
accumulate in macro-
phages → makes large multiple sclerosis (acquired)
spherical globoid cells zz All gangliosidosis are sphingolipidosis

Fabry’s Disease:
A dditional E dge
zz
 X- linked recessive, so only males affected
 Triad of peripheral neuropathy, Angiokeratoma Wolman’s disease (AR) or Cholesterol Ester Storage disease or
(see figure) and Angiokeratoma Lysosomal Acid Lipase deficiency
Hypohidrosis zz Not a sphingolipidosis

 Fabry’s crisis occurs zz A lysosomal storage disease

zz Enzyme Acid Lipase
(similar to sickle cell
zz Watery green diarrhoea –presents in 1st week of life
crisis) – pain and
zz Failure to thrive
swelling in proximal zz Relentless vomiting, Steatorrhoea, Abdominal distension,
joints. Hepatosplenomegaly, Lymphadenopathy
zz Sandhoff’s Disease: zz Calification of adrenals is pathogenomic feature
A GM2 gangliosidosis (like Tay Sach’s ) zz ↑↑Ch esters and TG

zz Tay Sach’s Disease: zz Hepatic dysfunction

 No hepatosplenomegaly
 Mental retardation
 Cherry red spots on macula
 Progressive neurodegeneration
zz GM1 gangliosidosis:

Bile Acids
T
zz Bile acids are more appropriately called bile salts as they H
are present in deprotonated form at physiological pH. E
Patient has mental retardation, Hepatosplenomegaly, zz Bile salts, made in liver and stored in gall bladder,
Angiokeratoma, cherry red spots. O
emulsify the dietary fat which increases the surface area.
R
Y
 zz Formed from cholesterol by modification of side chain. zz Least enterophatic circulation is for lithocholic acid.
zz Primary bile acids are cholic acid and chenodeoxycholic
234 acid (cholic acid is more abundant)
zz Primary bile acids are synthesized in liver (RLE is
7-a-Hydroxylase)
zz Conjugation of primary bile acid occurs in liver with
CRO BIOCHEMISTRY

glycine or taurine and these are stored in gall bladder.

zz Conjugated primary bile acid from liver reaches intestine
via common bile duct. In intestine, secondary bile acid is
formed by bacterial action.

Fig. 8.14: Regulation of Bile Acid and Cholesterol synthesis.

zz Hypothyroidism increases plasma cholesterol as T3, T4

activates 7-a– Hydroxylase.


zz Deconjugation and remonal of 7-a-hydroxy group occurs

zz 98-99% secondary bile acid undergo enterohepatic re-
circulation and reaches back liver. So, bile contains both
primary and secondary bile acid.

URSO-DEOXY CHOLIC ACID

An isomer of Primary Bile Acid i.e. Chenodeoxycholic acid
A metabolic by-product of Intestinal Bacteria
Found in very low concentration in bile
Bear bile contains ursodiol (Ursus means Bear)
zz It is hepato-protective: Improves bile flow, modifies bile acid pool by increasing non-toxic hydrophilic bile acid and decreasing formation
of hydrophobic bile acid
zz Cyto-protective
zz Anti-apoptotic
zz Immuno-modulatory
zz Delays cirrhosis and gastro oesophageal varices
Used in treatment of Liver Diseases:
zz Primary biliary cirrhosis
zz Obstetric cholestasis (to relieve itching)
zz Non-alcoholic fatty liver diseases
zz Cystic fibrosis associated liver diseases
zz Treatment of cholesterol gall stones non-surgically

Pearls of the Chapter

zz Fatty acid is Polar. Fat is non-Polar.
zz Ester bond in all Fats. But ceramide has amide bond
zz Glycero-phospholipids – parent alcohol is Glycerol (3C)
zz Sphingo-phospholipids – parent alcohol is Sphingosine
zz Phosphatidic acid is the simplest phospholipid and is also the precursor of all phospholipids
zz Lecithin is Phosphatidyl Choline and Cephalin is Phosphatidyl Ethanolamine
T
zz SAM (S-Adenosyl Methionine) is used in the synthesis of lecithin from Cephalin
H
zz Lecithin is the largest body store of choline
E
zz L/S ratio (Lecithin: Sphingomyelin ratio) > 2 indicate fetal lung maturity, which occurs at 32 weeks of gestation
O
zz Lipoprotein with maximum Phospholipid → HDL
R
zz A Phospholipid which can be antigenic is Cardiolipin
Y
zz Platelet Activating Factor (PAF) is a Glycerol-phospholipid
Contd…
zz Sphingosine is a 20 C amino alcohol, which is synthesized from serine and palmitate.
For sugar activation, UDP (Uridine Diphosphate) is used in carbohydrate synthesis. Similarly, In phospholipid synthesis, CDP (Cytidine
zz
235
Diphosphate) is used
zz Glycolipids or Glycosphingolipids are the source of ABO blood group antigens, where carbohydrate is the antigenic determinant.
zz Fabry’s Disease (alpha-Galactosylceramidase deficiency) is XR

CHAPTER 8  CHEMISTRY OF LIPIDS

zz Gaucher’s disease is the most common Lysosomal Storage Disease
zz Coconut and coconut oil has highest content of medium chain fatty acids
zz Number of hydrogens in case of a saturated fatty acids are double the number of carbons
zz Linoleic acid - 18 C, 2 double bonds. This fatty acid is the precursor of omega-6 category
zz Alpha-Linolenic Acid – 18 C, 3 double bonds. This fatty acid is the precursor of omega-3 category.

COMPOSITION OF PHOSPHOLIPIDS
PHOSPHOLIPIDS COMPOSED OF
Phosphatidic Acid (PA) Glycerol + 2 Fatty Acid+ Phosphate
Lecithin/ Phosphatidyl Choline Phosphatidic Acid + Choline
Cephalin/ Phosphatidyl Ethanolamine Phosphatidic Acid + Ethanolamine
Cardiolipin Glycerol attached to 2 Phosphatidic Acid
Sphingomyelin Sphingosine + Fatty Acid+ Phosphate + Choline
• Sphingosine and fatty acid (mostly long chair) is known as Ceramide as there is amide bond in this.

T
H
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 236 Multiple Choice Questions
Chemistry of Lipids 11. Which of the following processes yields arachidonic
1. Rate limiting enzyme in bile acid synthesis: acid in mammals? (Recent Question 2017)
 (PGMEE 2015) a. Elongation of stearic acid
a. Sterol 27- hydroxylase b. Chain elongation and one desaturation of linolenic
b. HMG CoA synthase acid
c. 7-α hydroxylase c. Chain elongation and two desaturations of linoleic
d. HMG- CoA reductase acid
2. Bile acids consist of all EXCEPT-  (PGMEE 2015) d. Chain elongation and one desaturation of linoleic
a. Deoxycholic acid acid
b. Lithocholic acid 12. Which of the following is most essential fatty acid?
c. Dideoxy cholic acid  (PGMEE 2015, 2017)
d. Chenodeoxycholic acid a. Alpha Linolenic acid
3. The main function of bile salts is:  (PGMEE 2009) b. Gamma Linolenic acid
a. Fat absorption c. Lauric acid
b. Water soluble d. Linoleic acid
c. Bowel motility control 13. C17 H31 COOH is formula of:
d. Bacteriostatic a. Linoleic acid b. Linolenic acid
4. Bile salts help in absorption of fat by: (PGMEE 2015) c. Arachidonic acid d. Timnodonic acid
a. Micelle formation 14. Products of complete hydrolysis of cardiolipin are:
b. Creation of concentration gradient a. 3 Glycerol, 4 fatty acids, 2 phosphates
c. Activation of transporter protein b. 3 Glycerol, 4 fatty acids, 1 phosphates
d. All of the above c. 3 Glycerol, 3 fatty acids, 2 phosphates
5. Bile acids are synthesized from-  (PGMEE 2015) d. 5 Glycerol, 4 fatty acids, 2 phosphates
a. Arachidonic acid 15. Which of the following is not a product of complete
M b. Linoleic acid hydrolysis of Sphingomyelin?
C c. Linolenic acid a. Choline b. Phosphate
Qs d. Acetyl CoA c. Ceramide d. Palmitic acid
Ans. 6. Products of cyclooxygenase pathway are all EXCEPT: 16. Ceramide contains which bond :
 (PGMEE 2013) a. Acid (COOH)
1. c b. Amide (CONH)
a. PGE2 b. LTB4
2. c c. Ester (COOR)
c. PGF2 d. PGD2
3. a d. Hydrogen bond
7. Retinitis pigmentosa patients doesn’t have:
4. a 17. Alcoholic is found in all EXCEPT:
 (Recent Pattern Jan 2019)
5. d a. Glucocerebroside
a. DHA
6. b b. DHA (Docosa Hexaenoic Acid)
b. Eicosapentaenoic acid
7. a c. Lecithin
c. Arachidonic acid
8. b d. Sphingomyelin
d. Timnodonic acid
9. a 18. Lipoxins belong to which of the following family of
8. Which of the following is an Essential fatty acids?
10. c chemical mediators of inflammation:
 (FMGE JUNE- 2018 )
11. c  (PGMEE 2016-17; PGMEE 2007, 2009)
a. Citric acid 
12. d a. Kinin system
b. Linoleic acid
13. a
c. Stearic acid b. Cytokines
14. a
d. Palmitic acid c. Chemokines
15. c
9. Essential fatty acids are helpful in controlling which d. Arachidonic acid metabolites
16. b
of the following? (Recent Question 2018) 19. A major lipid of mitochondrial membrane is:
17. b
a. Atherosclerosis b. Nephritis  (PGMEE 2008, 2013)
18. d
c. Diabetes Mellitus d. Oedema a. Inositol b. Plasmalogen
19. c
10. Which of the following has maximum medium chain c. Cardiolipin d. Lecithin
20. d
fatty acid content (Recent Question 2018) 20. Which of the following is not a steroid?
a. Sunflower oil b. Flaxeed oil  (PGMEE 2012)
c. Coconut oil d. Fish oil a. Vitamin D b. Cholic acid
c. Estrogen d. Leukotrienes
21. Which of the following is an omega 6 fatty acid? 33. Glycosphingolipid is made up of: (PGI Nov 2011)
 (PGMEE 2013-14) a. Glucose
a. DHA b. Linoleic acid b. Glycerol 237
c. Alpha-linolenic acid d. EPA c. Sphingosine
22. Which is an omega – 9 fatty acid-(PGMEE 2012,2014) d. Fatty acids
a. Cervonic acid b. Oleic acid e. Thromboxane A2
c. Linolenic acid d. Arachidonic acid 34. Monoprotic acids are: (PGI Nov 2011)
23. Which among the fatty acids are very harmful for a. Formic acid b. Carbonic acid
health:- (PGMEE 2016-17) c. Acetic acid d. Citric acid
a. Trans fatty acid e. Nitric acid
b. Cis- saturated fatty acid 35. False about Trans Fatty Acid (TFA): (PGI Nov 2009)
c. Mono unsaturated FA (MUFA) a. Increase risk of cardiovascular disease
d. Poly unsaturated FA (PUFA) b. Fried foods have high content of TFA
24. In Wolman’s disease there is accumulation of- c. Hydrogenation increases TFA
 (PGMEE 2013) d. Hydrogenation decreases TFA
a. Cerebrosides b. Triglycerides e. It ↑ LDL and ↓ HDL
c. Sphingosides d. Cholesterol
25. Which of the following is Simplest Ganglioside? Lipid Storage Diseases
a. GM1 Ganglioside b. GM2 Ganglioside 36. Enzyme deficient in Niemann Pick disease:
c. GM3 Ganglioside d. GM4 Ganglioside a. Glucocerebrosidase (Recent Question 2017)
26. Which of the following does not contain cholesterol? b. Sphingomyelinase
a. Membranes b. Vitamin D c. Beta galactosylceramidase
c. Estrogen d. Adrenaline d. Alpha galactosylceramidase
27. Cholesterol is not a precursor for synthesis of- 37. A 48 year old female presented with bony pain and
 (PGMEE 2015) hepatosplenomegaly. On examination of biopsy
a. Vitamin D b. Bile acids from spleen, crumpled tissue paper appearance is
c. Lipocortin d. Progesterone seen. Which of the following product is likely to have
28. True statement(s) about lipid digestion and accumulated?
absorption: (PGI May 2017) a. Ganglioside b. Sulfatide
a. Micelles play an important role in lipid absorption c. Sphingomyelin d. Glucocerebroside
b. Absorption of long-chain fatty acids is greatest in the M
38. Multiple sclerosis is characterized by loss of which
upper parts of the small intestine lipids: (PGMEE 2016-17) C
c. Bile acid has no role in fat absorption a. Phospholipids and sphingolipids Qs
d. Fatty acids after absorption are reesterified to b. Sphingolipids and ceramide Ans.
triglycerides in the enterocytes c. Phospholipids and cramide 21. b
29. Correct statement about membrane: (PGI May 2016) d. Sphingolipids and gangliosides 22. b
a. Phospholipids undergo rapid lateral diffusion 39. Tay Sach’s disease is because of increased : 23. a
b. Transverse movement of lipids across the membrane a. GM1 Gangliosides b. Glucocerebrosides 24. b
is faster than protein c. GM2 Gangliosides d. Galactocerebrosides 25. c
c. Hydrophobic core of the phospholipid bilayer 40. Enzyme deficient in Gaucher’s disease- 26. d
remains constantly in motion because of rotations  (PGMEE 2015) 27. c
around the bonds of lipid tails a. G-6PD b. Sphingomyelinase 28. a,b,d
d. Phospholipds that have one fatty acyl group, cannot c. Glucocerebrosidase d. Glucokinase 29. a,c,d
form the bilayer 30. b,c,e
41. Hexosaminidase A deficiency causes:~
30. Gangliosides contains: (PGI May 2008,2015) 31. b,c
 (PGI May 2015)
a. Phosphate b. Galactose 32. a,b,c
a. Niemann Pick b. Tay-Sachs disease
c. Long chain FA d. Serine 33. a,c,d
c. Hurler syndrome d. Gaucher’s disease
e. Sialic acid 34. a,c,e
e. Krabbe’s disease
31. Essential Fatty Acid is/are: (PGI May 2013) 35. d
42. Which of the following is/are not sphingolipidosis:
a. Palmitic acid b. Linoleic acid 36. b
 (PGI May 2013)
c. Linolenic acid d. Oleic acid 37. d
a. Tay Sach’s disease b. Sandhoff ’s disease
e. Free Fatty Acid 38. a
c. Krabbe’s disease d. Fabry’s disease
32. True statement about fatty acid: (PGI Nov 2011) 39. c
e. Wolman disease
a. Polyunsaturated FA is essential for membrane 40. c
43. True about Gaucher’s disease: (PGI Nov 2009)
structure 41. b
b. Biologically arachidonic acid is essential to life a. Due to deficiency of enzyme sphingomyelinases
b. Due to deficiency of enzyme β-Gluco-cerebrosidase 42. e
c. Hydrogenated vegetable oils contains trans fatty
c. Deposition of glucosyl ceramide 43. b,c
acid
d. Most of the naturally occurring unsaturated FA d. Foam cell deposition
exists as trans-isomer e. Crumpled tissue paper appearance
 44. Wrinkled paper appearance of macrophage in bone 48. Beta galactosidase deficiency causes: 
marrow aspirate is seen in: (PGMEE 2015) a. Gaucher’s disease (PGI May 2018)
238 a. Tay Sachs Disease b. Krabbe’s disease
b. Gaucher’s disease c. Fabry’s disease
c. Niemann-Pick disease d. Neimann Pick disease
d. Leukodystrophy e. Metachromatic Leukodystrophy
45. Accumulation of cerebral gangliosides occurs due to 49. Krabbe’s disease or globoid cell leukodystrophy is a
deficiency of:  (PGMEE 2015) fatal, degenerative disorder affecting myelin sheath.
a. β glucocerebrosidase Lipid accumulating in Krabbe’s disease is 
b. β-galactosidase  (PGMEE 2013-14)
c. Sphingomyelinase a. Sphingomyelin
d. Hexosaminidase A b. Glucosyl ceramide
46. Which of the following does not occur in lysosomal c. Galactosyl ceramide
storage disorders? (PGMEE 2013-14) d. GM2 ganglioside
a. Increased number and size of lysosomes 50. Correct match of disease and enzyme defect:
b. Accumulation of the polyubiquinated proteins  (PGMEE 2016-17)
c. No neurological deficit a. Krabbe’s – Galactosyl Ceramidase
d. Apoptosis defects b. Gaucher’s- galactocerebrosidase
47. Which is lipotropic factor:- (PGMEE 2016-17) c. Farber’s disease – Alpha galactosidase A
a. HDL b. Carnitine d. Sandhoff’s –Arylsulfatase A
c. Choline d. Insulin

M
C
Qs
Ans.
44. b
45. b
46. c
47. c
48. b
49. c
50. a
 Answers with Explanations
239
1. Ans. (c ) 7- α hydroxylase •• Omega-3 PUFAs include Cervonic acid/ DHA, Alpha
Linolenic, Timnodonic acid.

CHAPTER 8  CHEMISTRY OF LIPIDS

[Ref: Harper 30th /e pg.273] •• Omega-6 PUFAs include Gamma Linolenic acid,
•• Rate limiting enzyme in bile acid synthesis is Linoleic acid and Arachidonic acid.
7-α-hydroxylase. •• Omega-3 PUFAs are cardioprotective as they shift
the equilibrium towards beneficial anti- atherogenic
2. Ans. (c ) Dideoxy cholic acid state.
•• A lower omega-6/omega-3 ratio is more desirable in
[Ref : Harper 30th /e pg. 273]
reducing the risk of many of the chronic diseases of
•• Bile acids consists of Deoxycholic acid, Lithocholic high prevalence in western societies.
acid and Chenodeoxycholic acid.
9. Ans. (a) Atherosclerosis
3. Ans. (a) Fat absorption
[Ref: Lehninger 7th/e pg. Fig. 821]
[Ref: Harper 30th /e pg. 539] •• Essential Fatty Acids (EFA) are helpful in controlling
•• The main function of bile salts is digestion and atherosclerosis. Essential fatty acids are constituents
absorption of fat and fat soluble vitamins. of phospholipids. Phospholipids are present in lipo-
proteins. Lipoprotein with maximum phospholipid
4. Ans. (a) Micelles formation is HDL. HDL prevents atherosclerosis. So we can say
that EFA prevent atherosclerosis.
[Ref: Gangong 24th /e p. 465}
•• Bile salts help in absorption of fat by micelles 10. Ans. (c) Coconut oil
formation as bile salts are amphipathic in nature. [Ref: Lehninger 7th/e pg. Fig. 360]
Hydrophilic nonpolar lipids are in centre of micelle •• Maximum medium chain fatty acid content is in
and are absorbed. Coconut oil.
5. Ans. (d) Acetyl CoA 11. Ans. (c) Chain elongation and two desaturations of
th
[Ref: Harper 30 /e pg. 273] linoleic acid

•• Bile acids are synthesized from cholesterol, which is [Ref: Harper 30th/e pg. 238]
synthesized from Acetyl CoA. •• Arachidonic acid (20 C, 4 double bonds) is a semi-
•• Arachidonic acid synthesized prostaglandins and essential fatty acid belonging to omega-6 category.
This can be synthesized in body from the precursor
A
Leukotrienes
of omega-6 category i.e. Linoleic acid (18 C, 2 double N
6. Ans. (b) LTB4 bonds). So from Linoleic to Arachidonic, we have to S
convert 2 to 4 double bonds and 18C to 20C. W
[Ref: Harper 30th /e pg. 24] E
•• Products of cyclooxygenase pathway are prosta- 12. Ans. (d) Linoleic acid R
glandins (PGE2, PGF2 and PGD2). Prostacyclins and [Ref: Lehninger 7th/e pg. Fig. 362] S
thromboxanes
•• Most essential fatty acid is Linoleic acid (18-C, 2 WITH
7. Ans. (a) DHA double bonds).
•• Linoleic acid is the precursor of omega-6 category. E
[Ref : Harper 30th /e pg. 214] It can synthesize the other fatty acids of omega-6 X
category i.e. gamma Linolenic acid and Arachidonic P
•• Not all, but many studies have found that patients
acid. L
with Retinitis Pigmentosa tend to have lower blood
•• Precursor of omega-3 category is a-Linolenic acid. A
levels of DHA (Docosa Hexaenoic Acid), an omega -3
N
fatty acid found in the photoreceptor cells. 13. Ans. (a) Linoleic acid A
8. Ans. (b) Linoleic acid T
[Ref: Lehninger 7th/e pg. Fig. 362]
I
[Ref: Lehninger 7th/e pg. Fig. 362] •• In case of a saturated fatty acid, number of hydrogens O
•• Essential fatty acids are those fatty acids which are
are double the number of carbons. In this fatty acid, N
required in diet. These are also known as PUFAs.
number of carbons are 18. Number of hydrogens are S
 32. In case of saturated fatty acid, if carbons are 18, 19. Ans. (c) Cardiolipin
then Hydrogen must be 36 (double). So there is loss of
240 4 H (means 2 double bonds). So, this is Linoleic acid
[Ref: Harper 30th/e pg. 217]
•• Cardiolipin is a phospholipid which is present in
(18 C, 2 double bonds).
inner mitochondrial membrane. It is found only in
14. Ans. (a) 3 Glycerol, 4 fatty acids, 2 phosphates mitochondria and has complex structure. It can be
CRO BIOCHEMISTRY

antigenic. Decreased or altered cardiolipin can lead

[Ref: Harper 30th/e pg. 217]
to ageing, heart failure, hypothyroidism and Barth
•• Cardiolipin is made up of glycerol attached to two syndrome (cardioskeletal myopathy).
Phosphatidic acid. Each phosphatidic acid contains
one glycerol, two fatty acids and one phosphate. 20. Ans. (d) Leukotrienes
So in total, there are 3 glycerols, 4 fatty acids and 2
[Ref: Harper 30th/e pg. 502; Fig 41-2 and p.273]
phosphates.
•• Steroids have four – fused ring structure. Cholesterol
15. Ans. (c) Ceramide is the steroid in mammals.
•• Estrogen is a steroid hormone.
[Ref: Harper 30th/e pg. 215] •• Cholic acid is a bile acid formed from cholesterol by


•• Ceramide is a product of partial hydrolysis. In the modification of its side chain

question, complete hydrolysis is asked. •• Vitamin D is also a steroid synthesized from chole-
•• Sphingomyelin is made up of sphingosine alcohol, sterol.
one fatty acid, one phosphate and one choline.
•• Ceramide = Sphingosine + Fatty Acid 21. Ans. (b) Linoleic acid
•• The fatty acid can be palmitic acid
[Ref: Harper 30th/e pg. 214]
16. Ans. (a) Amide (CONH) •• Omega-6 fatty acids are gamma linolenic acid,
linoleic acid and arachidonic acid.
[Ref: Harper 30th/e pg. 216]
•• Ceramide, as the name indicates, it contains amide 22. Ans. (b) Oleic acid
bond.
[Ref: Harper 30th/e pg. 214]
17. Ans. (b) DHA (Docosa-Hexaenoic Acid) •• Oleic acid is a mono-unsaturated or monoenoic fatty
[Ref: Harper 30th/e pg. 217,218] acid i.e. 18 Carbons with one double bond at ω-9
position (ω-numbering starts from methyl end of
•• Glucocerebrosides and Sphingomyelin contains
Sphingosine alcohol. fatty acid)
•• Lecithin contains glycerol alcohol. 23. Ans. (a) Trans fatty acid
•• DHA (Docosa-Hexaenoic Acid) is a 22-C, 6 double
bond fatty acid. [Ref: Harper 30th/e pg. 213]
A
N •• Trans fatty acids are harmful for health because
18. Ans. (d) Arachidonic acid metabolites
S they increase LDL – Cholesterol, TGs, inflammation,
[Ref: Harper 30th/e pg. 240] cancer, infertility. They also decrease HDL – Chole-
W
E •• Leukotrienes and Lipoxins are formed by the sterol.
Lipoxygenase pathway. These are synthesized from
R
Arachidonic acid (20 C, 4 double bonds) and its 24. Ans. (b) Triglycerides
S metabolites.
[Ref: Harper 30th/e pg. 628]
WITH
•• Triglycerides >> cholesterol
E •• Wolman's disease, also known as lysosomal Acid
X Lipase deficiency, is a rare autosomal recessive
P disease. Triglycerides and cholesterol esters gets
L accumulated. They are accumulated in organs like
A liver, spleen, gut and blood vessels.
N
A 25. Ans. (c) GM3 Ganglioside
T
[Ref: Harper 30th/e pg. 218]
I
O •• The simplest ganglioside is GM3. It contains Ceramide
N + Glucose + Galactose + NANA (N-acetyl Neuraminic
S Acid).
26. Ans. (d) Adrenaline 32. Ans. (a); (b); (c)
[Ref: Harper 30th/e pg. 217, 218, 219] [Ref: Lehninger 7th/e pg. 362] 241
•• Cholesterol is present in membranes, Vitamin D
•• Most of the naturally occurring unsaturated FA exists
and Estrogen (steroid hormones). Cholesterol is not
as cis-isomer (not trans).
present in adrenaline.
•• Hydrogenation increases trans fatty acid.

CHAPTER 8  CHEMISTRY OF LIPIDS

•• Arachidonic acid is used for synthesis of prosta-
glandins and leukotrienes.

33. Ans. (a); (c); (d)

[Ref: Harper 30th/e pg. 217]

•• Glycosphingolipids are sphingolipids with carbohy-
drate attached. They have Sphingosine, Fatty acid
and Glucose or Galactose or oligosaccharide.
NANA is present in gangliosides, which are complex
glycosphingolipids.

34. Ans. (a); (c); (e)

[Ref: Harper 30th/e pg. 13]

27. Ans. (c) Lipocortin

[Ref: Harper 30th/e pg. 219]
•• Lipocortin or Annexin is a protein which is used as
one of the mechanism, by which glucocorticoids
inhibit inflammation.
•• Cholesterol is used for Bile acids/salts, membranes,
vitamin D and steroid hormones.

28. Ans. (a); (b); (d)

[Ref: Harper 30th/e pg. 273]
•• Bile acid has a very important role in fat absorption. A
•• Other options given in question are true. N
29. Ans. (a); (c); (d) S
W
[Ref: Harper 30th/e pg. 218] E
Refer Ques. 37. of concepts R
S
30. Ans. (b);(c); (e)
WITH
[Ref: Harper 30th/e pg. 218]
•• Gangliosides are complex glycolipids. Glycolipids E
do not contain Glycerol alcohol. They only contain X
sphingosine alcohol. They do not have phosphate P
and choline. L
•• Gangliosides do not have phosphate, base (Cholins) A
and glycerol. 35. Ans. (d) Hydrogenation decreases TFA N
[Ref: Harper 30th/e pg. 213] A
31. Ans. (b); (c) T
•• Partial hydrogenation of oils is done to increase
their shelf life and to prevent rancidity. But this I
[Ref: Harper 30th/e pg. 362]
increases trans fatty acids. They increase LDL, TG, O
•• 2 essential fatty acids are Linoleic (omega-6 category)
inflammation, cancer and decrease HDL- cholesterol N
and a-Linolenic (omega-3 category).
S
 36. Ans. (b) Sphingomyelinase 44. Ans. (b) Gaucher's disease

242 [Ref: Harper 30th/e pg. 251] [Ref: Robbins 9th/e pg. 141]
•• Enzyme deficient in Niemann Pick disease is Sphing- •• Wrinkled paper or crumpled tissue appearance of
omyelinase and Sphingomyelins accumulate. macrophage in bone marrow aspirate is seen in
Gaucher’s disease.
CRO BIOCHEMISTRY

37. Ans. (d) Glucocerebroside

[Ref: Harper 30th/e pg. 251] 45. Ans. (d) Hexosaminidase A
•• This is Gaucher’s disease (most common lysosomal [Ref: Robbins 9th/e pg. 151-154]
storage disease). Glucocerebrosides accumulate •• Accumulation of cerebral gangliosides occur in
in reticuloendothelial system. Macrophages have Tay Sach’s disease (deficiency of Hexosaminidose
typical crumpled tissue paper appearance/wrinkled
A). Patient has mental retardation but no hepa-
appearance.
tosplenomegaly
38. Ans. (a) Phospholipids and sphingolipids
46. Ans. (c) No Neurological deficit
[Ref: Harper 30th/e pg. 250]


•• Multiple sclerosis is a demyelinating disease in which [Ref: Robbins 9th/e pg. 149-150]
there is loss of phospholipids and sphingolipids from •• Lysosomal storage disorders are inborn diseases
white matter. So lipid composition of white matter due to lack or inactivity of lysosomal hydrolases,
resembles that of gray matter. CSF shows raised transporters or integral membrane proteins.
phospholipid levels. •• Usually LSD have severe neurological abnormalities.
There are large vacuoles and ubiquitinated
39. Ans. (c) GM2 Gangliosides aggregates, promoting degradation and also
induction of lysosome biogenesis. There occurs
[Ref: Lehninger 7th/e pg. 373 b ]
defects in apoptosis. Substrate accumulation in
•• Tay Sach’s disease is because of increased GM2 lysosomes increase the size of lysosomes.
Gangliosides.
•• Cherry red spot occurs in Tay Sach’s disease. 47. Ans. (c) Choline
40. Ans. (c) Glucocerebrosidase [Ref: Harper 30th/e pg. 261]
[Ref: Harper 30th/e pg. 179] •• Lipoprotic factors are those which are required for
•• Gaucher’s disease is due to deficiency of β-Gluco- synthesis of phospholipids. E.g.: EFAs, Choline,
sidase or β-Glucocerebrosidase and Glucosyl Methionine, Betaine.
ceramide gets accumulated.
48. Ans. (b) Krabbe’s disease
41. Ans. (b) Tay-Sachs disease
[Ref: Harper 30th /e pg.251]
A [Ref: Robbins 9th/e pg. 151table]
•• Beta galactosidase deficiency causes Krabbe’s
N •• Hexosaminidase A deficiency causes Tay-Sachs disease.
S disease.
W 49. Ans. (c) Galactosyl ceramide
42. Ans. (e) Wolman disease
E
R [Ref: Harper 30th/e pg. 628] [Ref: Harper 30th /e pg. 250]
•• Wolman disease is caused by deficiency of Lysosomal
S •• Lipid accumulating in Krabbe’s disease is Galacto-
Acid Lipase. Wolman disease is a lysosomal storage
sylceramide.
WITH disease but not sphingolipidosis. All other options
are sphingolipidosis. 50. Ans. (a) Krabbe's – Galactosyl Ceramidase
E
X 43. Ans. (b); (c); (e) [Ref: Harper 30th /e pg. 179]
P [Ref: Harper 30th/e pg. 251] •• Krabbe’s Disease is caused due to defect in Galactosyl
L •• In Gaucher’s disease, Gaucher cells are found which Ceramidase/ β- Galactosidase
A have appearance of crumpled tissue paper (not foam •• Gaucher’s Disease is caused due to defect in β-
N cells) Glucosidase or β-Glucocerebrosidase.
A •• Farber’s Disease is caused due to defect in Ceram-
T inidase.
I •• Sandhoff’s Disease is caused due to defect in Hexo-
O saminidase A and B
N
S
Lipoproteins 9
Overview of Chapter Proteins present in lipoprotein are known as apolipo-
proteins. Their synthesis occurs in RER (Rough Endoplasmic
•• Chylomicrons Reticulum) and golgi. They are Apo AI, AII, AIV, Apo B48, Apo
•• VLDL B100, Apo C-I, C-II, C-III and Apo E.
•• IDL Lipoproteins are
•• LDL
1. Chylomicron
•• HDL 2. Chylomicron Remnant
•• Hyperlipoproteinemia 3. VLDL (Very Low Density Lipoprotein)
•• Micelle and Lipoproteins 4. VLDL Remnant/IDL (Intermediate Density Lipoprotein)
5. LDL (Low Density Lipoprotein)
6. HDL (High Density Lipoprotein)
LIPID TRANSPORT
Lipid transport is a special topic in lipids (no such topic for
carbohydrates or proteins) because lipids are non-polar and
the transport medium in our body is blood, which is polar.
A non polar thing is insoluble in a polar medium. So how
to transport lipids in body from one place to another? For
this purpose, lipoproteins are synthesized in the body i.e.
lipid and protein is combined. Lipid portion (non-polar) is
placed in the core and protein portion (polar) is placed in the
periphery so that lipoprotein can easily move in blood and
Fig. 9.2: Order of lipoproteins
helps in the transport of lipids from one place to another.

Lipids Present in Lipoprotein are Lipoprotein Function Synthesized In

zz Triglycerides (TG) Chylomicrons Transports Exogenous Intestine
zz Phospholipids (PL) Triglycerides from Intestinal to
zz Free cholesterol (Unesterified) Peripheral tissues.
zz Cholesterol-ester (CE) VLDL Transports Endogenous Liver
Triglycerides from Liver to
Peripheral tissue.
LDL Transports Cholesterol from Liver
Liver to Peripheral tissue.
HDL Transports excess Cholesterol Liver
from Peripheral tissue to Liver
(Known as Reverse Cholesterol
Transport)

TRANSPORT OF LIPID
Exogenous Fat
Fig. 9.1: Neutral lipids lie in the core and amphipathic lipids lie in In diet, 90% lipids are triglycerides (TGs). From diet, TGs
the periphery. TGs and Cholesterol ester are neutral or totally non- reach intestinal lumen where they are broken down by
polar. Free Cholesterol and Phospholipids are amphipathic digestive lipases into three fatty acids and glycerol. These are
smaller molecules so they get absorbed into the intestinal
zz Cholesterol—a kind of amphipathic alcohol mucosal cells. Inside the cell, they rejoin to form TGs. Now
zz Cholesterol ester (Cholesterol + Fatty Acid) is non polar fat intestinal cells add Apo B-48 around the TGs, making them
 Chylomicrons. These chylomicrons move from intestinal periphery of these endogenous TGs, the resultant structure
cells into the lymphatic vessels. This milky appearing lymph formed is known as VLDL, which is secreted into the
244 is known as chyle. From lymphatic vessels they enter thoracic blood from liver. So VLDL transports endogenous TGs or
duct and then into the left subclavian vein, from where they endogenous lipids from liver to peripheral tissues.
enter blood. So chylomicrons carry exogenous lipid (or In blood, VLDL is acted upon by Lipoprotein Lipase
exogenous TGs) from intestine to peripheral tissues. (Fig. 9.3) and loses TGs and forms VLDL remnant (also known as
CRO BIOCHEMISTRY

IDL-Intermediate Density Lipoprotein). Then further IDL is

acted upon by Lipoprotein Lipase and loss of TGs occur till
the point all TGs are finished and just cholesterol left. That
structure is known as LDL (Low Density Lipoprotein).


Fig. 9.5: Lipoprotein Cascade Pathway. The conversion of VLDL to

IDL and then to LDL is known as Lipoprotein Cascade Pathway.
Fig. 9.3: Formation and release of chylomicron
LDL is then taken by liver with the help of special LDL
Then chylomicrons move throughout the body in blood
–receptors present on the surface of liver. LDL is taken by a
and they are acted upon by enzyme lipoprotein lipase. This
special mechanism – Receptor Mediated Endocytosis, using
enzyme breaks the TGs of chylomicrons into fatty acids and
Clathrin protein.
glycerol. These are smaller molecules so they enter peripheral
tissues. After losing many TGs and after taking Apo E and
Apo C (from HDL), chylomicron is converted to chylomicron
remnant. (Fig. 9.4)

Fig. 9.4: Conversion of chylomicron to chylomicron remnant

zz In circulation, HDL donates Apo C and Apo E to
chylomicron and VLDL so that they are converted to
chylomicron remnant and VLDL remnant. Role of Apo C
is to activate lipoprotein lipase and Apo E is required as a
ligand on the surface of remnant for uptake by liver.

H igh R eturn
Fig. 9.6: LDL-Receptor Mediated Endocytosis. LDL receptor is a
zz Chylomicron gets converted to chylomicron remnant. glycoprotein present on liver and extra-hepatic tissues
Chylomicron has mainly TG and very less cholesterol. So if
chylomicron increased in a patient then that means only TGs zz Entry of cholesterol in cells suppress LDL receptor synthesis by
are increased, and cholesterol is normal (because cholesterol SREBP (Sterol Regulatory Element Binding Protein)
is negligible in the presence of too many TGs). But chylomicron zz PUFAs and MUFAs decrease cholesterol levels as they
remnant are formed from chylomicron after losing some upregulate LDL cholesterol synthesis
T TGs. So if chylomicron remnant is increased in some patient
H means both TGs and cholesterol is increased (because in the
E presence of little TGs, the cholesterol molecules present now H igh R eturn
O are not negligible). zz Exogenous TG transported to peripheral tissues by:
R Chylomicron
Y Endogenous Fat zz Endogenous TG transported to peripheral tissues by: VLDL
Endogenous fats are synthesized by liver. Mainly TGs are zz Exogenous cholesterol transported to peripheral tissues by: LDL
synthesized. Then liver synthesize Apo B100 around the zz Endogenous cholesterol transported to peripheral tissues by: LDL
High-Density Lipoprotein (HDL)
It is synthesized and secreted by liver cells. It has maximum
phospholipids. It takes cholesterol from periphery to liver,
245
also known as reverse cholesterol transport. HDL converts
this cholesterol into cholesterol ester with the help of enzyme

CHAPTER 9  LIPOPROTEINS
LCAT–Lecithin Cholesterol Acyl Transferase. This enzyme
transfers fatty acid from lecithin (a phospholipid) to cholesterol.
Lecithin after losing fatty acid becomes lysolecithin. (Fig. 9.7)

Fig. 9.7: Conversion of cholesterol to cholesterol ester in HDL

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Fig. 9.8: HDL metabolism Y
 LCAT Deficiency  HDL-receptors → Apo A1

246 Table 9.1: Lipid and protein constituents of lipoproteins

Lipoprotein Lipid Protein/

Apoprotein
CRO BIOCHEMISTRY

Chylomicron TG Apo B-48

Chylomicron TG + Cholesterol Apo B-48 + Apo E
zz LCAT deficiency leads to increased cholesterol and Remnant
lecithin (substrates of reaction) VLDL TG Apo B-100
zz Free cholesterol gets deposited in cornea leading to
VLDL-Remnant/IDL TG + Cholesterol Apo B-100 + Apo E
opacification
zz There is decreased cholesterol ester and lysolecithin LDL Cholesterol Apo B-100 + Apo E
zz HDL is decreased as it is not properly formed HDL Cholesterol ester Apo-A, Apo-C and
Apo-E
zz Complete deficiency of enzyme is more severe, known as
Norum’s disease, which leads to hemolytic anemia and ESRD


(End Stage Renal Disease)

zz Partial deficiency is benign, known as fish eye disease.

Role of Apo-Proteins
1. Structural role – e.g. Apo B-48 and Apo B-100. They
provide a hydrophilic thing in the periphery of lipo-
protein. These cannot be removed.
2. Enzyme activators and inhibitors:
 Apo C-I and C-II → activate lipoprotein lipase (main
is C-II)
 Apo C-III → inhibits lipoprotein lipase
 Apo A-I → activate LCAT
 Apo A-II → inhibits LCAT
3. Ligands for receptors:
Fig. 9.9: Shown is the electrophoretic pattern of serum
 Ligand for remnants→Apo E
lipoproteins. Chylomicrons stay at the origin because they are
 LDL-receptors → Apo B100 (main receptor) and largest in size
Apo E

Do not get confused as sequence of lipoproteins is different in electrophoresis (than the sequence for density). Because electrophoresis
depends on multiple factors. Learn these two lists separately.

H igh R eturn
zz HDL is known as alpha-Lipoprotein or Lipoprotein–A
zz Lipoprotein ‘a’ → its structure is similar to plasminogen. It interferes with activation of Plasminogen to plasmin, thus leading to
intravascular thrombosis. It is associated with an increased risk for coronary artery disease. It contains LDL + Apo ‘a’
zz So Lipoprotein—A → prevent atherosclerosis (as it is HDL)
zz Lipoprotein—‘a’ → cause atherosclerosis (as it contains LDL)

Table 9.2: Fredrickson classification of hyperlipoproteinemia.

Type Defect Lipoprotein increased TGs Cholesterol Name

I Lipoprotein Lipase or Apo C- II Chylomicron ↑↑ VLDL ↑ ↑↑ Normal Familial Hyperchylomicronemia (AR)
T IIa LDL-receptor or Apo B100 LDL↑ ↑ Normal ↑↑ Familial Hypercholesterolemia (AD)
H IIb Unknown VLDL ↑ ↑ ↑ Familial combined Hyperlipoproteinemia
E LDL↑ (AR)
O
III Apo E Chylomicron Remnant ↑ ↑ ↑↑ ↑↑ Dysbeta-lipoproteinemia or Broad-Beta
R VLDL remnant ↑ disease (AR)
Y
Apo C is the activator of lipoprotein lipase. Apo B-100 is the main ligand of LDL receptor. Apo E is required for the metabolism
of chylomicron remnant and VLDL remnant/ IDL.
 A dditional E dge zz After heparin injection, if LPL levels are not raised in
circulation then it is diagnostic of LPL deficiency or
Sitosterolemia Apo C-II deficiency. So heparinized plasma is used in 247
zz It comes under Type II (a) Hyperlipoproteinemia diagnosis of Type I Hyperlipoproteinemia or familial
zz Patient has Hypercholesterolemia not responding to statins. Hyperchylomicronemia.
zz Defect: Mutation in ATP binding cassette proteins (G5 and

CHAPTER 9  LIPOPROTEINS
G8), which actively secrete plant sterols and cholesterol into Causes of 2o Hyperlipoproteinemia
intestine and bile.
zz Increased plant sterols in liver and intestine. This leads to 1. Excess carbohydrates in diet (VLDL ↑)
decrease transcription of LDL receptors because liver thinks 2. Nephrotic syndrome (VLDL ↑)
that it has too much sterol (cholesterol in LDL is a sterol). 3. Diabetes (VLDL ↑)
zz Increased blood LDL and cholesterol 4. Cushing’s syndrome (VLDL ↑)
zz Clinical features: Anisocytosis, Poikilocytosis, mega throm- 5. Alcohol (TG↑)
bocytes, increased risk of coronary heart disease, Tendon 6. Hypothyroidism (↑LDL) – because thyroid hormones
Xanthoma, Hemolysis, Splenomegaly increase expression of LDL receptor on liver.
7. Estrogen (↑HDL, VLDL)

Lipoprotein Lipase (Lpl) Hypolipoproteinemia

zz It is attached to the endothelium by GAGs, which are zz Abetalipoproteinemia – autosomal recessive
negatively charged (mainly Heparin Sulfate).)  Absent or negligible β-lipoproteins
 Mutation in MTP (Microsomal Triglyceride Transfer
Protein) which transfers TGs to chylomicrons and
VLDL in liver and intestine respectively.
 Decreased plasma TG, chylomicrons
 All β lipoproteins also decreased i.e. VLDL (pre β),
IDL (broad β), LDL (β)
 Also defective absorption of fat soluble vitamins
as chylomicrons are not formed. Bleeding
manifestations occur due to vitamin K deficiency
 Neurological deficit present
Interpretation
Clinical Feature
Tendon Xanthoma Cholesterol increased
Eruptive Xanthoma TG increased
Milky Plasma Chylomicron increased
Acute Pain Abdomen TG increased
zz Heparin releases LPL from Heparin Sulfate into the (Acute Pancreatitis)
circulation
Palmar and Tubero eruptive Chylomicron remnant, VLDL
zz Therefore, heparin is known as clearing factor (as it
Xanthoma remnant
clears lipemia).

Lipoprotein Lipase Hormone sensitive lipase LCAT ACAT

• Present in blood • Present inside the cells of • Lecithin Cholesterol Acyl • Acyl CoA Cholesterol Acyl
(Endothelium of blood Adipose tissues Transferase Transferase
vessels)
• This enzyme transfers fatty • This enzyme transfers fatty
• Activated by Insulin • Inhibited by Insulin acid from lecithin acid from Acyl CoA (a fatty
(a Phospholipid) to acid activated by coenzyme
T
• Belongs to anabolic enzyme • Belongs to catabolic enzyme H
cholesterol A) to cholesterol
category category E
• Present in HDL in blood • Present inside the cells of
• Helps in the storage of fats • Helps in lipolysis adipose tissue
O
(TGs) in the body R
Y
248
CRO BIOCHEMISTRY


Treatment of increased TG: Fibrates, omega -3 FA, high

zz
dose nicotinic acid
MICELLE AND LIPOPROTEINS
zz Treatment of increased cholesterol: Statins, increased zz Micelle and lipoproteins have one thing in common that
cholesterol absorption inhibitors, bile acid sequestrants, outer is polar and core is non polar.
selective removal of LDL from blood; known as LDL zz Name micelle is used for detergents and soaps or
apharesis. digestion of lipids.
Q. How high dose Nicotinic acid is used as anti- hyper- zz But lipoproteins are forms of lipid transport in blood.
lipidemic?
A. Nicotinic acid in high doses inhibits hormone sensitive
lipase (HSL), so TGs from adipose tissue are not broken
down and free fatty acids do not reach blood and liver.
This leads to decreased VLDL and LDL. Also, HDL levels
are raised by decreasing its catabolic rate.
# Also Niacin is contraindicated in diabetes as it increases
blood glucose levels.

Summary Box

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 SUMMARY TABLE 249
Lipoprotein Characteristic feature
Chylomicron • Largest size

CHAPTER 9  LIPOPROTEINS
• Maximum Lipid content
• Maximum TG content
• Transport exogenous lipid or TG from intestine to peripheral tissue
• Minimum phospholipid
• Minimum protein
• Lowest density
• Least (no) electrophoretic mobility, least plasma half life
VLDL • Maximum endogenous TG
• Transports endogenous lipid or TG from liver to peripheral tissues
LDL • Maximum cholesterol
• Was earlier called bad cholesterol
• Major source of cholesterol for peripheral tissues
HDL • Minimum lipid content
• Minimum TG content
• Smallest size
• Maximum phospholipid
• Maximum protein
• Maximum electrophoretic mobility
• Known as α-Lipoprotein or Lipoprotein ‘A’
• Was earlier called good cholesterol
• Transport excess cholesterol from peripheral tissues to liver and steroidogenic tissues in the form of cholesterol
ester.

Pearls of the Chapter

zz In circulation, HDL donates Apo C and Apo E to chylomicron and VLDL so that they are converted to Chylomicron remnant and VLDL
remnant
zz Lipoprotein Lipase (LPL): It is synthesized by adipose tissues, cardiac and skeletal muscles
zz Ligand for LDL-Receptor is Apo B100 and minor ligand is – Apo E
zz Apo C-I and C-II → activate Lipoprotein Lipase
zz Apo C-III → inhibits Lipoprotein Lipase
zz Apo A-I → activate LCAT
zz Apo A-II → inhibits LCAT
zz Apo C-2 is the activator of Lipoprotein Lipase
zz Apo E is required for the metabolism of Chylomicron remnant and VLDL remnant/ IDL
zz Activators of Hormone Sensitive Lipase (HSL) → Glucagon, Thyroxine, Catecholamines, ACTH, TSH, Glucocorticoids
zz Inhibitors of HSL → Insulin, high doses of Nicotinic acid, PGE
zz HSL can only break fatty acid from position number 1 and 3 of TGs. It cannot break fatty acid from position number 2
zz Insulin activates Lipoprotein Lipase but only in the capillary beds of adipose tissue.

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 250 Multiple Choice Questions
Lipoproteins 10. Chylomicrons core is composed of (PGMEE 2011)
1. Action of lipoprotein lipase  a. Triglyceride
 (PGMEE 2013, 2015) b. Free fatty acids
a. Activation of IP3-DAG c. Triglyceride, Cholesterol and Phospholipids
b. Hydrolysis of triglycerides in chylomicrons d. Triglyceride and Cholesterol
c. Coupling of DAG and MAG to form triglycerides 11. Lipoprotein x is an indirect estimate of:
d. Hydrolysis of dietary and endogenous TG  (PGMEE 2015)
2. Enzyme LCAT (Lecithin Cholesterol Acyl Transferase) a. Hepatitis b. Myocardial infarction
 (PGMEE 2011, 2010) c. Cholestasis d. Atherosclerosis
a. Hydrolyses triglyceride into glycerol and fatty acids 12. Apo (a) and Lp (a) resembles- (PGMEE 2015)
b. Is one of the enzymes involved in cholesterol a. Fibrinogen
biosynthesis b. LDL receptor
c. Helps in the conversion of cholesterol to cholesterol c. Plasminogen
ester d. Prothrombin
d. Is involved in the formation of chylomicrons 13. Dietary lipids are transported from intestine in:
3. HDL is synthesized and secreted from :  (PGMEE 2015)
 (Recent Question 2017) a. HDL b. VLDL
a. Liver c. Chylomicrons d. LDL
b. Peripheral tissue 14. Which cholesterol is good for heart?
c. Adipose tissue  (PGMEE 2016-17)
d. Muscle a. VLDL b. CDL
4. Which of the following apolipoprotein is synthesized c. HDL d. IDL
in the liver as part of the coat of very-low density 15. Hormone sensitive lipase doesn’t remove which
lipoproteins (VLDLs):- (PGMEE 2015, 2016-17) carbon from Triacylglycerol? (PGMEE 2012)
M a. 1st b. 2nd
C a. B48 b. B100 c. 3rd d. Removes all
Qs c. C11 d. A1 16. Which of the following is maximum in HDL?
Ans. 5. Dietary cholesterol is transported from intestine to  (AIIMS NOV 2016)
liver by a. Triglycerides
1. b
 (PGMEE 2007, 2013; FMGE June 2018) b. Apoproteins
2. c
a. Apo-A b. Apo-B c. Cholesterol
3. a
c. Apo-C d. Apo-E d. Fatty Acids
4. b
17. A person on fat free, carbohydrate rich diet, continues
5. d
6. Apoprotein – C: (PGMEE 2012-13, 2017) to grow obese. Which lipoprotein is increased ?
6. d
a. Activates lipoprotein lipase  (PGMEE 2007)
7. c
b. Facilitates triglyceride transport a. LDL c. HDL
8. d
c. Inactivates lipoprotein lipase b. VLDL d. Chylomicrons
9. a
d. All of the above 18. HDL3 is converted to HDL2 by:
10. d
7. LDL level in non diabetics should be below what a. CETP b. LCAT
11. c
value in mg/dl? (PGMEE 2015) c. Both d. PLTP
12. c
a. 50 b. 75 19. All are correct EXCEPT:
13. c
c. 100 d. 130 a. HDL-2 is more dense than HDL-3
14. c
8. HDL is called good cholesterol because- b. HDL-2 is larger than HDL-3
15. b
 (PGMEE 2015) c. Accumulation of more cholesterol ester in HDL-3 to
16. b
a. Activates lipoprotein lipase produce HDL-2
17. b
b. Stimulate hepatic TGs synthesis d. CETP is synthesized by liver and adipose tissue
18. b
c. Causes transport to extrahepatic tissues 20. Cholesterol is transported to arterial smooth muscle
19. a
d. Removes chole–sterol from extrahepatic tissues by:
20. a
9. In coronary artery disease the cholesterol level (mg/ a. LDL
dl) recommended is: (PGMEE 2015) b. HDL
a. Below 200 b. < 220 c. VLDL
c. < 250 d. < 280 d. IDL
21. Which of the following statements about high density Diseases of Lipoprotein Metabolism
lipoprotein (HDL) is CORRECT? 31. Which of the following is increased in LCAT
a. HDL lipoproteins are only synthesized in the liver deficiency?  (Recent Question Jan 2019 Q)
251
b. HDL lipoproteins transport triacylglycerols to a. HDL b. LDL
peripheral tissues c. VLDL d. Chylomicron
c. HDL lipoproteins pick up cholesterol and convert it 32. In case of LPL deficiency, which of the following will
to cholesterol ester increase after a fat rich diet ? 
d. HDL lipoproteins cannot be endocytosed by the  (Recent Question Jan 2019 Q)
liver a. Chylomicron b. HDL
22. Which is most potent HDL? c. Lipoprotein (a) d. LDL
a. Pre-beta HDL 33. Which of the following lipoproteins are increased in
b. HDL-2 type III Hyperlipoproteinemia? (PGI May 2017)
c. HDL-3 a. VLDL Remnant b. Chylomicron remnant
d. Discoidal HDL c. Triglycerides d. Cholesterol
23. Which enzyme can convert cholesterol to cholesterol e. IDL (Intermediate density lipoprotein)
esters ? 34. Which of the following is used in the treatment of
a. LCAT (Lecithin cholesterol acyl transferase) type I Hyperlipoproteinemia ? (PGI May 2017)
b. ACAT (Acyl CoA cholesterol acyl transferase) a. Omega -3 fatty acids
c. Both A and B b. Omega -6 fatty acids
d. Hormone sensitive lipase c. Saturated fatty acids
24. Which has maximum density?  (PGMEE 2009) d. MUFA (Monounsaturated fatty acids)
a. HDL b. LDL 35. A 10-yr old boy was admitted for surgery of cleft
c. VLDL d. Chylomicrons lip. Suddenly he had acute pain in abdomen. On
25. All are true about HDL EXCEPT: (PGMEE 2013-14) examination, he had Xanthomas. On investigation,
a. Carries cholesterol from peripheral tissues to liver we found milky plasma. Which lipoprotein is
b. Pre HDL is the inactive precusor, which later forms increased?
the active HDL a. VLDL Remnant
c. In electrophoresis, HDL is the lipoprotein which b. Chylomicron
moves farthest from starting point c. Triglycerides1
d. It has apoproteins ApoA, ApoC, ApoE d. Cholesterol
26. Which of the following liporoteins would contribute M
36. Familial hypercholesterolemia is due to defect in: C
to a measurement of plasma cholesterol in a normal  (PGMEE 2013) Qs
person following a 12-hour fast? (PGMEE 2016-17) a. Lipoprotein lipase b. Apo C
a. Chylomicrons Ans.
c. Apo A d. Apo B 100
b. VLDL 37. Familial Hypercholesterolemia is inherited as: 21. c
c. HDL  (PGMEE 2011) 22. a
d. LDL a. Autosomal dominant b. Autosomal recessive
27. True about Apo-proteins: (PGI May 2016) 23. c
c. X-linked recessive d. X-linked dominant 24. a
a. Constitute peripheral region of plasma lipoproteins 38. Absence of this apo lipoprotein is responsible 25. b
b. Divided into A, B, C only for the genetic disorder, familial type III hyper- 26. c
c. Apo A-I is the major protein component of high lipoproteinema (PGMEE 2015) 27. a,c
density lipoprotein (HDL) a. Apo B100
d. Apo A, B and C are further divided d,e
b. Apo B48 28. a
e. Role in enzyme activation c. Apo CII
28. Full form of LCAT is: (PGI May 2014, 2018) 29. b
d. Apo E 30. a
a. Lecithin Cholesterol Acyl Transferase 39. Which of the following biochemical changes is not 31. a
b. Lecithin Choline Acyl Transferase indicative of Hyperlipidemia Type IIA?
c. Lecithin Cholesterol Alkyl Ttransferase 32. a
(PGMEE 2016-17) 33. a,b,e
d. Lecithin Choline Alcohol Transferase a. Total cholesterol increased
e. Lecithin CoA Transferase 34. a
b. Triglycerides normal 35. b
29. Main transporter of cholesterol to peripheral tissue c. LDL increased
is: (PGI Nov 2011) 36. d
d. HDL increased 37. a
a. HDL b. LDL 40. Which vitamin is given in type IIB familial hyper- 38. d
c. IDL d. VLDL lipidemia? (PGMEE 2013) 39. d
e. Chylomicron a. Pantotheinc acid
30. Calculate LDL. TGs = 200, Total cholesterol= 300, 40. c
b. Riboflavin
HDL= 40.  (JIPMER Nov 2017) c. Nicotinic acid
a. 220 b. 260 d. Thiamine
c. 60 d. 100
 41. Type- I hyperlipoproteinemia is characterized by – 43. A patient was found to have high LDL, increased total
a. Elevated lipoprotein lipase (PGMEE 2014) cholesterol. But normal levels of LDL-Receptors.
252 b. Elevated HDL What is the most probable cause? (AIIMS May 2015)
c. Elevated chylomicrons a. Apo B100 mutation
d. Elevated LDL b. LCAT Deficiency
42. Tangier’s diseases is characterized by:(PGMEE 2012) c. Apo E defect
a. Low or absence of HDL d. Lipoprotein lipase Deficiency
b. Low LDL concentration 44. Deficiency of LCAT leads to:
c. Raised chylomicrons a. Decrease nascent discoidal HDL
d. Deficiency of LPL b. Increase nascent discoidal HDL
c. Decrease pre beta HDL
d. Increase HDL-2

Answers with Explanations

1. Ans. (b) Hydrolysis of triglycerides in chylomicrons 5. Ans. (d) Apo-E

[Ref: Harper 30th/e pg. 263] [Ref: Harper 30th/e pg. 255]
•• Lipoprotein lipase is a functional plasma enzyme, •• Dietary lipids (TG+ cholesterol) is transported
which is responsible for the hydrolysis of triglycerides from intestine to blood via chylomicron. They get
in chylomicrons and VLDL and convert them to chy- converted to chylomicron remnants.
lomicron remnant and VLDL remnant respectively. •• Apo-E is the ligand for chylomicron remnants for
uptake by liver.
2. Ans. (c) Helps in the conversion of cholesterol to
cholesterol ester 6. Ans. (d) All of the above

[Ref: Harper 30th/e pg. 263] [Ref: Harper 30th/e pg. 255]
M •• Enzyme LCAT (Lecithin Cholesterol Acyl Transferase) •• Apo C-II → activate Lipoprotein Lipase
C is present in HDL, which is responsible for adding •• Apo C-III → inhibits Lipoprotein Lipase
Qs one fatty acid in cholesterol to make cholesterol ester. •• Apo C help in TG transport as it activates Lipoprotein
Ans. This fatty acid is taken from lecithin (phospholipid) Lipase. So it moves TGs from circulation into the
adipose tissues.
41. c 3. Ans. (a) Liver
42. a 7. (c)100
43. a [Ref: Harper 30th/e pg. 258]
44. b [Ref: Harper 30th/e pg. 271]
•• HDL is synthesized and secreted from liver and
intestinal cells. Proteins are Apo -A, Apo -C and Apo •• LDL can be oxidized to form oxidized LDL which
-E. HDL takes cholesterol from peripheral cells to causes atherosclerosis. So, levels should be kept
liver (Reverse cholesterol transport). below 100 mg/dL
LDL Levels
In normal person <100 mg/dL
In diabetic or heart disease <130

8. Ans. (d) Removes cholesterol from extrahepatic

tissues

4. Ans. (b) B100 [Ref: Harper 30th/e pg. 271]

•• Good and Bad cholesterol terminology was used
[Ref: Harper 30th/e pg. 275]
earlier. HDL was called good cholesterol because of
•• Apo B 100 is synthesized by liver for VLDL. Apo B 48 is reverse cholesterol transport i.e. it takes cholesterol
synthesized by intestine for Chylomicrons. from peripheral tissue to liver.
 9. Ans. (a) Below 200 15. Ans. (b) 2nd

[Ref: Tietz: Fundamentals of Clinical Chemistry 6/e [Ref: Harper 30th/e pg. 263] 253
p419]
•• Hormone sensitive lipase, doesnot remove 2nd
Total cholesterol <200 desirable carbon from Triacylglycerol. It can remove FA from
>240 High risk of heart disease C1 and C3 only.

CHAPTER 9  LIPOPROTEINS
HDL ≥ 60 desirable
<40 High risk 16. Ans. (b) Apoproteins

LDL <100 – desirable

•• Maximum proteins are present in HDL and minimum
>200 – High risk proteins are present in chylomicron.
TG <150 – Desirable 17. Ans. (b) VLDL
10. Ans. (d) Triglyceride and Cholesterol [Ref: Harper 30th/e pg. 257]
[Ref: Harper 30th/e pg. 257] •• Carbohydrate rich diet will lead to increase in Acetyl
CoA, which will synthesize fats in body. These fats are
•• Chylomicrons contain both Triglycerides and Chole- endogenous fat, which are transported in the form of
sterol (But cholesterol is very less) VLDL. Hence, VLDL is increased.
•• Option d is better than option a.
18. Ans. (b) LCAT
11. Ans. (c) Cholestasis
[Ref: Harper 30th/e pg. 847]
[Ref: Harper 30th/e pg. 271]
•• LCAT converts nascent discoidal HDL into HDL-3
•• Lipoprotein x (Lp-x) is an abnormal lipoprotein (spherical HDL). Further, HDL-3 is converted to HDL-
found in LCAT deficiency & in cholestatic states 2 by LCAT (Lecithin Cholesterol Acyl Transferase).
e.g. primary biliary cirrhosis, primary sclerosing •• CETP transfers cholesterol esters from cholesterol
cholangitis. It is rich in amphipathic lipids ester-rich HDL2 to VLDL and LDL. PLTP
(cholesterol & phospholipids) and is poor in neutral (Phospholipid Transfer Protein) transfer cholesterol
lipids (cholesterol esters & TGs). It gives an onion ester from Apo B containing lipoproteins to HDL.
like appearance or vesicular appearance in electron
microscopy. Its density is between VLDL & LDL. 19. Ans. (a) HDL-2 is more dense than HDL-3
Major protein in Lp-x is albumin and small amount
[Ref: Harper 30th/e pg. 847]
of variable apoproteins are present.
•• HDL-2 is less dense than HDL-3. HDL-2 is larger than
12. Ans. (c) Plasminogen HDL-3. Accumulation of more cholesterol ester in
HDL-3 produce HDL-2. CETP is synthesized by liver
[Ref: Harper 30th/e pg. 217] and adipose tissue. A
•• Lipoprotein ‘a’ has its structure similar to Plasm- N
inogen. It interferes with activation of plasminogen 20. Ans. (a) LDL S
to plasmin, thus leading to intravascular thrombosis. W
[Ref: Harper 30th/e pg. 845]
It is associated with an increased risk for coronary E
•• Cholesterol is transported to arterial smooth muscle
artery disease. It contains LDL + Apo ‘a’ R
by LDL. HDL remove cholesterol from other tissues
(such as arterial walls) to transport it back to liver. S
13. Ans. (c) Chylomicrons
WITH
[Ref: Harper 30th/e pg. 263] 21. Ans. (c) HDL lipoproteins pick up cholesterol and
convert it to cholesterol ester E
•• Chylomicrons transport dietary or exogenous lipids
X
or TGs from intestine to peripheral tissue. [Ref: Harper 30th/e pg. 847]
P
14. Ans. (c) HDL •• HDL is synthesized in the liver and intestine. HDL L
lipoproteins transport cholesterol from peripheral A
[Ref: Harper 30th/e pg. 271] tissues to liver and convert this cholesterol to N
cholesterol ester by adding fatty acid. HDL can be A
HDL is cardioprotective. endocytosed by the liver. (ligand is apo A1) T
I
O
N
S
 22. Ans. (a) Pre-beta HDL 30. Ans. (a) 220

254 [Ref: Harper 30th/e pg. 847] [Ref: Harper 30th/e pg. 249]
•• Pre-beta HDL is the most potent HDL in taking •• Friedwald’s equation:
cholesterol from peripheral tissues. Total cholesterol = HDL+LDL+VLDL
= HDL+ LDL+ TG/5
CRO BIOCHEMISTRY

23. Ans. (c) Both a and b (we do not measure VLDL, we measure TG)
LDL= Total cholesterol –HDL-TG/5 (mg/dl)
[Ref: Harper 30th/e pg. 847]
LDL= Total cholesterol – HDL – TG/2.2 (mmol/L)
•• Both LCAT and ACAT esterify cholesterol (i.e. add LDL chol = total chol – HDL chol – TG/5
fatty acid to cholesterol). LCAT takes cholesterol from = 300 – 40 – 200/5
lecithin and ACAT takes cholesterol from Acyl CoA (a      = 300 – 40 – 40
fatty acid attached to Coenzyme A).        = 220
24. Ans. (a) HDL 31. Ans. (a) HDL


[Ref: Harper 30th/e pg. 847] [Ref: Harper 30th/e pg. 847]
•• HDL has maximum density, that’s why the name. •• LCAT is Lecithin Cholesterol Acyl Transferase, which
Chylomicron has least density. converts HDL- cholesterol to HDL-cholesterol esters.
25. Ans. (b) Pre HDL is the inactive precusor, which •• It converts nascent discoidal HDL to HDL 2 and HDL
later forms the active HDL 3.
•• Complete LCAT deficiency is called Norum’s disease.
[Ref: Harper 30th/e pg. 263] •• There is increase in nascent discoidal HDL,
containing free cholesterol. This free cholesterol gets
•• Pre- β HDL is most potent. HDL is smallest in size, so
deposited in cornea leading to corneal opacification
moves fastest in electrophoresis.
•• HDL 2 and HDL 3 are not formed.
26. Ans. (c) HDL
32. Ans. (a) Chylomicron
[Ref: Harper 30th/e pg. 257]
[Ref: Harper 30th/e pg. 270]
•• Some journals say that HDL is best parameter to
•• Lipoprotein lipase deficiency is also called
measure plasma cholesterol after a 12 hour fast. Many
Familial Hyperchylomicronemia or Type I Hyper-
studies say that total cholesterol/ HDL cholesterol
lipoproteinemia. Chylomicrons are formed after a fat
ratio is a better predictor of plasma cholesterol as
rich diet and this patient cannot metabolize them.
compared to LDL/HDL ratio.
•• Best answer is c here 33. Ans. (a); (b); (e)
A
N 27. Ans. (a); (c); (d); (e) [Ref: Harper 30th/e pg. 263; Harper 30th/e pg. 846, 851]
S
W [Ref: Harper 30th/e pg. 255] •• In Type III Hyperlipoproteinemia, chylomicron
E remnant, VLDL remnant (IDL) are increased. So
•• Apo-proteins are divided into A, B, C, D, E and H (F
R lipids increased are TG and cholesterol both. This is
and G have very minor role)- Option (b) is wrong
also known as Dysbeta Hyperlipoproteinemia. All 5
S options are increased in this disease. But question is
28. Ans. (a) Lecithin Cholesterol Acyl-Transferase
WITH asking Lipoproteins, so mark only lipoproteins (TG
[Ref: Harper 30th/e pg. 249] and cholesterol are lipids, not Lipoproteins)
E
•• Full form of LCAT is Lecithin Cholesterol Acyl
X 34. Ans. (a) Omega -3 fatty acids
Transferase.
P
[Ref: Harper 30th/e pg. 263]
L 29. Ans. (b) LDL
A •• Type I Hyperlipoproteinemia, also known as Familial
N [Ref: Harper 30th/e pg. 845] Hyperchylomicronemia, is deficiency of lipoprotein
A •• Major transporter of cholesterol from liver to peri- lipase or Apo C defect (as Apo C is the activator of
T pheral tissue is LDL. lipoprotein lipase). Omega -3 fatty acids are used in
I the treatment of type I Hyperlipoproteinemia as they
O reduce blood TG levels. They do not have any effect
N on LDL or HDL - cholesterol.
S
35. Ans. (b) Chylomicron 41. Ans. (c) Elevated chylomicrons

[Ref: Harper 30th/e pg. 263] [Ref: Harper 30th/e pg. 275] 255
•• Milky plasma means chylomicrons are increased.   Refer Table 9.2
Chylomicrons contains TGs. But TG is a lipid and •• Type –I is known as Familial Hyperchylomicronemia.

CHAPTER 9  LIPOPROTEINS
chylomicron is a lipoprotein. This is a picture of Type There is defect in Lipoprotein Lipase. It is characte-
I Hyperlipoproteinemia, which is due to the defect rized by elevated chylomicrons VRDL and TGs.
in lipoprotein lipase enzyme. Question in asking
Lipoprotein, so answer is chylomicron. 42. Ans. (a) Low or absence of HDL

36. Ans. (d) Apo B 100 [Ref: Harper 30th/e pg. 275]
•• In Tangier’s disease, there is defect in ABC-A1
[Ref: Harper 30th/e pg. 275]
receptors which helps in giving cholesterol from
•• Familial hypercholesterolemia is due to defect in Apo peripheral tissue to HDL. In this defect, HDL is not
B 100 or defect in LDL receptor. Apo B 100 is the main properly formed, so HDL is decreased and other
ligand for LDL receptor. lipoproteins are normal. Also Apo A1 decreases.
Cholesterol accumulates (especially in Reticulo
37. Ans. (a) Autosomal dominant Endothelial system). Patient has peripheral
[Ref: Harper 30th/e pg. 275] neuropathy, hepatosplenomegaly and large orange/
yellow tonsils.
•• Familial hypercholesterolemia is autosomal do-
minant. Other Hyperlipoproteinemias are AR (Auto- 43. Ans. (a) Apo B100 mutation
somal Recessive).
[Ref: Harper 30th/e pg. 275]
38. Ans. (d) Apo E •• This is Familial defective Apo B100, also known
[Ref: Harper 30th/e pg. 275] as Autosomal Dominant Hypercholesterolemia.
There is a mutation in the gene encoding Apo B100,
•• Apo E is defective and remnants are increased as the specifically in LDL-receptor binding domain of Apo
only ligand for remnants is Apo E. B100.
39. Ans. (d) HDL increased 44. Ans. (b) Increase nascent discoidal HDL
[Ref: Harper 30th/e pg. 275] [Ref: Harper 30th/e pg. 847]
•• In Type II- A, LDL receptor or Apo B- 100 is defective. •• LCAT converts nascent discoidal HDL into HDL-3
So, cholesterol is raised, TG normal and no change or (spherical HDL).
decreased HDL. •• Deficiency of LCAT leads to increase in nascent A
discoidal HDL containing free cholesterol. Refer Fig. N
40. Ans. (c) Nicotinic acid 9.8. S
[Ref: Harper 30th/e pg. 275] W
E
•• High dose of Niacin act as anti-hyperlipidemic.
R
S
WITH

E
X
P
L
A
N
A
T
I
O
N
S
 10 Metabolism
of Lipids
Overview of Chapter Enzymes of Fatty Acid Synthesis
•• Fatty Acid Synthesis Acetyl CoA Carboxylase
•• Ketone Bodies Metabolism This enzyme converts Acetyl CoA to Malonyl CoA, by adding
•• Beta Oxidation of Fatty Acids one CO2. Enzyme is Carboxylase. Like any other carboxylase,
•• Cholesterol Biosynthesis this enzyme also uses biotin as coenzyme and also ATP. As
ATP is used, name of enzyme is also Malonyl CoA Synthetase.
This is the rate limiting enzyme. It is allosterically inhibited by
FATTY ACID SYNTHESIS fatty acyl CoA and activated by citrate.

A dditional E dge
H igh R eturn Acetyl CoA Carboxylase – a multienzyme protein contain-
zz Nature of pathway: Anabolic ing: Biotin (bound to lysine residue on the enzyme), Biotin
zz Occurs in Fed state Carboxylase, Biotin Carboxyl Carrier Protein (BCP), Carboxyl
zz Activated by hormone: Insulin transferase, a regulatory allosteric site
zz Organelle: Cytoplasm (any Anabolic pathway occurs in
cytoplasm)
zz Organ/cell: Liver, lactating mammary glands and to a limited Carboxylation Reaction Occurs in 2 Steps
extent in adipose tissue 1. Carboxylation of biotin (uses ATP)
zz Starting material – Acetyl CoA
2. Transfer of carboxyl group to Acetyl CoA
zz Rate limiting enzyme of fatty acid synthesis – acetyl CoA
Carboxylase
zz Main enzyme of fatty acid synthesis – fatty acid synthase
Fatty Acid Synthase Complex
complex Also known as palmitate synthase as the end product of fatty
acid synthesis is mostly palmitic acid in humans.
Fatty acid synthesis occurs in fed state. Carbohydrates from This is a multienzyme complex, containing 6 enzymatic
diet are used to produce energy (ATP) but when carbohydrates activities. This enzyme is a dimer, made up of two identical
are in excess then the extra Acetyl CoA is diverted to fatty acid monomers/subunits. The shape of this enzyme is X-shape.
synthesis. Each subunit is attached to Acyl Carrier Protein (ACP), having
–SH (sulfhydryl) group with phosphopantetheine. Fatty acid
synthase complex is the main enzyme of fatty acid synthesis.

Fig. 10.1: Sequence of enzymes in the primary structure of one monomer of fatty acid synthase complex shown. X-shape of fatty acid
synthase homodimer. ACP (Acyl Carrier Protein) is attached here with the enzyme
 257

CHAPTER 10  METABOLISM OF LIPIDS

Fig. 10.2: Complete structure of fatty acid synthase complex shown. In this figure, upper and lower subunit (above and below the
horizontal dotted line)

Cysteine and PAN ends: Each unit is attached to ACP Synthesis of fatty acid on fatty acid synthase
(Acyl Carrier Protein) from which phosphopantetheine is complex:
protruding, so known as ‘PAN’ end. On the other side of the
subunit, Cysteine is present, so this is known as ‘Cysteine 1. 
Loading Step: Loading of Acetyl CoA and Malonyl CoA
end’. Each subunit has PAN end and Cysteine end. occurs. Acetyl CoA is loaded at Cysteine end and Malonyl
The two subunits (upper and lower) are joined by CoA is loaded at PAN end. Malonyl-Acetyl Transacylase
disulphide bond between SH group of Cysteine end of one enzyme is involved
subunit with the SH group from PAN end of other subunit and 2. 
Condensation Step: In this step, one CO2 is removed
vice-versa on the other side. from Malonyl CoA and 2C left at lower monomer (PAN
end) gets joined to the 2C of upper monomer (Cys end),
leading to formation of a fatty acid with 4C and having
a keto group, known as ketoacyl (4C). This ketoacyl is
attached at the lower monomer PAN end (known as
temporary holding site) and now the upper monomer is
empty. Enzyme involved here is ketoacyl synthase.
T
Reduction step → Occurs for the removal of keto group.
3. 
This step requires 3 enzymes –Ketoacyl reductase, H
Hydratase and Enoyl reductase. There are two reductases E
involved here and each requires NADPH. So two NADPH O
are used at one reduction step. R
Y
 where new Malonyl CoA (3C) can join. This is next loading
step. Then again condensation and reduction steps will occur.
258 So loading, condensation and reduction steps will keep
repeating until there is formation of a 16 C palmitic acid.

H igh R eturn
CRO BIOCHEMISTRY

zz Total 7 cycles of loading, condensation and reduction occur to

synthesize 16-C fatty acid
zz Synthesis of one palmitate requires 14 NADPH

Then the last enzyme Palmitoyl thioesterase/Deacylase

cleaves the synthesized fatty acid attached with thioester
bond at the cysteine end.

H igh R eturn
In first cycle of loading, condensation, reduction:


zz We load 5 C (in the form of Acetyl and Malonyl CoA)

zz But only 4 C are added into the fatty acid
In next all cycles of loading, condensation, reduction:
zz We load 3 C (only Malonyl CoA loaded)
zz But only 2 C are added into the fatty acid

Citrate Shuttle (Fig. 10.3)

Fatty acid synthesis occurs in cytoplasm and the starting
material is Acetyl CoA. But Acetyl CoA is synthesized in
mitochondria and it cannot cross IMM. So there is a need
to bring this acetyl CoA from mitochondria to cytoplasm.
For this citrate shuttle is needed. In mitochondria, pyruvate
forms Acetyl CoA by enzyme pyruvate dehydrogenase (link
reaction). Then this Acetyl CoA joins oxaloacetate to form
citrate by enzyme Citrate Synthase (first step of TCA cycle).
Then this citrate can cross IMM (by tricarboxylate transporter)
and reaches cytoplasm, where it is again broken down into
same constituents i.e. oxaloacetate and acetyl CoA. Enzyme
involved is ATP citrate lyase. Acetyl CoA can take part in fatty
acid synthesis in cytosol. Oxaloacetate gets converted to
malate by cytoplasmic malate dehydrogenase. Then malate
is converted to pyruvate by malic enzyme, which is a minor
After this step, the 4C fatty acid is shifted to upper
source of NADPH. Pyruvate formed can enter mitochondria
Cysteine end- this is now the permanent holding site of
and the cycle can continue.
the synthesized fatty acid. So, now lower PAN end is empty

T
H
E
O
R
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 259

CHAPTER 10  METABOLISM OF LIPIDS

Fig. 10.3: Citrate shuttle

Regulation of Fatty Acid Synthesis (Fig. 10.4) Fates of fatty acids

Rate limiting enzyme is Acetyl CoA Carboxylase. This enzyme zz Chain elongation to produce Very Long Chain FA
is inactive in dimer form. In the presence of Citrate, it gets (VLC FA)
converted to its polymer, which is active. Then the activated zz Desaturation to produce unsaturated FA
enzyme converts acetyl CoA to malonyl CoA. The second zz Formation of TGs
enzyme, fatty acid synthase complex, mainly uses Malonyl zz Esterified into cholesterol ester
CoA to synthesize fatty acid. This fatty acid (end product
of fatty acid synthesis) inhibits the citrate transporter and Chain Elongation to Produce Very Long Chain FA
also makes Acetyl CoA Carboxylase inactive, by converting (The Microsomal System) (Fig. 10.5)
it into inactive dimer form. This inactivation is done by zz Very long chain fatty acids cannot be synthesized by FA
phosphorylation. Synthase complex
Acetyl CoA Carboxylase is active in dephosphorylated zz This occurs in smooth ER and mitochondria (less active
state and inactive in phosphorylated state. Insulin in mitochondria)
dephosphorylate the enzyme and makes it active in fed state. zz Enzyme – Fatty Acid Elongase system
Glucagon and Epinephrine phosphorylate the enzyme and zz Both saturated and unsaturated fatty acids above C-10
inactivate it. can be elongated by this system
zz NADPH preferred but NADH can also be used
zz E.g. C-22 and C-24 fatty acids (very long chain fatty acids)
are required for sphingolipids, required in brain. So,
elongation increases in brain during myelination

T
H
E
Fig. 10.5: Elongation occurs same like the FA synthesis of O
Fig. 10.4: Regulation of Fatty Acid Synthesis cytoplasm (addition of 2C each time, derived from Malonyl CoA) R
via loading, condensation and reduction steps. But this system
requires all separate enzymes rather than a multienzyme complex Y
 Desaturation to produce unsaturated FA So, from this knowledge, we can conclude that adipose tissue
Occurs in smooth ER strictly depends on glucose uptake (that occurs via GLUT-
260 zz
zz Mainly in Liver 4) for TG synthesis. Because there is only one enzyme in
adipose tissues for the synthesis of Glycerol-3-P and that is
zz Membrane bound enzyme – Fatty Acyl CoA Desaturase
Glycerol-3-P Dehydrogenase which synthesize Glycerol-3-P
zz Most common Desaturase – ∆9 Desaturase (double
from DHAP. This DHAP is derived from glucose. So glucose
CRO BIOCHEMISTRY

bond between C9 and C 10). Humans have ∆9, ∆6, ∆5,

∆4 desaturase activity only. Humans cannot introduce entry is must in adipose tissues for TG synthesis.
double bond beyond ∆9 position. And GLUT-4 is Insulin Dependent, so in fasting when
there is lack of glucose and insulin, then adipose tissues
cannot synthesize Glycerol-3-P and thus TGs de novo.


Formation of TGs
Triglycerides are made by joining three fatty acids with
one molecule of glycerol. For this purpose, Glycerol-3-P is
required initially on which three fatty acids are added.

Sources of Glycerol-3-P (Fig. 10.6)

1. DHAP (Di-Hydroxy Acetone Phosphate) gets convert-
ed to Glycerol-3-P
 Enzyme – Glycerol-3-P Dehydrogenase
 Present in both liver and adipose tissues
2. Glycerol gets converted to Glycerol-3-P
 Enzyme – Glycerol-kinase Fig. 10.6: Comparison of how synthesis of Glycerol-3-P can occur
in liver and adipose tissue
 Present only in Liver

T
H
E
O
R Fig. 10.7: Glycerol released in adipose tissue (by TG degradation) cannot form TGs in adipose tissue. This glycerol travels via blood to liver
Y where it can be converted to glycerol-3-P by glycerol kinase. This glycerol-3-P can be either used to form TG in liver, or converted to DHAP
(by reversal of Glycerol-3-P Dehydrogenase reaction). This DHAP can be either used in glycolysis or gluconeogenesis
KETONE BODY METABOLISM
Liver constantly synthesize minimal amounts of ketone 261
bodies normally but the amounts are significantly raised
during fasting and starvation.
Ketone bodies are:

CHAPTER 10  METABOLISM OF LIPIDS

zz
1. Acetoacetate
2. β-hydroxy butyrate (or 3-hydroxy butyrate)
3. Acetone
zz Concentration in blood (normal person) = 0.2 mmol/L
zz Renal threshold – 3 mg/dL
zz In normal person, ratio of β-Hydroxy Butyrate/Aceto-
acetate = 1:1
zz In Ketosis, Ratio of β-Hydroxy Butyrate/Acetoacetate =
6:1

Advantages of use of ketone bodies in starvation

zz Use of ketone bodies spare glucose
zz They are soluble in aqueous solution unlike lipids
zz They are used in proportion to the concentration in
blood
Fig. 10.8: Sources of Acetyl CoA during fasting
zz As ketone bodies are not linked to CoA, they can easily
cross IMM (Inner Mitochondrial Membrane) zz Ketone bodies synthesis occurs during:
 Starvation
Q. Why Acetyl CoA cannot directly be used by brain?  Severe uncontrolled diabetes mellitus
What is the need of ketone bodies?
T A. Acetyl CoA cannot travel in blood and fatty acid
H cannot enter brain. So, fatty acid are broken in liver
I
forming Acetyl CoA which synthesize ketone bodies.
N
K These ketone bodies travel via blood to brain and
gets broken down to Acetyl CoA which gives energy
in brain.

Table 10.1: Differences and similarities between Ketone Body

Synthesis and Ketone Body Utilization

Ketone body synthesis Ketone body utilization

• In mitochondria • In mitochondria
• Inhibited by insulin • Inhibited by insulin
• Occurs in liver • In brain, heart and skeletal
muscles

Ketone Body Synthesis (Fig. 10.9)

zz Reasons of K.B. Synthesis:
 Increased lipolysis
 Increased Acetyl CoA or Decreased OAA (Oxaloac-
etate) Fig. 10.9: Ketone Body Synthesis
Because OAA is used in gluconeogenesis and also Primary ketone body – Acetoacetate
zz T
converted to Malate. Due to lack of OAA, Acetyl CoA cannot Secondary ketone bodies- β-Hydroxy Butyrate and
be used up for TCA and therefore this increased Acetyl CoA
zz H
Acetone E
is diverted to KB synthesis. Also Acetyl CoA comes from beta zz First ketone body synthesized - Acetoacetate
oxidation of fatty acids and from breakdown of ketogenic O
zz Most common ketone body in blood and urine during R
amino acids. ketosis - β-OH-Butyrate
Y
 zz Acetone is volatilized in lungs. It does not act as a fuel in
body. It is just released from breath and has a diagnostic
262 role.
zz Rate limiting step: HMG CoA Synthase. This enzyme is
common to both KB synthesis and Cholesterol Synthesis
CRO BIOCHEMISTRY

H igh R eturn
zz Thiolase: It is a common enzyme to following four pathways:
 KB Synthesis
 KB Utilization
 Cholesterol Synthesis
 Beta-oxidation of FA
zz Pathways where HMG CoA is intermediate Fig. 10.10: Ketone Body Utilization. First enzyme is Thiophorase,
also known as Succinyl CoA – Acetoacetate CoA Transferase. This
 KB synthesis
enzyme is a special enzyme as it does not use ATP but it creates a
 Cholesterol synthesis
high energy CoA bond. This Coenzyme A is taken from Succinyl CoA
 Leucine catabolism (intermediate of TCA cycle). This enzyme is absent in liver. Liver can


only produce ketone bodies but cannot use ketone bodies.

Ketone Body Utilization (Fig. 10.10)
This pathway occurs during starvation and can occur only in
brain, heart and skeletal muscles. These organs can extract
ATPs (energy) from ketone bodies.

Fig. 10.11: Ketone body synthesis in liver and utilization in peripheral tissues like muscle
Energetics from Acetoacetate zz Therefore 20 + 2.5 = 22.5 ATP
zz 2 Acetyl CoA = 20 ATP (each Acetyl CoA gives 10 ATPs in zz 1 less because Succinyl CoA → Succinate does not yield
TCA cycle) any ATP
zz 1 less because Succinyl CoA to Succinate conversion zz So energy yield from β-hydroxy butyrate = 22.5 – 1 = 21.5
does not yield any ATP ATP
zz So energy yield from Acetoacetate = 20 – 1 = 19 ATP
Tests for Detection of Ketone
Energetics from β-hydroxy Butyrate Bodies
T
H Rothera’s Test
E Ketone bodies react with sodium
O nitroprusside (in the presence of
R NH3) to form a purple colored ring
Y zz Positive test indicate
zz 1 NADH formed = 2.5 ATP
acetoacetate and acetone
zz 2 Acetyl CoA = 20 ATP (from Acetoacetate)
Gerhardt’s Test 20 C) require this to be transported. Short chain (2-4 C)
Ferric chloride added → wine color and medium chain (6-12 C) fatty acids do not require
zz
zz Positive test indicate acetoacetate and not acetone
Carnitine. Three enzymes are required in this shuttle: 263
1. CPT-I→Carnitine Palmitoyl Transferase–I (or CAT-I
zz Test for β-Hydroxy Butyrate – as this is the main ketone body i.e. Carnitine Acyl Transferase-I)
found in blood and urine of patients during ketoacidosis 2. CPT-II→Carnitine Palmitoyl Transferase–II (or CAT-

CHAPTER 10  METABOLISM OF LIPIDS

zz No specific test available for β-Hydroxy Butyrate II i.e. Carnitine Acyl Transferase-II)
zz So it is oxidized to acetoacetate first 3. Translocase
zz Then we can perform Rothera’s test. CPT-I is present in OMM (Outer Mitochondrial Membrane).
CPT-II and Translocase are present in Inner Mitochondrial
H igh R eturn Membrane (IMM).
So, CPT-I is the outermost enzyme, which decides the entry of
zz Why Liver cannot use KB?- because of absence of Thiophorase. activated fatty acid into the mitochondria. Source of carnitine
zz Why RBCs cannot use KB?- because they lack Mitochondria. is diet or endogenous synthesis. Skeletal muscles contain 97%
zz Starting point of Ketone Body Synthesis - Acetyl CoA of all carnitine present in body.
zz First enzyme of Ketone Body Synthesis – Thiolase
zz First enzyme of Ketone Body Utilisation – Thiophorase Carnitine synthesis: Occurs in Liver
zz Immediate precursor of Acetoacetate – HMG CoA zz Lysine required
zz SAM is the Methyl donor (so Methionine required)
zz Vit C also required
BETA OXIDATION OF FATTY ACID
Deficiency in Carnitine Shuttle
H igh R eturn zz Severe Hypoglycemia, which is non-ketotic
zz Nature of pathway: Catabolic
zz Occurs in fasting state/starvation zz Any defect in β-oxidation leads to non-ketotic hypoglycemia
zz Activated by hormone: Glucagon
zz Muscle weakness with myoglobinemia following prol-
zz Organ– Liver, Adipose tissue, Muscles
onged exercise
zz Organelle: Mitochondria (any catabolic pathway occurs in
mitochondria)
Treatment
Steps zz Avoid fasting
zz Activation of fatty acid zz Diet high in carbohydrates and low in fat
zz Transfer of activated FA from cytoplasm to mitochondria zz Medium chain TGs given (short and medium chain fatty
zz Actual beta-oxidation in mitochondria acids do not require carnitine shuttle)

Activation of FA
Enzyme is Acyl CoA Synthetase/Thiokinase, which is present
in outer mitochondrial membrane. It belongs to ligase
category. ATP gets converted to AMP. This is equivalent to
two ATPs used at this step. This reaction occurs in cytoplasm.

Transfer of Activated FA from cytoplasm to

mitochondria T
zz Acyl CoA is produced in cytoplasm but beta oxidation H
occurs in mitochondria. So, there is a need for the E
transport of fatty acid into the mitochondria O
zz Carnitine is a protein/carrier which is required for the R
transport. This is rate limiting transport process known Y
Fig. 10.12: Cartinine shuttle
as carnitine shuttle. Only long chain fatty acids (i.e. 14-
 Activated fatty acid reaches intermembrane space with the Actual beta-oxidation in mitochondria
help of CPT-I, it gets converted to Acyl Carnitine and CoA
264 is regenerated. Then translocase (Carnitine-Acyl Carnitine
zz In fasting state, beta oxidation (proper) occurs in mito-
chondria. It mainly occurs in liver cells. The resulting
Translocase) enzyme transports this acylcarnitine into the
Acetyl CoA has three fates: either it enters TCA cycle to
mitochondrial matrix and it also transports free carnitine out
provide energy to liver cells, or this Acetyl CoA is used
from mitochondrial matrix. Then CPT-II transfers Acyl group
CRO BIOCHEMISTRY

to CoA, making acyl CoA and regenerates free carnitine. to synthesize Ketone Bodies (for energy requirement of
This Acyl CoA can take part in beta oxidation proper in vital organs during starvation) or this Acetyl CoA is used
mitochondria. to activate the first step of gluconeogenesis (Pyruvate
Carboxylase).
Reciprocal Regulation zz Beta carbon is C3. Beta oxidation means cleavage occurs
between C2 and C3, so that each time one molecule of
In fed state, fatty acid synthesis occurs and Malonyl CoA is Acetyl CoA (2C) is cleaved from the fatty acid. For each
formed. Malonyl CoA (3C) is the inhibitor of CPT-I (outermost cleavage, four enzymes are required: dehydrogenase,
enzyme required for the transfer of activated fatty acid into hydratase, dehydrogenase and thiolase (cleaves the
the mitochondria in beta oxidation). So in fed state, beta Acetyl CoA to be released). First dehydrogenase produces


oxidation is inhibited. FADH2 and second dehydrogenase produces NADH. So

In fasting state, fatty acid synthesis does not occur. So from each cleavage, 4 ATPs are generated.
Malonyl CoA is not formed. So the inhibition to beta oxidation
is lost. So in fasting state, beta oxidation can occur. Table 10.2: ATP Production from β-Oxidation of Fatty Acids
This is known as reciprocal regulation i.e. two opposite
From Palmitic Acid (16 C) → 7 Cleavages
pathways don’t occur simultaneously. This is to prevent futile → 8 Acetyl CoA
cycle.
Enzyme Mode of ATP Net ATP produced
production
1st Dehydrogenase Forms FADH2 1.5 ATP
2nd Dehydrogenase Forms NADH 2.5 ATP
So, ATP per cleavage (two 4 ATP
Dehydrogenases)
ATPs from 7 cleavages (β-Oxidation 7 7 x 4 ATP
cycles) = 28 ATP
ATPs from 8 Acetyl CoA (TCA) 8 x 10 ATP per cycle
= 80 ATP
Total ATP produced 80 + 28 ATP
= 108 ATP
ATP consumed in activation of fatty acid -2 ATP
Fig. 10.13: Reciprocal regulation
Total ATP from 16C Palmitic Acid 108 – 2 ATP
C linical B ox = 106 ATP

Sulfonyl ureas inhibit CPT-I of β-oxidation. So Acetyl CoA is not Similarly, β-Oxidation of 18C Stearic Acid 120 ATP
obtained which is activator of gluconeogenesis. So decreased
gluconeogenesis decreases blood glucose. zz So from cleavage, ATPs obtained are 28 and from Acetyl
CoA, number of ATPs obtained are 80. So when Acetyl
Q. Muscle does not synthesize fatty acids. Then why CoA is not entering TCA, then these 80 ATPs will not be
muscles cells have the rate limiting enzyme of fatty obtained and this occurs during ketone body synthesis.
T acid synthesis i.e. Acetyl CoA Carboxylase?
H A. Muscles have Acetyl CoA Carboxylase because this
T I
H enzyme produces Malonyl CoA which can take part
N
E K in the regulation of beta oxidation of fatty acids
O whenever required.
R
Y
 265

CHAPTER 10  METABOLISM OF LIPIDS

Fig. 10.14: β-oxidation of fatty acid in liver mitochondria; beta oxidation occurring in liver mitochondria. Cleavage occurs between
C2 and C3, to release 2-C Acetyl CoA each time. For each cleavage, 4 enzymes are required. Two of them are dehydrogenases. First
dehydrogenase produces FADH2 and second dehydrogenase produces NADH

Dehydrogenases of beta-oxidation are of two types depen-

Jamaican Vomiting Sickness
ding on chain length
1. Long Chain Acyl CoA Dehydrogenase: Starts breaking a zz It occurs after ingestion of unripe fruit of akee tree, which
long chain fatty acid. It can breaks up to 12 C. contains a toxin named- Hypoglycin. It inhibits Fatty
Acyl CoA Dehydrogenase
2. Medium chain acyl CoA dehydrogenase: It works after
long chain acyl CoA dehydrogenase. And this enzyme zz Sudden vomiting occurs 2-6 hours after ingestion and
severe hypoglycemia
can do all the cleavages below 12 C.
zz It can be severe to cause convulsions, coma and death
also
MCAD Deficiency
zz Most common metabolic disorder associated with fatty Table 10.3: Ketotic and Non-Ketotic Hypoglycaemia. Any defect
acid metabolism. in beta oxidation pathway or ketone body synthesis pathway
cannot synthesize ketone bodies so it will lead to non-ketotic
zz Patient is normal in fed state. But during starvation,
hypoglycaemia. But when hypoglycaemia occurs due to fat
patient has decreased β-oxidation:
utilization in body, then it will lead to ketotic hypoglycaemia.
zz Low ketone bodies
zz Hypoglycemia (Non-Ketotic Hypoglycemia) Nonketotic Hypoglycemia Ketotic Hypoglycemia

C oncept B ox • Insulinemia
• Defect in beta oxidation of
• E.g. Von Gierke’s disease
• Alcoholism
This occurs because very few ATPs released in liver from the fatty acid • When fasting, Hypoglycemia
cleavage of beta oxidation. As fatty acid can be broken only till • Defect in KB pathway in body, then fats are used
12 C. So all the fatty acids (Acetyl CoA) will be used to provide T
ƒƒ MCAD deficiency
energy in liver. Acetyl CoA will not be available for entry into ƒƒ Jamaican vomiting H
ketone body synthesis or activation of gluconeogenesis. And the sickness E
12 C fatty acids, accumulates and are excreted in urine. As Beta • Carnitine, CPT-I or CPT-II O
Oxidation is not occurring, these fatty acids (12 C) are diverted deficiency R
to omega oxidation and leads to dicarboxylic acidosis. Treatment Y
includes avoidance of fasting.
 Table 10.4: Differences between fatty acid synthesis and beta Propionyl CoA (3C) is converted to D-Methyl Malonyl
oxidation of fatty acids CoA (4C) by carboxylase. Any Carboxylase requires biotin
266 i.e. Vitamin B7. Then D-methyl Malonyl CoA is converted
Fatty Acid Synthesis Beta Oxidation of Fatty Acid
to its L-isomer by enzyme Racemase (Racemase always
• Occurs in fed state when • Occurs in fasting state when interconverts D and L isomers). Then L-Methyl Malonyl CoA
insulin present glucagon present (4C) is converted to Succinyl CoA (4C) by enzyme mutase
CRO BIOCHEMISTRY

• Mainly in liver • In liver and muscle (Mutase is an isomerase and it always transfers groups within
a molecule: here it transfers methyl group). For methyl
• In cytoplasm • In mitochondria
malonyl CoA mutase, deoxyadenosylcobalamin is required.
• Carrier required for • Carrier required for Succinyl CoA is an intermediate of TCA cycle. So propionyl
transport between transport between CoA is glucogenic (i.e. can form glucose) as it forms an
mitochondria and cytoplasm mitochondria and cytoplasm intermediate of TCA cycle
is citrate is carnitine
• NADPH required for • NAD/FAD required for E nergetics B ox
reduction oxidation
Generally fats can never be converted to carbohydrates. But two
• Fatty Acid is synthesized • Fatty Acid is cleaved into EXCEPTIONS are:


from the carbons of Acetyl Acetyl CoA zz Glycerol

CoA zz Propionic acid

• Inhibitor —Palmitoyl CoA • Inhibitor—Malonyl CoA In Vitamin B12 deficiency, L-Methyl Malonic Acid
• Product of pathway- • Product of pathway- Acetyl and Propionic Acid are excreted in urine. In B7 deficiency,
Palmitoyl CoA CoA Propionic Acid is excreted in urine.
• 3-steps repeated — loading, • 4-enzymes repeated-
condensation and reduction- dehydrogenase, hydratase, Energetics of Odd Chain FA
for each 2 C to be added dehydrogenase, thiolase for
each cleavage Table 10.5: 17C Acetyl CoA and one melecule of Propionyl CoA

Beta Oxidation of Odd Chain Fatty Acids • Activation of FA Enzyme - Acyl CoA -2 ATP
Synthetase
Propionyl CoA is derived from the beta oxidation of odd chain
fatty acids and also from some branched chain amino acids. • 7 cleavage 4 ATPs from each 7×4=
zz Only one Propionyl CoA obtained from odd chain FA, so cleavage 28 ATP
this is not a significant contribution to glucose. • 7 Acetyl CoA 10 ATPs per Acetyl CoA 7 × 10 =
zz Odd chain fatty acid are present in ruminant fats e.g. from TCA 70 ATP
milk, ghee • Propionyl CoA Uses 1 ATP -1 ATP
Carboxylase
• Conversion of D- No ATP used or produced 0 ATP
Methyl Malonyl CoA to
Succinyl CoA
• Succinyl CoA to Succinate Thiokinase 1 ATP
Succinate produces 1 ATP
• Succinate to Fumarate Succinate 1.5 ATP
Dehydrogenase produces
1 FADH2, which produces
1.5 ATP
• Fumarate of Malate No ATP used or produced 0 ATP
• Malate to Pyruvate One NADPH produced 0 ATP

• Pyruvate to Acetyle Pyruvate Dehydrogenase 2.5 ATP

T CoA enzyme produces one
H NADH
E
Acetyl CoA (TCA) 10 ATPs per Acetyl CoA 10 ATP
O from TCA
R
Total ATPs 110 ATPs
Y
Fig. 10.15: Metabolism of Propionyl CoA But in short you can easily calculate like this consider 4 ATPs
used from one Propionyl CoA to Acetyl CoA and add this Acetyl
CoA also.
Oxidation of VLCFA (Very Long Chain Fatty Acids)
Induced by high fat diet
zz
zz For C-20, C-22 fatty acids
267
zz Synthetase enzyme for activation of VLCFA lies in
peroxisomes

CHAPTER 10  METABOLISM OF LIPIDS

zz It occurs in peroxisomes upto Octanoyl CoA and rest in
mitochondria
zz No ATP produced

Defect in Oxidation of VLCFA in Peroxisomes Fig. 10.16: Two disorders occur due to defect in peroxisomes
zz Zellweger syndrome
 Also called Cerebro-Hepato-Renal syndrome Omega – oxidation
 Rare, Autosomal Recessive disorder zz Occurs in ER (Endoplasmic Reticulum)
 Absence of peroxisomes in all tissues zz It is a minor pathway
 Defect in beta oxidation of very long chain fatty acids zz Occurs at methyl terminal of fatty acid
 Accumulation of polyenoic acids in brain (con- zz Result in the production of short chain dicarboxylic acids
taining C > 22) zz Occurs by Cytochrome P450 (a Mixed Function Oxidase)
 Severe neurological symptoms, Impaired neuronal
migration, Hypomyelination, hepatomegaly, Renal
cysts occurs
 Most patients die within 1st year of life
zz X-linked Adrenoleuko Dystrophy (XALD): Defect
in the transport of VLCFA across the peroxisomal Fig. 10.17: A dicarboxylic acid
membrane. Demyelination of CNS and degradation of
adrenal function. Accumulation of VLCFA in blood and CHOLESTEROL BIOSYNTHESIS
tissues occurs. zz Nature of pathway: anabolic
Beta Oxidation of Unsaturated FA zz Occurs in fed state
zz Activated by Hormone: Insulin
zz Occurs in mitochondria
zz Organelle: Cytoplasm (any anabolic pathway occurs in
zz Normal beta oxidation occurs till the double bond cytoplasm)
zz The first dehydrogenase step is bypassed. So FADH2 zz Organ/cell: liver, mammary lactating glands and to a
not produced (Because unsaturated fatty acids are limited extent in adipose tissue
less reduced than saturated fatty acids, less reducing zz Starting material – Acetyl CoA
equivalent produced during their oxidation) zz Rate Limiting Enzyme of fatty acid synthesis – HMG CoA
zz So energy yield is 1.5 ATP less/double bond Reductase
Minor Oxidation Pathways Cholesterol is required for
zz Alpha-Oxidation Membranes, Steroid Hormones, Lipoproteins, Vitamin D and
zz Omega- Oxidation Bile acids

Alpha – Oxidation
zz Occurs in Peroxisomes and ER
zz For branched chain FA e.g. Phytanic acid (product of
chlorophyll metabolism)
zz Beta oxidation cannot occur for phytanic acid as the
methyl group is on beta carbon. So alpha oxidation
occurs i.e. cleavage between C1 and C2.
zz No ATPs produced T
zz Defect in alpha oxidation in peroxisomes → Refsum’s H
disease – autosomal recessive. In this disease, Phytanic E
acid not oxidized, so accumulated in plasma and tissues. O
zz Enzyme affected- Phytanoyl CoA Oxidase or Hydroxylase R
zz CNS affected Fig. 10.18: Cholesterol structure –27 C and –OH group at C3. Y
There are 4 fused hydrocarbon rings (A to D), which is known as
zz Treatment is restriction of dairy products and green leafy Steroid nucleus. A branched hydrocarbon chain is attached at C-17
vegetables, which are rich in phytanic acid
268
CRO BIOCHEMISTRY

Fig. 10.19: Structure of cholesterol ester: at C3 fatty acid is attached to cholesterol, making it cholesterol ester

Cholesterol Synthesis
It occurs in all tissues. Acetyl CoA is the starting material and NADPH is used for reductive biosynthesis. ATP is also used.


Fig. 10.20: Cholesterol synthesis shown in cytoplasm as compared to ketone body synthesis in mitochondria
First two steps of cholesterol synthesis are same as ketone body synthesis but cholesterol synthesis occurs in cytoplasm
whereas ketone body synthesis occurs in mitochondria. The third step in cholesterol synthesis is done by enzyme HMG CoA
Reductase (conversion of HMG CoA to Mevalonate), which is the Rate Limiting Enzyme. This is inhibited by statins group of
drugs.

Pearls of the Chapter

zz No ATP produced by oxidation of VLCFA and alpha oxidation of zz Immediate precursor of acetoacetate – HMG CoA
FA zz RLE of KB synthesis – HMG CoA synthase
T zz Starting point of ketone body synthesis – Acetyl CoA zz RLE of cholesterol synthesis – HMG CoA Reductase
H zz First enzyme of ketone body synthesis – Thiolase zz Odd chain Fatty Acids can form glucose but even chain fatty
E zz First enzyme of ketone body utilisation–Thiophorase acids cannot.
O zz Neutral KB – Acetone
R
Y
SITES OF FATTY ACID SYNTHESIS AND FATTY ACID OXIDATON
Pathway Location Special point 269
Fatty Acid Synthesis Cytoplasm Synthesis of saturated fatty acid upto 20 C
Elongation of Fatty Acid ER Elongation above 20 C

CHAPTER 10  METABOLISM OF LIPIDS

+
Mitochondria
Desaturation of Fatty Acid ER Creating double bond in Fatty Acid
β- oxidation Mitochondria Major catabolic pathway for Fatty Acid
Activation of Fatty Acid for β oxidation Cytoplasm Activated Fatty Acid travels from Cytoplasm to
Mitochondria via Carnitine system
α- oxidation Peroxisomes For Branched Chain Fatty Acid e.g. Phytanic Acid
Defect leads to Refsum’s disease
ω- oxidation ER Leads to formation of Dicarboxylic Acids
Oxidation of Very Long Chain Fatty Acid Peroxisomes Very Long Chain Fatty Acid have a modified β -oxidation
first in peroxisomes upto Octanoyl CoA, then remaining in
mitochondria

T
H
E
O
R
Y
 270 Multiple Choice Questions
Lipid Metabolism 11. Co-factors required for fatty acid synthesis in humans
1. Major metabolism of saturated fatty acids in the are? (PGMEE-2016)
mitochondria is called as: (PGMEE 2015) a. ATP b. NADPH
a. a – oxidation b. β- oxidation c. Biotin d. Pyridoxine
c. w – oxidation d. None of the above e. Pantothenic acid
2. How many cycles of beta oxidation are required for 12. In beta oxidation of palmitic acid, if final product is
complete oxidation of palmitic acid into acetyl CoA? acetoacetate, then net gain of ATP is:
 (PGMEE 2016) a. 21 b. 26
a. 7 b. 8 c. 106 d. 129
c. 9 d. 2 13. Which one of the following tissues can metabolize
3. Citrate used in fatty acid synthesis use which enzyme- glucose, fatty acids, and ketone bodies for ATP
 (PGMEE 2012-13) production?
a. Malic enzyme b. ATP citrate lyase a. Liver b. Muscle
c. Aconitase d. Citrate synthase c. Brain d. Red blood cells
4. Formation of HMG-CoA in liver mitochondria is 14. In MCAD deficiency, during starvation, which of the
inhibited by: (PGMEE 2012-13) following is correct? (Recent Question 2018)
a. Insulin a. Normal blood glucose
b. Glucocorticoid b. Ketoacidosis
c. Glucagon c. Dicarboxylic acidosis
d. Epinephrine d. Hyperchylomicronemia
5. Which of the following is not a component of fatty 15. When liver is actively synthesizing fatty acids in fed
acid synthase complex? (AIIMS Nov 13) state, then there is concomitant decrease in beta
a. Enyol reductase oxidation of fatty acids due to: (PGMEE 2010)
b. Acetyl transacylase a. Inhibition of CPT-I by Malonyl CoA
M c. Acetyl-CoA carboxylase b. Inhibition of translocation of fatty acid from cytosol
C d. Ketoacyl synthase to mitochondria
Qs 6. Refsum’s disease is due to defect in: (PGMEE 2013) c. Inhibition of translocation between the cellular
Ans. a. Phytanic acid oxidase compartments
b. Phytanic acid lyase d. All
1. b 16. Most abundant source of fuel in starvation: 
c. Phytanic acid isomerase
2. a  (PGMEE 2015)
d. Phytanic acid reductase
3. b a. Liver glycogen b. Blood glucose
7. In Zellweger syndrome, which of the following is
4. a c. Muscle glucose d. Adipose tissue
absent? (Recent Question Jan 2019 Q)
5. c 17. Fuel utilized by brain in starvation- 
a. ER b. Golgi apparatus
6. a  (PGMEE 2015, 2011, 2009)
c. Mitochondria d. Peroxisomes
7. d a. Ketone bodies b. Fatty acid
8. Which type of cholesterol is present in gall stones?
8. d c. Glycogen d. Amino acids
 (Recent Question Jan 2019 Q)
9. d 18. During starvation, which of the following shows the
a. Amorphous cholesterol dihydrate
10. c most marked increase in plasma concentration:
b. Amorphous cholesterol monohydrate
11. a,b,c  (PGMEE 2016-17)
c. Crystalline cholesterol dihydrate
12. b a. Glycogen b. Glucose
d. Crystalline cholesterol monohydrate
13. b c. Ketone bodies d. Free fatty acids
9. Major product of fatty acid synthesis is:
14. c 19. In a person fasting overnight with carnitine defi-
 (Recent Question Jan 2017,2018)
15. d ciency, following chemicals increase in quantity in
a. Acetyl CoA b. ATP
16. d blood: (PGMEE 2012-13)
c. Citrate d. Palmitate
17. a a. Ketone bodies b. Fatty acids
10. In cerebrohepatorenal syndrome, which of the
18. c c. Amino acids d. Glucose
following accumulate in brain? (AIIMS Nov 2018)
19. b 20. Fatty acids are not used by which organ-
a. Pyruvate
20. a  (PGMEE 2015)
b. Short chain Fatty acid
c. Very long chain fatty acid a. Brain b. Liver
d. Acetyl CoA c. Muscle d. Heart
21. Which of the following is not seen in 12 days of 34. Allosteric inhibitor of fatty acid synthase is:
fasting- (PGMEE 2015)  (PGMEE 2015)
a. Ketogenesis b. Lipolysis a. NAD b. Long chain acyl CoA 271
c. Gluconeogenesis d. Glycogenesis c. ATP d. Citrate
22. In diabetes and starvation, acidosis occur due to 35. Beta-oxidation of odd-chain fatty acids produces:
 (PGMEE 2011)  (PGMEE 2016-17; PGMEE 2007)
a. Glycogen b. Ketone bodies a. Malonyl CoA b. Propionyl CoA
c. Glucose d. Sphingolipids c. Acetyl CoA d. Succinyl CoA
23. In diabetes, the factor limiting synthesis of trigly- 36. Beta Oxidation in peroxisome generate:
cerides in adipose tissue is: (PGMEE 2011)  (PGMEE 2015)
a. NADPH b. ATP a. H2O2
c. Acetyl CoA d. Glycerol-3-P b. NADPH
24. Building block for fatty acid biosynthesis is: c. Long Chain fatty acid
 (PGMEE 2010) d. FADH2
a. Acetate b. Acetyl CoA 37. a-oxidation of fatty acid takes place in:
c. Acyl-CoA d. NADH  (PGMEE 2011)
25. Acetyl CoA Carboxylase is allosterically activated by: a. Mitochondria b. Golgi apparatus
 (PGMEE 2015) c. ER d. Peroxisomes
a. Citrate b. Glucagon 38. Dietary fiber reduces atherosclerosis by:
c. Acetoacetate d. Malonyl CoA  (PGMEE 2008)
26. 1st acetyl group donor in fatty acid synthesis is – a. Binding to cholesterol
 (PGMEE 2012-13) b. Decreasing VLDL
a. Citrate b. Palmitate c. Forming antioxidants
c. Acetyl CoA d. Malonyl CoA d. Increasing fluid retention
27. Long chain fatty acid transported into inner mito- 39. “Squalene” is the intermediate product during
chondrial membrane by:- (PGMEE 2016-17) synthesis of: (PGMEE 2013)
a. Acyl carrier protein a. Cholesterol b. Lanosterol
c. LDL d. Tachysterol
b. Energy mediated
40. Number of ATPs formed by beta oxidation of one
c. Acyl carnitine
molecule of Stearic acid is: (PGMEE-2015)
d. Simple diffusion M
a. 106 b. 116
28. Reducing agent used in lipogenesis is derived from: C
c. 118 d. 120
 (PGMEE 2009) Qs
41. Enzyme common to cholesterol synthesis and ketone
a. Pentose phosphate pathway Ans.
body synthesis:
b. Glycolysis
a. HMG CoA Reductase 21. d
c. Gluconeogenesis
b. HMG CoA Synthase 22. b
d. TCA cycle
c. Thiolase 23. d
29. Triglyceride synthesis in increased by: (PGMEE 2011) d. Thiophorase
a. Glucagon b. Insulin 24. b
42. A 30-year-old man has been fasting for religious 25. a
c. Growth hormone d. Cortisol reason for several days. His brain has reduced the
30. Acetyl CoA directly gives rise to all EXCEPT: 26. c
need for glucose by using which of the following 27. c
 (PGMEE 2009) substance as an alternate source of energy?
a. Fatty acid b. Ketone 28. a
a. Fatty acids 29. b
c. Cholesterol d. Glucose b. Glycerol
31. Multienzyme complex in humans: (PGMEE 2017,16) 30. d
c. Alanine 31. c
a. Adenosine phospho-ribosyltransferase d. Beta- hydroxy butyrate
b. Malonyl CoA carboxylase 32. c
43. A 9-year-old girl is brought to emergency by her 33. a
c. Fatty acid synthetase parents with complaints of severe polyuria and
d. Carbamoyl phosphate synthetase 34. b
polydipsia. Laboratory examination reveals ketones 35. c
32. The vitamin present in the fatty acid synthase com- in her urine. Which of the following is the most likely 36. a
plex is: (PGMEE 13) source of these ketones? (Recent Question 2017) 37. d
a. Pyridoxine b. Folate a. Protein breakdown 38. a
c. Pantothenate d. Thiamine b. Fatty acid breakdown 39. a
33. Rate limiting enzyme in the synthesis of cholesterol c. Gluconeogenesis 40. d
is: (PGMEE 2015) d. Side chain of cholesterol 41. b
a. HMG CoA reductase 44. Which of the following is used by RBCs for metabolism 42. d
b. Acetyl CoA synthetase during fasting state? (AIIMS May 2015) 43. b
c. Acetyl CoA carboxylase a. Glucose b. Alanine 44. a
d. HMG CoA synthetase c. Ketone body d. Fatty acid
 45. Which of the following enzyme is not a component of 52. Odd chain fatty acids can form glucose by which
fatty acid synthase complex? pathway? (JIMPER May 2018)
272  (AIIMS November 2013) a. By forming Propionyl CoA
a. Ketoacyl synthase b. By forming Glycerol
b. Thioesterase c. Acetyl CoA entering TCA cycle
c. Acetyl-CoA carboxylase d. By lactic acid formation
d. Enoyl reductase 53. Active metabolite form in synthesis of fatty acid is:
46. All are features of Refsum’s disease EXCEPT:  (Recent Question Jan 2018)
 (PGI Nov 2014) a. Acetyl CoA b. Malonyl CoA
a. Defect of a- oxidation c. Stearate d. Palmitate
b. Defect in β oxidation 54. Which organ cannot use ketone bodies:
c. Accumulation of phytanic acid  (PGI May 2018)
d. Peripheral neuropathy a. Brain b. RBC
e. Treated by removing phytanic acid precursors from c. Muscle d. Heart
diet e. Liver
47. In prolong fasting glycerol is formed from 55. First enzyme of ketone body utilisation:
triglyceride. Which of the following statement (s) is/ a. Thiophorase
are true regarding glycerol: (PGI May 2015) b. Thiolase
a. Used in synthesis of chylomicron c. HMG CoA Reductase
b. It is directly used by tissues for energy needs d. HMG CoA Synthase
c. It is formed due to increased activity of lipoprotein 56. Immediate precursor of acetoacetate:
lipase a. HMG CoA
d. It is formed due to increased activity of hormone b. Acetoacetyl CoA
sensitive lipase c. Acetyl CoA
e. Glycerol acts as a substrate for gluconeogenesis in d. Malonyl CoA
the liver 57. Which of the following is true about ketone bodies?
48. In which organelle(s) of hepatocyte, the elongation  (PGMEE 2016)
of long chain fatty acid takes place: (PGI Nov 2008) a. Acetoacetate is most common ketone body present
a. Endoplasmic reticulum (ER) in blood
b. Golgi body b. Beta hydroxy butyrate is the first ketone body to be
M c. Mitochondria
C synthesized
d. Lysosomes c. Thiophorase is absent in liver
Qs e. Ribosome d. Muscle can not utilize ketone bodies
Ans. 49. What is role of insulin in lipid metabolism: 58. Ketone body that cannot be detected by Rothera’s
 (PGI Nov 2008) test? (PGMEE 2013-14)
45. c a. Activate lipoprotein lipase a. Acetoacetate
46. b b. Increase lipolysis b. Acetone
47. d,e c. Activate hormone sensitive lipase
48. a,c
c. Beta hydroxy butyrate
d. Activate Acetyl CoA carboxylase d. All the above can be detected
49. a,d,e e. Decrease free fatty acid level
50. a
59. Severity of ketosis assessed by:
50. During starvation, muscle uses: a. Measurement of ketone bodies in blood
51. d a. Fatty acids
52. a
b. Measurement of ketone bodies in urine
b. Ketone bodies
53. b
c. Measurement of free fatty acids in blood
c. Pyruvate
54. b d. Measurement of free fatty acids in urine
d. None
55. a 60. Fatty acid is synthesized from:
51. HMG CoA directly converted into all; EXCEPT:
56. a a. Acetyl CoA
 (Recent Question June 2018)
57. c b. Malonyl CoA
a. Acetyl CoA
58. c c. Both a and b
b. Acetoacetate
59. a d. Citrate
c. Mevalonate
60. a d. Acetoacetyl – CoA
 Answers with Explanations
273
1. Ans. (b) β - oxidation 7. Ans. (d) Peroxisomes

CHAPTER 10  METABOLISM OF LIPIDS

[Ref: Harper 30th/e pg. 225] [Ref: Harper 30th /e p 237]
•• Major metabolism of saturated fatty acids in the •• Zellweger syndrome was earlier named Cerebro-
mitochondria is called as β-oxidation. Refer Fig. 10.14 Hepato- Renal syndrome.
•• It is a Peroxisome Biogenesis Disorder (PBD), which
2. Ans. (a) 7 is very rare
•• Most severe PBD is Zellweger syndrome in which
[Ref: Harper 30th/e pg. 225] there is absence of peroxisomes in all tissues. So there
•• 16 C Palmitic acid is broken into 2 C – Acetyl CoA. is defect in oxidation of very long chain fatty acids &
So, 7 cleavages are required. Last cleavage gives two also defect in alpha oxidation as both these pathways
molecules of Acetyl CoA. occur in peroxisomes.
Formula is:
No. of C–atoms 8. Ans. (d) Crystalline cholesterol monohydrate
No. of cycles or cleavage = –1
2
No. of C–atoms [Ref: Harper 30th /e p 264]
No. of Acetyl CoA obtained from β–Oxidation =
2 Gall stones have typical plate like cholesterol mono-
3. Ans. (b) ATP citrate lyase hydrate crystals

[Ref: Harper 30th/e pg. 223]

Citrate in FA synthesis uses ATP Citrate Lyase

4. Ans. (a) Insulin

[Ref: Harper 30th/e pg. 269]
Synthesis of HMG CoA occurs in both mitochondria
and cytoplasm. In mitochondria, it is used for Ketone
body synthesis (inhibited by Insulin). In cytoplasm, it is
used for cholesterol biosynthesis (activated by Insulin)
9. Ans. (d) Palmitate
5. Ans. (c) Acetyl-CoA Carboxylase
[Ref: Harper 30th/e pg. 234] A
[Ref: Harper 30th/e pg. 234] •• Major fatty acid which gets synthesized in body is N
•• Acetyl CoA Carboxylase is rate limiting enzyme of Palmitic acid. That’s why enzyme fatty acid synthase S
Fatty acid synthesis. Fatty Acid Synthase complex complex is also known as Palmitate Synthase.
W
is main enzyme of fatty acid synthesis. Acetyl CoA •• Major product of fatty acid breakdown is Acetyl CoA.
E
carboxylase is not a component of FA synthase R
complex. Refer Fig. 10.1 10. Ans. (c) Very long chain fatty acid
S
[Ref: Harper 30th /e p 237]
6. Ans. (a) Phytanic acid oxidase
WITH
For explanation, refer Ques 7.
[Ref: Harper 30th/e pg. 231] E
•• Refsum disease is a rare, autosomal recessive, neuro- 11. Ans. (a) ATP; (b) NADPH; (c) Biotin X
logical disorder. It is a phytanic acid storage disease [Ref: Harper 30th/e pg. 234, 236] P
due to deficiency of enzyme phytanic acid oxidase. L
•• Phytanic acid is found in meat, beef, ruminant fat •• Two enzymes are involved in fatty acid synthesis A
and dairy products. Phytanic acid is thought to have 1. Acetyl CoA Carboxylase – This is the rate limiting N
effects on membrane function, protein prenylation enzyme in fatty acid synthesis and is required for A
and gene expression. the conversion of acetyl CoA to malonyl CoA. As T
•• Clinical features include night blindness, retinitis carboxylation requires biotin and ATP as their I
pigmentosa, deafness, dry scaly skin, shortened cofactors so this enzyme acetyl CoA carboxylase O
fingers or toes, anosmia and ataxia. requires ATP & biotin. N
2. Fatty Acid Synthase complex - requires NADPH S
 12. Ans. (b) 26 19. Ans. (b) Fatty acids

274 •• In beta oxidation of Palmitic acid, we obtain 28 ATPs [Ref: Harper 30th/e pg. 214]
from beta oxidation and 80 ATPs from Kreb’s cycle. In carnitine deficiency, ketone bodies cannot be raised
But if Acetyl CoA is forming acetoacetate (ketone as β oxidation is not occurring. So fatty acids will be
body), that means Acetyl CoA is not entering Kreb’s increased.
CRO BIOCHEMISTRY

cycle. So those 80 ATPs are not obtained. Now net

gain of ATP is 28-2 (2 ATPs used in activation of fatty 20. Ans. (a) Brain
acid). So answer here is 26. [Ref: Harper 30th/e pg. 150]
13. Ans. (b) Muscle Fatty acid cannot cross blood brain barrier.
•• Muscles can use glucose, fatty acids and ketone
21. Ans. (d) Glycogenesis
bodies.
•• Liver cannot use ketone bodies, because of deficiency [Ref: Harper 30th/e pg. 149]
of Thiophorase (first enzyme of ketone body After 12 days of fasting, an anabolic pathway like
utilization). glycogenesis cannot occur.


•• Brain cannot use FA because FA cannot cross blood

brain barrier. 22. Ans. (b) Ketone bodies
•• RBCs cannot use fatty acids and ketone bodies, [Ref: Harper 30th/e pg. 597; Table 48–5]
because they lack mitochondria. Ketoacidosis occurs in diabetes and starvation.
14. Ans. (c) Dicarboxylic acidosis 23. Ans. (d) Glycerol-3-P
•• In MCAD deficiency (Medium chain acyl CoA
[Ref: Harper 30th/e pg. 261]
Dehydrogenase), during starvation, patient will be
having low ketone bodies and hypoglycemia. But in Glucose entry via GLUT-4 is needed in adipose tissue
fed state, patient will be almost normal. And 12 C for triglyceride synthesis. But in diabetes, GLUT-4 is not
fatty acid gets accumulated, leading to dicarboxylic activated due to lack of Insulin.
acidosis.
24. Ans. (b) Acetyl CoA
•• Hyperchylomicronemia (Option d) is not possible
during starvation, as chylomicrons are formed from [Ref: Harper 30th/e pg. 2]
diet. Building block for fatty acid biosynthesis is Acetyl CoA

15. Ans. (d) All 25. Ans. (a) Citrate

•• This is reciprocal regulation of β-oxidation of
[Ref: Harper 30th/e pg. 237]
Fatty acid and Fatty acid synthesis. Activated Fatty
A acid needs to be transported from Cytoplasm to •• Acetyl CoA is allosterically activated by citrate and
N Mitochondria for β- oxidation by CPT-I: In fed state, inhibited by Fatty Acyl CoA
S Malonate (3 C) from Fatty acid synthesis inhibits
CPT-I. Refer Fig. 10.13. 26. Ans. (c) Acetyl CoA
W
E 16. Ans. (d) Adipose tissue
[Ref: Harper 30th/e pg. 225]
R •• Only 1st acetyl group donor in fatty acid synthesis is
S [Ref: Harper 30th/e pg. 231] Acetyl CoA.
Fats are the main fuel in fasting and starvation. Refer •• Rest all acetyl group donors in fatty acid synthesis is
WITH
Table 1.1 Malonyl CoA.
E
X 17. Ans. (a) Ketone bodies 27. Ans. (c) Acyl carnitine
P [Ref: Harper 30th/e pg. 149] [Ref: Harper 30th/e pg. 224]
L
A Brain and heart (vital organs) use ketone bodies during •• Long chain fatty acid is transported into inner
N starvation. Refer Table 1.1 mitochondria by acyl carnitine. Refer Fig. 10.12
A
18. Ans. (c) Ketone bodies 28. Ans. (a) Pentose phosphate pathway
T
I [Ref: Harper 30th/e pg. 149] [Ref: Harper 30th/e pg. 237]
O Ketone bodies are increased only in starvation, not in •• HMP forms NADPH which is used in fatty acid
N fasting. Free fatty acids are increased in both fasting and synthesis.
S starvation.
29. Ans. (b) Insulin 37. Ans. (d) Peroxisomes

[Ref: Harper 30th/e pg. 237] [Ref: Harper 30th/e pg. 225] 275
•• Insulin is anabolic hormone which increases fat •• α-oxidation of fatty acid takes place in peroxisomes
synthesis in body.
38. Ans. (a) Binding to cholesterol

CHAPTER 10  METABOLISM OF LIPIDS

30. Ans. (d) Glucose
[Ref: Harper 30th/e pg. 264]
[Ref: Harper 30th/e pg. 226]
•• Dietary fiber reduces atherosclerosis by binding to
•• Acetyl CoA is never glucogenic but it is starting cholesterol
material for Ketone Bodies, Fatty Acids and
Cholesterol. 39. Ans. (a) Cholesterol

31. Ans. (c) Fatty acid synthetase [Ref: Harper 30th/e pg. 264]

[Ref: Harper 30th/e pg. 233] •• Squalene is the intermediate product during synth-
esis of cholesterol.
•• Fatty Acid synthetase complex is a multienzyme
complex. It is main enzyme of fatty acid synthesis. 40. Ans. (d) 120
Refer Fig. 10.1
[Ref: Harper 30th/e pg. 214]
32. Ans. (c) Pantothenate
Number of ATPs formed by beta oxidation of one
[Ref: Lippincott’s illustrated biochemistry pg. 130] molecule of Stearic acid is 120 ATPs. Number of ATPs
The vitamin present in the fatty acid synthase complex formed by beta oxidation of one molecule of Palmitic
is Pantothenate. acid is 106 ATPs.

33. Ans. (a) HMG CoA reductase 41. Ans. (b) HMG CoA Synthase

[Ref: Harper 30th/e pg. 268] [Ref: Lippincott 4th/e pg. 197]
•• Rate limiting enzyme in the synthesis of cholesterol is Enzyme common to cholesterol synthesis and ketone
HMG CoA reductase. body synthesis is HMG CoA Synthase and Thiolase. Best
•• Inhibitors of this enzyme are Statins, Cholesterol and option here to be marked is HMG CoA Synthase.
Bile acids. Refer Fig. 10.20.
42. Ans. (d) Beta- hydroxy butyrate
34. Ans. (b) Long chain acyl CoA
[Ref: Harper 30th/e pg. 149]
[Ref: Harper 30th/e pg. 234] A
•• Beta hydroxy Butyrate is a ketone body. Ketone bodies N
•• Allosteric inhibitor of fatty acid synthase is long chain
serve as alternative fuel for brain during prolonged S
acyl CoA. Allosteric activator of fatty acid synthase
fasting or starvation. Ketone bodies can only be used W
complex is citrate.
by heart, brain and skeletal muscles. E
35. Ans. (c) Acetyl CoA •• Fatty acids due to long hydrophobic chain cannot R
cross blood brain barrier.
[Ref: Harper 30th/e pg. 226] S
•• Acetyl CoA > Propionyl CoA 43. Ans. (b) Fatty acid break down WITH
•• β-oxidation of odd chain fatty acid produces many
[Ref: Harper 30th/e pg. 278] E
Acetyl CoA and only one molecule of Propionyl CoA.
So the major end product is Acetyl CoA. •• Fatty acid breakdown provides Acetyl CoA that serves X
as a precursor for ketone bodies. In diabetes mellitus, P
36. Ans. (a) H2O2 glucose utilization is impaired due to absolute or L
relative insulin deficiency. A
[Ref: Harper 30th/e pg. 226] N
•• Fatty acid breakdown occurs to provide energy and
•• Beta Oxidation in peroxisome generate hydrogen A
the resultant excessive Acetyl CoA enters the pathway
per-oxide (H2O2) T
of ketogenesis.
I
O
N
S
 44. Ans. (a) Glucose 49. Ans. (a); (d); (e)

276 [Ref: Harper 30th/e pg. 150] [Ref: Harper 30th/e pg. 148]
•• RBCs always use glucose (fed, fasting or starvation).
•• Insulin activates Lipoprotein lipase and shift
45. Ans. (c) Acetyl-CoA carboxylase triglycerides and fatty acid of blood into adipose
CRO BIOCHEMISTRY

tissues.
[Ref: Harper 30th/e pg. 234]
•• Components of Fatty Acid Synthase Complex are: 50. Ans. (a) Fatty acids
Ketoacyl Synthase, Malonyl CoA, Hydratase, Enoyl [Ref: Harper 30th/e pg. 662]
reductase, Ketoacyl Reductase, ACP and Thio-
esterase. •• Muscles can use both fatty acids and ketone bodies.
Best option is fatty acids here. But muscles can also
46. Ans. (b) Defect in β oxidation use ketone bodies.

[Ref: Harper 30th/e pg. 231] 51. Ans. (d) Acetoacetyl – CoA
For explanation Refer Ques. 6


[Ref: Harper 30th/e pg 149]

47. Ans. (d); (e) •• Acetyl CoA, Acetoacetate and Mevalonate can be
[Ref: Harper 30th/e pg. 255] formed from HMG- CoA.
•• Glycerol is formed due to increased activity of HSL •• Acetoacetate, acetyl CoA are ketone bodies formed
and it acts as a substrate for gluconeogenesis in the by ketone body synthesis pathway.
liver. •• Mevalonate is formed from the cholesterol synthesis
pathway.
48. Ans. (a); (c)

[Ref: Harper 30th/e pg. 236]

•• The elongation of long chain fatty acid takes place
in endoplasmic reticulum and mitochondria of
hepatocytes.

A
N
S
W
E
R
S
WITH

E
X
P
L
A
N 52. Ans. (a) By forming Propionyl CoA 53. Ans. (b) Malonyl CoA
A
T [Ref: Lippincott 4th/e pg. 192] [Ref: Harper 30th/e pg. 288]
I Odd chain fatty acid forms Propionyl CoA which is Acetyl CoA is converted to malonyl CoA which is mainly
O glucogenic. used for fatty acid synthesis. So , active metabolite form
N is malonyl CoA.
S
54. Ans. (b) ; (e) use ketone body. Liver has to produce ketone body for
Brain, Heart and Skeletal muscles.
[Ref: Lippincott 4th/e pg. 197] 277
•• Brain, muscle and heart can use ketone bodies during 58. Ans. (c) Beta hydroxy butyrate
starvation [Ref: Harper 30th/e pg. 232]
•• In RBC, there is no mitochondria and ketone body

CHAPTER 10  METABOLISM OF LIPIDS

utilization pathway takes place in mitochondria. So Rothera’s test can only detect Acetoacetate and Acetone.
RBC cannot utilize ketone bodies
59. Ans. (a) Measurement of ketone bodies in blood
•• In liver there is absence of thiophorase, first enzyme
of ketone body utilization pathway. [Ref: Harper 30th/e pg. 229]
55. Ans. (a) Thiophorase In moderate ketonemia, the loss of ketone bodies in
urine is only a few percent of the total ketone body
[Ref: Harper 30th/e pg. 222] production and utilization. Since there are renal
•• The first enzyme of ketone body utilization is Thio- threshold like effects that vary between species and
phorase. individuals, measurement of ketonemia and not
•• The first enzyme of ketone body synthesis is Thiolase. ketonuria, is the preferred method of assessing the
severity of ketosis.
56. Ans. (a) HMG CoA Free fatty acids are the precursors of ketone bodies in
liver.
[Ref: Harper 30th/e pg. 222]
Immdediate precursor of acetoacetate (ketone body) is 60. Ans. (a) Acetyl CoA
HMG CoA, not Acetoacetyl CoA. [Ref: Harper 30th/e pg. 225]
57. Ans. (c) Thiophorase is absent in liver •• Fatty acid is entirely synthesized from the carbons
of Acetyl CoA. Malonyl CoA is used during fatty acid
[Ref: Harper 30th/e pg. 222] synthesis but all the carbons are derived from Acetyl
Thiophorase is the first enzyme of ketone body CoA. The extra carbon of malonyl is lost during
utilization. This is absent in liver so that liver cannot condensation step of fatty acid synthase complex.

A
N
S
W
E
R
S
WITH

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L
A
N
A
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I
O
N
S
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UNIT
V

MOLECULAR BIOLOGY
Unit Outline
Chapter 11 Chemistry and Metabolism of
Nucleotides
Chapter 12 Basics in Genetics
Chapter 13 Replication, Transcription, Translation
Chapter 14 Techniques in Molecular Biology
Chemistry and
Metabolism of
Nucleotides
11
Overview of Chapter zz Deamination of 5-Methyl Cytosine forms Thymine.
zz Methylation of Uracil forms Thymine
•• Nitrogenous Bases
•• Sugar Purines Pyrimidines
•• Synthesis of Purine Nucleotides • Have two rings • Have one ring
•• Purine catabolism • Sugar is attached at N9 • Sugar is attached at N1
•• Pyrimidine Synthesis • Three amino acids used • Two amino acids used are:
•• Functions of Nucleotides and their derivatives are: Glycine, Aspartate, Aspartate, Glutamine.
Glutamine
Molecular biology refer to the discussion of all the molecules • Product of catabolism– • Product of catabolism are
of life which take part in the transfer of genetic information. poorly soluble uric acid highly soluble CO2, NH3
i.e. DNA, RNA and Proteins (histone and non-histone) (purine ring not broken into and beta alanine. But for
zz NucleoSide = Base + (Deoxy)Ribose (Sugar) pieces during excretion) Thymine–they are CO2, NH3,
beta amino iso butyrate
zz NucleoTide = Base + (Deoxy)Ribose + PhosphaTe; linked
by 3’-5’ phosphodiester bond.
Sources of N and C of Purines
NITROGENOUS BASES
zz N1 → from Aspartate
They are of two types: Purines and Pyrimidines zz N3 , N 9→ from amide nitrogen of Glutamine
zz C 4, C5, N7→ from Glycine
zz C2→ N10 Formyl THF
zz C6→ from CO2
zz C8→ N5 N10methenyl THF

Fig.: 11.1: Sources of atoms of purines

Sources of N and C of Pyrimidines

zz Deamination of Cytosine forms Uracil zz C4, C5, C6, N1 → from Aspartate
zz Deamination of Adenine forms Hypoxanthine zz N3 → from Glutamine
zz Deamination of Guanine forms Xanthine zz C2 → from CO2
 SUGAR
282 Sugar in nucleic acids is always a Pentose. This pentose can
be ribose or deoxy-ribose.

Ribose
CRO BIOCHEMISTRY

This is a 5C sugar, so exists in furanose form. At 2’ position,

OH group is present.

Fig. 11.2: Thymine Deoxy-ribose

Oxygen is removed form 2’ position. So it is known as 2’-deoxy
F undamental B ox ribose.
zz THIAMINE is a vitamin – Vitamin B1
zz THYMINE – a pyrimidine nitrogenous base Q. Why a special mark put on the number like 2’ or 3’?
zz Thiamine (Vitamin B1) has a Pyrimidine like ring in its structure T A. When a special mark is present on the number that


H means this number is of sugar moiety. If number is

I written simply without any mark on it, then it means
N this is number of nitrogenous base.
N–base Nucleoside Nucleotide K
Adenine Adenosine AMP, ADP, ATP
Guanine Purine Guanosine sine
GMP, GDP, GTP

Cytosine Cytidine CMP, CDP, CTP

Uracil Pyrimidines Uridine dine UMP, UDP, UTP
Thymine Thymidine
TMP, TDP, TTP

Full form of nucleotide always uses the name of nucleoside

(not nitrogenous base) for e.g.:
zz ATP— Adenosine tri phosphate (not Adenine tri
phosphate)
zz CTP— Cytidine tri phosphate
zz TTP— Thymidine tri phosphate Conversion of Ribose to its Deoxyribose form
This conversion occurs at the level of diphosphate.
H igh R eturn
zz Guanidine – is a protein denaturing agent.
zz Guanosine – nucleoside of Guanine (i.e. Guanine + Sugar)
zz Most abundant free nucleotide in mammalian cells is ATP

Absorption of UV-light

T
H
E zz Nucleic acids absorb UV light at 260 nm due to
O Nitrogenous-base. This absorption is more for Purines
R as compared to pyrimidines. Fig. 11.3: Conversion of Ribonucleoside Diphosphate to its
Y zz Similarly, Proteins also absorb UV light at 280 nm due to deoxy form: enzyme is Ribonuceotide Reductase, which requires
vitamin B12. Thioredoxin is the donor of reducing equivalents
Aromatic amino acids. This absortion is maximum for
but the ultimate donor is NADPH. dATP is the inhibitor of
tryptophan. Ribonucleotide Reductase
 Q. In ATP (energy currency) which sugar is present? zz From Mono to Di Phosphate forms: Enzymes required
A. Ribose here are base specific e.g. specific for adenine and a
If dATP, then sugar is Deoxyribose. different enzyme for guanine nucleotide. 283

SYNTHESIS OF PURINE NUCLEOTIDES

CHAPTER 11  CHEMISTRY AND METABOLISM OF NUCLEOTIDES

Mainly in liver. Free purine ring is not synthesized. Directly
nucleotide is formed. There are two pathways:
1. Denovo pathway
2. Salvage pathway
Denovo Pathway
zz It is high energy consuming pathway. Around 15–20 zz From Di-Phosphate to Tri- Phosphate forms: The
steps occur in this pathway. This pathway starts from enzymes involved here have broad specificity. So, one
Ribose-5-Phosphate (source is HMP) and the first step is single enzyme for all i.e. Nucleoside Diphosphate Kinase
conversion to PRPP (Phospho-Ribosyl Pyro Phosphate),
by enzyme PRPP Synthetase. This step uses ATP and
forms AMP. This step is not committed for purine
synthesis as PRPP has other fates in body also.
zz The second step in denovo pathway is done by
enzyme PRPP Glutamyl Amido Transferase. This is the
committed and the rate limiting step of purine synthesis.
In this step –NH2 group is transferred from Glutamine to
PRPP, making PRA (Phospho Ribosyl Amine). Uses of PRPP
zz After the formation of PRPP, many steps occur and finally
there is formation of IMP i.e. Inosine Mono Phosphate. zz Purine synthesis
IMP is the first purine nucleotide to be formed. IMP can zz Pyrimidine synthesis
be converted to AMP and GMP and then Diphosphate zz Histidine synthesis
and Triphosphate forms can be synthesized. zz Tryptophan to Niacin conversion

Salvage Pathway
zz This pathway requires less energy. It occurs in RBCs,
WBCs, brain and bone marrow. “Salvage’’ means - saved
from degradation.
zz Out of the purines which are undergoing catabolism,
some of them are saved from degradation and are used
in salvage pathway to synthesize nucleotides.

Fig. 11.4: Denovo Pathway of synthesis of purine nucleotides

T
H
E
Fig. 11.6: Salvage pathway and simplified form of catabolic
pathway shown. When purine nucleotides are undergoing O
catabolism, then some of the N-bases are taken up back into the R
body and reused to make nucleotide (by adding just Sugar and Y
phosphate). This is known as salvage pathway, which is just a single
step pathway.
Fig. 11.5: Synthesis of AMP and GMP from IMP
 Hypoxanthine Guanine are taken by enzyme HGPRT
(Hypoxanthine and Guanine Phospho Ribosyl Transferase), Complete deficiency of Partial deficiency of
HGPRT
284 Phosphate and Ribose are transferred to them, making HGPRT
their nucleotides. From Hypoxanthine, IMP (Inosine Mono • Lesch Nyhansyndrome • Kelley Seegmiller syndrome
Phosphate) is formed. From Guanine, GMP (Guanosine
Mono Phosphate) is formed.
• Gout + self mutilation • Only gout
CRO BIOCHEMISTRY

• PRPP accumulation occurs to • PRPP levels are raised but

Lesch Nyhan Syndrome a level which is neurotoxic mild, which is not neurotoxic
Complete deficiency of HGPRT is Lesch Nyhan syndrome One more enzyme of salvage pathway is → APRT –
(X-linked recessive), in which patient has gout – because all Adenine Phospho Ribosyl Transferase for the conversion of
pyrimidines are going into catabolic pathway finally forming Adenine to AMP
uric acid (orange sand sodium- urate crystals in diaper).
This hyperuricemia is the result of combined mechanism
(both increased synthesis of uric acid and decreased
purine reutilization). Uric acid gets deposited in joints
and soft tissues and uric acid stones are found in kidneys.


Neuropsychiatric behavior of self mutilation also appears

which is due to accumulation of PRPP, which is neurotoxic.
Patient has tendency to bite lips and nails. PRPP is a substrate
for HG PRTase, so it accumulates.

Adenosine is the only purine nucleoside which can be

salvaged i.e. Adenosine gets converted to AMP by Adenosine
kinase.
Treatment
Allopurinol, alkalinization of urine and high fluid intake
given. Neuropsychiatric symptoms persists.

T
H
E
O
R
Y
PURINE CATABOLISM
zz Organ: Liver (mainly) 285

CHAPTER 11  CHEMISTRY AND METABOLISM OF NUCLEOTIDES

Fig. 11.7: Purine Catabolism

T
Fig. 11.8: ADA (Adenosine Deaminase Enzyme) Conversion of Adenosine to Inosine by enzyme ADA. This enzyme is very important in B- H
and T- lymphocytes and Natural Killer (NK) cells. This ADA enzyme can be measured in lab in any fluid of the body like blood, CSF, Pleural
fluid, Ascitic fluid etc. If this ADA is found to be increased in some patient, it is highly suggestive of Tuberculosis. E
O
ADA Deficiency: It leads to AR-SCID (Autosomal Recessive Severe Combined Immuno-Deficiency) as both B and T arms of R
immunity are affected. ADA degrades both adenosine and deoxyadenosine. Increased deoxyadenosine inhibits ribonucletide Y
reductase and stops production of deoxy nucleotides in lymphocytes (cell cannot divide due to lack of DNA synthesis)
 PYRIMIDINE SYNTHESIS
H igh R eturn Organ: Liver (mainly)
286 Table 1: Causes of hyperuricemia Organelle: Cytoplasm
Increased UA formation Decreased UA excretion Steps are: (Figure)
zz CPS-II → Carbamoyl Phosphate Synthetase –II:
• HGPRT deficiency • Diabetes insipidus
CRO BIOCHEMISTRY

Glutamine, 2ATP and CO2 combine to form Carbamoyl

• PRPP Synthetase over activity • Hypertension
• Hemolysis • Lactic acidosis phosphate. Then Carbamoyl phosphate gets converted
• Lymphoproliferative disease • DKA to Carbamoyl Aspartate by enzyme Aspartate Trans-
• Myeloproliferative diseases • Starvation ketosis Carbamoylase (ATC). Then after 1-2 steps Orotate/
• Exercise • Lead intoxication Orotic acid is formed.
• Obesity • Hyperparathyroidism zz Orotic acid is a kind of pyrimidine nitrogenous base.
• Purine rich diet • Hypothyroidism Now sugar and phosphate added to this base, making a
• Toxemia of pregnancy nucleotide known as OMP (Orotidine Mono Phosphate).
• Down syndrome The enzyme is OPRT – Orotate Phospho Ribosyl
• Diuretics
Transferase as this enzyme transfers phosphate and
• Many drugs
ribose from PRPP to Orotate.


Combined mechanism of hyperuricemia (increased formation of zz OMP undergoes decarboxylation to form UMP –
uric acid as well as decreased excretion): (Uridine Mono Phosphate) OPRT and Decarboxylase
zz Von Gierke’s disease
are bifunctional enzymes (two enzymatic activities on
zz Aldolase B deficiency
a single protein). This enzyme/protein is also known as
zz Shock
UMP Synthase
zz Alcohol
zz HGPRT deficiency → increased uric acid OPRT – Orotate Phospho Ribosyl Transferase (an enzyme)
zz Xanthine oxidase deficiency → decreased uric acid OMP – Orotidine Mono Phosphate (a nucleotide)
zz PRPP Synthetase deficiency → decreased uric acid

T
H
E
O
R
Y

Fig. 11.9: Pyrimidine Synthesis

 Q. Why ‘Orotate’ is used in OPRT while ‘Orotidine’ zz First purine nucleotide to be synthesized is IMP (Inosine Mono
Phosphate)
used in OMP?
T A. OPRT is an enzyme. For naming the enzyme, name zz First pyrimidine nucleotide to be synthesized is OMP (Orotidine
287
H of substrate is used. Substrate of this enzyme is Mono Phosphate)
I Pyrimidine synthesis requires Aspartate and Glutamine
Orotic acid/ Orotate. OMP is a nucleotide. For zz
N
Denovo Purine synthesis requires aspartate, glycine, gluta-

CHAPTER 11  CHEMISTRY AND METABOLISM OF NUCLEOTIDES

K naming a nucleotide, name of nucleoside is used. For zz

pyrimidines “dine” suffix is used. So it is Orotidine. mine and THF.

Rate Limiting Step of Pyrimidine Synthesis FUNCTIONS OF NUCLEOTIDES AND THEIR

zz In eukaryotes → CPS-II (Carbamoyl Phosphate Synthe-
DERIVATIVES
tase –II) zz UDP Sugars: used in synthesis of glycogen, galactose and
zz In prokaryotes → Aspartate Trans-Carbamoylase glycoproteins - serve as carrier of activated intermediates
e.g. UDP-Glucose, CDP-Choline
Orotic Aciduria zz UDP-Glucuronic acid: For conjugation reactions (like
zz Rare bilirubin), synthesis of proteoglycans
zz AR (Autosomal Recessive) zz SAM (S-Adenosyl Methionine): as methyl donor
zz Most common metabolic error in pyrimidine synthesis zz PAPS (Phospho Adenosyl Phospho Sulfate): as sul-
zz Type I – Both enzymes deficient i.e. OPRT and phate donor
Decarboxylase (it is a bifunctional enzyme)
zz ATP: as energy currency of cell
zz Type II – Only one enzyme deficient (mostly decar-
boxylase) zz Cyclic AMP and Cyclic GMP: as second messengers
zz C/F → Megaloblastic anemia (due to deficiency of zz As coenzymes: NAD, NADP, FAD, FMN, Coenzyme A
pyrimidines) as DNA synthesis is impaired
zz Growth failure, developmental delay, neurological deficit Nucleotides- Acting as Coenzymes
zz Present within 1st year of life NAD- Nicotinamide Adenine Dinucleotide
zz Treatment → Give only uridine because other two
pyrimidines can be synthesized from uridine.
zz Urea cycle not affected.
Diagnosis
zz Megaloblastic anemia with normal serum B12 and Folate
zz Confirmatory diagnosis by enzyme assays in RBCs

Other causes of raised Orotic Acid

zz Reye’s syndrome (defect in mitochondria so that
Carbamoyl phosphate is not used up)
zz OTC deficiency (Urea cycle enzyme)– distinguished by
NADP-Nicotinamide Adenine Dinucleotide Phosphate
Hyperammonemia.

Drugs which may precipitate Orotic Aciduria

zz Allopurinol (competes with orotic acid for the enzyme
OPRT)
zz 6-Azauridine (competes with enzyme decarboxylase)

H igh R eturn
zz Xanthine Oxidase – Rate limiting enzyme of purine catabolism
zz PRPP Glutamyl Amido Transferase- Rate limiting enzyme of T
purine synthesis
H
zz Parent purine nucleotide for AMP and GMP - IMP (Inosine
FMN- Flavin Mono Nucleotide E
Mono Phosphate)
O
Contd… R
Y
 FAD – Flavin Adenine Dinucleotide
Dinucleotide means two nucleotides: first nucleotide has Flavin as base and second nucleotide has Adenine as the base. So the
288 name mentions both the nitrogenous bases- Flavin and Adenine
CRO BIOCHEMISTRY

Structure of Acetyl CoA and Coenzyme A



Pearls of the Chapter

zz Nucleic acid is polymer of nucleotides
zz Nucleotide = Nitrogenous base + Sugar + Phosphate
zz Nucleoside = Nitrogenous base + Sugar
zz THYMINE is a pyrimidine nitrogenous base. THIAMINE is a vitamin – Vitamin B1. Thiamine (Vitamin B1) has a pyrimidine like ring in its
structure
zz Guanidine is a protein denaturing agent but Guanosine is a nucleoside of Guanine (i.e. Guanine + Sugar)
zz Most abundant free nucleotide in mammalian cells is ATP
zz Conjugated double bonds in the rings is responsible for absorption of UV light
zz Sugar in nucleic acids is always a pentose. This pentose can be ribose or deoxy-ribose
zz NADPH is the ultimate donor of reducing equivalents for the conversion of Ribonucleoside diphosphate to its Deoxy Ribonucleoside
Diphosphate
zz PRPP is used in purine synthesis, pyrimidine synthesis, histidine synthesis and for the conversion of tryptophan to niacin
zz Salvage pathway of purine occurs in RBCs, WBCs, brain and bone marrow
zz Adenosine is the only purine nucleoside which can be salvaged i.e. Adenosine gets converted to AMP by Adenosine kinase
zz Low Purine Diet is Milk (specially cow’s milk), Yoghurt, Cheese and Vit C.
zz Xanthine Oxidase: Rate limiting enzyme of purine catabolism
T zz PRPP Glutamyl Amido Transferase: rate limiting enzyme of Purine synthesis
H zz Parent purine nucleotide for AMP and GMP: IMP (Inosine Mono Phosphate)
E zz First purine nucleotide to be synthesized is IMP (Inosine Mono Phosphate)
O zz First pyrimidine nucleotide to be synthesized is OMP (Orotidine Mono Phosphate)
R zz Rate limiting step of pyrimidine synthesis in eukaryotes is CPS-II (Carbamoyl Phosphate Synthetase –II)
Y zz Rate limiting step of pyrimidine synthesis in prokaryotes is Aspartate Trans Carbamoylase
 Multiple Choice Questions 289

1. The enzyme deficient in Lesch Nyhan syndrome is: 11. Uric acid is converted to allantoin in: (PGMEE 2015)
 (Recent Question Jan 2018) a. Catabolism of pyrimidines
a. Adenosine Deaminase b. Catabolism of purines
b. PRPP Synthetase c. Synthesis of pyrimidines
c. HGPRTase d. Synthesis of purines
d. Xanthine Oxidase 12. Which of the following is the rate limiting enzyme in
2. C4, C5, N7 in purine ring are derived from pyrimidine synthesis? (PGMEE 2012-13)
 (PGMEE 2015) a. Asparate Transcarbamoylase
a. CO2 b. Glutamine b. Reductase
c. Asparate d. Glycine c. Dihydro-orotase
3. Nitrogen atoms in purines are derived from all d. UMP kinase
EXCEPT: 13. The purines salvage pathway is for: (PGMEE 2008)
a. Aspartate b. Glutamine a. Hypoxanthines and Xanthine
c. Glutamate d. Glycine b. Hypoxanthines and Adenine
4. Inosine is biological precursor of: (PGMEE 2013) c. Adenine and Guanine
a. Orotic acid and Uridylic acid d. Xanthine and Guanine
b. Uracil and Thymine 14. Which of the following does not contribute to the ring
c. Adenylic acid and Guanylic acid of thymine?
d. Purines and Thymine a. Aspartate b. Glutamine
5. Guanidine is a: c. THF d. Bicarbonate
a. Purine 15. A person was diagnosed with gout. You will suggest
b. Pyrimidine the patient to avoid which of the following food
c. Nucleoside product in his diet? (Recent Question 2017)
d. Denaturing agent of proteins a. Whisky and Beer
6. Which of the following statements about nucleotide b. Spinach and Mushrooms M
metabolism is NOT CORRECT? c. Meat and Fish C
a. An early step in purine biosynthesis is formation of d. All Qs
PRPP (Phospho Ribosyl Pyro Phosphate) 16. Rate limiting enzyme of purine nucleotide synthesis Ans.
b. IMP (Inosine Monophosphate) is a precursor of is : (Recent Question 2017)
both CTP and TMP a. Xanthine oxidase 1. c
c. Orotic acid is an intermediate in pyrimidine b. HGPRT 2. d
nucleotide biosynthesis c. ADA 3. c
d. Ribonucleotide reductase converts ribonucleoside d. PRPP Glutamyl Amido-Transferase 4. c
diphosphates to the corresponding deoxy-ribo- 17. Common end product of pyrimidine metabolism: 5. d
nucleoside diphosphates. a. Beta alanine 6. b
7. End product of purine catabolism in non-primates: b. Uric acid 7. b
 (PGMEE 2013) c. Urea 8. b
a. Uric acid b. Allantion d. Xanthine 9. a
c. Beta alanine d. Ammonia 18. A female child presented with growth retardation, 10. d
8. Salvage pathway of purine nucleotide synthesis is anemia, lethargy and her urine showed elevated 11. b
used by all EXCEPT: (PGMEE 2013) levels of orotic acid. Treatment given should be : 12. a
a. Leukocytes b. Liver a. Uridine b. Thymine 13. c
c. RBC d. Brain c. Cytosine d. Adenosine 14. c
9. About purine and pyrimidine nucleotides, all are 19. Orotic aciduria is due to deficiency of: (PGMEE 2013) 15. d
true EXCEPT: (PGMEE 2015) a. Decarboxylase 16. d
a. Adenosine is a nucleotide b. Isomerase 17. c
b. Adenine is present in both DNA and RNA c. Tyrosinase 18. a
c. Uracil is not found in DNA d. Homogentisate oxidase 19. a
d. Thymine is not found in RNA 20. Which of the following nucleotide bases constitute 20. b
10. TRUE about cAMP and cGMP: (PGMEE 2015) Hypoxanthine and Ribose?
a. Act on membrane receptors a. Adenosine
b. Act by post-translational modification b. Inosine
c. Second messengers c. Guanosine
d. All of the above d. Cytidine
 21. What is the other name for 6-oxopurine? 24. A child who was normal at birth, at 4 years age
a. Hypoxanthine he developed delayed motor development and
290 b. Adenine involuntary movements. CT brain was done and was
c. Guanine normal. The child has self- mutilating behavior of
d. Cytosine head banging, biting of lip and fingers and arthritis
22. Which of the following is a suicide inhibitor of and later on he dies of renal failure. Enzyme
Xanthine Oxidase? responsible for this is:
a. Allopurinol a. Phenylalanine hydroxylase
b. Azauridine b. HGPRTase
c. 5-Phosphoribosyl Pyrophosphatee (PRPP) c. Hexosaminidase A
d. 5-Fluorouracil d. Adenine deaminase
23. Hyperuricemia is/are associated with: 25. All of the following abbreviations are TRUE EXCEPT:
 (PGI May 2016)  (PGI Nov 2011)
a. HGPRT deficiency a. AMP – Adenosine monophosphate
b. HGPRT over activity b. CMP – Cytidine monophosphate
c. PRPP synthetase deficiency c. GMP – Guanosine monophosphate
d. Glucose 6- phosphatase deficiency d. TMP – Thymine monophosphate
e. Glucose-6- phosphate dehydrogenase deficiency e. UMP – Uracil monophosphate

M
C
Qs
Ans.
21. a
22. a
23. a,d
24. b
25. d,e
 Answers with Explanations
291
1. Ans. (c) HGPRTase 6. Ans. (b) IMP (Inosine Monophosphate) is a pre-
cursor of both CTP and TMP

CHAPTER 11  CHEMISTRY AND METABOLISM OF NUCLEOTIDES

[Ref: Harper 30th/e pg. 354]
•• Enzyme deficient in Lesch Nyhan syndrome is [Ref: Harper 30th/e pg. 348-353]
HGPRTase i.e. Hypoxanthine Guanine Phospho •• IMP (Inosine Monophosphate) is a precursor of both
Ribosyl Transferase. This enzyme is involved in AMP and GMP (Not CTP and TMP)
salvage pathway. Clinical features include gout & self
Mutilation behaviour. 7. Ans. (b) Allantion

2. Ans. (d) Glycine [Ref: Harper 30th/e pg. 353]

•• End product of purine catabolism in non-primates
[Ref: Harper 30th/e pg. 340]
mammals is Allantoin. But in primates it is uric acid.
•• In purines, N1 is derived from Aspartate, C2 from N10    (Refer Fig. 11.7)
Formyl Tetrahydrofolate, N3 & N9 from Glutamine,
C4, C5, N7 from Glycine, C6 from CO2, C8 from N5, N10 8. Ans. (b) Liver
Methenyl THF
[Ref: Harper 30th/e pg. 350]
   (Refer Fig. 11.1)
•• Salvage pathway of purine nucleotide synthesis
3. Ans. (c) Glutamate occurs in brain, RBCs, WBCs & bone marrow.
These cells lack the enzyme PRPP Glutamyl Amido
[Ref: Harper 30th/e pg. 340]
Transferase and depends on exogenous purine
•• Nitrogen atoms in purines are derived from amino nucleosides to form nucleotides.
acids - Aspartate, Glutamine and Glycine. But •• Liver is the major site for denovo purine nucleotide
Glutamate is not involved here. synthesis and provide purine nucleosides which
N1 from Aspartate, N3 & N9 from Glutamine, N7 from travel to brain, RBCs, WBCs and bone marrow for
Glycine salvage and for utilization by tissues incapable of
•• Carbon atoms in purines are derived from: their biosynthesis.
•• C2 from N10 Formyl Tetrahydrofolate, C4 and C5 from
Glycine, C6 from CO2, C8 from N5,N10 Methenyl THF 9. Ans. (a) Adenosine is a nucleotide
   (Refer Fig. 11.1)
[Ref: Harper 30th/e pg. 341]
4. Ans. (c) Adenylic acid and Guanylic acid •• Adenosine is a nucleoside i.e. nitrogenous base + A
sugar (not nucleotide as phosphate is not present) N
[Ref: Harper 30th/e pg. 348]
•• In DNA there four nitrogenous bases A T C G. In RNA S
•• IMP (Inosine Mono Phosphate) is the first purine there are four nitrogenous bases A U C G. Adenine is
W
nucleotide to be formed. IMP (Inosine Mono present in both DNA and RNA (option b). Uracil is not
E
Phosphate) is the parent nucleotide which synthesize found in DNA. Instead of uracil, Thymine is present
R
AMP and GMP. (Refer Fig. 11.5) in DNA (option c). Thymine is not found in RNA,
Uracil is present in RNA(option d) S
5. Ans. (d) Denaturing agent of proteins
WITH
10. Ans. (d) All of the above
[Ref: Harper 30th/e pg. 314]
[Ref: Harper 30th/e pg. 344]
E
•• Guanidine is a chemical, used for denaturing X
proteins. It resembles guanidino group of Arginine in •• cAMP and cGMP are cyclic nucleotides derived P
structure, thats why this name was given. It is neither from ATP & GTP respectrively. Kinases are usually L
a purine, nor a pyrimidine associated with them and they regulate various A
•• Guanidinium chloride is one of the strongest de- biochemical pathways by post translational N
naturants used in physiochemical studies of protein modifications. They act as intracellular second A
folding. Guanidine Hydrochloride also has the ability messengers as they can move numerous signaling T
to decrease enzyme activity (by denaturation) and informations quickly throughout the cytoplasm. G I
increase the solubility of hydrophobic molecules. protein coupled receptors are involved in cAMP& O
cGMP signaling. N
S
 11. Ans. (b) Catabolism of purines 16. Ans. (d) PRPP Glutamyl Amido-Transferase

292 [Ref: Harper 30th/e pg. 353] [Ref: Harper 30th/e pg. 348]
•• Uric acid is converted to allantoin in non primates by •• Rate limiting enzyme of purine nucleotide synthesis
enzyme uricase. This occurs in purine catabolism. is PRPP Glutamyl Amido-Transferase I. It catalyzes
•• In pyrimidine catabolism, the end products are more
CRO BIOCHEMISTRY

the conversion of PRPP to Phospho Ribosyl Amine

than one i.e. ammonia, CO2& beta-alanine. In case (PRA).Rate limiting enzyme of purine nucleotide
of thymine, it is beta amino iso butyrate (not beta catabolism is xanthine oxidase.
alanine).
17. Ans. (c) Urea
12. Ans. (a) Asparate Transcarbamoylase
[Ref: Harper 30th/e pg. 355]
[Ref: Harper 30th/e pg. 353]
The end products of pyrimidine catabolism are NH3, CO2
•• Rate limiting step of pyrimidine synthesis: In & Beta-Alanine. For Thymine, they are NH3, CO2 & beta
eukaryotes → CPS-II (Carbamoyl Phosphate amino iso butyrate. NH3 is the common end product,
Synthetase –II). In prokaryotes → Aspartate which can form Urea. Also CO2 and bicarbonate is the


Transcarbamoylase common end product of pyrimidines.

13. Ans. (c) Adenine and Guanine 18. Ans. (a) Uridine
[Ref: Harper 30th/e pg. 350] [Ref: Harper 30th/e pg. 356]
•• The purine salvage pathway is for Adenine, Guanine Diagnosis is Orotic Aciduria: Orotic Aciduria is a rare,
and Hypoxanthine. Adenine is converted to AMP autosomal recessive disorder. It is the most common
using enzyme Adenine Phospho Ribosyl Transferase. metabolic error in pyrimidine synthesis. Treatment is
Guanine converts to GMP using Hypoxanthine administration of uridine (results in the improvement
Guanine Phospho Ribosyl Transferase and of anemia and decreased excretion of Orotate). All
Hypoxanthine uses Hypoxanthine Guanine Phospho pyrimidines are not getting formed in these patients but
Ribosyl Transferase and gets converted to IMP. only uridine is given as other pyrimidines (C & T) will
be synthesized from uridine because those enzymes are
14. Ans. (c) THF- Tetra Hydro Folate not deficient.
[Ref: Harper 30th/e pg. 340] 19. Ans. (a) Decarboxylase
•• In Thymine (Pyrimidine) ring, sources of N & C are:
[Ref: Harper 30th/e pg. 356]
N3 from Glutamine, C2 from CO2 (or Bicarbonate), N1,
C4, C5, C6 - from Aspartate. THF contributes a Carbon •• There are two types of Orotic acidurias:
A which is out of the ring of Thymine. (Refer Fig. 11.2) ƒƒ Type I: Both enzymes deficient i.e. OPRT
N (Orotate Phosphoribosyl Transferase) and OMP
S 15. Ans. (d) ALL Decarboxylase (It is a bifunctional enzyme)
W ƒƒ Type II: Only one enzyme deficient (mostly OMP
[Ref: Lippincott 4th/e pg 301]
E Decarboxylase)
R •• Foods rich in Purine are :
1. Vegetarian-Spinach, Peas, Cauliflower, Mushrooms 20. Ans. (b) Inosine
S 2. Non-vegetarian- Meat, Liver, Fish (tuna).
[Ref: Lehninger 7th/e p282, 899]
WITH Also alcohol & heavy exercise should be avoided in
patients of gout, because they causes Lactic acidosis & •• Catabolism of AMP leads to the formation of Inosine
E Hyperuricemia. by Adenosine deaminase. From Inosine, sugar
X •• Foods low in purine diet are milk, yoghurt, cheese, Ribose is removed and results in the formation of
P Vit C. All these help in the excretion of uric acid from Hypoxanthine, Nitrogenous Base (Inosine = Ribose +
L kidneys. Hypoxanthine)
A
N
A
T
I
O
N
S
21. Ans. (a) Hypoxanthine which inhibits Xanthine Oxidase. So this is suicidal
inhibition. Allopurinol is used for the treatment of
[Ref: Harper 30th/e pg. 342] gout and hyperuricemia. 293
23. Ans. (a); (d)

CHAPTER 11  CHEMISTRY AND METABOLISM OF NUCLEOTIDES

[Ref: Harper 30th /e pg 354, 355]
•• HGPRT deficiency is Lysch Nyhan syndrome in
which patient has hyperuricemia and gout. Glucose
6- Phosphatase deficiency is Von Gierke’s disease in
which hyperuricemia and lactic acidosis occur. Over
activity of enzyme PRPP Synthetase (not deficiency)
leads to hyperuricemia.

24. Ans. (b) HGPRTase

[Ref: Harper 30th /e pg 354]

•• This is Lesch Nyhan syndrome which is characterized
by hyperuricemia and self mutilation. Enzyme
deficient is Hypoxanthine Guanine Phospho Ribosyl
Transferase (HGPRTase).

25. Ans. (d); (e)

[Ref: Harper 30th /e pg 350]

22. Ans. (a) Allopurinol
•• For the full form of a nucleotide, use the name of
[Ref: Harper 30th /e pg 355] nucleoside (not nitrogenous base). Nucleosides
of purines are Adenosine and Guanosine.
•• Allopurinol, a structural analogue of hypoxanthine,
Nucleosides of pyrimidines are Cytidine, Uridine and
competitively inhibits Xanthine Oxidase activity.
Thymidine. TMP has full abbreviation as Thymidine
Xanthine Oxidase converts Allopurinol to Oxypurinol,
Monophosphate (not Thymine) and UMP is Uridine
Monophosphate (not Uracil). So, (option d & e) are
not correct.

A
N
S
W
E
R
S
WITH

E
X
P
L
A
N
A
T
I
O
N
S
 12 Basics in Genetics

Overview of Chapter BASIC TERMINOLOGY

•• Basic Terminology zz Genes → A stretch of DNA that determines a particular
•• Nucleic Acids trait or produces a functional product (RNA or Protein
or both). Each chromosome contains many genes.
•• DNA Markers
A specific gene is located at same location on same
•• Genetic disorders chromosome in all individuals. E.g. Gene for eye color.
•• DNA Recombination There are two sets of chromosomes called homologous
pairs.

zz Allele → Alternative forms of gene, present at same place  Dominant allele can express in homozygous as well
on homologous chromosomes. as heterozygous state.
 For each gene, there can be many different alleles.  Recessive allele can express only in homozygous
Each individual carries only 2 alleles of a particular state.
gene.  Codominance: Two different alleles present and
both are able to express. E.g. blood group genes.
zz Individuals can be homozygous or heterozygous for a
gene:
 Homozygous Dominant → e.g. AA i.e. 2 copies of
same dominant allele.
 Homozygous Recessive → e.g. aa i.e. 2 copies of
same recessive allele.
 Heterozygous → e.g. Aa i.e. 2 different alleles of a
gene
zz Polymorphism → Occurrence of different alleles at the
same time in the genome of a population or species.
zz Haploid (n) → One set of chromosomes i.e. 23 in humans
e.g. gametes. It consists of approx. 3 billion base pairs.
 E.g. Gene for eye color has various alleles: allele for zz Diploid(2n)→Two sets of chromosomes i.e. 46 e.g. Somatic
black eye color, allele for blue eye color, allele for cells have 46 chromosomes or 23 pairs of chromosomes.
brown eye color. (23 from mother and 23 from father). Diploid genome
contains two alleles of each gene.
 295

CHAPTER 12  BASICS IN GENETICS

23 Pair of Chromosomes

Fig. 12.1: Somatic and germ cell

T
H
E
O
R
Y
296
CRO BIOCHEMISTRY


zz Genotype → Genetic make-up of an individual. (b) Regulated genes: expressed only in special
zz Phenotype → Physical appearance of an individual. circumstances. E.g. genes which synthesize enzymes
Genotype determines the phenotype. of gluconeogenesis are not required in normal
zz Euploidy → When a cell has a multiple of haploid situation (fed state). So these genes are inhibited in a
number of chromosomes normal situation. But if required (e.g. during fasting/
zz Aneuploidy → When a cell’s chromosomes are not in starvation), these genes are activated.
multiple of n (abnormality in number of chromosomes) These genes may be expressed in all cells or only in
zz Loss of function mutation → When there occurs few cells where this role is required. E.g. genes for
underactivity of a gene due to mutation. gluconeogenesis enzymes are only expressed in liver and
zz Gain of function mutation → When there occurs over kidney, whenever required.
activity of a gene due to mutation. zz Earlier One Gene One Protein Theory was discovered.
zz Pleitropy → When one gene influences more than one But later they found that genes are 20,000 to 25,000 only
trait i.e. more than one organ affected due to defect in but proteins are more than 2.5 lakh in human body. This
one particular gene. is explained by exceptions in ‘One Gene One Protein
zz Gene expression refers to the way how genes express Theory’- mainly Alternate RNA Processing. They are:
themselves to do the desired work in the cell. Gene 1. Alternate Splicing
expresses by forming RNA and then forming proteins. So, 2. RNA Editing or Chemical Modification of RNA.
Gene expression = Transcription + Translation 3. Alternate Promoter Utilization
zz Transcription is formation of RNA from DNA. 4. Alternative Polyadenylation
zz Translation is formation of proteins from RNA.
zz Regulation of Gene Expression Basics: H igh R eturn
Regulation of gene expression means the way products zz Average size of a gene – 27,000 bp
of a gene are controlled in kind, quantity or in location. zz Protein coding genes – 19000 to 20000 only (earlier thought
In prokaryotes, transcription is the main site of regulation to be 1 lakh)
of gene expression. zz Percentage of exons = 1-2 % of human genome
 But in eukaryotes, in addition to transcription, zz Percentage of Adenine in human genome = 54 % (So this is the
regulation of gene expression also occurs at the level of most abundant base)
post transcription and post translation (very complex in zz Longest/ largest gene = DMD gene, which codes for muscle
Eukaryotes) protein Dystrophin
T Not all genes are regulated. There are actually two types zz Largest protein – Titin
H of genes: zz Dystrophin is not the largest protein (DMD gene is largest)
E (a) Constitutive/housekeeping genes: always active – because the mRNA formed from this gene is smaller in size.
O zz Most abundant proteins in mammals – Collagen
they form products (proteins/ enzymes) required
zz Number of proteins in human body – more than 2.5 lakh
R for basic cell function, which are needed all the time
zz Number of SNPs in human genome = 3 million (106)
Y in the cells. These genes are expressed at a constant
zz Maximum number of genes is present for – Olfactory receptors
rate.
Barr Bodies Difference between DNA and RNA
zz A small dense peri-nuclear structure in cells of females
DNA RNA 297
which is condensed inactive X-Chromosome
zz Seen during interphase • Double stranded • Single stranded
zz Number of Barr bodies = Number of X- chromosome – 1
• So Purines always equals • As it is single stranded, so

CHAPTER 12  BASICS IN GENETICS

zz Lyon’s Hypothesis states that only one X- chromosome is
active in an individual, other is inactive. This inactivation Pyrimidines Purines are not necessarily
is because of Xist gene which causes DNA methylation equal to Pyrimidines
and inactivation • Adenine, Thymine, • Adenine, Uracil, Cytosine and
zz This inactivation occurs at the level of blastocyst Cytosine and Guanine Guanine bases (AUCG) are
zz This inactivation is equal for maternally or paternally bases (ATCG) present present
derived X-chromosome
• Sugar is Deoxyribose • Sugar is Ribose

A dditional E dge
Q. Why T (Thymine) instead of U (Uracil) in DNA?
A. This is to prevent mutations. Cytosine gets converted to
uracil very spontaneously in DNA, by enzyme cytosine
deaminase. But uracil being not present in DNA, is
quickly removed and this defect repaired. So mutation
rate is decreased.

NUCLEIC ACIDS
DNA and RNA are two nucleic acids found in cells. These are
polymers of nucleotides.
DNA is present in nucleus of eukaryotes and also in
mitochondria. Prokaryotes contain single chromosome (in
non membrane bound region known as nucleoid) but may
also contain extra chromosomal material in the form of
plasmids. This plasmid can replicate which may or may not
be synchronized with chromosomal division.
Nucleosomes = DNA + Proteins
DNA Structure
zz Double stranded
zz Helical (right handed)
zz The two strands are Antiparallel – means one strand is in
5’ → 3’ direction and the other is in 3’ → 5’ direction. Or
5’ end of one strand is paired with 3’ end of other.

Types of DNA
B-DNA A-DNA Z-DNA
Base pairs/turn 10 11 12 T
Right/left Right Right Left (plus zigzag sugar-
H
handed phosphate backbone) E
O
Function Major Dehydrated Regulation of
R
DNA in DNA or in gene expression,
cells DNA-RNA particularly in GC Y
Fig. 12.2: Nucleosomes which resemble beads on a string. Beads
regions sequence (alternating are DNA wrapped around histones and string is linker DNA in
purine-pyrimidine) between two beads
 Histone octamer contains H2A, H2B, H3 and H4 histone Facultative Heterochromatin
proteins. Linker/ spacer DNA is the DNA present in between Heterochromatin – means transcriptionally inactive
298 two histone octamers. It is associated with linker histones i.e.
zz
zz Facultative means this chromatin has the ability to
H1 and H5. become transcriptionally active again
DNA is negatively charged due to phosphates. Histones zz This may be packaged as euchromatin in another cell
are proteins which are rich in basic amino acids (lysine,
CRO BIOCHEMISTRY

zz An example of facultative heterochromatin is X

arginine). These amino acids have positive charge so histone chromosome inactivation in female mammals: one X
proteins have positive charge. Due to opposite charge on chromosome is packaged as facultative heterochromatin
DNA and histone proteins, they get tightly bound forming and silenced, while the other X chromosome is packaged
nucleosomes. This binding between DNA and protein is as euchromatin and expressed
influenced during regulation of gene expression. If DNA
is tightly bound to histone then this piece of DNA is not
Facultative Constitutive heterochromatin
free or transcriptionally inactive. If DNA is separated from heterochromatin
histone then it is free for transcription to occur on it so it is
transcriptionally active. • Transcriptionally inactive, • Repetitive
PTMs of Histones: These are reversible covalent but can be activated • Always inactive


modifications. N-terminal of histone proteins is modified by • Provides structural role

• Usually in centromere &
adding or removing a group. This helps in regulation of gene
telomere
expression.

A dditional E dge Acetylation and phosphorylation of histones lead to increased

Various ways of histone modifications are: euchromatin formation → gene activation.
� Acetylation � Phosphorylation
� Methylation � ADP Ribosylation The interconversion of Euchromatin and Heterochroma-
� Mono-ubiquitylation � Sumoylation tin is known as Chromatin Remodelling.

Acetylation and Phosphorylation gives a negative charge

to histones, which are normally positively charged. So now
negative charge on histone as well as on DNA makes them
repel each other. So this DNA is now separated from histones,
making this DNA transcriptionally active now.

H igh R eturn
zz So Acetylation and Phosphorylation of histones leads to gene
activation
zz Similarly Deacetylation and Dephosphorylation of histones
leads to gene inactivation Bonds in Protein –DNA Interaction
Protein and DNA are bound by weak bonds (like hydrogen,
Chromatin ionic, vanderwaals) and never by covalent bond (strong
bonds). Because regulation of gene expression uses DNA
DNA is present in compact form along with histones and protein separation to make the gene active, if it is bound
this is known as chromatin, which exists only in eukaryotes. by covalent bond, then DNA can never be separated from
During mitosis, it is heavily condensed. But during interphase, histones so it can never be activated. As there are many genes
chromatin is less condensed. kept in cells whose function is normally not required (so they
are kept in inactive state) but if needed then they can be used,
Heterochromatin v/s Euchromatin by gene activation.
Typically, interphase chromatin is named as either
Euchromatin or Heterochromatin, depending on its level of Bonds in DNA
compaction. Euchromatin has a less compact structure, and DNA is a polymer of deoxy-ribonucleoside monophosphates.
T
heterochromatin is more compact. The bonds present are hydrogen bonds, 3’ 5’ phosphodiester
H bonds and beta-N-glycosidic bonds.
E
O Euchromatin Heterochromatin
R • Loose DNA • Tightly bound DNA
Y • Genes are active • Genes are inactive
 299

CHAPTER 12  BASICS IN GENETICS

Fig. 12.3: Bonds in DNA ‘N’ stands for nitrogenous base. S and P stands for sugar and phosphate respectively. Backbone is made up of
sugar and phosphate. Nitrogenous base protrudes out from the backbone. H- bond is a weak bond, so it is relatively easy to separate
the two strands, as the two strands are joined by these H-bonds present between the N-bases. Bond between N-base and sugar is beta
N-glycosidic bond. 3’5’ phosphodiester bond is present between sugar and phosphate

3’ 5’ Phosphodiester Bond

Fig. 12.4: 3’5’ Phosphodiester Bond

3’5’ phosphodiester bond is formed between the 3’ –OH Q. If at 5’ end Adenine is present and at 3’ end Guanine
of previous nucleotide with the 5’ phosphate of the incoming is present. Which out of the following is correct?
nucleotide. So direction of synthesis is from 5’ to 3’ direction. T a. 5’ AG 3’   b. 3’ GA 5’  c. Both
zz At 5’ end, there is always a free phosphate present H A. (c) Both
zz At 3’ end, a free –OH group is always present I As direction is written in the options, both are correct.
N If we are mentioning the direction, then we can write
zz New nucleotide is added at the 3’ end K T
either way. But information should be correct. 3’AG 5’
zz We always read and write in 5’ → 3’ direction is not correct.
H
zz Synthesis always occurs in 5’ → 3’ direction Q. If at 5’ end Adenine is present and at 3’ end Guanine E
is present. Which out of the following is correct? O
a. AG     b. GA     c. Both R
A. (a) AG Y
Here direction is not given in the options. Whenever
direction is not written that means left side is 5’ and
right side is 3’. So 5’AG3’ or AG is correct. But 5’ GA 3’
(option B) is wrong.
 Nucleases DNA MARKERS
Enzymes which breaks down nucleic acid. They break
300 the phosphodiester bonds present between the sugar Polymorphisms in DNA can be used as DNA markers.
and phosphate. They are of two types: Exonucleases and
Endonucleases. 1. SNPs (Single Nucleotide Polymorphism)
CRO BIOCHEMISTRY

Most common type of polymorphism. Around 10 million

Exonuclease SNPs are found in humans. A single nucleotide at a particular
Cut the nucleic acid from the sides either from 5’ or from 3’ end location is variable in different population (Polymorphism
1. 5’ → 3’ Exonuclease (starts cutting from 5’ end and means variability in population).
moves from 5’ to 3’ direction)
2. 3’ → 5’ Exonuclease (starts cutting from 3’ end and
moves from 3’ to 5’ direction)

Endonuclease/Exci-nuclease
They can cut anywhere in between . One of the category


of endonuclease cut the nucleic acid at a specific site i.e.

Restriction endonuclease. They cut at the palindromes (same
sequence on opposite strands). Restriction endonucleases
are also known as Molecular Scissors.

H igh R eturn
Q. Which of the following is a Palindrome?
a. GGCC b. GACC c. TAAT 2. Repeat Length Polymorphism
A. (a) GGCC Tandem repeats of a particular length of DNA occurs and the
5' G G C C 3' given DNA strand
number of these repeats are variable in different population.
3' C C G G 5' Complementary DNA strand
1. STR: Short tandem repeats- also known as Micro
Read both strands in 5' → 3' direction
satellites. Repeat size is 2 – 6 base pairs.
So upper strand is G G C C
Lower strand is G G C C (Read in 5' → 3' direction)
2. VNTR: Variable number tandem repeats- also known
So palindrome is same sequence on both strands as Mini satellites. Repeat size is 15- 70 base pairs.

CG- Islands/ CG-Sites

zz CpG or CG islands have >50% GC content and >200 base
pairs
zz In mammals, 70-80 % of CpG cytosines are methylated

zz Majority of gene promotors have high CG content

zz So regulation of gene expression can be done
T
zz In non-hematopoietic cells, globin chain is not required.
H
So DNA methylation is done to inhibit transcription of
E globin chain genes
O zz In hematopoietic cells, these genes are hypomethylated
R
Y
3. RFLP (Restriction Fragment Length Polymorphism) (Discussed in Techniques)
GENETIC DISORDERS 301

CHAPTER 12  BASICS IN GENETICS

Chromosomal Disorders (b) Monosomy
Numerical Defects in Chromosomes
zz Known as Aneuploidy (chromosomes are not in
multiples of n)
Reasons of numerical defects:
1. Anaphase Lag → results in a daughter cell deficient of
one chromatid i.e. n-1
2. Meiotic Non-Dysfunction →
 Mostly occurs at Meiosis-I
 There is unequal distribution of chromosomes
H igh R eturn
zz Most dangerous numerical defect of chromosomes –
Autosomal monosomy (lethal)
zz Most common Trisomy – Trisomy 16 (lethal)
zz Only Monosomy in humans – Turner Syndrome (X0)
zz Most common chromosomal disorder in humans – Down
Syndrome
zz Most common inheritable cause of mental retardation – Down
Syndrome
zz Diseases due to increase maternal age – Down Syndrome,
(a) Trisomy Meiotic Non- Dysjunction.
zz Diseases due to increased paternal age – Marfan Syndrome,
Brittle Bone disease, Neurofibromatosis, Achondroplasia
zz Disease which can occur due to increased age of either parent
–Klinefelter Syndrome
zz Most common cause of primary amenorrhoea – Turner
Syndrome
zz Most common cause of secondary amenorrhoea – Pregnancy T
zz Most common genetic cause of infertility – Klinefeltor H
Syndrome
E
Structural defects in Chromosomes O
(a) Structural Chromosomal Mutations (Macro-alterations) or R
Y
Large Scale Mutations
Amount of genetic material is less or more or there is change
in position of genes.
 1. Insertion Isochromosomes

302 2. Duplication
CRO BIOCHEMISTRY


5. 
Translocation: Genetic material exchanged between 2
different chromosomes.
3. Deletion

4. Inversion
Robertsonian Translocation

T
H
E
O
R
Y
(b) Gene Mutations (Micro-alterations) or Small Scale Point Mutations
Mutations Transition and Transversion
303

CHAPTER 12  BASICS IN GENETICS

Three types of point mutations are – silent, missense and non sense mutation.

Insertion
A dditional E dge
zz Frameshift mutation: Addition or deletion of one or two
nucleotides alter the reading frame

Deletion
T
H
zz Splice site mutation: This can interfere in the removal of E
introns from the primary transcript. O
R
Y
 Classical Mendelian Inheritance Disorders X-Linked Disorders
No male to male transmission
304 Autosomal Dominant (AD) zz

(1) X-Linked Recessive (XR)

zz Most common sex linked pattern of inheritance (out of
CRO BIOCHEMISTRY

XD, XR & Y- linked inheritance)

zz X- linked genes usually code for enzymes
zz Males are usually affected
zz Transmitted through unaffected females
zz 100% incidence of affected sons from an affected mother
suggests XR inheritance.
(a) If Carrier/Heterozygous mother → 50% sons are affected
and 50% daughters are carriers.


zz Both male and female equally affected in autosomal

disorders
zz One parent of the patient is always affected
zz There is atleast one affected member in each generation;
known as Vertical Inheritance
zz Usually involve structural proteins
zz Can arise from fresh mutations
zz Transmission stops if a generation arises in which no one
is affected
zz Majority of individuals have loss of function mutations
zz These disorders have incomplete penetrance i.e. all If Affected/Homozygous mother → 100% sons are
(b) 
individuals with defective gene do not have the disease affected and 100% daughters are carriers.
as it also depends on environmental factors
zz These disorders have variable expressivity i.e. some
patients have mild disease, others have severe
Autosomal Reccesive (AR)

(c) If Affected Father → All daughters are carriers and All
sons are normal.

zz Usually more severe than AD disorders

zz Usually involve genes coding for enzymes
zz Patients often present in childhood as both genes are
defective (homozygous state) and disease has complete
T penetrance
H zz Increased risk in consanguineous marriages
E zz The trait can skip generations
O zz Parents are usually unaffected and siblings have high
R chances to be affected; known as Horizontal Inheritance.
Y
(2) X-linked Dominant (XD)
Autosomal Dominant Autosomal Recessive disorders
zz Males as well as females can be affected but often with an disorders (AD) (AR)
excess of females 305
zz Females are less severely affected than males • Albinism
zz Mother transmits to 50% daughters and 50% sons
• Phenylketonuria
• Wilson's disease
Father transmits to all daughters and no sons

CHAPTER 12  BASICS IN GENETICS

zz
• Hemochromatosis
zz Usually involve structural proteins • AR Polycystic Kidney disease
(a) If mother affected: • Orotic Aciduria
• Glycogen Storage Diseases
• Mucopolysaccharidosis (EXCEPT
Hurler’s)
X-Linked Dominant X- Linked recessive trait (XR)
Trait (XD)
• Vitamin D • Color blindness
resistant Rickets / • Duchenne’s Muscular Dystrophy
Hypophosphatemic • Becker’s Muscular Dystrophy
Rickets • Glucose-6-Phosphate
• Charcot: Marie Tooth Dehydrogenase deficiency(G6PD)
disease (Hereditary • Testicular Feminization
Sensory and Motor • Fragile X-Syndrome
(b) If father affected: Neuropathy) • Hunter’s Syndrome
• Incontinentia Pigmenti • Fabry’s disease
• Ornithine Transcarboxylase
deficiency
• Lesch Nyhan syndrome
• Hemophilia A and B
• Agammaglobulinemia
# Alport Syndrome has 3 patterns of inheritance → XR, AR, XD
(80% mutations are X-linked)

Non-Mendelian Inheritance
(a) Epigenetics and Genomic Imprinting
Epigenetics
zz Epigenetics is information in the genome which alter the
Y- Linked Inheritance pattern of gene expression. But it is not change in DNA
zz Also called Holandric inheritance code. This is chemical modification of DNA or proteins
zz Only males affected zz These changes are transferred to next generation
zz Male to male transmission zz Unlike mutations, these are reversible changes
zz E.g. of trait – hair on pinna, webbed toes zz Can lead to gene activation or inhibition
Table: Examples of Classical Mendelian Inheritance Disorders Epigenetic Changes are:
1. DNA Methylation of cytosine of CG sites: By enzyme –
Autosomal Dominant Autosomal Recessive disorders DNA Methyl Transferases (DNMTs)
disorders (AD) (AR) 2. PTMs of histones (Acetylation or Deacetylation):
• Familial • Ataxia Telangiectasia Enzymes are HAT – Histone Acetyl Transferase for
Hypercholesterolemia • Cystic fibrosis histone acetylation, HDAC – Histone Deacetylases – for
• Huntington's disease • Familial Hypertrophic histone deacetylation. It occurs mostly on Lysine residue
• Lactose Intolerance Cardiomyopathy on proteins. Less common is histone methylation, which
• Marfan Syndrome • Sickle Cell disease can be done by Histone Methyl Transferases. T
• Myotonic Dystrophy • Thalassemia 3. Changes in higher order of chromosome structure like H
• AD Polycystic Kidney • Maple Syrup Urine disease Heterochromatin or Euchromatin E
Disease • Homocystinuria O
• Porphyria Variegate • Alkaptonuria R
Y
Contd…
 Genomic Imprinting Causing Diseases
(1) Prader Willi Syndrome (PWS)
306
There occurs a particular gene on chromosome 15, for which
maternal allele is normally imprinted and paternal copy is
active. But if paternal allele is deleted, then it leads to PWS.
CRO BIOCHEMISTRY

Causes of PWS
1. Deletion of paternal copy of allele (most common)
Fig. 12.5: CG site shown. C and G present on same strand of DNA 2. Maternal uniparental disomy → both copies of maternal
and this C of CG site can be methylated to inactivate the gene chromosome are present. Maternal alleles are already
imprinted in this case, so not expressed.

(2) Angelman Syndrome

There occurs another gene on chromosome, for which
normally paternal copy of allele is imprinted and maternal


copy is active. But if this maternal allele is deleted, then it

leads to this syndrome.
Causes of Angelman syndrome
1. Deletion of maternal copy of allele (most common)
Epigenetic Changes Causing Diseases 2. Paternal uniparental disomy
zz Cancer: promoter hypermethylation of tumor suppres- zz Prader Will syndrome → Paternal deleted (maternal
sor genes can lead to cancer normally imprinted)
zz Ageing zz Angelman syndrome → Maternal deleted (paternal
zz Fragile X-syndrome mormally imprinted)
Uses of Epigenetic changes
zz Regulation of gene expression (b) Mitochondrial DNA or mtDNA
zz Defence mechanism Circular Double Stranded (like prokaryotic DNA)
zz X-chromosome inactivation/Facultative heterochroma-
tin
zz Genomic Imprinting

Genomic Imprinting
zz An epigenetic phenomenon which helps in regulation of
gene expression
zz ‘Imprinting’ means inhibited. This is inhibition at the
level of transcription i.e. gene is inhibited or silenced
zz Results in mono-allelic expression. Allele from one
parent is expressed (either maternal or paternal)
zz Has high rate of mutations as compared to nuclear DNA
zz It is a normal phenomenon, occurs in <1% genes.
because:
zz Occurs before fertilization (in egg/sperm). It is removed
 No introns (introns prevent mutations)
at fertilization and re-established during embryogenesis
 No repair enzymes
zz Paternal Imprinting: means paternal allele is inhibited
(only maternal allele is functioning)  Exposed to OFR (Oxygen Free Radicals) from ETC
zz Maternal Imprinting: means maternal allele is inhibited zz Contain around 16,000 base pairs (which is around 1% of
(only paternal allele is functioning) total cell DNA)
zz Contain 37 genes coding for 2rRNA, 22 tRNA and 13
Mechanisms of Genomic Imprinting: protein subunits of respiratory chain
zz DNA Methylation at CG sites (Most common) zz There are total 67 proteins of respiratory chain, out of
T zz Histone deacetylation Cause gene which 13 are from mitochondrial DNA. So, mitochondrial
H zz Histone methylation silencing DNA encodes for approximately 19% (13/67) proteins of
E Both PTMs of histones and DNA methylation can be ETC
O reversed. zz Mitochondria contain their own ribosome (55S) and
R their own unique circular ds DNA. Most mitochondrial
Y proteins are encoded by nuclear DNA, synthesized
completely in cytoplasm and targeted to mitochondria.
zz Mitochondria has unique genetic code
 Codon Universal Code Mitochondrial Code (c) Trinucleotide Repeat Expansion
A sequence of three bases that is repeated in tandem will
UGA Stop Tryptophan
become amplified in number, so that too many copies of the
307
AGA & AGG Isoleucine Stop triplet occur.
AUA Arginine Methionine

CHAPTER 12  BASICS IN GENETICS

Diseases related to mutations in mt DNA
Mostly affect oxidative phosphorylation and thus there
is decreased ATP in cells. Mostly eyes are affected. Lactic
acidosis occurs because of anaerobic glycolysis in cells.
zz MELAS: Mitochondrial Encephalopathy, Lactic Acidosis,
and Stroke like episodes. Diseases due to Trinucleotide Repeat Expansion
zz Kearns-Sayre syndrome: There is a large deletion of mt Usually have neurodegenerative manifestations
DNA. Patient has ophthalmoplegia and retinopathy.
zz Leber hereditary optic neuropathy: Optic nerve affect-
Disease Trinucleotide Repeat
ed leading to vision loss. Huntington disease CAG repeats
zz Leigh syndrome: Brain affected leading to developmen- Fragile X syndrome CCG or CGG
tal delay, muscle weakness.
Friedrich Ataxia GAA
Mitochondrial Inheritance Myotonic Dystrophy CTG
zz Mitochondria are transmitted only through mother. So,
this inheritance is also called Maternal Inheritance zz Huntington disease is an example of gain of function
zz Mother transmit disease to all offspring mutation. There is formation of excessive amount of
protein, Huntington which is neurotoxic.
zz Fathers do not transmit disease
zz Fragile X Syndrome
zz An individual is affected only when mother is affected
 FMR-1 gene (Fragile X mental retardation gene) is
zz This inheritance shows variable expressivity within
present on long arm of X-chromosome, which codes for
population or individual (in different cells of same
individual) Fragile X-mental retardation protein which is required
normally for the development of the connection between
neurons.

zz The number of repeats increases from generation to (d) Germline Mosaicism/Gonadal Mosaicism
generation until an affected individual is produced.
zz Parents do not have any defective gene but child has
This process is called Genetic Anticipation i.e. clinical
defective genes
features increase in severity in generations. T
zz This is a post zygotic mutation, affecting only germ
Fragile X-Syndrome Characteristics H
cells (Somatic cells are normal), so mutation is not
zz Fragile X syndrome is second most common cause of phenotypically expressed in individual E
inherited mental retardation (first is Down Syndrome). O
zz This mutation is significant enough that it can be now
zz Patients have mild to moderate mental retardation, long passed on to the next generation. R
narrow face, large protuberant ears, macroorchidism Y
(enlarged testicles).
 DNA RECOMBINATION Types of DNA Recombination
308 Evolution could not happen without genetic recombination. 1. Homologous Recombination (HR)
DNA recombination is the exchange of DNA strands to When exchange of genetic information occurs between
produce new nucleotide sequence arrangements. similar or homologous chromosomes. It occurs during
It occurs by breaking and rejoining DNA segments. This meiosis and requires alignment of homologous metaphase
CRO BIOCHEMISTRY

is responsible for genetic diversity and for maintainence of chromosomes. Usually equal crossing over occurs but
genomic integrity. sometimes unequal can also occur but its rare.

2. 
Non-Homologous Recombination or Site specific recombination (SSR)
It is rarer than homologous recombination. In this, specific recognition protein is used. It is error prone but preferred over ‘no’
repair in many circumstances.


Fig. 12.6: Homologous v/s Non-Homologous Recombination [E. coli]

3. Transposition
zz Transposition is a highly specialized form of
recombination in which a segment of DNA moves from
one location to another, either on the same chromosome
or a different chromosome
zz Transposable elements are also known as jumping
genes as these are DNA sequences that can move from
one chromosome locus to another. Two types:
1.  Transposons move by means of a DNA intermediate.

T
H
E
O
R
Y
 Retrotransposons move by means of an RNA
A dditional E dge
2. 
intermediate.
Viral integration in DNA of bacterial host is a form of recombination. 309
zz If bacteriophage has homologous DNA sequence to host
sequence, then it occurs like homologous recombination.
zz If no homologous sequence, then bacteriophage synthesize

CHAPTER 12  BASICS IN GENETICS

proteins which bind specific sites on bacterial chromosome
and it occurs like non homologous end joining. Integration
occurs at a particular site and is said to be site specific.

T
H
E
O
R
Y
310 Multiple Choice Questions
1. Which model of DNA was discovered by Watson and 11. Most lethal karyotype is:
Crick? (PGMEE 2015, 2012-13) a. 45, Y0 b. 45, X0
a. A-DNA b. B-DNA c. 47, XXY d. 48, XYYY
c. C-DNA d. Z-DNA 12. Karyotype in klinefelter’s syndrome is:
2. About DNA, true is: (PGMEE 2016-17)  (PGMEE 2011, 2013)
a. Two strands are held together by peptide bonds a. 47 XXY b. 45XO
b. Non-covalent bonds in sugar-phosphate backbone c. 46XXY d. 45XXX
c. Most common DNA is Z- DNA 13. The chromosomal karyotype in Patau syndrome is:
d. Melting point of DNA is closely related to Cytosine  (PGMEE 2013-14)
Guanine content of DNA a. 46XX/47XX, + 18 b. 47XX +21
3. Number of barr bodies in XXY males is?  c. 45XX, der (14:21) d. 47XX, +13
 (PGMEE 2012) 14. Pedigree Chart- (PGMEE 2013-14)
a. 1 b. 2 a. Used for growth monitoring
c. 3 d. None b. To assess side effect during chemotherapy
4. Denaturation of DNA breaks: (Recent Question 2018) c. To assess developmental delay in infant
a. Hydrogen bonds d. Used to see genetic transmission
b. 3’ 5’ Phosphodiester bonds 15. Male to male transmission is seen in -
c. Beta- N Glycosidic bonds  (PGMEE 2012-13)
d. O-Glycosidic bonds a. Autosomal dominant disease
5. True about Mitochondrial DNA: b. X- linked recessive
a. Single stranded c. X- linked dominant
b. Linear, double stranded d. Mitochondrial disease
c. Circular, double stranded 16. Inheritance of Huntington’s chorea is:
 (PGMEE 2012-13)
M d. Can be all depending on cell type
a. AD b. AR
C 6. When a dideoxy nucleotide is added in PCR, what will
c. XR d. XD
Qs happen? (Recent Question 2018)
17. Which of the following is an autosomal dominant
Ans. a. DNA synthesis stops
metabolic disorder? (PGMEE 2016)
b. Fluorescence occurs
1. b a. Hereditary hypercholesterolemia
c. DNA synthesis is faster
2. d b. Tay-Sachs disease
d. DNA synthesis is slower
3. a c. Tyrosenemia
7. Which of the following represents the most
4. a d. Gaucher’s disease
characteristic function of type II restriction enzymes:
5. c 18. Mother is affected, father is not. Disease is autosomal
 (PGMEE 2012)
6. a dominant, what is the chance in children?
a. Prevent folding of proteins  (PGMEE 2012-13)
7. d
b. Remove formed DNA a. 50% affected b. 25% affected
8. b
c. Prevent super-coiling c. 75% affected d. All affected
9. d
10. a
d. Cut DNA at palindromic sites 19. Consanguinous marriages increase the risk of
11. a
8. Which mutation is seen in Down’s syndrome: following diseases: (PGMEE 2015)
12. a
 (PGMEE 2016-2017, 2015) a. Environmental disease
13. d a. Trisomy 13 b. Trisomy 21 b. Mitochondrial disorders
14. d c. Trisomy 22 d. Monosomy X c. X linked dominant diseases
15. a 9. Which of the following is a marker for Down’s d. Autosomal recessive disease
16. a Syndrome: (PGMEE 2015) 20. Niemann- Pick disease is: (PGMEE 2013-14)
17. a a. Increased maternal estriol a. AR b. X-linked
18. a b. Decreased HCG c. AD d. Mitochondrial
19. d c. Increased CA-125 21. Type of inheritance in Wilson’s disease:
20. a d. Decreased maternal serum alpha-fetoproteins  (PGMEE 2013-14)
21. b 10. In Turner syndrome, number of chromosomes are: a. X-linked recessive
 (PGMEE 2012-13, 2015) b. Autosomal recessive
a. 45 b. 42 c. Autosomal dominant
c. 46 d. 47 d. X-linked dominan
22. Which of the following is X- linked recessive- 33. % of offspring affected by colour blindness, of a healthy
 (PGMEE 2013-14) male and heterozygous female:  (PGMEE 2015)
a. G- 6 -PD deficiency b. Thalassemia a. 100% b. 25% 311
c. Alkaptonuria d. Neurofibromatosis c. 50% d. None
23. Which of the following is inherited as autosomal 34. Frame shift mutation doesn’t occur in multiples of:
recessive form? (PGMEE 2010, 2015)  (PGMEE 2012)
a. Sickle cell anemia a. 2 b. 3
b. Hemophilia c. 4 d. 5
c. Hereditary Spherocytosis 35. Genetic factor is associated with which of the
d. Glucose 6-Phosphate Dehydrogenase deficiency following- (PGMEE 2013-14)
24. If both parents have sickle cell anemia, then the a. Rheumatic heart disease
likelihood of children (offsprings) having the disease b. Pellagra
is- (PGMEE 2015, 2012-13) c. Autism
a. 10% b. 25% d. All of the above
c. 50% d. 100% 36. Type of inheritance in MELAS- (PGMEE 2013-14)
25. Which of the following is autosomal recessive
inherited cancer syndrome? (PGMEE 2015) a. AD b. AR
a. Ataxia telangiectasia b. Cowden syndrome c. Mitochondrial d. X-linked
c. HNPCC d. Retinoblastoma 37. All the following are mitochondrial disorders
26. In a family, mother is normal while father has a EXCEPT  (PGMEE 2014, 2015)
genetic disease. All their daughters are carriers and a. MELAS b. Kearns sayre syndrome
sons are normal, what is the pattern of inheritance of
c. NARP syndrome d. Incontinentia Pigmenti
disease? (PGMEE 2016)
38. Frameshift mutation occurs due to: (PGMEE 2016)
a. X-linked recessive b. AD
a. Transition b. Point mutation
c. X-linked dominant d. AR
c. Insertion d. Transversion
27. Single gene disorder is – (PGMEE 2012-13)
39. An example for gain of function mutation is:
a. Glycogen storage disease
 (PGMEE 2015)
b. Retinoblastoma
a. Osteogenesisimperfecta
c. Both a and b
b. Marfan syndrome M
d. Diabetes mellitus
c. Ehlerdanlos syndrome C
28. True statement about inheritence of an X linked
recessive trait is- (PGMEE 2012-13) d. Huntington’s disease Qs
40. Microdeletion is seen in: (PGMEE 2012) Ans.
a. 50% of boys of carrier mother are affected
b. 50% of girls of diseased father are carrier a. Beta thalassemia b. Di George syndrome
c. Marfan’s syndrome d. All 22. a
c. Father transmits disease to the son 23. a
d. Mother transmits the disease to the daughter 41. Li Fraumani syndrome is due to mutation of which
gene- (PGMEE 2012-13) 24. d
29. Parents are carrier of an autosomal recessive 25. a
disorder. Chances of offspring to get affected are: a. p43 b. p53
26. a
 (PGMEE 2013-14) c. p21 d. p41
27. c
a. 1:2 b. 1:3 42. Mammalian genome contains maximum genes that
28. a
c. 1:1 d. 1:4 code for receptors of: (AIIMS May 2016)
29. d
30. What is the chance of an offspring being affected with a. Odorants b. Interleukins 30. a
an affected mother and normal father, in an X-linked c. Immunoglobulins d. Growth factors 31. c
recessive condition? (PGMEE 2012) 43. Point mutations include all EXCEPT: 32. d
a. All of daughters are carriers  (AIIMS November 2012) 33. b
b. 50% of the off springs are carriers a. Deletion b. Insertion 34. b
c. 50% of sons are asymptomatic c. Substitution d. Inversion 35. c
d. Males will never be affected 44. Which is true regarding mitochondrial DNA: 36. c
31. Hypophostemic Vit D Resistant Rickets is? a. 3 × 109 base pairs 37. d
 (PGMEE 2016) b. Few mutations than nuclear DNA 38. c
a. AR b. AD c. 23 pairs inherited from each parent 39. d
c. XD d. XR d. Less than 20 % respiratory chain proteins coded 40. b
32. Mode of inheritance of Duchene muscular dystrophy: from mitochondrial DNA 41. b
 (PGMEE 2015) 45. Restriction endonuclease cleaves : (PGI 2017) 42. a
a. Autosomal dominant a. ds DNA b. RNA 43. d
44. d
b. X-linked dominant c. Histone d. Protein
45. a
c. Autosomal recessive e. ssDNA
d. X-linked recessive
 46. Function of endonucleases:- (PGMEE 2016-17) 56. Melting temperature of DNA is: (JIPMER May 2018)
a. Cut DNA at specific DNA sequences a. Proportional to AT pairs
312 b. Enhancers b. Proportional to GC pairs
c. To find out antibiotic resistances c. No relation with base pair composition
d. To point out the coding regions d. Directly proportional to length of DNA
47. cDNA from RNA is synthesized by- (PGMEE 2015) 57. If content of A is 15%, what is the amount of G in DNA
a. Topoisomerase according to Chargraff’s rule: (PGMEE 2015)
b. Helicase a. 35% b. 15%
c. Reverse transcriptase c. 85% d. 70%
d. DNA dependent DNA polymerase 58. Jumping genes are known as:- (PGMEE 2016-17)
48. Bacteria A has an adenine content of 33% in its a. Intron b. Transposons
genome. The expected content of thymine in the c. Plasmids d. Exon
genome of A is: 59. Leucine zipper motif is a mediator for:
a. 22% b. 33%  (PGMEE 2016-17)
c. 44% d. 66% a. Cyclic GMP
49. In a sample of DNA, if Adenine content is 32%, what b. Ligand membranes
will be the amount of Guanine? c. Binding of regulatory proteins to DNA
a. 18% b. 32% d. Membrane attack complexes
c. 36% d. 64% 60. If Codon no 302 UAG is replaced by UAA, then this
50. Which is the distance spanned by one turn of B-DNA? mutation is: (Recent Question 2016)
a. 2nm b. 2.4 nm a. Missense b. Silent
c. 3.2 nm d. 3.4 nm c. Nonsense
51. Which of the following is NOT TRUE about histones? d. Given information is not sufficient to identify
a. These are positively charged proteins 61. Which of the following is the correct sequence of
b. They undergo covalent modification severity of damage for a mutation in DNA ?
c. H3 and H4 are highly conserved proteins a. Missense > Nonsense >Frameshift
d. Their amino acid sequence are highly variable b. Nonsense > Missense >Frameshift
among organisms c. Frameshift>Missense > Nonsense
52. In a DNA double helix, the CpT sequences are as d. Frameshift> Nonsense > Missense
M frequent as the sequence of: 62. Main difference between DNA and RNA is :
C a. GpA b. ApG a. Sugar b. Nitrogenous base
Qs c. GpU d. GpC c. Vitamin d. None
Ans. 53. In RNA, there are unequal number of complementary 63. The number of base pairs in human diploid genome
bases i.e. it’s ‘A’ content unequals ‘U’ content and its are:- (PGMEE 2015)
46. a a. 2 billion base pairs (bp)
‘G’ content unequals its ‘C’ content. It is possible in
47. c b. 3 billion base pairs (bp)
case of
48. b c. 5 billion base pairs (bp)
a. Double stranded molecule
49. a d. 6 billion base pairs (bp)
b. Double stranded molecule with mutations
50. d 64. The estimated number of human genes are:
c. Single stranded molecule
51. d
d. Single stranded and partial double stranded  (PGMEE 2015)
52. b
molecule a. 10 thousand b. 20-25 thousand
53. d
54. In two bacterial DNA samples X and Y, Adenine c. 50 thousand d. 1 lakh
54. b
content is 32% and 17% respectively. One of these 65. Human mitochondrial genome encodes:
55. c
species was isolated from a hot spring (64°C). Which a. 37 genes b. 47 genes
56. b
species is most likely the thermophilic bacterium? c. 57 genes d. 67 genes
57. a
a. X 66. Double stranded RNA exists in:
58. b
b. Y a. A-DNA like conformation
59. c
c. Both b. B-DNA like conformation
60. b
d. None c. Z-DNA like conformation
61. d
55. Which of the following results in the structural d. None of these
62. a
stability of double helix of DNA? 67. UGA in mitochondrial genome codes for:
63. d
a. Hydrogen bonding between adjacent purine bases a. Methionine b. Tryptophan
64. b
b. Hydrogen bonding between purine and pyrimidine c. N-formyl Methionine d. None
65. a
bases of two chains 68. Which of the following play a predominant role in
66. a
67. b c. Hydrogen bonding between stacked purine and driving RNA secondary structure formation?
68. d pyrimidine bases. a. Vander Waals Interaction
d. The energy released due to the turning of nucleic b. Hydrophobic interaction
acid of molecule c. Hydrophilic repulsion
d. Formation of complementary base pair regions
69. What is the approximate number of base pairs 82. Which of the following does not favor permissive
associated with a single nucleosome? euchromatin due to changes occurring at cytosine residues
a. 73 b. 146 at CpG islands in DNA? (AIIMS May 2018) 313
c. 292 d. 334 a. Methylation
70. In humans, unmethylated CpG islands are found in: b. Phosphorylation
a. Operator b. Promoter c. Alkylation
c. Introns d. Exons d. Sumoylation
71. Which of the following protein is a major nuclear 83. One of the following disorders is due to maternal
sperm protein? disomy of chromosome 15- (PGMEE 2012-13)
a. Histone b. Protamine a. Prader Willi syndrome
c. Polyamine d. None b. Angelman syndrome
72. Prenatal diagnosis of Down Syndrome is done by- c. Hydatidiform mole
 (PGMEE 2013-14) d. Klinefelter’s syndrome
a. Fetal ultrasonography b. Triple test 84. Genomic imprinting is seen in- (PGMEE 2012-13)
c. Karyotyping d. All of the above a. Praderwilli syndrome b. Marfan syndrome
73. Among the following, the most sensitive USG finding c. EDS d. Osteogenesis imperfecta
of Trisomy 21 is: (PGMEE 2015) 85. Paternal 15 chromosome deletion is seen in-
a. Absent nasal bone b. Thickenend nuchal folds  (PGMEE 2013-14)
c. Echogenic bowel d. Short femur a. Turner syndrome b. Prader Willi syndrome
74. Y chromosome is: (PGMEE 2015)
c. Angelman syndrome d. Down syndrome
a. Telocentric b. Submetacentric
86. In genomic imprinting, DNA is modified by:
c. Acrocentric d. Metacentric
a. Acetylation b. Methylation
75. Which of the following test is not used in epigenetics?
c. Phosphorylation d. Deamination
a. HPLC
b. Chip on chip Anticipation
c. Bi-sulfate sequencing
87. The phenomenon where subsequent generations are
d. Methylation sensitive restriction enzymes digestion
at risk of earlier and more severe disease is known as:
76. Epigenetic factors refers to:  (PGMEE 2015)
 (PGMEE 2015)
a. Histone methylation b. Histone phosphorylation
a. Anticipation b. Dominance
c. DNA methylation d. All of the above M
c. Pleiotropy d. Dominance C
77. Epigenetics is a: (PGMEE 2015)
88. Anticipation phenomenon is seen in: (PGMEE 2015) Qs
a. Chemical modification of DNA
a. Limb girdle dystrophy
b. Normal variation of nucleotides Ans.
b. Becker muscular dystrophy
c. Change in nucleotide sequence 69. b
c. Duchenne muscular dystrophy
d. Irreversible modification of DNA 70. b
78. Heritable changes in gene expression that are not d. Fragile X-syndrome
89. Anticipation is a feature of: (PGMEE 2016, 2012-13) 71. b
caused by alterations in DNA sequence is: 72. d
 (PGMEE 2015) a. Trinucleotide repeats
73. b
a. Bioinformatics b. Genomics b. Mosaicism
74. c
c. Epigenetics d. Proteomics c. Trisomy
75. a
79. CG region is involved in:  (AIIMS Nov 2016) d. Genomic imprinting 76. d
a. Acetylation b. Methylation 90. Trinucleotide repeat disorder is: (PGMEE 2016) 77. a
c. Phosphorylation d. DNA Replication a. Mitochondrial myopathy 78. c
80. Methylation of cytosine leads to: (AIIMS May 2014) b. Myotonia dystrophica 79. b
a. Increased expression of gene c. Inflammatory myopathy 80. b
b. Decreased expression of gene d. Duchene’s dystrophy 81. c
c. No effect on gene expression 91. 20 year old male presents with mental retardation, 82. a
d. Mutation large mandible, large everted ears and large testes. 83. a
81. The differential expression of a gene based on What is the most likely diagnosis? (PGMEE 2015) 84. a
chromosomal inheritance from maternal or paternal a. Patau syndrome 85. b
origin is: (PGMEE 2015) b. Down syndrome 86. b
a. Germline mosaicism b. Mosaicism c. Fragile X syndrome 87. a
c. Genomic imprinting d. Pleiotropy d. Klinefelter syndrome 88. d
89. a
90. b
91. c
 Answers with Explanations
314
1. Ans. (b) B-DNA 3. Ans. (a) 1
CRO BIOCHEMISTRY

[Ref: Harper 30th/e pg. 361] [Ref: Robbins 9th/e pg. 165]
•• Watson and Crick in early 1950s proposed a model of •• Number of barr bodies is always one less the total
the double helical structure of B-form of DNA. number of X-chromosomes. Barr body is the inactive
X-chromosome in a female somatic cell. If two
2. Ans. (d) Melting point of DNA is closely related to
X-chromosomes present, then number of Barr bodies
Cytosine Guanine content of DNA
present is 1.
[Ref: Harper 30th/e pg. 360-361]
4. Ans. (a) Hydrogen bonds
•• In DNA, the two strands are held together by hydrogen
bonds. There are covalent bonds (3’ 5’ phosphodiester [Ref: Harper 30th/e pg. 360]


bonds) in sugar-phosphate backbone. Most common

•• DNA is double stranded (in both prokaryotes and
DNA is B- DNA. Melting point of DNA is closely
eukaryotes). Denaturation or Melting of DNA is a
related to Cytosine Guanine content of DNA as C and
process of separation of two strands of DNA. The
G have three bonds, as compared to the two bonds
two strands of DNA are joined by hydrogen bonds (2
present between A and T.
bonds between A and T and 3 bonds between G and
C). These bonds are broken on denaturation of DNA.

5. Ans. (c) Circular, double stranded

[Ref: Harper 30th/e pg. 378]

•• Eukaryotic DNA is linear, double stranded. Prokaryotic DNA is circular, double stranded.
•• Mitochondrial DNA is present in eukaryotes, but it resembles prokaryotic DNA i.e. it is circular, double stranded.

6. Ans. (a) DNA synthesis stops

[Ref: Harper 30th/e pg. 457]

•• When a dideoxy nucleotide is added in PCR, the dideoxy nucleotide is added in the growing chain. But it will not allow
the next normal nucleotide (deoxy nucleotide) to be added because there is no free 3'OH group in dideoxy nucleotide.
A So DNA synthesis stops. (Refer Fig. 12.3)
N
S
W
E
R
S
WITH

E
X
P 7. Ans. (d) Cut DNA at palindromic sites
L
A [Ref: Harper 30th/e pg. 452]
N •• Type II restriction enzymes recognize a palindromic nucleotide sequence in DNA and cut the DNA at that site, in predictable and
A specific manner. Palindrome is same sequence on opposite strands of DNA read in 5’→ 3’ direction.
T
I 8. Ans. (b) Trisomy 21
O
[Ref: Nelson 20th/e pg. 611]
N
S •• There are three trisomies affecting autosomes i.e. Trisomy 13 Patau syndrome, Trisomy 18 is Edward syndrome,
Trisomy 21 is Down syndrome. Most common trisomy in humans is Trisomy 16, but it is lethal.
 9. Ans. (d) Decreased maternal serum alpha- 16. Ans. (a) AD
fetoproteins
[Ref: Harper 30th/e pg. 460] 315
[Ref: Ghai 8th/e pg. 646] •• Huntington’s chorea is an Autosomal Dominant (AD)
•• There are 4 markers for screening of Down syndrome disorder.

CHAPTER 12  BASICS IN GENETICS

decreased AFP, decreased estriol, increased HCG and
increased Inhibin-α. Radiological screening can be 17. Ans. (a) Hereditary hypercholesterolemia
used to see for Nuchal Translucency. [Ref: Robbins 9th/e pg. 140]
10. Ans. (a) 45 •• Hereditary hypercholesterolemia or Familial hyper-
cholesterolemia is Autosomal Dominant (AD).
[Ref: Robbin’s 8th/e pg. 165-166] Wheras Tay Sachs disease, Tyrosenemia, Gaucher’s
•• In Turner syndrome, one of the X-chromosome is disease all are Autosomal Reccessive (AR).
missing in a female. This is the only example of a 18. Ans. (a) 50% affected
monosomy compatible with life. Genotype is X0.
There is no Barr body. [Ref: Robbin’s 8th/e pg. 141]

11. Ans. (a) 45, Y0 •• Mostly AD diseases have heterozygous trait.

[Ref: Robbins 9th/e pg. 160]

•• Monosomy affecting autosomes is lethal but in this
question options are monosomy or trisomy in sex
chromosomes. Monosomy is more dangerous than
trisomy. Turner Syndrome(X0) is only monosomy
compatible with life. 45,Y0 karotype is fetal in utero.

12. Ans. (a) 47, XXY

[Ref: Robbins 9th/e pg. 165]

•• In Klinefelter’s syndrome, there are two or more
X-chromosome in a male. In XXY karyotype (most
common in Klinefelter), there is one Barr body and 19. Ans. (d) Autosomal reccesive disease
phenotype is male. Number of chromosomes are 47. [Ref: Ghai 8th/e/c 22]
13. Ans. (d) 47XX, +13 •• Consanguinous marriages increase the risk of A
autosomal recessive diseases. N
[Ref: Robbins 9th/e pg.162]
S
•• Patau syndrome is trisomy 13, so number of 20. Ans. (a) AR
W
chromosomes are 47. It can be in male or female. [Ref: Harper 30th/e pg 251] E
R
14. Ans. (d) Used to see genetic transmission •• Most Inborn Errors of Metabolism are inherited in
Autosomal Recessive (AR) fashion. S
[Ref: Nelson’s 20th/e/pg. 583] WITH
21. Ans. (b) Autosomal recessive
•• Pedigree chart is a diagram of genetic history of a
E
family over several generations. Various symbols are [Ref: Robbin’s 8th/e pg. 141]
X
used. Pedigree analysis can be used for counselling
•• Wilson disease, Cystic Fibrosis and Thalassemia have P
parents about the chances of having genetic disease
autosomal recessive (AR) pattern of inheritance. L
in siblings of an affected individual. A
22. Ans. (a) G- 6 -PD deficiency N
15. Ans. (a) Autosomal dominant disease
[Ref: Harper 30th/e pg 693]
A
[Ref: Robbins 9th/e pg. 140] T
•• X-linked recessive inheritance disorders are: G-6- PD I
•• Male to male transmission cannot occur in X- linked deficiency, Fabry’s disease, Hunter disease, Lesch O
disease and mitochondrial diseases. Nyhan syndrome, Hemophillia A & B. N
S
 23. Ans. (a) Sickle cell anemia 31. Ans. (c) XD

316 [Ref: Robbin 9th/e pg. 141 table 5-2 2] [Ref: Robbins 9th/e pg. 142]
Refer to Ans 24 •• Hypophostemic Vit D Resistant Rickets is X-linked
Dominant
24. Ans. (d) 100%
CRO BIOCHEMISTRY

32. Ans. (d) X-linked recessive

[Ref: Robbins 9th/e pg. 141]
[Ref: Robbins 9th/e pg. 140-142]
•• Q 23 & 24 Sickle cell anemia has AR inheritance.
If both parents are affected means both parents •• Duchene Muscular Dystrophy is X-linked dominant
are homozygous for the trait as recessive allele is disorder.
expressed only in homozygous state. So, all offsprings
33. Ans. (b) 25%
will inherit the disease.
[Ref: Robbins 9th/e pg. 140-142]
25. Ans. (a) Ataxia telangiectasia
•• Color Blindness has XR inheritance. If mother


[Ref: Robbins 9th/e pg. 141] is carrier, then 50% of sons are affected and 50%
daughters are carriers. Out of all offsprings (sons as
•• Ataxia telangiectasia is autosomal recessive inherited well as daughters), 25% offsprings are affected and
cancer syndrome.
they are male offsprings.
26. Ans. (a) X-linked recessive
34. Ans. (b) 3
[Ref: Robbins 9th/e pg. 140-142] [Ref: Harper 30th/e pg. 417]
•• The picture depicts X-linked recessive pattern of •• Frameshift mutation occurs due to addition or
inheritance. deletion of one or two nucleotides which alter the
reading frame. It does not occur in multiples of 3.
27. Ans. (c) Both a and b

[Ref: Robbin’s 9th/e pg. 142] 35. Ans. (c) Autism

•• Single gene disorder or Mendelian disorder [Ref: Nelson 20th/e/ch 30]

means a particular gene is causing the disease e.g. •• Autism is a genetic disorder (also has environmental
Inborn errors of metabolism, Sickle cell disease, influences).Whereas rheumatic heart disease is due
Retinoblastoma etc. Whereas Diabetes Mellitus and to untreated or under-treated streptococcal infection
Hypertension are multifactorial i.e. they have both and pellagra is due to niacin deficiency.
genetic and environmental influences.
A 36. Ans. (c) Mitochondrial
N 28. Ans. (a) 50% of boys of carrier mother are affected
[Ref: Nelson 18th/e pg. 172]
S
[Ref: Robbin’s 8th/e pg. 142] •• Diseases due to mutation in Mitochondrial DNA are
W
E •• In XR inheritance, if mother is carrier then 50% of MELAS, NARP Syndrome, Leigh disease, Leber Optic
sons are affected and 50% of daughters are carriers. Neurpathy & Kearns-Sayre Syndrome.
R
If mother is affected, then 100% sons are affected &
S 37. Ans. (d) Incontinentia Pigmenti
100% daughters are carriers. If father is affected, then
WITH all daughters are carriers and all sons are normal. [Ref: Robbins 9th/e pg. 171]
E 29. Ans. (d) 1:4 •• Incontinentia Pigmenti is a rare, XD disorder affecting
X CNS and many organs including skin.
P [Ref: Robbins 9th/e pg. 140-142]
L •• 25% offsprings have chance to be affected if both 38. Ans. (c) Insertion
A parents are carriers for AR disorder. So, one in four [Ref: Robbins 9th/pg 160; 8th/pg 160]
N offspring have chances to be affected.
A •• Frameshift mutation can occur due to insertion or
T 30. Ans. (a) All of daughters are carriers deletion of nucleotides in numbers which are not in
I multiple of 3, so these mutations alter genetic reading
[Ref: BRS Genetics by Ronald W. Dude pg. 31-33]
O frame.
N •• If mother is affected with XR disorder, then all sons
S are affected and all daughters are carriers.
39. Ans. (d) Huntington’s disease mt DNA encodes for 13 protein subunits of respiratory
chain (total 67) ; 13/67 = 19 %. It is inherited only from
[Ref: Internet] mother. There are no introns in mtDNA so it has more 317
Gain of function mutation means there is overactivity mutations than nuclear DNA.
of gene. E.g. Huntington’s disease leads to formation
of excessive Huntington protein, which is toxic for 45. Ans. (a) ds DNA

CHAPTER 12  BASICS IN GENETICS

neurons. [Ref: Harper 30th/e pg. 176]
Loss of function mutation leads to underactivity of
genes. •• Restriction endonuclease cleaves double stranded
DNA at palindromic sites.
40. Ans. (b) Di George syndrome
46. Ans. (a) Cut DNA at specific DNA sequences
[Ref: Harrison 18th/e Chapter 62]
[Ref: Harper 30th/e pg. 452]
•• Di George syndrome is due to deletion of small
•• Endonuclease cut in between the DNA sequence
segment of chromosome 22 (microdeletion).
but exonuclease cut from the sides either from 5’
30–40 genes are deleted from the middle of this
or from 3’ side. A special endonuclease – known as
chromosome. It involves multiple systems and
Restriction endonuclease cleaves double stranded
patients have congenital heart defects.
DNA at palindromic sites.
41. Ans. (b) p53
47. Ans. (c) Reverse transcriptase
[Ref: Robbin’s 8th/e pg. 275]
[Ref: Harper 30th/e pg. 405]
•• Li Fraumani is a rare, AD disorder due to mutation •• RNA can be converted to DNA by reverse transcriptase.
in the tumor suppressor gene – p53. Patient is This DNA is cDNA (complementary DNA).
predisposed to development of multiple cancers.
48. Ans. (b) 33%
42. Ans. (a) Odorants
[Ref: Lehniniger 7th/e pg. 286]
[Ref: Harper 30th/e pg. 522]
•• According to Chargaff’s rule, the content of adenine
•• Odorant receptor superfamily contains maximum should be equal to content of thymine. So thymine
genes in mammalian genome. content in the genome should also equal 33%.
43. Ans. (d) Inversion 49. Ans. (a) 18%
[Ref: Harper 30th/e pg. 417] [Ref: Lehniniger 7th/e p286]
•• Point mutations are silent mutation, missense A
•• According to chargaff’s rule in a ds DNA, the total
mutation, non-sense mutation, insertion, deletion, N
purine content is equal to the pyrimidine content.
transition (substitution of Purines with Purine Since [A] =[T], so [T] = 32% S
or Pyrimidine with Pyrimidine), transversion [A]+[T] = 32 = 32 = 64% W
(substitution of Purine with Pyrimidine or vice-versa). [C]+[G] = 100–([A]+[T])= 100–64% =36% E
Whereas Inversions are large scale chromosomal As [C] = [G], so [G] = 36/2 = 18% R
mutations ( and there occurs alteration in position of S
genes on a chromosome). 50. Ans. (d) 3.4 nm
WITH
44. Ans. (d) Less than 20 % respiratory chain proteins [Ref: Harper 30th /e pg 361]
E
coded from mitochondrial DNA •• A single turn of B-DNA about the long axis of the X
molecule contains 10 bp. The distance spanned by P
[Ref: Harper 30th/e pg. 378]
one turn of B-DNA is 3.4nm (34A°). The width (helical L
•• Mitochondrial DNA (mtDNA) is a double-stranded, diameter) of the double helix in B-DNA is 2nm (20A°) A
circular molecule of 16,569 bp and contains 37 genes N
coding for two rRNAs, 22 tRNAs and 13 polypeptides. A
T
I
O
N
S
 54. Ans. (b) Y
318 [Ref: Lehniniger 7th/e pg. 296]
•• For any double stranded DNA, A=T and G=C.
Bacteria X: [A] = 32% therefore [T]=32%
CRO BIOCHEMISTRY

[A]+[T] = 32+32 = 64%

[G]+[C] = 100 – ([A]+[T]) = 100 – 64 = 36%
BacteriaY: [A]=17% therefore [T]=17%
[A]+[T] = 17+17 = 34%
[G]+[C] = 100 – ([A]+[T]) = 100 – 34 = 66%
The higher the G +C content of a DNA molecule,
the higher the melting temperature (Bacteria Y)
and this bacteria can withstand high temperature
(thermophilic).

55. Ans. (c) Hydrogen bonding between stacked purine



and pyrimidine bases

51. Ans. (d) Their amino acid sequence are highly
[Ref: Lehniniger 7th/e pg. 287]
variable among organisms
•• Although the structural stability of any molecule
[Ref: Lehniniger 7th/e pg 939] is a combination of all kinds of forces, the major
•• Histones are rich in basic amino acids i.e arginine and contributor to the helical structure of duplex DNA is
lysine (positive charge). So, histones are positively the hydrogen bonding between stacked purine and
charged proteins. Histones are highly conservative pyrimidine bases.
proteins. H1 histones (Linker histone) are least
tightly bound and are therefore, easily removed with 56. Ans. (b) Proportional to GC pairs
a salt solution, after which chromatin becomes more [Ref: Harper 30th/e pg. 361]
soluble. The core histone proteins are subjected to
atleast six types of covalent modifications or post •• Denaturation or melting of two DNA strands means
translational modifications. separation of the two strands of DNA. In this process,
hydrogen bonds between the complementary base
52. Ans. (b) ApG pairs are broken, leading to separation
•• Melting temperature (Tm) of DNA – that temperature
[Ref: Harper 30th/e pg. 360] at which half of the DNA duplex gets dissociated to
•• The DNA sequence is always read from 5’ → 3’ become single stranded. Tm of DNA is between 85-
A direction, Therefore CpT means 5’-C-p-T-3’. 95°C. Primers with melting temperatures in the range
N 5’-C-p-T-3’ of 52-58°C generally produce the best results. Tm is
S 3’-G-p-A-5’ proportional to GC pairs because there are three
W Since the two strands are antiparallel, the
 hydrogen bonds between G & C, as compared to the
E complementary sequence would be 3’-G-p-A-5’. two hydrogen bonds between A & T. So more GC
R While writing we will write ApG i.e. from 5’→ 3’ content gives thermostability to DNA.
S direction For every 10 % increase in GC content, melting
temperature raises by 5°C.
WITH 53. Ans. (d) Single stranded and partial double stranded
molecule 57. Ans. (a) 35%
E
X [Ref: Lehniniger 7th/e pg. 286] [Ref: Harper 30th/e pg. 382]
P •• According to Chargraff’s rule, A is equal to T and G
•• The Chargaff’s Rule i.e. G=C and A=U or A=T is true
L is equal to C. If A is 15 % then T is also 15%. This is
only in case of fully double stranded molecules due
A total 30 %. Now G+C= 60%. So G is 30%. Best option
to complementary pairing. The discordance in the
N is A here.
complementary bases i.e. bases being unequal in
A
number could happen in case of molecule which is
T 58. Ans. (b) Transposons
single stranded or partially double stranded. More
I
are the equal number of complementary bases, more [Ref: Harper 30th/e pg. 380]
O
is the double stranded portion in the nucleic acid.
N •• Transposons are also known as jumping genes as
S these are DNA sequences that can move from one
 chromosome locus to another. Transposition is a 65. Ans. (a) 37 genes
highly specialized form of recombination in which a
segment of DNA moves from one location to another, [Ref: Harper 30th/e p378] 319
either on the same chromosome or a different Features of Mitochondrial DNA:
chromosome •• Circular double stranded (like Prokaryotic DNA)
•• Constitutes about 1% of total cellular DNA

CHAPTER 12  BASICS IN GENETICS

59. Ans. (c) Binding of regulatory proteins to DNA •• Rate of Mutation is high as there is no proof reading
[Ref: Harper 30th/e pg. 445] •• Multiple copies are present in the same cell
•• Contains 37 genes: 13 genes for proteins of ETC, 22
•• Leucine zipper motif is a kind of protein modification tRNA genes, 2 rRNA genes
of regulatory protein which binds to the response •• Maternally inherited
element on DNA. •• Genetic code of mitochondria is different. It contains
4 stop codons.
60. Ans. (b) Silent Mutation

[Ref: Harper 30th/e pg. 416] 66. Ans. (a) A-DNA like conformation

•• In this mutation, stop codon – UAG is replaced by [Ref: Lehninger 7th/e pg. 965]
another stop codon- UAA. So, this is silent mutation. •• Double stranded RNA exists in A- DNA like confor-
In silent mutations, there is no change in primary mation. Usually RNA is single stranded, DNA is
structure of protein. double stranded. But in few species, double stranded
RNA is found e.g. Rotavirus.
61. Ans. (d) Frameshift> Nonsense > Missense
67. Ans. (b) Tryptophan
[Ref: Harper 30th/e pg. 417]
•• Frameshift mutation is deletion or insertion of a [Ref: Harper 30th/e pg. 414]
number of nucleotides not divisible by 3, resulting in •• UGA is a universal stop codon. But in mitochondria,
misleading of all nucleotides downstream. it acts as a codon for Tryptophan.
•• Nonsense mutation means change of a codon
with a stop codon (UAA, UAG, UGA). This leads to 68. Ans. (d) Formation of complementary base pair
premature termination of Protein synthesis. regions
•• Missense mutation means a codon is replaced by
[Ref: Harper 30th/e pg. 366]
another codon, coding for a different amino acid.
•• RNA can form various secondary structures because
62. Ans. (a) Sugar of the base pairing between the complementary
regions. E.g. RNA duplexes, Hairpin/Stem loop,
[Ref: Lehninger 7th/e pg. 965] Bulges
A
•• Out of sugar and base, main difference is of sugar as N
three nitrogenous bases (adenine, cytosine, guanine) S
are same . but sugar is always ribose in RNA and it is
W
always deoxyribose in DNA.
E
63. Ans. (d) 6 billion base pairs (bp) R
S
[Ref: Harper 30th/e p472; Robbin’s 9th/e pg. 160]
WITH
•• Human haploid genome consists of 3 billion base
pairs. E
 Therefore, the diploid genome consists of = 3 × 2 = 6 X
69. Ans. (b) 146
billion base pairs (billion is 109). P
[Ref: Harper 30th/e p373] L
64. Ans. (b) 20-25 thousand A
•• Nucleosome has dimensions: 10nm (width) × 5 nm
[Ref: Harper 30th/e p368,448) (height)
N
•• DNA with approximately 146 bp length wraps 1.75
A
•• Earlier One Gene One Protein Theory was discovered. T
But later they found that genes are 20,000 to 25000 times around core histones.
•• DNA connecting 2 nucleosomes is known as Linker
I
only but proteins are more than 2.5 lakh in human O
DNA (30 bp in length)
body. N
S
 73. Ans. (b) Thickenend nuchal folds

320 [Ref: Robbins 9th/e pg. 161]

•• The most sensitive USG finding of Trisomy 21 is
thickenend nuchal folds.
CRO BIOCHEMISTRY

74. Ans. (c) Acrocentric

[Ref: Emery Genetics]

•• Y-chromosome is acrocentric, means centromere lies
close to the end of chromosome. X- chromosome is
sub- metacentric, means centromere lies between
middle and end of chromosome. Majority of human
chromosomes are sub-metacentric.

75. Ans. (a) HPLC



70. Ans. (b) Promoter Tests done in epigenetics are:

[Ref: Harper 30th/e pg 735] •• Na bisulfite method – detects DNA methylation.
•• ChIP – Chromatin Immuno Precipitation- it detects
•• CpG dinucleotides are mostly present in the post translational modifications of histones
promoter region and are unmethylated and reversible ƒƒ ChIP is used together with its large-scale variants
methylation of these CpG sites serves a way of turning ChIP-on-chip and ChIP-Seq
off/on the gene expression. •• Fluorescent in situ hybridization
•• Methylation-sensitive restriction enzymes

76. Ans. (d) All of the above

[Ref: Robbins 9th/e pg. 3-4]

•• Epigenetic Changes are: DNA Methylation at
cytosine of CG sites, PTMs of histones (acetylation
or deacetylation) and changes in higher order of
chromosome structure like heterochromatin or
euchromatin

A 77. Ans. (a) Chemical modification of DNA

N [Ref: Robbins 9th/e pg. 140-142; Harper 30th/e pg. 440]
71. Ans. (b) Protamine
S
W [Ref: Harper 30th/e pg. 427] Refer to answer 78
E
•• Protamines are major nuclear sperm proteins and 78. Ans. (c) Epigenetics
R
binds to DNA in spermatid. They are small, arginine-
S rich proteins that replace histones late in the haploid
[Ref: Robbins 9th/e pg. 140-142] [Ref: Harper 30th/e pg.
440]
WITH phase of spermatogenesis and are believed essential
for sperm head condensation and DNA stabilization. •• Q77, 78 Epigenetics is information in the genome
E which alter the pattern of gene expression. But it is not
X 72. Ans. (d) All of the above change in DNA code. This is chemical modification
P of DNA or proteins. These changes are transferred
L [Ref: Ghai 8th/e pg. 645]
to next generation. Unlike mutations, these are
A •• Down syndrome is trisomy 21. Triple test for reversible changes and can lead to gene activation or
N screening detects levels of Alpha Feto Protein (AFP) inhibition.
A which is decreased, increased hCG and decreased
T 79. Ans. (b) Methylation
estriol. Fetal USG shows nuchal translucency.
I
Refer to answer 80
O
N
S
80. Ans. (b) Decreased expression of gene 86. Ans. (b) Methylation
[Ref: Harper 30th/e pg. 440] [Ref: Robbins 10 /e pg 174,175] 321
•• Q 79, 80 DNA Methylation of cytosine of CG sites •• Most common mechanism of genomic imprinting is
occurs by enzyme – DNA Methyl Transferases DNA methylation at CG sites and this leads to gene

CHAPTER 12  BASICS IN GENETICS

(DNMTs). It leads to decreased expression of gene. inactivation.
•• The cytosine residue of CG site is methylated.
81. Ans. (c) Genomic imprinting
87. Ans. (a) Anticipation
[Ref: Robbins 9th/e pg. 172-173]
•• Genomic imprinting is an epigenetic phenomenon [Ref: Robbins 9th/e pg. 168]
which helps in regulation of gene expression. •• Anticipation is when the genetic defect increase from
‘Imprinting’ means inhibited. This is inhibition at the generation to generation until an affected individual
level of transcription i.e. gene is inhibited or silenced. is produced.
It results in mono-allelic expression i.e. allele from
one parent is expressed (either material or paternal). 88. Ans. (d) Fragile X-syndrome

82. Ans. (a) Methylation [Ref: Robbins 9th/e pg. 140]

Refer to answer 90
[Ref: Harper’s illustrated biochemistry, 30th ed., pg. 560]
•• DNA methylation leads to gene inhibition & 89. Ans. (a) Trinucleotide repeats
formation of heterochromatin. This mostly occurs at
[Ref: Robbins 9th/e pg. 168]
cytosine residue in DNA.
Refer to answer 90
83. Ans. (a) Prader Willi syndrome
90. Ans. (b) Myotonia dystrophica
[Ref: Robbin’s 8th/e pg. 172]
[Ref: Robbins 9th/e pg. 140]
•• Maternal uniparental disomy occurs in Prader Willi
syndrome. Paternal uniparental disomy occur in Q88, 89, 90 Anticipation occurs in Fragile X syndrome
Angelman syndrome. where there is expansion of CGG triplet repeat. A carrier
of the disease has 50-200 repeats. But these repeats
84. Ans. ( a) Prader willi syndrome increase from generation to generation (anticipation).
And an affected individual has more than 200 repeats.
[Ref: Robbin’s 8th/e pg. 172-173]
Myotonic dystrophy is also a trinucleotide repeat
•• Disease caused due to Genomic Imprinting are disorder.
Prader Willi Syndrome and Angelman Syndrome. A
91. Ans. (c) Fragile X syndrome N
85. Ans. (b) Prader Willi syndrome
[Ref: Robbins 9th/e pg. 170] S
[Ref: Robbins 9th/e pg.172] W
•• Fragile X-Syndrome is a trinucleotide repeat expan- E
•• Deletion of paternal copy of allele is seen in Prader sion disorder in which patient has large face, large R
Willi syndrome. Deletion of maternal copy is seen in mandible, large testis, large everted ears and tall
Angelman syndrome. stature. It is the second most common cause of S
mental retardation (first is Down Syndrome). WITH

E
X
P
L
A
N
A
T
I
O
N
S
 13 Replication,
Transcription,
Translation
Overview of Chapter
•• DNA Replication
•• Proofreading and Repair
•• Types of RNA
•• Transcription
•• Operon mode
•• Codons
•• Translation

Central Dogma of Molecular Biology

The flow of information from DNA to RNA and then to
proteins is known as central dogma of molecular biology.
Fig. 13.1: Ori (one in prokaryotes and multiple in eukaryotes)
Some viruses have RNA as genetic material and they undergo
reverse transcription.
Steps in Replication
The first requirement is to separate the two strands, also
known as unwinding. The temperature at which half of the
helical structure is lost is known as melting point (Tm).
High GC content of DNA raises Tm. Enzymes are given in a
sequence.
1. Helicase:
 Causes strand separation
 Uses ATP
 Creates positive supercoils (overwinding is known
Reverse transcription: Synthesis of DNA from RNA. Enzyme
as positive supercoiling)
is RNA dependent DNA polymerase. This reverse trans-
2. Topo-isomerases: To relieve supercoils
cription is involved in:
 They cut and reseal DNA and relieve positive
zz Retroviruses e.g. HIV virus
supercoils
zz Transposons/jumping genes (Transposons are DNA
 Do not use ATP. They gain energy from the cleavage
sequences which are transcribed into RNA. These RNAs
of phosphodiester bond.
are used as a template for DNA synthesis by reverse
transcription and this DNA is randomly inserted into the  Type I Topo-isomerases–cut one strand
genome)  Type II Topo-isomerases–cut two strands
zz Telomerase DNA gyrase: A counterpart of Topoisomerase-II in prokaryotes.
This enzyme introduces negative supercoils and uses ATP.
DNA REPLICATION (underwinding is known as negative supercoiling)

zz Occurs in S-phase of cell cycle  Helicase and Topo-isomerases work in tandem to

zz Semiconservative in nature (one of the parental strand is do strand separation
conserved each time) 3. SSBs (Single Strand DNA Binding Proteins):
zz Bidirectional (means replication moves in both directions  Prevent reannealing or rejoining of the two
from the origin of replication) separated strands as they have high affinity for single
strand. SSB proteins are not enzymes.
  They provide the single strand required by  This same enzyme fills this gap with DNA but only
polymerase and also protect from nuclease attack. on the Lagging strand.
 SSBs → in prokaryotes 7. DNA Ligase: Acts only on lagging strand 323
 RPA (Replication Protein A) → in eukaryotes  Creates 3’ 5’ phosphodiester bond
Helicase, Topo-isomerases and SSBs → together known as  Uses ATP (ATP gets converted to AMP)

CHAPTER 13  REPLICATION, TRANSCRIPTION, TRANSLATION

Unwinding Proteins
Telomere and Telomerase
4. Primase: Synthesize primers
 Primers synthesized are RNA. But the template Telomeres are ends of chromosomes. They have tandem
used is DNA. So, this enzyme can also be called repeats of a transcriptionally inactive hexameric sequence
DNA dependent RNA Polymerase. Later these RNA (TTAGGG)n with proteins also attached here. This G rich
primers are removed. sequence is base paired to complementary region, which is
 In eukaryotes → α-Polymerase synthesize primer C-rich sequence. G-rich strand is longer than C-rich strand.
 In prokaryotes → DNA-G protein synthesize primer This provides an overhanging ssDNA present at the 3’ end,
 One primer is required for leading strand but which is approximately 100 nucleotides in length. This ssDNA
multiple primers are required for lagging strand. gets folded along with proteins and is stabilized.
(Fig. 13.2)

Benefits of Telomeres
zz Provide structural support to the chromosome
zz Prevent attack by nucleases
zz Distinguish a true end of chromosome from a break in
dsDNA
Fig. 13.2: Replication fork. Replication occurs in 5’ → 3’
direction. The template or the parent strand is read from 3’ to 5’ Telomere Shortening
direction. The upper daughter strand in the figure is synthesized
at one stretch, so known as leading strand. But the lower strand With each replication, some of this telomeric sequence is
is synthesized in short fragments. These fragments are known as lost (genes are not lost) because there is no template for this
okazaki fragments and this strand is known as lagging strand. DNA. Once telomeres are shortened beyond a critical length,
5. DNA Polymerase III: the cell cannot divide further. This occurs in case of somatic
 It synthesize both leading and lagging strands. cells. Due to this somatic cells have limited number of cell
Substrate for this enzyme is Deoxyribonucleotide divisions. This loss of DNA gradually leads to ageing
Triphosphates. Deoxyribonucleoside Monophos-
phate is added and pyrophosphate is released. So Telomerase/Telomere Terminal Transferase
two high energy phosphates are used to add one
Deoxyribonucleotide. The free OH at 3’ end of prim- zz In case of germ cells and stem cells, unlimited number
er is the acceptor of deoxyribonucleotides added by of cell divisions can occur because this telomere loss
this polymerase. does not occur. Telomerase enzyme can synthesize these
 If 2’ 3’ dideoxy ribose is used, then DNA synthesis sequences.
stops. Or if any of the 4 substrates (dATP, dTTP, dCTP, zz This enzyme contains RNA + protein (act as enzyme).
dGTP) is not available then DNA synthesis stops. RNA portion acts as a template on which DNA synthesis T
6. DNA Polymerase I: occurs, so this enzyme also known as RNA dependent H
 Remove RNA primers from both leading and lagging DNA polymerase or reverse transcriptase. E
strand. It has 5’ → 3’ exonuclease activity, means it zz Telomerase is not a ribozyme. Its activity increases in O
starts cutting the primer from 5’ end and move in cancer and decreases in ageing. Its activity is more in R
5’→ 3’ direction. germ cells as compared to stem cells. Y
 zz Telomerase increases longevity of cells
zz If telomerase activated in somatic cells, then it leads to cancer.
324 zz Premature ageing (Progeria) occurs if telomerase decreases.
CRO BIOCHEMISTRY


Fig. 13.3: Telomere and Telomerase

Klenow Fragment PROOFREADING AND REPAIR

zz Klenow fragment is a large fragment produced by Subtilisin
mediated proteolytic cleavage of E.Coli DNA polymerase I. Proofreading Repair
Proteolysis removes the 5’ → 3’ exonuclease activity from
Correction Correction during Correction after synthesis
N-terminal.
synthesis
Enzymatic 3' → 5' exonuclease Mostly endonuclease, 5'
activity activity → 3' exonuclease activity
sometimes
Enzyme in DNA polymerase II DNA polymerase II
prokaryotes
Enzyme in All DNA β-Polymerase (main)
eukaryotes Polymerases can do ε-Polymerase (minor role)
proofreading except
alpha and beta
zz The 5' → 3' exonuclease activity of DNA polymerase I makes it polymerase
unsuitable for many applications, so Klenow fragment, which
Phase of cell S-phase Most repairs occur in G1
lacks this activity, can be used for various purposes:
cycle phase
1. Remove 3’ overhang
2. Fills 5’ overhangs (or Filling of receded 3' ends of DNA
fragments)
3. Synthesis of double-stranded DNA from single-stranded
templates
4. Preparation of radioactive DNA probes
5. Was used in PCR, before being replaced by thermostable
T Taq polymerase
H
E
O
R
Y
Comparison of Prokaryotic and Eukaryotic DNA Polymerases
E.coli (Prokaryotes) Eukaryotic Function
325
DNA Polymerase I Nucleus Mitochondria Remove primer and fill the gap
• RNase H • RNase H

CHAPTER 13  REPLICATION, TRANSCRIPTION, TRANSLATION

• FEN-1 (Flap endonuclease) • FEN-1 (Flap endonuclease)
• δ−polymerase (minor role)
DNA Polymerase II DNA proof reading and repair
β DNA repair
γ Mitochondrial DNA synthesis
DNA Polymerase III ε Leading strand synthesis
δ Lagging strand synthesis
DNA-G protein α Primase (synthesize Primers)

DNA Repair (Single Strand Break Repair)

Most DNA repairs occur in G1 phase of cell cycle but only mismatch repair occurs in G2 phase of cell cycle.

Type of repair Phase of cell cycle Damage Cause Disease

Nucleotide excision G1 Thymine-Thymine dimers UV radiation Xeroderma pigmentosa
repair (T-T dimers)
Base excision repair G1 mainly but can Cytosine deaminated to Spontaneous, heat, infra red MUTYH associated polyposis
occur in any phase uracil rays, viral infection, Nitrous
Oxide
Mismatch repair G2 Mismatched base Proofreading error HNPCC (Hereditary Non
Polyposis Colorectal Cancer)
zz HNPCC also known as Lynch syndrome → leads to increase risk for developing colon cancer.
zz Xeroderma Pigmentosa increase the chances of skin malignancy.

Double Strand Break Repair

All these types of DNA damage mentioned above have defect in one of the strand and so they can use other strand as a template
and can repair the damaged DNA. But in case of double strand break repair, these repair mechanisms cannot be used. So
there are separate repair systems for double strand break repair. They are Non homologous End Joining (NHEJ) repair and
Homologous Repair/Recombination (HR)

Non homologous end joining (NHEJ) repair Homologous repair/recombination (HR)

• Error prone and thus mutagenic • Error free
• DNA breaks directly ligated without any need for homologous • Homologous DNA used as a template
template
• During G1/G0 phase of cell cycle • Late S, G2 and M phase of cell cycle
• Defect in this repair system can lead to SCID (Severe Combined • Defective HR can lead to Bloom syndrome, Werner syndrome,
Immuno Deficiency) and radiosensitive SCID Rothmund-thomson syndrome, Breast cancer (susceptibility 1
and 2), Ataxia Telangiectasia-like disorders, Nijmegen breakage
syndrome

DNA Repair Method Source of information for repair T

H
• Single strand break repair Complementary DNA strand E
• Homologous repair Sister chromosome O
• NHEJ No source R
Y
 TYPES OF RNAs
326 Type Percentage Characteristic feature Synthesis
rRNA 80 % Most abundant Nucleolus
tRNA 15 % Smallest, Nucleus
CRO BIOCHEMISTRY

Maximum modified bases

mRNA 5% Most heterogenous Nucleus


Fig. 13.4: tRNA (Clover leaf shape)

T
H Fig. 13.5: Ribosomes
E
O
R mRNA which is formed from more than one gene is known as Polycistronic, which is characteristic of Prokaryotes. In eukaryotes, each
mRNA comes from a single gene (known as monocistronic mRNA)
Y
Introns
zz Are intervening sequences in between the coding 327
sequences/ exons
zz Present only in eukaryotes
zz Coding DNA sequence/Exons are only 1–2% of all the

CHAPTER 13  REPLICATION, TRANSCRIPTION, TRANSLATION

genome. Rest is non coding.
zz Presence of introns prevent mutation
zz They are transcribed but not translated

Eukaryotic DNA Prokaryotic DNA

Have introns No introns
Prevented from mutations Can easily mutate

Nuclear DNA Mitochondrial DNA

rRNA as Ribozyme
Replication Nucleus Mitochondria
zz 23 s rRNA in Prokaryotes
Transcription Nucleus Mitochondria
zz 28 s rRNA in Prokaryotes
Translation Cytoplasm Mitochondria zz Peptidyl Transferase enzyme
zz Role in elongation and termination of translation
Non-Coding RNA (ncRNA)
Ribozyme Having Role in Splicing
Coding RNA is only mRNA. Rest all are non-coding RNAs.
These are transcribed but not translated. Pi RNA is most Self splicing introns Spliceosomal introns
common small ncRNA. Intron (RNA) itself catalyze snRNA acts as ribozyme. Introns
cannot catalyze
Divided into two groups- Not assigned a group number
group I and II
No ATP used ATP is required for assembly
of spliceosome but no ATP
required for RNA cleavage/
ligation reactions
Lariat formation occurs in Lariat formation occurs same
group II introns but not in like group II introns
group I
Not limited to eukaryotes Limited to eukaryotes
(also found in bacterias)

What is 5’splice site, 3’ splice site and a branch site?

Ribozyme Exon intron junction towards the 5’ end is known as 5’ splice
zz Ribozyme means RNA acts as enzyme site. Exon intron junction towards the 3’ end is known as 3’
zz Substrate of Ribozyme → is mostly RNA splice site. Sometimes a branch site is present in the intron
which mostly contains Adenine.
zz No ATP is used
zz It catalyze the breakage and synthesis of phosphodiester
bond
Similarity of Protein Enzyme and Ribozyme T
zz Specificity for the substrate H
zz Accelerate rate of reaction E
zz Kinetic behaviour O
zz Can be competitively inhibited R
Y
 Trans-esterification Reaction Spliceosomal Introns
At 5 ‘end, phosphate is present and a free –OH group is present
328 at the 3’ end. To break exon-intron junction, 3’ OH of guanosine
zz Intron is not self splicing
zz It requires ribozyme – snRNPs (small nuclear ribonucleo
acts as a nucleophile and attacks the phosphodiester bond at
protein particles)
the junction of intron and exon.
zz snRNA- small nuclear RNA – U1, U2, U4, U5 and U6 are
CRO BIOCHEMISTRY

involved. They remove introns by forming base pairs

with the consensus sequences at each end of the intron.
zz snRNPs bind and align exons in such a way that two
transesterification reactions occur with lariat formation
(like group II introns)
zz Intron released can be degraded or act as a precursor of
ncRNA


C linical B ox Subunit Nuclear Function

In SLE (Systemic Lupus Erythmatosus), antibodies are produced α, ω alpha and omega Required for enzyme assembly
against snRNPs. This is an autoimmune disease. β beta Has catalytic activity
β’ beta’ For template binding
TRANSCRIPTION σ Sigma For initiation (recognize
T promotor)
Transcription is synthesis of RNA from DNA. Enzyme
H
involved is RNA Polymerase. Unlike DNA Polymerase, it
E α2 β β’ ω makes the core enzyme and together
does not requires a primer and cannot do proofreading.
O represented as ‘E’ as these mainly help in elongation. Sigma
So transcription has a higher error rate than replication.
R subunit helps in initiation. E + sigma subunit makes the
Polymerase activity of this enzyme is also in 5’ → 3’ direction.
Y holoenzyme.
Holoenzyme of RNA Polymerase contains 5 different subunits
α2 β β’ ω σ
RNA Polymerase Types  TATA box → known as TATA because there are many
T and A in this (easy to do unwinding as compared to
Prokaryotes have a single RNA Polymerase GC rich region). It is present at -10 position on DNA. 329
But there are many types in eukaryotes: In prokaryotes, it is known as Pribnow box whereas
in eukaryotes, TATA box is known as Hogness box.
Eukaryotic RNA polymerase RNA synthesized  –35 sequence → only in prokaryotes, 35 base pairs

CHAPTER 13  REPLICATION, TRANSCRIPTION, TRANSLATION

Type I All rRNA except 5s rRNA upstream to the transcription start site.
Type II mRNA, mi-RNA, inc-RNA, few
 CAAT box or –75 sequences → only in eukaryotes
snRNA and snoRNA
(present 75 base pairs upstream to the transcription
start site).
Type III 5s rRNA, tRNA, few snRNA
and snoRNA →B y convention, the regulatory sequences are present on coding
strand in 5’ → 3’ sequence.
Mitochondrial RNA Polymerase Mitochondrial RNA
→ Promoter present upstream to gene
→ Promoter present on DNA template strand.
Template and Non-Template Strand
Both strands can serve as template for transcription but for
a given gene, only one serve as template. So each time one is
template strand and other is non template strand, depending
on the gene.

Fig. 13.6: The first base at the transcription start site is given
+1 number. There is no base which is designated as “0”. Any
base towards the 5’ end or to the left of transcription start site is
assigned – (negative) number or called upstream. Base towards
the 3 ‘ end or to the right of transcription start site is assigned +
(positive) number, or called downstream.
zz Elongation: Substrate for this enzyme is ribonucleotide
triphosphates. Ribonucleoside monophosphate is added
and pyrophosphate is released. So two high energy
phosphates are used to add one ribonucleotide
zz Termination:
 Rho (ρ) independent: A hairpin structure formed at
the end which helps in separation of new RNA from
DNA
H igh R eturn  Rho dependent: A protein ‘rho’ is required, which
uses ATP and has helicase activity. This protein
zz Template strand → also known as antisense/non-coding/
release RNA at termination site.
minus strand
zz Non template strand → also known as sense/coding/plus
strand (Direction and codons of new RNA match with this
strand)
Enhancers of Transcription
zz They increase the rate of initiation
zz They lie on same chromosome but they may lie on either
Steps of Transcription strand of DNA
zz They may be located either upstream or downstream to T
zz Initiation: There occurs binding of RNA Polymerase with
the transcription start site H
the promotor region. These promotor sequence helps in
initiation, but not transcribed. These sequences are : zz May lie close to or away from promotor. E
O
R
Y
 for energy). This is done for all the mRNAs except for
the mRNA of histone proteins. Number of ‘A’ added
330 range from 40 to 200. This tail stabilizes the mRNA by
preventing it from the attack by 3’ exonucleases and also
help the mRNA exit from nucleus. In cytoplasm, this Poly
A tail is shortened.
CRO BIOCHEMISTRY

U7-snRNA is involved in the modification of 3’ end of

Histone mRNA (Poly A tail not added in this case)
Splicing: It is done by U1, U2, U4, U5 and U6 snRNA.
3. 
They are rich in Uracil (that’s why alphabet ‘U’ is used).
Splicing is explained in detail in ribozyme topic.


Fig. 13.7: Enhancers of transcription

A dditional E dge
Inhibitors of Transcription
� Actinomycin D � Rifampicin � α- Amanitin
A dditional E dge
Alternate Splicing: Alternative splicing, or differential splicing, is
a regulated process during gene expression that results in a single
Post Transcriptional Modifications- Occur in gene coding for multiple proteins. In this process, particular exons
Nucleus of a gene may be included within or excluded from the final,
processed messenger RNA (mRNA) produced from that gene.
rRNA and tRNA of both prokaryotes and eukaryotes are Alternative use of splice sites leads to different combination of
obtained from post transcriptional modification of a single exons which in turns results in variable number of proteins from
pre- rRNA and pre-tRNA , by their cleavage by ribonucleases. the same mRNA.
tRNA is further modified differently in different species. CCA
sequence is added at the 3’ end of tRNA by enzyme nucleotidyl
transferase and base modification is done. There is not much
post transcriptional modification occurring in the mRNA of
prokaryotes. But eukaryotic mRNA is extensively modified.
Three post transcriptional modifications occur in all the
eukaryotic mRNA. These are also known as RNA Processing.
These are cap at 5’ end, removal of introns and a tail at the 3’
end.
1.  Cap: This is the first modification done. 7-Methyl
Guanosine cap is added at the 5’ end of mRNA by enzyme
Guanylyl Transferase. Methyl group is donated by SAM
(S-Adenosyl Methionine) by enzyme Guanine-7-Methyl
T Transferase. This cap facilitate initiation of translation
H and stabilize mRNA by preventing it from attack by 5’
E exonucleases. Cap is attached to 5’ end of mRNA by an
O unusual 5’ 5’ triphosphate linkage.
R 2.  Tail: At 3’ end, a Poly A tail is added. Poly A tail is not 4. There is one special post transcriptional modification
Y transcribed from DNA, but it is added by enzyme which occurs only at few places and it is known as
Polyadenylate Polymerase (uses ATP as substrate, not Differential RNA
 Processing/RNA Editing/Chemical modification of RNA: RNA Editing is a mechanism to produce a diverse set of
proteins from a limited set of genes.
331

CHAPTER 13  REPLICATION, TRANSCRIPTION, TRANSLATION

Fig. 13.8: Apo B100 gene present in both liver and intestinal cells. In intestinal cell, gene is Apo B100 gene but protein synthesized is Apo
B 48. This is because of RNA editing.

Type of RNA Location of post transcriptional

modification
mRNA In nucleus (but methyl group on guanosine is
added by SAM in cytoplasm)
tRNA In nucleus (but amino acid is attached in
cytoplasm)
rRNA In nucleolus

Protein Motifs and Response Elements

The binding between DNA and regulatory protein has to be Fig. 13.9: Contd...
very specific as this will regulate many genes. So the areas on
DNA and protein are very well specified to make this gene Types of Protein Motifs
regulation very specific. zz Helix turn helix: regulate genes of development
Protein motif is that modified portion of a regulatory protein zz Zinc finger: steroid hormone receptors T
which binds to response element on DNA. zz Helix loop helix: regulate immune system genes H
Response element is a DNA sequence which gives a site of zz Leucine zipper: regulate cell division
protein binding. This sequence gives response to a particular
E
regulatory protein. At this site of DNA, protein motif binds. O
R
Y
332
CRO BIOCHEMISTRY


Fig. 13.9: Protein motifs

OPERON MODEL normally gets energy from glucose as glucose is mostly

present in the environment of E. coli. There is no need for
zz Operon: A functioning unit of DNA containing a this bacterial cell to use lactose. So these genes (which
cluster of genes, under the control of a single promoter. synthesize enzymes of lactose metabolism) are inhibited
Operons are usually found in prokaryotes. Lac operon is (repressed). But if required, these genes can be activated
a negatively controlled inducible operon. Tryptophan (induced).
operon is a negatively controlled repressible operon.
zz O-site: Operator – lies downstream to promotor. Rep- How these Genes are Inhibited? (Fig. 13.10)
ressor tetramer binds here
zz CAP site: lies upstream to promotor. CAP-cAMP complex  ac I is a house keeping gene, i.e. it is always active. So, this
L
binds here gene always produces its protein i.e. repressor tetramer. This
zz Repressor Tetramer: binds to operator repressor tetramer goes and binds with operator site on
zz Inducer: Lactose (it can bind repressor tetramer and can DNA. Upstream to operator, promotor is present where RNA
remove repressor tetramer from operator site) Polymerase binds. Repressor tetramer gives hinderance in
the path of RNA Polymerase, which has to move downstream
to transcribe these genes for lactose metabolism. So, these
Lac Operon/Lactose Operon
genes are inhibited.
zz Occurs in E. coli. There are 3 genes which synthesize
T enzymes of lactose metabolism. But bacterial cell
H
E
O
R
Y
 333

CHAPTER 13  REPLICATION, TRANSCRIPTION, TRANSLATION

Fig. 13.10: Z,Y and A are structural genes that give rise to enzymes of lactose metabolism. Lac I gene has its own promotor and is not a
part of Lac Operon (this thing is not shown in diagram). CAP-cAMP complex is a trans acting protein.

How these Genes are Activated? zz Lactose is converted to its isomer – Allolactose,
zz These genes are activated if glucose decreases in the which binds to repressor tetramer and removes
environment of E. coli and if lactose is present. So when repressor tetramer from operator site (by changing its
glucose decreases then cAMP is increased. This cAMP conformation). So, there is no hinderance of repressor
binds with CAP (Catabolic Activator Protein also known tetramer and RNA Polymerase is active. Now now genes
as CRP i.e. cAMP Regulatory Protein) and makes cAMP- are active and transcription occurs and lactose can be
CAP complex. This complex binds to CAP site (also used by bacterial cell.
known as cAMP Response Element i.e. CRE) on DNA.
This binding activates RNA polymerase.

T
H
E
O
•• Lac Z – codes for Beta Galactosidase – breaks Lactose to Galactose and Glucose R
•• Lac Y – codes for Permease – facilitate permeation of Lactose into the cell Y
•• Lac A – codes for Transacetylase – acetylates Lactose
 Situation
334 I Glucose decreased → activate enzyme
Transcription occurs
Lactose present → remove repressor tetramer
II Glucose decreased → activate enzyme
Lactose absent → repressor tetramer not removed No Transcription
CRO BIOCHEMISTRY

III Glucose present → enzyme inactive/very slow

No or very slow Transcription
Lactose present → remove repressor tetramer
IV Glucose present → enzyme inactive /very slow
No Transcription
Lactose absent → Repressor tetramer not removed

CODONS Other features of genetic code are:

Codons are nucleotide triplets. That means three nucleotides • Unambiguous i.e. each codon is specific for its amino acid
make one codon. Total 4 nucleotide bases are used to produce e.g. AUG will always code for methionine
three base codons i.e. A, U, C, G. • Universal (genetic code has been well conserved during


So total number of codons possible is: 4 × 4 × 4 = 64 (Three evolution with only slight differences)
are stop codons) • Non overlapping and commaless means that the codon is
read from a fixed starting point as a continuous sequence
Stop Codons of bases, taking three at a time, without any punctuation
between codons. e.g. AUCGGGACUGCA will be read as
Stop Codon Other Name Codes for AUC GGG ACU GCA
UAA Ochre —
Codon Recognition by tRNA
UAG Amber Pyrrolysine (22nd Amino Acid
zz Antiparallel binding between codon and anticodon:
UGA Opal/Umber Selenocysteine (21st Amino Acid) mRNA codon is read 5’ → 3’ by an anticodon pairing in
These stop codons do not code for any amino acid. There are opposite (3’ → 5’) direction.
no tRNA with anticodon complementary to stop codon.
So for amino acids, number of codons present are 61. There
are 20 amino acids which are having 61 codons. So average is
that each amino acid is having 3 codons. (this is just an average.
It can be less than 3 codons or more than 3 codons.) This is a
property of codons known as degeneracy or redundancy of
codons. i.e. each amino acid has more than one codon.

H igh R eturn
zz 2 Amino acids do not show degeneracy:
 Methionine
 Tryptophan
zz Means they have only 1 codon

zz Wobble hypothesis: Number of codons are 61 for amino

acids but tRNA are not 61. This hypothesis describes
how a tRNA can recognize more than one codon for a
specific amino acid. Wobble hypothesis says that codon-
anticodon pairing follows traditional Watson Crick rules
(A pairs with U, G pairs with C) for the first two bases, but
this rule can be less stringent for the last base.

T
H
E
O
R
Y
TRANSLATION
Location of translation is Ribosomes (can be free or bound)
335
Free Ribosomes/ Cytosolic Ribosomes Bound Ribosomes/ Rough Endoplasmic Reticulum (RER)
Free ribosomes present in cytoplasm (not attached to ER) Ribosomes attached to cytosolic side of ER

CHAPTER 13  REPLICATION, TRANSCRIPTION, TRANSLATION

Translation: mRNA is translated in 5’ to 3’ direction and This enzyme (amino acyl tRNA synthetase) is the only
protein is synthesized from amino terminal (N- terminal) to point of proofreading in translation. So this enzyme is
carboxy terminal (C- terminal). responsible for fidelity of protein synthesis.
All the amino acids should be present at the time of translation. Two functions of amino acyl tRNA Synthetase enzyme:
Even if one amino acid is missing, then translation will stop at zz Synthesis of amino acyl tRNA
that codon, specifying for that amino acid. So all the essential zz Proofreading or editing activity which can remove a
amino acids are required in sufficient quantities in diet for wrong amino acid if attached and replace it with a correct
protein synthesis to be continued. amino acid.

Steps of Translation Initiation

Three steps of Initiation, Elongation and Termination occurs Initiation codon is AUG, which codes for Methionine in
in translation. But Activation of Amino acid occur before all eukaryotes and N-formyl Methionine in prokaryotes and
the steps. in mitochondria. In both prokaryotes and eukaryotes, this
N-terminal Methionine is usually removed before translation
Activation of Amino Acid or Amino Acylation or is completed.
Charging of tRNA

Each of the 20 amino acids are recognized by a family of

20 specific Aminoacyl-tRNA synthetase. Each member of this
family recognize a specific amino acid and all the anticodons
of tRNA which correspond to that one amino acid. All these Fig. 13.11: Initiation in Eukaryotes (ATP and GTP required)
tRNAs are known as Isoaccepting tRNA. E.g. see figure below
with leucine amino acid. Initiation in Prokaryotes (See Figure)
Shine Dalgarno sequence (SD sequence) is present in
prokaryotes on mRNA for initiation. This is purine rich
sequence which has complementary bases with 16S rRNA
(present in 30S subunit of ribosome). First pre-initiation
complex is formed by taking 30S ribosomal subunit, mRNA and T
N-formyl methionine attached to tRNA. Codon of mRNA and H
anticodon of tRNA joins due to complementary base pairing. E
mRNA joins with 30S ribosome due to complementary base O
pairing between Shine Dalgarno sequence and 16S rRNA. R
Y
 Then initiation complex is formed when 50S subunit of ribosome also joins. Now here two sites are formed:
zz P site: always lies towards left side. ‘P’ means Polypeptide site i.e. polypeptide will be released from this site
336 zz A site: always lies towards right side. ‘A’ is Acceptor site, i.e. all amino acids are accepted here. EXCEPT – 1st Methionine
CRO BIOCHEMISTRY


T
H Fig. 13.11: Translation (Initiation, Elongation and Termination) in Prokaryotes
E
O
R
Y
Elongation Termination
Elongation Factors When a stop codon is encountered on mRNA, then no amino
acid will come. Instead releasing factors approach i.e. RF1,
337
Prokaryotic Eukaryotic Function RF2 and RF3. There are three releasing factors in prokaryotes
Factor Factor but only a single eukaryotic releasing factor (eRF) in

CHAPTER 13  REPLICATION, TRANSCRIPTION, TRANSLATION

EF-Tu eEF1α A delivery of Aminoacyl RNA to
eukaryotes.
ribosome Releasing factors do not release the polypeptide. The
name ‘releasing’ is a misnomer. They only recognize the stop
EF-Ts eEF1βy Aiding the recycling of EF-Tu or
codon.
eEF1α
EF-G eEF2 Translocation •• Peptidyl Transferase releases the polypeptide from P-site.
So peptidyl transferase has a role in elongation as well as in
Addition of amino acids to the carboxy end of growing chain termination.
occurs. •• GTP is required for termination in both eukaryotes and
Prokaryotes
Now second amino acid with its tRNA enters A site. For
entry into A site, one GTP is used. 23S & rRNA has ribozyme
activity i.e. Peptidyl Transferase, which will cut the amino Factors in Translation in Prokaryotes and
acid at P site, brings it to A site and make a peptide bond. Eukaryotes
This shifting of amino acid from P site to A site is known
as Transpeptidation. No ATP or GTP is used by Peptidyl Prokaryotes Eukaryotes
Transferase. Then Translocation occurs, which is movement
of ribosome on mRNA (Ribosome moves three nucleotides Initiation IF 1,2 , 3 eIF 1, 2, 3, 4F
towards the 3' end of mRNA). Translocation requires one Elongation EF- Tu, Ts, G eEF 1α, 1β γ, 2
GTP and Elongation factor – G (EF-G). Now the peptide is Termination RF 1, 2, 3 eRF
shifted to P site due to the movement in translocation and
new A site is formed. Next amino acid with tRNA can come to Q. How many ATP and GTP used to add 1 amino acid
this new A site formed. This way elongation continues until a in a growing polypeptide chain?
termination codon is encountered. T A. Two ATP and two GTPs are used to add one amino
H acid in a growing polypeptide chain
I
N •  Two ATPs are used during the activation. One GTP
K is used for entry of amino acid at A –site. One GTP
is used during translocation.
• Additional ATP and GTP are required for initiation
in eukaryotes, whereas an additional GTP is
required for termination in both eukaryotes and
prokaryotes. As initiation and termination is a
one time process so, energetics of these are not
considered here.

Co-translational Modification
Occurs in case of Selenocysteine and Pyrrolysine (21st and
22nd amino acids)

Post Translational Modifications

zz Cleavage of a sequence in protein
zz Covalent modifications → e.g. Phosphorylation, Gly-
cosylation, Hydroxylation, Acetylation, Vitamin K
Fig. 13.12: Polysome or Polyribosome dependent Carboxylation of Glutamate, Biotin covalently
For those mRNA which are lengthy, more than one bound to carboxylases. T
ribosome at a time can do translation. This is known as Polysome zz Glycosylation is the most frequent post translational H
or Polyribosome i.e. one mRNA and many ribo-somes. modification of protein. E
O
H igh R eturn R
zz Small ribosome subunit → binds mRNA and determines Y
accuracy of translation by correct base pairing
zz Large ribosome subunit → catalyse formation of peptide bond
 Pearls of the Chapter
338 zz Prokaryotes have circular double stranded DNA whereas eukaryotes have linear double stranded DNA
zz Helicase, Topo-isomerases and SSBs → together known as Unwinding Proteins
zz DNA dependent RNA Polymerase in replication is Primase
CRO BIOCHEMISTRY

zz Telomerase is not a ribozyme because here RNA is not used as enzyme. It is used as a template for DNA synthesis.
zz Proofreading occurs in S-phase
zz Most repairs occur in G1 phase of cell cycle. But mismatch repair occurs in G2 phase.
zz Mitochondrial DNA: Its same like prokaryotic DNA i.e. circular double stranded.
zz Ribozymes are rRNA, splicing ribozyme and Ribonuclease P
zz Ribozymes are rRNA, splicing ribozyme and Ribonuclease P
zz Splicing ribozyme → self splicing introns and spliceosomal introns
zz Lariat formation occurs in group II self splicing introns and spliceosomal introns
zz snRNA which takes part in splicing → U1, U2, U4, U5, U6
zz Telomerase and RNase H are not Ribozymes


zz Transcription is synthesis of RNA from DNA

zz 7-Methyl guanosine cap in added at the 5' end of mRNA and poly A tail added at 3' end
zz UAA, UAG, UGA are the stop codons
zz Activation of amino acid in translation occurs by enzyme amino acyl tRNA synthetase

T
H
E
O
R
Y
 Multiple Choice Questions 339

DNA Replication 10. UV radiation:  (PGMEE 2012-13)

1. Telomerases are: (PGMEE 2012-13; 2014) a. Purine dimers
a. DNA dependent DNA Polymerase b. Stimulates formation of Pyrimidine dimers
b. RNA dependent DNA Polymerase c. Prevents formation of Pyrimidine dimers
c. DNA dependent RNA Polymerase d. None
d. RNA dependent RNA Polymerase 11. Which of these is mismatch repair defect?
2. Telomerase has action of:   (PGMEE 2013)
 (Recent Question June 2018, 2015) a. Burkitt’s lymphoma
a. DNA Polymerase I b. Xeroderma pigmentosa
b. DNA Polymerase II c. Hereditary non- polyposis colorectal cancers
c. DNA Polymerase III d. None
d. Reverse Transcriptase 12. Which of the following is a DNA repair defect?
3. TRUE about telomerase is:   (PGMEE 2010)
 (PGI May 2018, JIPMER May 2018) a. Bloom syndrome
a. DNA dependent RNA polymerase b. Aplastic anemia
b. RNA dependent DNA polymerase c. Tuberous sclerosis
c. Reverse transcriptase enzyme d. Incontinentia pigmenti
d. Increased telomerase activity is seen in somatic cells 13. Primers are removed by all EXCEPT:
e. Telomerase increases the longevity of cells a. Delta Polymerase b. RNase H1
4. Enzyme responsible for unwinding of DNA is: c. FEN1 d. None
 (PGMEE 2013, 2011, 2015) 14. DNA Polymerase has: (PGI)
a. DNA primase b. Helicase a. 5’→ 3’ Polymerase activity
c. Ligase d. DNA polymerase b. 3’→ 5’ Polymerase activity
5. DNA replication occurs in which phase of cell cycle? c. Endonuclease activity M
 (PGMEE 2011) d. 3’→ 5’ Exonuclease activity C
a. G1 phase b. S phase e. 5’→ 3’ Exonuclease activity Qs
c. G2 phase d. M phase 15. In SCID, what type of DNA repair is defective?
6. Function of exonuclease: (PGMEE 2015)
Ans.
 (AIIMS Nov 2017)
1. b
a. Termination a. Double strand break repair
2. d
b. Proof reading b. Nucleotide excision repair 3.  ,
b
c. Chain elongation c. End joining repair c,e
d. Transketolase d. Mismatch repair 4. b
7. Function of DNA ligase: (PGMEE 2015) 16. A child presents with skin bullae on sun exposure. 5. b
a. DNA polymerization Irregular dark spots on skin are also found. The 6. b
b. Cutting of DNA at specific site defect is of which of the following mechanism: 7. c
c. To seal and nick okazaki fragments  (AIIMS November 2012) 8. d
d. Preventing negative supercoil a. Thymine dimers b. Trinucleotide repeats 9. a
8. What are okazaki fragments:  (PGMEE 2015) c. Sugar changes d. DNA methylation 10. b
a. Long pieces of DNA on leading strand e. Nucleotide excision repair 11. c
b. Short pieces of DNA on leading strand 17. ATM and Rad51 proteins repairs double stranded 12. a
c. Long pieces of DNA on lagging strand breaks by: 13. d
d. Short pieces of DNA on lagging strand a. Homologous recombination and repair 14. a,c,
9. All of the following statements are true EXCEPT: b. Non-homologous end joining d,e
a. Heterochromatin is the relatively uncondensed c. Base excision repair 15. c
portions of DNA d. Nucleotide excision repair 16. a,e
b. Euchromatin represents the transcriptionally active 18. Which is incorrect about DNA Polymerase? 17. a
portions of the DNA  (Recent Pattern Jan 2018) 18. a
c. Histones are positively charged because of the high a. Not required in bacteria
content of basic amino acids in them b. Role in primer removal
d. Methylation of DNA will increase the condensed c. Fill gaps between okazaki fragments
portions d. Involved in DNA replication
 19. TRUE about DNA Polymerase III: 30. A segment of eukaryotic gene that is not represented
 (Recent Question Jan 2018) in the mature mRNA is known as: 
340 a. Required for translation  (PGMEE 2007)
b. Has DNA repair function a. Intron b. Exon
c. Forms okazaki fragments & needs RNA primer c. Plasmid d. TATA box
d. Has no proof-reading activity 31. Splicing is done by: (PGMEE 2009)
20. Klenow fragment is formed by loss of fragment a. tRNA b. mRNA
having which activity? (AIIMS May 2018) c. snRNA d. rRNA
a. 5’ → 3’ polymerase 32. Intron is NOT found in which DNA:  (PGMEE 2009)
b. 3’ → 5’ exonuclease a. Nuclear DNA
c. 5’ → 3’ exonuclease b. Mitochondrial DNA
d. 3’ → 5’ polymerase c. B DNA
21. Klenow fragment does not contain: d. Z DNA
a. 5’→ 3’ polymerase 33. Post transcriptional modifications include all
b. 3’→ 5’ exonuclease EXCEPT: (PGMEE 2013-14)
c. 5’ → 3’ exonuclease a. 5’ capping b. 3’ polyadenylation
d. None c. Splicing d. Glycosylation
22. All are TRUE statements regarding Okazaki frag- 34. Which arm binds the aminoacylt RNA to the
ments EXCEPT:  (PGI May 2018) ribosomal surface? (PGMEE 2013-14)
a. Requires DNA polymerase a. Acceptor arm b. Anticodon arm
b. Formed on leading strand c. DHU arm d. Pseudouridine arm
c. Formed on lagging strand 35. Which of the following DOES NOT need a primer?
d. Requires helicase for opening a. DNA Polymerase I b. RNA Polymerase
e. Requires RNA primer c. DNA Polymerase II d. DNA Polymerase III
23. Which is the most processive DNA polymerase? 36. TRUE about RNA Polymerase is:
a. DNA Polymerase I b. DNA Polymerase II a. Requires primers
c. DNA Polymerase III d. None b. RNA dependent DNA Polymerase
24. Which of the following is NOT TRUE about eukaryotic c. No proof reading activity
DNA ligase? d. Recognizes Shine dalgarno sequence
M
a. Catalyses the formation of phosphodiester bond 37. The base sequence of the strand of DNA used as the
C
b. NAD+ is energy source template for transcription has the base sequence
Qs
c. ATP is the energy source GATCTAC. What is the base sequence of RNA
Ans. product? (Recent Question 2016)
d. Can act only on dsDNA
19. c 25. Defect in Ataxia Telangiectasia (PGMEE 2015) a. CTAGATG b. GTAGATC
20. c a. Defective translation c. GAUCUAC d. GUAGAUC
21. c b. Defective DNA replication 38. 7-Methyl Guanosine is added as a cap on mRNA. This
22. b methyl group is donated by:
c. Defective triphosphate
23. c a. S-Adenosyl methionine (SAM)
d. Defective DNA repair
24. b b. Methenyl THF
25. d Transcription c. Tetra hydro Folate (THF)
26. a
26. Signal recognition particle contains: d. Vit B12
27. a
a. 7S rRNA b. 10S rRNA 39. Apo B48 and Apo B100 are synthesised from same
28. b
c. 28S rRNA d. 60S rRNA mRNA due to difference in:
29. a
27. Which of the following is ribozyme? a. Splicing and chemical modification
30. a
31. c  (PGMEE 2012-13) b. Splicing
32. b a. Peptidyl Transferase b. Primase c. RNA processing
33. d c. Elongation factor 2 d. RNA Polymerase d. All
34. d 28. Function of sigma factor in transcription: 40. RNA Polymerase III synthesizes:
35. b  (PGMEE 2015) a. Fragment 28S of rRNA
36. c a. Termination of transcription. b. Fragment 23S of rRNA
37. d b. Binding of RNA polymerase to promoter region c. Fragment 5S of rRNA
38. a c. Elongation of RNA primer d. tRNA
39. c d. All of the above e. mRNA
40. c 29. DNA replication and transcription occurs in which 41. How many nucleotides does tRNA molecule cons-
41. b direction? (PGMEE 2015) titute?
a. 5’ → 3’ b. 5’→ 5’ a. 52-64 nucleotides b. 74-95 nucleotides
c. 3’→ 5’ d. 3’→ 3’ c. 96-110 nucleotides d. >110 nucleotides
42. Which of the following is a post transcriptional 53. Which of the following DOES NOT favor permissive
modification of tRNA? euchromatin due to changes occurring at cytosine
a. Poly A tail at the 3’end residues at CpG islands in DNA? (AIIMS May 2018) 341
b. 7-methyl guanosine cap at the 5’end a. Methylation b. Phosphorylation
c. CCA at 3’end c. Alkylation d. Sumoylation
d. CCA at 5’end 54. Pribnow Box in eukaryotes is equivalent to:
43. Which termination protein recognizes the signal for a. Hogness box b. GC box
the termination of synthesis of RNA molecule? c. CAAT box d. None
a. Alpha (a) factor b. Sigma (s) factor 55. The segment of DNA which resembles mRNA except
c. Rho (r) factor d. None of these for T/U is called:
44. The bacterial ribosome contains: a. Coding strand
a. 2 subunits b. 3 subunits b. Initiator Methionine domain
c. 4 subunits d. 5 subunits c. Translation unit
45. Which of the following mechanism is responsible for d. Template strand
multiple proteins from a single gene? 56. A mutation in which of the following sequence in
a. RNA interference eukaryotic mRNA will affect the process by which
b. mRNA capping poly-A tail is added to 3’end of mRNA?
c. Mutations in the promoter of the gene a. CCA b. CAAT
d. Alternate Splicing c. AAUAAA d. GU…A…AG
46. Initiation and termination of a sequence of intron: 57. Promotor is recognized by eukaryotic transcription
a. GU, AG b. AG, GU factor:
c. GA, GU d. UG, AG a. Sigma b. Rho
47. Which is a template-independent enzyme acting on c. TFIID d. Pribnow box
DNA? 58. The cis-element which typically resides adjacent to
or overlaps with many prokaryotic promoters is:
a. Poly A Polymerase
a. Regulatory Gene b. Operator
b. CCA adding enzyme
c. Repressor d. Structural gene
c. Terminal deoxynucleotidyl transferase
59. When will the ZYA region of lac operon be maximally
d. Polynucleotide kinase
expressed?
48. Which of the following is not a ribozyme?
a. If Cyclic AMP levels are low M
 (Recent Question Jan 2019)
b. Glucose and Lactose are both available C
a. Transpeptidase
c. CAP site is occupied Qs
b. Ribonuclease
d. The attenuation stem-loop is able to form Ans.
c. Peptidyl Transferase 60. The concept of Lac operon was first described by
d. Poly A Polymerase 42. c
Francois Jacob and Jacques Monod in E.coli. CAP in
49. Reverse transcription causes synthesis of: 43. c
Lac operon is an example of: (PGMEE 2013-14)
 (Recent Question June 2018) 44. a
a. Positive regulator
a. RNA from DNA template 45. d
b. Attenuation 46. a
b. DNA from RNA template c. Constitutive expression
c. RNA from RNA template 47. c
d. Negative Regulator 48. d
d. DNA from DNA template
49. b
50. Sigma (s) subunit is present in: Translation
50. b
 (Recent Question June 2018) 61. Termination of translation is caused by all EXCEPT: 51. a
a. 80S subunit  (PGMEE 2013) 52.  ,
b
b. RNA polymerase a. 48S complex b. UAA d,e
c. DNA polymerase II c. RF-1 d. Peptidyl Transferase 53. a
d. DNA polymerase III 62. Codon is a sequence of three nucleotides present in: 54. a
51. Which of the following does not require 5’ capping?  (PGMEE 2010) 55. a
 (AIIMS May 2018) a. rRNA b. tRNA 56. c
a. tRNA of alanine b. U2 snRNA c. mRNA d. DNA 57. c
c. mRNA for histone d. siRNA 63. Poly A translates into: (PGMEE 2009) 58. b
52. TRUE statement regarding tRNA is: (PGI May 2018) a. Poly proline b. Poly lysine 59. c
a. Contains codon c. Poly alanine d. Poly glycine 60. a
b. Contains anti – codon 64. Chemical nature of enzyme peptidyl transferase is: 61. a
c. Contains blunt ends  (PGMEE 2009) 62. c
d. Acts as acceptor for amino acids in protein synthesis a. Carbohydrate b. Lipid 63. b
e. Gets attached to ribosomes c. Nucleic acid d. Protein 64. c
 65. Degeneracy of codon means:  75. Which of the following is/are true about translation?
 (PGMEE 2016-17; 2008) a. N-formyl methionine is the first amino acid in
342 a. More than one codon for a single amino acid prokaryotes
b. More than one amino acid for a single codon b. Uses energy in the form of GMP
c. No punctuation in codons c. Elongation factor EF-1 and EF-2 used in prokaryotes
d. Termination of protein synthesis d. Elongation factor P is used in eukaryotes
66. Protein synthesis occurs in: (Recent Question 2016) e. Three initiation factors are required in prokaryotes
a. RER (Rough endoplasmic reticulum) 76. TRUE about genetic code:
b. SER (smooth endoplasmic reticulum) a. Follows mendelian law
c. Both a and b b. It is total number of chromosomes in the body
d. Nucleus c. It is nucleotide sequence which codes for amino
67. High energy phosphates required to form one peptide acids
bond: (Recent Question 2016) d. It codes for DNA.
a. 2 b. 4 77. How many base pairs are encoded by a prokaryotic
c. 6 d. 8 polypeptide of 250 amino acids?
68. Aminoacyl tRNA is required for all EXCEPT: a. 250 bp b. 500 bp
 (PGMEE 2013) c. 750 bp d. 1000 bp
a. Lysine b. Cysteine 78. Which of the following is NOT a synonymous codon
c. Methionine d. Hydroxyproline pair?
69. A tRNA molecule that is supposed to carry cysteine a. CAU and CAC
(tRNAcys) is mischarged. So that it actually carries b. AUU and AUC
alanine (ala-tRNAcys). Assuming no correction c. AUG and AUA
occurs, what will be the fate of this alanine residue d. AAU and AAC
during protein synthesis? 79. Anticodon 5’-I-G-C-3’ can recognize all of the follo-
a. It will be incorporated into a protein in response to wing codons EXCEPT:
an alanine codon. a. GCA b. GCG
b. It will be incorporated into a protein in response to a c. GCC d. GCU
cysteine codon. 80. Initiator tRNA during translation binds to which
c. It will remain attached to the tRNA as it cannot be site?
M used for protein synthesis. a. P site b. A site
C d. It will be chemically converted to cysteine by c. E site d. None
Qs cellular-enzymes 81. Which of the following is NOT TRUE for P bodies?
Ans. 70. 43S Preinitiation complex include all EXCEPT: a. These are processing bodies
65. a a. IF3 b. IF1A b. They are the sites of translation repression
66. a c. IF2B d. IF4F c. They are the sites of protein synthesis
67. b 71. The hydrolytic step leading to the release of a d. They are present in the cytoplasm
68. d polypeptide chain from a ribosome is catalyzed by 82. In prokaryotic translation, mRNA binds with
69. b which of the following? (Recent Question 2016) ribosome by interaction of Shine-Dalgarno sequence
70. d a. Dissociation of ribosomes of mRNA with the:
71. b b. Peptidyl transferase a. 5S rRNA
72. d c. Release factors b. 5.5S rRNA
73. d d. Stop codons c. 16S rRNA
74. a,c 72. Silent mutations occurs because codon is: d. 23S rRNA
75. a,e a. Non overlapping b. Commaless 83. Which one of the following inhibitor of translation
76. c c. Universal d. Degenerate causes premature chain termination?
77. c 73. Which of the following enzyme (s) participate in a. Erythromycin
78. c protein synthesis?  (PGI Nov 2014) b. Puromycin
79. b a. DNA ligase c. Cycloheximide
80. a b. DNA Helicase d. Diptheria toxin
81. c c. Peptidase 84. A 20 year old boy with microcytic anaemia is found to
82. c d. Peptidyl transferase have abnormal form of β-globin (haemoglobin con-
83. b e. DNA polymerase stant spring) that is 172 amino acids long, rather than
84. d 74. Component of 50S subunit is/are:  (PGI Nov 2011) 141 found in normal protein. Which of the following
a. 23S b. 28S point mutations results in this abnormality?
c. 5S d. 5.8S a. CGA → UGA b. GAU → GAC
e. 16S c. GCA → GAA d. UAA → CAA
 Answers with Explanations
343
1. Ans. (b) RNA dependent DNA Polymerase 9. Ans. (a) Heterochromatin is the relatively uncon-
densed portions of DNA

CHAPTER 13  REPLICATION, TRANSCRIPTION, TRANSLATION

Refer ans 3.

2. Ans. (d) Reverse Transcriptase [Ref: Harper 30th/e pg. 383]

Refer ans 3. •• Heterochromatin is the condensed DNA which is
transcriptionally inactive. However, euchromatin
3. Ans. (b); (c); (e) is the uncondensed, loose DNA, which is trans-
criptionally active.
[Ref: Harper 30th/e pg. 374,474] •• Methylation of DNA leads to gene inactivation or
•• Q1,2 and 3 Telomerase is the enzyme present in heterochromatin formation.
germ cells and stem cells, which prevent telomere •• Histones are positive due to basic amino acids. DNA
shortening in these cells. is negative due to phosphates.
•• This enzyme has a protein with RNA attached in its
structure. This RNA act as template on which DNA is 10. Ans. (b) Stimulates formation of Pyrimidine dimers
synthesized, so it is RNA dependent DNA polymerase [Ref: Robbins 9th/e pg. 324]
or reverse transcriptase.
•• It is responsible for the longevity of cells and is UV radiation exposure leads to the formation of
present in germ and stem cells. Its activity increase in Thymine-Thymine dimers. This is repaired by
cancerous cells and decreases during ageing. nucleotide excision repair during G1 phase of cell cycle.
Defect in this repair leads to Xeroderma Pigmentosa,
4. Ans. (b) Helicase which increases the chances of skin malignancy.
[Ref: Harper 30th/e pg. 474] 11. Ans. (c) Hereditary non- polyposis colorectal
Helicase is the first enzyme in DNA replication which cancers
causes strand separation (unwinding). It uses ATP and
[Ref: Robbin’s 8th/e pg. 275]
it creates positive supercoils.
Mismatch repair occurs during G2 phase of cell cycle.
5. Ans. (b) S phase Damage is a mismatched base. Defect in this repair
leads to HNPCC (Hereditary Non-Polyposis Colorectal
[Ref: Harper 30th/e pg. 371]
Cancer).
S means synthesis phase, in which synthesis of both
DNA and histones occur. 12. Ans. (a) Bloom syndrome A
N
6. Ans. (b) Proof reading [Ref: Harper 30th/e pg. 390]
S
Bloom Syndrome (BS) occurs due to defective homo- W
[Ref: Harper 30th/e pg. 383]
logous repair. E
Proof reading is done by enzyme DNA Polymerase II. R
This activity is 3’ → 5’ exonuclease activity. Repair is 13. Ans. (d) None
S
mostly endonuclease but sometimes it can be 5’ → 3’
[Ref: Harper 30th/e pg. 383]
exonuclease activity. WITH
During DNA synthesis, primers are RNA so they should
7. Ans. (c) To seal and nick okazaki fragments be removed. Primers are removed by RNaseH, FEN-1 E
and Delta Polymerase. So answer here is none. X
[Ref: Harper 30th/e pg. 383] P
DNA ligase joins DNA fragments by making 3’5’ 14. Ans. (a); (c); (d); (e) L
phosphodiester bond. It uses ATP. It acts only on lagging A
[Ref: Harper 30th/e pg. 383] N
strand. This enzyme is also used in repair.
DNA Polymerase has: A
8. Ans. (d) Short pieces of DNA on lagging strand •• 5’ → 3’ Polymerase activity (synthesis occurs in 5’ → T
3’ direction) I
[Ref: Harper 30th/e pg. 382] O
•• Endonuclease activity (Repair)
Okazaki fragments are short fragments of DNA •• 3’ → 5’ Exonuclease activity (Proofreading) N
synthesized in 5’ → 3’ direction on the lagging strand. •• 5’ → 3’ Exonuclease activity (Repair and Primer S
Later these fragments are joined by enzyme DNA ligase. removal).
 15. Ans. (c) End joining repair Klenow fragment is a large fragment produced by
Subtilisin mediated proteolytic cleavage of E.Coli
344 [Ref: Harper 30th/e pg. 390] DNA polymerase I. Proteolysis removes the 5’ → 3’
•• Non-Homologous End Joining Repair (NHEJ) defect exonuclease activity from N-terminal.
leads to SCID
•• Homologous Repair (HR) defect leads to Bloom 21. Ans. (c) 5’ → 3’ exonuclease
CRO BIOCHEMISTRY

syndrome, Werner syndrome, Breast cancer All prokaryotic DNA polymerases have 5’ → 3’
•• Nucleotide excision Repair (NER) defect leads to Polymerase (Synthesis) and 3’ → 5’ exonuclease
Xeroderma Pigmentosa (Proofreading) activity. Only DNA Polymerase I contain
•• Mismatch Repair (MMR) defect leads to Hereditary 5’ → 3’ exonuclease activity (primer removal activity).
Non- Polyposis Colorectal Cancer (HNPCC).
22. Ans. (b) Formed on leading strand
16. Ans. (a); (e)
[Ref: Harper 30th/e pg. 382]
[Ref: Harper 30th/e pg. 390] •• During DNA synthesis two strands are formed,
This picture is of Xeroderma Pigmentosa which occurs leading strand and lagging strand.


due to defect in Nucleotide Excision Repair. This repair •• Leading strand is continuous whereas lagging
normally occurs to correct Thymine. Thymine dimers strand is discontinuous and is formed of
which are formed due to UV light exposure. fragments of DNA. These fragments are known
as okazaki fragments.
17. Ans. (a) Homologous recombination and Repair •• Replication of DNA requires DNA polymerase
for the synthesis of these two strands.
[Ref: Harper 30th/e pg. 390]
•• Helicase is required for the unwinding of parent
ATM (Ataxia-Telangiectasia Mutated) DNA damage strand.
signal transducer is a protein kinase that phosphorylates •• RNA primers are also required for both strands.
the key proteins that initiate activation of DNA damage Leading strand requires only one primer
checkpoint. whereas multiple primers are required for the
Rad51 is Recombinase enzyme involved in homologous lagging strand.
recombination of DNA during double stranded break
repair. 23. Ans. (c) DNA Polymerase III

18. Ans. (a) Not required in bacteria [Ref: Harper 30th/e pg. 383]
•• Processive enzymes are those that remain bound to
[Ref: Harper 30th/e pg 383]
the end of the growing chain and keeps on adding
•• DNA polymerase I is not required in bacteria substrates. E.g. Glycogen Phosphorylase, DNA
A •• DNA Polymerase I RNA primers from both leading polymerases
N and lagging strand. •• Processivity of DNA Polymerase is an expression of
S •• It has 5’ → 3’ exonuclease activity, means it starts number of nucleotides added to the nascent chain
W cutting the primer from 5’ end and move in 5’→ 3’ before the polymerase disengages from the template.
E direction. •• Of all the prokaryotic polymerases, DNA Polymerase
R •• This same enzyme fills this gap with DNA but only on III catalyses the highest rate (20-50 nucleotides/sec)
the lagging strand. of chain elongation and is the most processive (0.5
S
Mb/cycle).
19. Ans. (c) Forms okazaki fragments & needs RNA •• Of all the eukaryotic polymerases, DNA polymerase d
WITH
primer is the most processive.
E
[Ref: Harper 30th/e pg 390]
X 24. Ans. (b) NAD+ is energy source
P •• DNA Polymerase III synthesizes both leading and
L lagging strands (Okazaki fragments). [Ref: Harper 30th/e pg. 383]
A •• Repair and Proofreading in prokaryotes is done •• NAD+ is the energy source for prokaryotic DNA ligase
N by enzyme DNA Polymerase II. In eukaryotes all •• ATP is the energy source for eukaryotic DNA ligase
A enzymes can do proofreading except alpha and beta Role of DNA ligase:
T polymerase. •• Involved in sealing the single stranded nick during
I replication and repair.
O 20. Ans. (c) 5’→ 3’ exonuclease •• In recombinant DNA technology, ligase is used to
N join two different DNA molecules.
[Ref: Lehninger principles of biochemistry, 7th ed., Pg
S 994 and Lippincott’s illustrated reviews 6th ed., Pg 74-75]
25. Ans. (d) Defective DNA repair •• Introns are only present in eukaryotic nuclear DNA.
They are not present in prokaryotes. Mitochondrial
[Ref: Harper 30th/e pg. 390] DNA is same like prokaryotic DNA i.e. it is circular 345
•• Ataxia Telangiectasia is due to defect in homologous double stranded and no introns.
DNA repair, which is one of the repair mechanism for
double strand breaks. 33. Ans. (d) Glycosylation

CHAPTER 13  REPLICATION, TRANSCRIPTION, TRANSLATION

[Ref: Harper 30th/e pg. 409]
26. Ans. (a) 7S RNA
•• Glycosylation is a post translational modification, not
[Ref: Harper 30th/e pg. 614] a post transcriptional modification.
•• 4.5S rRNA and 7S rRNA both are part of Single
Recognition Particle (SRP). SRP recognizes signaling 34. Ans. (d) Pseudouridine arm
amino acid sequence in the N-terminus of growing [Ref: Harper 30th/e pg. 365]
polypeptide chain. Upon signal recognition,
ribosome is attached to endoplasmic reticulum •• Pseudouridine arm of tRNA binds A-site of ribosome
so that the protein, made by ribosome, enters the and helps in initiation and elongation.
secretory pathway.
35. Ans. (b) RNA Polymerase
27. Ans. (a) Peptidyl Transferase
[Ref: Harper 30th/e pg. 434]
[Ref: Harper 30th/e pg. 71] •• DNA Polymerase needs RNA primer, which is
•• Ribozymes are Peptidyl Transferase, snRNA, Ribo- synthesized by Primase. Unlike DNA Polymerase,
nuclease P (these RNA act as enzymes). RNA Polymerase does not needs a primer.

28. Ans. (b) Binding of RNA polymerase to promoter 36. Ans. (c) No proof reading activity
region
[Ref: Harper 30th/e pg. 434]
[Ref: Harper 30th/e pg. 396] •• RNA Polymerase does not requires primers. It
•• Sigma factor in transcription is for initiation of cannot do proofreading. It is DNA dependent RNA
transcription. It recognize the promoter and bind Polymerase. Shine Dalgarno sequence is recognized
to it. by protein synthesizing machinery i.e. Ribosome.

29. Ans. (a) 5’→3’ 37. Ans. (d) GUAGAUC

[Ref: Harper 30th/e pg. 396] [Ref: Harper 30th/e pg. 361]

•• DNA Replication of both leading and lagging strand •• DNA Template strand is 5’ GATCTAC 3’
A
occurs in 5’ → 3’ direction. Also transcription (RNA •• So, Sense strand (complementary DNA strand) is 3’
N
synthesis) occurs in 5’ → 3’ direction. CTAGATG 5’
•• Therefore, RNA is 3’ CUAGAUG 5’ or 5’ GUAGAUC 3’. S
30. Ans. (a) Intron W
38. Ans. (a) S-Adenosyl methionine (SAM) E
[Ref: Harper 30th/e pg. 377] R
[Ref: Harper 30th/e pg. 345]
•• Introns are intervening sequences present in between S
exons. Introns are transcribed but they are not •• 7-Methyl Guanosine cap is a post-transcriptional
modification in mRNA. The methyl group here WITH
translated. Introns are present in immature mRNA
but then these are removed by post transcriptional is donated by S-Adenosyl methionine (SAM) in E
modification. So these are not present in mature the cytoplasm. Rest other post transcriptional X
mRNA. modifications occur in nucleus. P
L
31. Ans. (c) snRNA 39. Ans. (c) RNA Processing
A
[Ref: Harper 30th/e pg. 367] [Ref: Harper 30th/e pg. 348] N
•• Apo B48 and Apo B100 are synthesised from same A
•• Splicing (removal of introns) is done by snRNA (small mRNA due to difference in RNA Editing (also known T
nuclear RNA). as Chemical Modification of RNA, or Differential I
RNA Processing). O
32. Ans. (b) Mitochondrial DNA
•• Splicing (removal of introns) is same. So Splicing is N
[Ref: Harper 30th/e pg. 378; TABLE 35–3] not the answer. Therefore answer is RNA Processing. S
 40. Ans. (c); (d) 45. Ans. (d) Alternate Splicing

346 [Ref: Harper 30th/e pg. 434] [Ref: Harper 30th/e pg. 408
Types of Eukaryotic RNA Polymerases: •• Earlier one gene one protein theory was invented.
Type I → rRNA But later on it was found that genes are less (25,000
Type II → mRNA, miRNA, some snRNA, Inc. RNA protein coding genes) and proteins are more (2.5 lakh
CRO BIOCHEMISTRY

Type III → tRNA, 5S rRNA, some snRNA. proteins). So there are exceptions to one gene one
protein theory:
41. Ans. (b) 74-95 nucleotides 1. Alternate Splicing
2. RNA editing
[Ref: Harper 30th /e pg. 416]
•• Mutations in the promoter of the gene (option C) can
•• tRNA is the simplest and smallest non-coding RNA affect gene expression by affecting transcription of
having only 74-95 nucleotides length. The length of the gene, as promoter region is involved in initiation
tRNA is fairly conserved within a species but may of transcription.
show some heterogeneity between different species.
•• Cytoplasmic translational system contains 31 tRNA 46. Ans. (a) GU,AG


species whereas Mitochondria translation system

[Ref: Harper 30th/e pg. 407; Fig. 36.14]
possess 22 tRNA species. tRNA contains significant
proportion of nucleosides with unusual bases •• 5’ GU…….A……..AG 3’ is the conserved sequence of
such as Dihydrouridine, Pseudouridine, Inosine, introns i.e. all introns start with GU & ends with AG.
Ribothymidine. Sometimes A is present at the branch site of intron.

42. Ans. (c) CCA at 3’end

[Ref: Harper 30th/e pg. 409, 416]

•• The specific CCA sequence is present at the 3’ end of
tRNA which is also known as the Acceptor Arm. This
CCA sequence acts as an acceptor of the amino acids
on the tRNA molecule.(Option c)
•• Poly A tail at the 3’end (Option a) and 7-methyl
Guanosine cap at the 5’ end (Option b) are the
characteristics of post transcriptional modifications
of mRNA
•• Option d has been added to confuse as CCA sequence
is always present at the 3’end of t RNA.
A
N 43. Ans. (c) Rho (r) factor 47. Ans. (c) Terminal deoxynucleotidyl Transferase
S [Ref: Harper 30th/e pg. 398] [Ref: Harper 30th/e pg. 452]
W
E •• Termination of transcription is brought by a hair pin •• Terminal Transferase is a template-independent
R structure formed at the end which helps in separation enzyme that catalyses the addition of random nucle-
of new RNA from DNA. At other sites, termination otides to the 3’-hydroxyl group of single or dsDNA.
S of the synthesis of RNA molecule is signalled by a •• Terminal deoxynucleotidyl Transferase (TdT) plays
WITH sequence in the template strand of DNA molecule – a crucial role in N-nucleotide addition during V(D)J
a signal that is recognized by a termination protein, recombination of gene segments of antibodies.
E the rho (r) factor. Rho uses ATP and helicase activity. •• Poly A Polymerase and CCA adding enzyme are also
X This protein releases RNA at the termination site. template independent. But they act on RNA.
P
L 44. Ans. (a) 2 subunits 48. Ans. (d) Poly A Polymerase
A
[Ref: Lehniniger 7th/e pg. 1091] [Ref: Harper 30th/e pg. 409]
N
A The mammalian as well as bacterial ribosome contain •• Ribozyme is RNA having enzymatic activity e.g.
T two subunits. In eukaryotes, the two subunits are 60S Ribonuclease P, Peptidyl Transferase or Trans-
I and 40S, while the prokaryotic ribosomes have 50S and peptidase, RNase H and snRNA (small nuclear RNA).
O 30S. •• Poly A Polymerase is not a ribozyme. This enzyme
N adds poly A tail in mRNA.
S
49. Ans. (b) DNA from RNA template RNA polymerase. It is the DNA segment from which the
primary mRNA transcript is copied or transcribed.
[Ref: Harper 30th/e pg. 364] •• Non-template/Coding/Sense/Plus strand refers to 347
•• Reverse transcriptase is synthesis of DNA using RNA the other strand of DNA, where sequence resembles
as template. mRNA except for T/U (T in DNA & U in RNA).
•• The direction as well as sequence of new RNA

CHAPTER 13  REPLICATION, TRANSCRIPTION, TRANSLATION

50. Ans. (b) RNA polymerase resembles the non template strand.
[Ref: Harper 30th/e pg 397] 56. Ans. (c) AAUAAA
•• RNA polymerase is an enzyme involved in
[Ref: Harper 30th/e p 403]
transcription. It is a holoenzyme composed of 6
subunits, which are two identical a subunits, b, b’, W •• AAUAAA is the polyadenylation signal present at the
and s subunit. 3’ end of the eukaryotic heterogenous nuclear RNA
•• s subunit is for initiation of transcription and others (hnRNA), also called immature mRNA.
are for elongation during transcription.
•• b is main catalytic subunit and b’ is subunit for
template binding.

51. Ans. (a) tRNA of alanine

[Ref: Nature Reviews Molecular Cell Biology, volume 8,

Pg. 209-220 (2007)
•• tRNA & rRNA do not require 5’ cap, as their sequences
are not translated. Only mRNA require cap and Poly
A tail. But mRNA for histone does not has Poly A tail.
•• Function of 5’ cap → recognize start codon —
AUG, stabilize mRNA, prevent attack from 5’
exonuclease.
•• U2 snRNA and siRNA also have cap at 5’ end.

52. Ans. (b); (d); (e) •• CCA (Option A) is a sequence present at the 3’end of
all tRNAs
[Ref: Lippincott 4th/e pg. 437] •• CAAT (Option B) is a concensus sequence located 75
•• tRNA is transfer RNA and is required for translation. to 80 bp (base pairs) upstream of the Transcription
It accepts the amino acids in protein synthesis. tRNA Start Site (TSS). It is one of the proximal promoter
+ ribosomes + mRNA complex is machinery for elements. It binds to CAAT transcription factors A
protein synthesis present in cytoplasm. It is clover (CTFs). N
leaf shaped where 3’ end contains an unpaired CCA •• GU…A…AG (option D) is the conserved sequence of S
sequence, so its end is not blunt. It has anticodon introns i.e. all introns start with GU and ends with AG, W
which is complementary to codon of mRNA. A mostly present at branch site. E
57. Ans. (c) TFIID R
53. Ans. (a) Methylation
S
[Ref: Harper’s illustrated biochemistry, 30th ed., pg. 560] [Ref: Harper 30th/e p 423, Lehninger 7th/e pg. 1044]
WITH
DNA methylation at CG sites favors formation of •• TFII is the transcription factor for RNA Polymerase
heterochromatin or condensed regions. II in eukaryotes. TFIID is required for initiation at E
promoters lacking the TATA box. X
54. Ans. (a) Hogness box Subunits of RNA Polymerase P
TATA box helps in initiation of transcription by acting as L
promoter. TATA box is called pribnow box in prokaryotes General Function A
and hogness box in eukaryotes. Transcription N
Factor A
55. Ans. (a) Coding strand TFIIA Stabilise the Pre-initiation complex by T
binding to TBP of TFIID I
[Ref: Harper 30th/e p360] O
TFIIB Binds to RNA Polymerase
•• Template strand/Antisense/Minus/Non-coding strand N
refers to the sequence of DNA that acts as a template for S
 binding of lactose/Allolactose to the repressor
General Function
Transcription ƒƒ CAP binding site should be occupied by CAP-
348 Factor cAMP complex, which is formed due to decreased
glucose. CAP-cAMP complex is essential for
TFIID Acts as a bridge in binding RNA effective functioning of RNA polymerase.
Polymerase ƒƒ If any of the above event doesn’t happen, ZYA
CRO BIOCHEMISTRY

TFIIE Recruits TFIIH genes won’t be expressed.

TFIIF Recruitment of RNA Polymerase to 60. Ans. (a) Positive regulator
Promoter Site
TFIIH Helicase Activity – Unwinds the [Ref: Harper 30th/e pg. 431]
promoter region •• CAP is catabolic activator protein, which is a positive
Kinase Activity – Phosphorylates regulator. Binding CAP with cAMP makes a complex
the C-terminal domain (CTD) of RNA which binds at CAP site on DNA which activates RNA
Polymerase which leads to Promoter
Polymerase. So transcription is activated. So this is
Clearance (escape)
example of positive regulation.


•• Sigma factor (option a) recognizes the Pribnow box in

Prokaryotes. It forms a complex with RNA polymerase 61. Ans. (a) 48S complex
(a2bb’w) to form the holoenzyme. Different types of [Ref: Harper 30th/e pg. 423]
subunits recognize different promoters. •• During termination of translation, releasing factors
•• Rho (option b) can be either Rho factor or Rho recognize the stop codon. Stop codons are UAA, UAG
GTPase. Rho factor is an ATP-dependent hexameric and UGA, which do not code for any amino acid.
helicase involved in transcription termination in Peptidyl Transferase is responsible for the release of
prokaryotes. Rho family of GTPases are involved in polypeptide chain from the P-site of ribosome. 48S
vesicular trafficking. complex is involved in initiation.
•• Pribnow box (option d) is the consensus sequence of
six nucleotides (TATAAT) that forms the prokaryotic 62. Ans. (c) mRNA
promoter.
[Ref: Harper 30th/e pg. 414]
58. Ans. (b) Operator •• Codon is present on mRNA and anticodon is on
•• Cis acting elements are the regions of DNA located tRNA.
near the structural gene to which trans acting factors 63. Ans. (b) Poly lysine
bind.
•• Operator region is located between the prokaryotic [Ref: Harper 30th/e pg. 418]
promoter and the structural genes. •• AAA is the codon for lysine. So Poly A translates into
A Poly lysine.
N
S 64. Ans. (c) Nucleic acid
W [Ref: Harper 30th/e pg. 363]
E •• Peptidyl transferase is a ribozyme i.e. RNA acting as
R enzyme. So, it is a nucleic acid. DNA & RNA are two
S nucleic acids found in cells.
WITH 65. Ans. (a) More than one codon for a single amino acid
E [Ref: Harper’s 30th/e pg. 414]
X •• Degeneracy or redundancy is a property of codon
P Trans acting elements are usually proteins like
transcription factors. They are produced from the that each amino acid has more than one codon. Two
L amino acids do not show degeneracy i.e. Methionine
A region of DNA which is far away from (may be from
different chromosome) the structural gene. That is why and Tryptophan. They have only one codon.
N
A they are trans acting.
66. Ans. (a) RER (Rough endoplasmic reticulum)
T
59. Ans. (c) CAP site is occupied [Ref: Harper’s 30th/e pg. 419]
I
O •• For Lac operon to be expressed i.e. ZYA genes to be Protein synthesis occurs in ribosomes - either free or
N expressed, following 2 events should happen: bound to ER (known as RER). But SER is responsible for
S ƒƒ Operator site should be free from repressor by the synthesis of lipids.
67. Ans. (b) 4 DNA ligase, DNA helicase, DNA polymerase are
involved in replication.
[Ref: Harper’s 30th/e pg. 419] 349
2 ATP for activation of each amino acid (amino acid 74. Ans. (a) 23S; (c) 5S
added on tRNA), 1 GTP required for entry at A-site of
[Ref: Harper 30th/e pg. 367]
ribosome during elongation of translation and 1 GTP

CHAPTER 13  REPLICATION, TRANSCRIPTION, TRANSLATION

required for translocation (movement of ribosome on 50S is the larger subunit of the 70S ribosome of
mRNA). So, 2 ATPs and 2 GTPs (Total 4 high energy prokaryotes. It includes the 5S ribosomal RNA and 23S
phosphates) required to add 1 amino acid in a growing ribosomal RNA.
polypeptide chain. 75. Ans. (a); (e)
68. Ans. (d) Hydroxyproline [Ref: Harper 30th/e pg. 419]
[Ref: Harper 30th/e pg. 60] •• N-formyl Methionine is the first amino acid in
Aminoacyl tRNA is required for all twenty amino acids prokaryotes and methionine is the first amino acid
but Hydroxyproline is a derived amino acid. Aminoacyl in eukaryotes. Translation uses energy in the form of
tRNA is not required for any derived amino acid. GTP and ATPs.
•• Prokaryotic Initiation Factors are → IF-1, IF-2, IF-3
69. Ans. (b) It will be incorporated into a protein in •• Elongation factors in prokaryotes → EF-Tu, EF-Ts,
response to a cysteine codon EF-G
•• Elongation factors in eukaryotes → EF-1A, EF-2
[Ref: Harper 30th/e pg. 286]
Aminoacyl tRNA synthetase enzyme is the only proof 76. Ans. (c) It is nucleotide sequence which codes for
reading step in translation. If wrong amino acid gets amino acids
attached during this step, then it will be added in the
[Ref: Harper 30th/e pg. 412]
protein.
Genetic code is nucleotide sequence which codes for
70. Ans. (d) IF4F amino acids ( not DNA). Inheritance follows mendelian
laws.
[Ref: Harper 30th/e pg. 419]
•• During initiation of translation, first pre-initiation 77. Ans. (c) 750 bp
complex is formed, which requires initiation factors
[Ref: Lehniniger 7th/e, p1081]
IF-1, IF-2 and IF-3.
•• Then initiation complex is formed, which requires A prokaryotic DNA does not contain any introns and the
initiation factors IF-4F. base sequence is exactly collinear with the amino acid
sequence of proteins. Each amino acid is represented
71. Ans. (b) Peptidyl transferase by a 3 base codon, therefore 250 amino acid protein will
A
require 750 base pairs (250 x 3 = 750).
[Ref: Harper 30th/e pg. 363] N
•• The hydrolytic step leading to the release of a 78. Ans. (c) AUG and AUA S
polypeptide chain from a ribosome is catalyzed by W
[Ref: Lehniniger 7th/e, p1081 E
Peptidyl transferase (not by releasing factors).
•• Peptidyl transferase is a ribozyme (i.e. RNA having •• Synonymous codons are the same-sense codons i.e. R
enzymatic activity). they code for same amino acid. E.g. GGG, GGC, GGU S
and GGA codes for the same amino acid i.e Glycine.
72. Ans. (d) Degenerate •• Only Tryptophan(UGG) and Methionine (AUG) are WITH
the two amino acids coded by a single codon. All E
[Ref: Harper 30th/e pg. 416]
other amino acids are coded by multiple codons X
•• Silent mutations occurs because codon is degenerate. (degeneracy).
Degeneracy or Redundancy is a property of codons P
•• Degeneracy is explained by wobbling phenomenon. L
that each amino acid is having more than one codons. Flexible base pairing between 1st base of anticodon
•• Silent mutations means a codon is replaced by A
and 3rd base of codon is referred to as wobbling. N
another codon, which is coding for same amino acid.
A
79. Ans. (b) GCG
73. Ans. (d) Peptidyl transferase T
[Ref: Lehninger 7th /e p1084 fig 27-8] I
[Ref: Harper 30th/e pg. 363} O
•• Wobble Hypothesis: Movement of first base of
Peptidyl transferase is involved in elongation and N
Anticodon (towards 5’ end on tRNA) can occur
termination of translation (protein synthesis). Whereas S
 to allow non traditional base pairing with the last ƒƒ A site: always lies towards right side. ‘A’ is Acceptor
base of codon (towards the 3’ end on mRNA). This site, i.e. all amino acids are accepted here. EXCEPT
350 movement is known as wobble, which allows a single – 1st Methionine or Formyl methionione
tRNA to recognize more than one codon.
•• Wobble position is the 3rd position of the codon and 81. Ans. (c) They are the sites of protein synthesis
the first position of Anticodon which is in 5’→ 3’
CRO BIOCHEMISTRY

[Ref: Harper 30th/e p425]

direction.
•• Here in this question, the first base of Anticodon is •• P bodies (Processing bodies) are the cytoplasmic
Inosine which is the Wobble base of tRNA. organelles involved in mRNA metabolism. mRNA-
•• Inosine is the Wobble base which can pair with C, A miRNA protein complex is stored in P bodies.
and U but not with G. •• P bodies are the sites of translation repression and
mRNA decay. mRNA stored in P-bodies can either
Anticodon 5’ – I – G – C – 3’
undergo degradation or can go back for translation.
Codon 3’ – A – C – G – 5’
U–U 82. Ans. (c) 16S rRNA
C
[Ref: Lehninger 7th /e p1101


•• In prokaryotes, there is no 5’ cap to help in ribosomal

recognition in bacterial mRNA. Recognition of start
site of translation is aided by base pairing between
the Shine Dalgarno sequence of mRNA.
•• Shine dalgarno sequence (SD sequence) is present
in prokaryotes on mRNA for initiation. This is purine
rich sequence which has complementary bases with
16S rRNA (present in 30S subunit of ribosome).

83. Ans. (b) Puromycin

[Ref: Harper 30th/e p426]

•• Puromycin is a structural analog of tyrosinyl-tRNA . It
is incorporated via the A-site on the ribosome into the
•• Codon also written in 5’ → 3’ carboxy terminal position of a peptide but causes the
•• Therefore, Anticodon 5’-I-G-C-3’ can code for the premature release of the polypeptide. It effectively
codon GCA,GCC and GCU and not GCG inhibits protein synthesis in both prokaryotes and
eukaryotes.
80. Ans. (a) P site
A
84. Ans. (d) UAA → CAA
N [Ref: Harper 30th/e p421]
S •• Polypeptide is abnormally long which means the
•• Initiator complex forms 2 sites: protein synthesis was not terminated at the stop
W
ƒƒ P site: always lies towards left side. ‘P’ means codon. Hence, there must be mutation in the stop
E
Polypeptide site i.e. polypeptide will be released codon. UAA, UGA, UAG are the stop codons.
R
from this site. •• Option a (CGA → UGA) will result in premature
S Initiator tRNA (Met-tRNA in eukaryotes or F-Met- termination as a codon gets replaced by stop codon
tRNA in prokaryotes) is the only aminoacyl tRNA – UGA.
WITH
to directly enter the P site of ribosome.
E
X
P
L
A
N
A
T
I
O
N
S
Techniques in
Molecular Biology
14
Overview of Chapter PCR – Polymerase Chain Reaction

•• Mutation detection techniques zz PCR is amplification of DNA

•• Methods to detect aneuploidy Components of PCR :

•• Methods to detect Genomic Imprinting 1. ds-DNA
•• Methods of gene transfer 2. Two primers
3. Thermostable Polymerase- e.g. Taq Polymerase
•• Nucleic acid amplification techniques 4. Deoxy-ribonucleotides
•• RNA interference 5. Mg++
•• CRISPR
H igh R eturn
•• Sample taken for prenatal diagnosis. If dideoxy nucleotide is used in PCR, then DNA synthesis will stop,
as it will be incorporated in the growing DNA chain and it will not
allow the next normal nucleotide to be added.
MUTATION DETECTION TECHNIQUES
(MOLECULAR CYTOGENETICS)
Steps of PCR:
zz Polymerase chain reaction (PCR) and its versions
1. Denaturation: Two DNA strands separated (94 °C)
zz Ligase Chain Reaction (LCR)
2. Annealing: Addition of primers (one for each strand)
zz Multiplex Ligation-dependent Probe Amplification
(60 °C)
(MLPA)
3. Extension/Elongation: Polymerization or synthesis of
zz Restriction Fragment Length Polymorphism (RFLP)
DNA (72 °C)
zz Blotting
These cycles of Denaturation, Annealing and Extension are
zz Conventional Karyotyping
repeated again and again to synthesize large amount of DNA.
zz Fluorescence In Situ Hybridization (FISH)
zz Microarray
zz Comparative Genomic Hybridization (CGH)
zz DNA Sequencing

Fig. 14.1: Step of PCR

 zz Taq Polymerase lacks proof reading.
zz Thermostable DNA Polymerase enzymes with proof
352 reading function in PCR:
1. Pfu: Pyrococcus furiosus
2. Pwo polymerase: Pyrococcus woesei
CRO BIOCHEMISTRY

3. Tli: Thermococcus litoralis – also known as Vent

Poly-merase
Best is pfu polymerase, but it is slow in action. So best is to use
Pfu in conjunction with Taq polymerase to obtain the fidelity
of Pfu with the speed of Taq polymerase activity.

PCR Applications

PCR is used in
Structural analysis, DNA typing, Disease detection, Cloning,


Mutation analysis, Detection of gene expression, Mapping,

Site-directed mutagenesis and Sequencing.

Real Time PCR / Quantitative Fluorescence PCR (QF-PCR) Fig. 14.2: Reverse transcription PCR
/q PCR
RT PCR qPCR
zz Products are detected by fluorescence during (not after)
the PCR reaction (in real time), not end time PCR Qualitatively detect gene Quantitatively measure
expression amplification of DNA
zz In addition to these five components (ds-DNA, two
Primers, Taq Polymerase, Deoxy-ribonucleotides, Mg)
Quantitative Fluorescent Polymerase Chain Reaction
SYBR green dye is added. This dye gives fluorescence
(QF-PCR)
when bound to ds DNA. When DNA synthesis is
occurring then gradually ds DNA is increased in amount
so fluorescence is also increased. This fluorescence can
be measured and then amount of DNA can be calculated
from that. Therefore it is known as Quantitative PCR
(q PCR).
zz Performed in specialized thermal cyclers with fluorescent
detection systems

Dyes used in qPCR

zz SYBR Based Detection: Uses SYBR Green dye (ds-DNA

binding dye) – not sequence specific.
zz TaqMan Based Detection: Uses a fluorogenic probe
specific to target gene to detect target, as it accumulates
during PCR.

RT-PCR (Reverse Transcription PCR)

zz Used to detect RNA expression
zz Sample here is RNA. This RNA is converted to cDNA by
enzyme reverse transcriptase.
T zz Tth polymerase (Thermus thermophilus HB-8) has
H Polymerase as well as reverse transcriptase activity when
E mixed with manganese ions and can thus be used for
O the amplification of RNA to cDNA. This polymerase is
R thermostable but does not has proofreading activity. So
Y it is usually combined with a proofreading enzyme.
Fig. 14.3: Quantitative Fluorescent Polymerase Chain Reaction
(QF-PCR)
PCR Advantages
PCR is specific, simple, rapid, relatively inexpensive, sensitive and Flexible. It amplifies from low quantities. It works on 353
damaged DNA

Ligase Chain Reaction (LCR) or Ligation Amplification Reaction (LAR)

CHAPTER 14  TECHNIQUES IN MOLECULAR BIOLOGY

zz A method of DNA amplification similar to PCR.
zz Probe is amplified in this technique. Two probes are used for each DNA strand and are ligated together to form a single
probe.
zz Enzymes used: Thermostable DNA polymerase and thermostable DNA ligase (Taq DNA ligase and Taq DNA polymerase)
zz DNA probes also required

Steps of LCR
zz Steps of LCR are Denaturation, Annealing of probes to target DNA and Ligation
zz The separated ligated unit becomes target for next cycle. These steps are repeated through several cycles. This way there
is rapid amplification of a specific target fragment of DNA.
zz Can be used to detect mutation (point mutation mainly)

Fig. 14.4: Ligase chain reaction

Multiplex Ligation-Dependent Probe Amplification (MLPA)

zz This is a kind of multiplex PCR technique that can simultaneously analyze multiple genomic regions.
zz Amplification occurs which is made dependent on a ligation step so that this amplification occurs only if target DNA is
present in the sample

Advantages
zz Small sample requirement
zz Multiple probes are added in a single reaction, so multiple targets can be amplified.
zz It can detect copy number variations.
zz Can detect deletion and duplication of any size (small or large)
T
H
E
O
R
Y
354
CRO BIOCHEMISTRY


RFLP – Restriction Fragment Length Polymorphism tendency to reanneal. This is known as hybridization, which
is used for visualization of the sample DNA.
Probe: is a DNA sequence, which is complementary to
The fragments formed after digestion by restriction gene of interest or only a small portion on gene of interest. This
endonuclease enzyme are known as Restriction fragments. probe is labeled with a radioactive material or a fluorescent
dye.
Limitations of RFLP
zz Only single mutation can be detected.
zz Can detect only those mutations which affect Pallin-
drome.
zz Lengthy procedure (after RFLP, electrophoresis is needed
to separate the fragments and then blotting is needed to
visualize the fragments).

T
H
E
O
R
Y
Blotting /Hybridization
DNA strand can be separated by denaturation but if
complementary single strands are present, then they have a Fig. 14.5: Basic Principle of Blotting
 H igh R eturn Technique Sample Analyzed Probe

Q. Single gene expression analysis can be done by : Western Blot/ Protein Antibody 355
   a. Northern blotting Immuno Blot
   b. Western blotting South Western Blot Protein-DNA DNA
   c. Both a and b Microarray mRNA or cDNA DNA

CHAPTER 14  TECHNIQUES IN MOLECULAR BIOLOGY

   d. Southern blotting
A. (c) Both a and b ELISA Protein or Antibody
Gene expression = Transcription + Translation.
Transcription is formation of MRNA and Translation is Karyotyping
formation of Protein. mRNA can be detected by northern zz Study of chromosome
blotting and protein can be detected by western blotting. zz Best technique for Monosomy/Trisomy
So answer is both a and b zz But can be only done during Metaphase chromosome
zz Sample used: Blood, Bone marrow, Amniotic fluid,
Technique Sample Analyzed Probe Placental tissue
Southern Blot/ DNA DNA zz Fixative used is Carnoy’s fixative (Methanol and Glacial
DNA-DNA Acetic acid in ratio 3:1)
Hybridization zz Most common stain used is Geimsa (so known as
G-banding)
Northern Blot/RNA RNA DNA
Contd… zz Limitations: Cannot be done in any phase of cell
-DNA Hybridization
cycle. It cannot detect amplifications, microdeletions
Contd... and complex translocations and has low resolution i.e.
subchromosomal alterations.

(a) (b)

Fig. 14.6: (a) Preparation of chromosomes for Karyotyping (b) Karyogram

FISH (Fluorescent In-Situ Hybridization)

zz Rapid/ fast technique (results can be obtained within 24 hours, if necessary), better resolution
zz Principle is same as Southern Blot
zz Can be done in any phase of cell cycle
zz In situ – means can be done in a morphologically intact cell/tissue/ organ
zz Detects chromosomal regions of greater than 100 Kilobases in size
FISH methodology: Hybridizing a probe with a fluorescent tag T
zz Fluorescent labeled probe is taken for a particular gene of interest. Probe can be DNA or RNA. H
zz Three types of probes can be used: Whole chromosome painting probes, repetitive sequence probes and locus specific E
probes. O
R
Y
356
CRO BIOCHEMISTRY

Uses
FISH can detect Microdeletions, Amplifications, Complex translocations, Can detect Monosomy/ Trisomy (Aneuploidy) and
tells gene location on a chromosome to identify and enumerate specific microbial groups


A dditional E dge
zz FRAP→ Fluorescence Recovery After Photobleaching. This technique detects movement of proteins from one compartment of cell to
another and lateral diffusion of lipids and proteins in the membrane.
zz FRET→ Fluorescence Resonance Energy Transfer. It is a phenomenon in which an excited donor transfers energy (not electron) to an
acceptor by radiationless energy transfer process which occurs when they are in close proximity.

Note: FISH requires previous knowledge of that chromosomal region which is altered in the sample. But detection of genomic
abnormalities without previous knowledge can be done by Microarray and CGH.

Microarray/Chip/ Bioarray/ Gene Array

Thousands of small DNA fragments/probes are immobilized on a solid support e.g. glass slide, in an ordered fashion. DNA,
RNA or protein can be detected.

T zz Microarray can detect mutiple mutations, mutiple gene expression analysis, global pattern of gene expression, SNPs and
H copy number variations, genetic transfer of disease, Protein-DNA interactions, methylation patterns (epigenetic status)
E zz It cannot detect aneuploidy: monosomy/trisomy
O zz Applications include genotyping, clinical genetic testing, forensic analysis, cancer mutations and genetic linkage analysis
R
Y Comparative Genomic Hybridization (CGH) or Chromosomal Microarray Analysis (CMA)
A cytogenetic method for analysis of copy differences between two genomes (Test and Reference), without the need for
culturing cells.
 If hybridization is done on microarray plate (instead of using metaphase chromosomes) then it is known as Microarray-
based Comparative Genomic Hybridization (aCGH). It has high resolution as compared to normal CGH. It is better than FISH
in genome wide screening 357
Methodology of CGH
zz DNA taken from test sample (test DNA), denatured and labeled with a specific fluorescent dye color.

CHAPTER 14  TECHNIQUES IN MOLECULAR BIOLOGY

zz DNA also taken from a normal control (reference DNA), denatured and labeled with a different dye color.
zz These two DNA samples (single stranded as they are denatured) are mixed together and applied to microarray slide, on
which single stranded probes are applied. Test and reference DNA both hybridize with probes on microarray slide.
zz The fluorescence ratio of the test and reference hybridization signals is determined at different positions along the
genome, and it provides information on the relative copy number of sequences in the test genome as compared to the
normal genome (Fig. 14.7)

Fig. 14.7: Comparative genome hybridization (CGH). CGH uses two genomes, a test and a control, which are differentially labeled and
competitively hybridized to metaphase chromosomes. The fluorescent signal intensity of the labeled test DNA relative to that of the
reference DNA can then be linearly plotted across each chromosome, allowing the identification of copy number changes
DNA Sequencing Methods 2. RT-PCR:Reverse Transcriptase
1. Sanger’s or chain termination method or Dideoxy 3. QF-PCR: Quantitative Fluorescence PCR or Real time
method (Dideoxy nucleotides act as terminators of DNA PCR.
synthesis) 4. MLPA: Multiplex ligation-dependent Probe Ampli-
fication
2. Maxam and Gilbert chemical degradation method
(chemical used which cut dsDNA at specific nucleotide All these methods avoid the generation of cultured cells
positions) and can rapidly detect (within 1 or 2 days) the aneuploidy.
Microarray cannot defect aneuploidy.
3. Pyro-sequencing method – based on ‘sequencing by
synthesis’ principle as addition of deoxunucleotide is
accompanied by release of flash of light METHOD TO DETECT GENOMIC IMPRINTING
4. Second generation sequencing or Next generation
zz Na bisulfite method: detects DNA methylation. T
Sequencing (NGS) – This is massive parallel sequencing
zz ChIP: Chromatin Immuno Precipitation- it detects post H
which is cheaper and faster and entire genome can be
translational modifications of histones E
sequenced in less than one day.
zz ChIP is used together with its large-scale variants ChIP- O
on-chip and ChIP-Seq
METHODS FOR DETECTION OF zz Fluorescent in situ hybridization
R
CHROMOSOMAL ANEUPLOIDY Y
zz Methylation-sensitive restriction enzymes
1. FISH: Fluorescent In Situ Hybridization zz PCR
zz Microarray
 ChIP-Seq (Fig. 14.8)
358 Chromatin immunoprecipitation (ChIP) assays are combined with sequencing, It can be used to map global binding sites
precisely for any protein of interest like transcription factors.

H igh R eturn
CRO BIOCHEMISTRY

zz ChIP: Chromatin Immuno Precipitation – detects Protein DNA interactions and histone modifications
zz Chip – also known as Microarray


Fig. 14.8: ChIp (Chromatin Immunoprecipitation)

ChIP-on-Chip
Also known as ChIP-chip. This technique combines chromatin immunoprecipitation with DNA Microarray. It can detect
T protein-DNA interactions in vivo.
H Chromatin IP is done using an antibody against a protein or protein modification of interest. The resulting purified DNA
E sample is fluorescently labeled and co-hybridized to a microarray. The input sample (DNA prior to IP) is also fluorescently
O labeled using a complementary fluorescent dye. Both sets of labelled DNA are combined and hybridized microarrays,
R corresponding to defined domains, loci or even whole genomes. Based on the ratios of signals from ChIP DNA and control
Y DNA, enriched regions can be identified.
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CHAPTER 14  TECHNIQUES IN MOLECULAR BIOLOGY

Fig. 14.9: Chromatin immunoprecipitation on chip (Cy3 and Cy5 → are fluorescent dyes (Cyanine dye))

zz Foot Printing Assay identify those sites of DNA where protein UV cross linking unlike formaldehyde cross linking are:
binds. It is used to study protein-DNA interactions both outside zz Irreversible
and within the cell. It is also known as DNAse I footprinting zz Not formed between two proteins
Principle: This enzyme digests DNA that is not bound by zz More specific - They bind RNA with protein that are in
proteins. The protected DNA region which is left is called the close proximity
foot print.
METHODS OF GENE TRANSFER
RIP (RNA Immunoprecipitation)
1. Transformation: uptake and incorporation of naked
zzIt detects binding of protein to RNA. DNA into bacteria from external environment
 “RIP and CLIP” are two fundamental approaches for 2. Transfection: transfer of non viral genetic material into
analyzing RNA-protein interactions. RIP is the older eukaryotic cells (fungi, plant or animal cells). The uptake
technique from which CLIP was derived. of chimeric DNA takes place by transfection.
3. Transduction: introduction of genetic material using
RIP CLIP
viruses (bacteriophage)
•• RNA Immunoprecipitation •• Cross Linking 4. Conjugation: a cell contact- dependent DNA transfer
•• Binding of protein to RNA Immunoprecipitation mechanism T
•• Uses formaldehyde-cross •• Binding of protein to RNA 5. Lipofection: introduction of genetic material using H
linking to induce covalent •• Uses UV-cross linking to liposomes. E
attachment of proteins to induce covalent attachment 6. Electroporation: Electric field is applied which increases O
RNA of proteins to RNA
cell membrane permeability, allowing DNA, drugs or R
chemicals to be introduced into the cell. Y
360
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

NUCLEIC ACID AMPLIFICATION TECHNIQUES Vectors

zz PCR (in vitro) Vectors are DNA molecules which can carry a foreign DNA to
zz Branched DNA (bDNA) Signal Amplification. be cloned for recombinant DNA technology.
zz LCR Table 14.1: Cloning Capacities of common Cloning Vectors
zz MLPA
zz Nucleic Acid Sequence based Amplification (NASBA) Vector DNA insert size (kb)
zz DNA cloning/Recombinant DNA technology (cell based Plasmid 0.01-10
in vivo technique) Lambda Phage 10-20
Recombinant DNA Technology Cosmids (Plasmids with features of 35-20
Steps plasmids and phages)
zz DNA to be cloned taken (usually small quantity present) Bacterial artificial chromosome (BAC) 50-250
zz Ligate DNA into a nucleic acid vector which can YAC (Yeast artificial chromosome) 500-300 (Largest)
automatically replicate in a host. This is known as
recombinant vector.
zz Transfer recombinant vector into host (mostly bacteria). Site-Specific Recombinase
zz Grow large quantity of colony having recombinant These enzymes catalyse DNA exchange which is direction
vector. sensitive, between short target site sequences (30-40
zz Either recombinant proteins can be obtained by inducing nucleotides), that are specific to each recombinase. Example
gene expression or large amount of cloned DNA can be of recombinase enzymes with their site of recognition given
used for various applications. in the table. (See the below table)

Recombinase enzyme Site of recognition

CRE recombinase Bacterial Lox P site

λ Phage encoded INT protein Bacteriophage ATT Site

Yeast FLP recombinase Yeast FRT site

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CHAPTER 14  TECHNIQUES IN MOLECULAR BIOLOGY

Fig. 14.10: Cre/Lox recombination

Blue-white Assay
zz Screening technique to detect successful ligation of DNA of interest into vector of recombinant DNA technology
zz Principle: α- complementation of β- galactosidase
zz β-galactosidase is a protein from Lac Z gene of Lac operon. The active protein is a homotetramer having ω and α-peptides.
zz In this assay, a mutant E coli Lac Z Δ M 15 is taken which can only produce ω peptide which is non functional as no
α-peptide.
zz A plasmid is taken which has gene only for α peptide and has internal multi cloning site (MCS)
zz Functional β-galactosidase is only formed when plasmid is successfully ligated or transformed into host genome.
zz Rescue of function of mutant protein by α-peptide is known as α-complementation.

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 RNA INTERFERENCE/GENE KNOWN DOWN/ gene is present but its function is suppressed. Also known as
silencing technique (gene present but silent). This is known
362 SILENCING TECHNIQUE as RNA interference because this is interference at the level
This is a biological phenomenon by which RNA molecules of RNA i.e. mRNA.
inhibit translation. Synthetic dsRNA can be introduced into cells which can
This is a method to inhibit gene but inhibition is done at selectively induce suppression of specific genes of interest.
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the level of translation i.e. gene is able to be transcribed

and mRNA is formed. But this mRNA is degraded so that zz Gene knock ‘out’ → gene moved or taken ‘out’
the protein is not formed. miRNA - micro RNA (a short non zz Gene knock ‘in’ → gene inserted
coding RNA approx. 22 nucleotide in length) is used for zz Gene knock ‘down’ → gene present but function is suppressed
such purpose. So this is also known as gene knock down i.e.


Fig. 14.11: RNA Interference. Primary micro RNA gene is present in nucleus, which is transcribed by RNA Polymerase to form primary
micro RNA. then ‘drosha’ (an endonuclease) cut this primary micro RNA to form pre- micro RNA. This pre micro RNA can escape nuclear
pore and reaches cytoplasm, where it is acted upon by another endonuclease i.e. ‘Dicer' which cut this pre micro RNA to form ds RNA
(around 20 nucleotides in length). Unwinding of the two strands is done and out of the two strands, only one strand is selected. The
strand which is selected is known as guide or antisense strand. This single stranded micro RNA is attached to proteins and the complex
formed is known as RISC (RNA induced silencing complex). This RISC can bind to target mRNA and can either just suppress it or degrade it
by enzyme slicer (also known as Argonaute/ Ago).
zz Drosha works with DGCR8, also known as PASHA (to rhyme with Drosha)
zz Silencing RNA (si RNA) or small interfering RNA- This is involved in silencing technique. This siRNA is synthesized from
cytoplasmic RNA (e.g. tRNA or viral RNA) and this siRNA can bind anywhere on the target mRNA. But miRNA can only
bind to 3’ end of target mRNA.

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CHAPTER 14  TECHNIQUES IN MOLECULAR BIOLOGY

CRISPR –CAS 9 SYSTEM
zz Simple, cheap, genome editing tool
zz This system is present in prokaryotes. CRISPR and Cas endonuclease acts together to degrade the target DNA
zz CRISPR– Clustered Regularly Interspersed Short Pallindromic Repeats
zz Cas-9 → CRISPR associated endonuclease enzyme.
zz This is like an immune system in bacteria, which can destroy bacteriophage DNA is transmitted to the progeny.

Uses
zz Has been adapted to be used in eukaryotes for various things such as, Double strand breaks (gene deletion), Single strand
breaks, To know the function of a gene, Multigene editing, Gene additions (at exactly the place where we want), Altering
gene transcription and regulation, Insertion of exogenous gene and Create SNPs
This system can be used in eukaryotes for double strand or single strand breaks, knock in, knock-out, to assess gene
function, altering gene regulation, create SNPs etc.

Advantages over older techniques of DNA breaks

zz Cheap, simple, rapid, more accessible, highly efficient and can target a specific gene.
zz Guide RNA- (gRNA)cas 9 endonuclease near to specific target DNA, which can be cut

SAMPLE TAKEN FOR PRENATAL DIAGNOSIS

zz Amniotic fluid: known as Amniocentesis – done after 16 weeks of gestation
zz Chorionic villus sampling (CVS): done after 11 weeks of gestation
zz Fetal blood sampling: done after 18 weeks of gestation
zz Adult samples which can be taken for DNA testing – blood (WBCs), cheek swab, hair.

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 Pearls of the Chapter
364 zz CRISPR – Clustered Regularly Interspersed Short Pallindromic Repeats
zz Cas-9 → is an endonuclease
zz Known as RNA guided Nuclease system
CRO BIOCHEMISTRY

zz This system is in prokaryotes, to destroy bacteriophage DNA

zz Cas→ CRISPR associated proteins
zz CRISPR and cas endonuclease acts together to degrade the target DNA
zz This is RNA-based targeting of DNA, to bring cas 9 endonuclease near to that specific DNA, which can be cut.
zz Most DNA methylation is removed at fertilization and re-established during embryogenesis
zz Imprinted genes keep their parental pattern of methylation giving rise to parental patterns of expression
zz Patterns of histone modifications parallel DNA methylatL.;ion
zz Methylated gene regions are genetically inactive, highly condensed and special histone modifications
zz Praderwilli syndrome – only paternal allele working
zz Angelman syndrome – only maternal allele working


zz Euchromatin is transcriptionally active

zz Heterochromatin is transcriptionally inactive
zz Epigenetic changes are transferred to next generation and are reversible
zz Acetylation and Phosphorylation of histones lead to gene activation
zz X chromosome inactivation in females is correlated with extensive CG island methylation on one chromosome, condensation,
inactivation and Barr body formation
zz Alterations in gene and CG island methylation patterns are seen in aging and in cancer

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 Multiple Choice Questions 365

Molecular Techniques 11. Restriction Fragment Length Polymorphism is used

1. Which of the following is not required in PCR? for:  (Recent Question 2016)
 (AIIMS May 2012, 2013, 2017) a. Analysis of DNA fragments
a. dNTPs b. DNA estimation
b. Primer c. Synthesis of nucleic acid
c. Radio-labelled DNA probe d. Detecting proteins in a cell
d. Taq polymerase 12. Which of the following methods is most suited to
2. Which of the following test is not used in epigenetics? assess function of a gene? (Recent Question 2016)
a. HPLC a. Southern blot
b. ChIP on chip b. Northern blot
c. Bi-sulfate sequencing c. Gene knockout animals
d. Methylation sensitive restriction enzymes digestion d. Transgenic animals
3. Test for RNA:  (JIPMER May 2018) 13. Which of the following techniques is used for
a. Northern blot b. Southern blot detection of variations in DNA sequence and gene
c. Immuno blot d. South-Western blot expression?
4. Which of the following will be more towards the a. Northern blot b. Southern blot
negative pole in gel electrophoresis? (PGI May 2018) c. Western blot d. Microarray
a. 5 bp 14. Which of the following tests is not used for detection
b. 50 kbp of specific aneuploidy?
c. 160 bp a. FISH b. RT-PCR
d. 550 bp c. QF-PCR d. Microarray
e. 50000 bp 15. Steps of PCR in sequence are: 
5. Sanger’s reagent is:  (PGMEE 2015, FMGE Nov 2018)
a. Dinitrobenzene b. Dichlorobenzene a. Extend DNA, Anneal Primers, Denature DNA
c. Tetra-nitrobenzene d. Tetrachlorbenzene b. Anneal Primers, Extend DNA, Denature DNA M
6. Prenatal diagnosis of hemophilia is best done by: c. Denature DNA, Anneal Primers, Extend DNA C
a. PCR b. Linkage analysis d. Denature DNA, Extend DNA, Anneal Primers Qs
c. Cytometry d. Microarray 16. Denaturation of DNA in PCR is carried out by heating Ans.
7. SYBR Green Dye is used for (AIIMS May 2007) to a temperature of : (PGMEE 2013-14)
a. 76°C b. 60°C 1. c
a. HPLC b. Immuno-fluorescence
c. 40 °
C d. 94 °
C 2. a
c. PCR d. ELISA
8. Methods of introducing gene in target cells are all 17. Which one of the following enzymes is obtained from 3. a
EXCEPT: (AIIMS Nov 2010) Thermophillus aquaticus bacterium which is heat 4. b,e
a. Electroporation stable and used in PCR at high temperature? 5. a
b. Transfection (PGMEE 2012, 2013) 6. a
a. DNA polymerase III b. Endonuclease 7. c
c. Site directed recombination
c. Taq polymerase d. DNA gyrase 8. d
d. FISH
9. Which of the following technique can be used for 18. Movement of protein from nucleus to cytoplasm can 9. a
exact localization of a genetic locus? be seen by: (PGMEE 11) 10. b
a. FISH b. FRAP 11. a
 (AIIMS Nov 2013, May 2013)
c. Confocal Microscopy d. Electron microscopy 12. c
a. Chromosome Painting
b. Western blot 19. Most common enzymatic tool in genetic engine- 13. d
c. Comparative genomic hybridization ering: (PGMEE 2015) 14. d
a. DNA Ligase 15. c
d. FACS
b. Halicase 16. d
c. Topoisomerase 17. c
10. Rapid method of chromosome identification in inter-
d. Restriction endonuclease 18. b
sex is: (AIIMS May 2007)
a. PCR 20. Which is not a method of gene therapy? 19. d
b. FISH  (PGMEE 2016-17) 20. a
c. Karyotyping a. FISH b. Transfection
d. Single-strand conformational polymorphism c. Electroporation d. Bacteriophage
(SSCP)
 21. Which of the following procedures as routine 33. Real-time PCR is used for:
technique for karyotyping using light microscopy- a. DNA detection only
366  (PGMEE 2007, 2009, 2010, 2015) b. RNA detection only
a. Q-banding b. G-banding c. Protein detection only
c. R-banding d. Brd V-staining d. For monitoring the amplification target DNA
22. Karyotyping is done in which phase of cell cycle? 34. Which of the following is a primarily RNA based
 (PGMEE 2012-13) technique?  (AIIMS May 2018)
a. Anaphase b. Metaphase a. Next generation sequencing
c. Telophase d. S phase b. RT PCR
23. Klineflter Syndrome is diagnosed by: c. Sanger’s technique
 (PGMEE 2012-13) d. Western blotting
a. USG abdomen b. Echocardiography 35. RFLP was used in-order to identify the five different
c. Triple test d. Karyotyping species of staphylococci in a surgical ICU. Which of
24. For karyotyping, the dividing cells are arrested by the the following site does the restriction enzyme act?
addition of colchicine in the following mitotic phase:  (AIIMS May 2017)
 (PGMEE 2015) a. TA GA TA b. AT GG AC
a. Telophase b. Metaphase AT CT AT TA CG TG
c. Anaphase d. Prophase c. AA TA TA d. GA TA TC
25. The best suited nucleated cell for chromosomal TA TA AT CT AT AG
study: (PGMEE 2015) 36. RNAi causes the following to a gene:
a. Polymorphs b. Lymphocytes  (AIIMS May 2015)
c. Langerhan’s cell d. Epithelial cells a. Knock in b. Knock out
26. Microarray is: (PGMEE 2012-13) c. Knock down d. Knock up
a. Study of multiple genes 37. miRNA binding site for gene knock down:
b. Study of blood group  (Recent Question June 2018)
c. Study of disease a. 3’ untranslated region
d. Study of organisms b. 5’ untranslated region
27. Protein-encoding genes comprise what percentage c. Introns
of genome? (PGMEE 2015) d. Both 3’ and 5’ untranslated region
a. 98.5% b. 1.5% 38. Test to differentiate the chromosome of normal and
M c. 5% d. 95% cancer cells: (AIIMS November 2012)
C 28. Commonest form of DNA variation is?(PGMEE 2015) a. PCR
Qs a. Single Nucleotide Polymorphism (SNPs) b. Comparative genomic hybridization
Ans. b. Copy Number Variations (CNVs) c. Western Blotting
21. b c. Transposons d. Southern Blotting
22. b d. Mutations 39. DNA estimation can be done by: (AIIMS May 2012)
23. d 29. Submicroscopic deletions of any size can be detected a. Spirometer
24. b by: (PGMEE 2015) b. Spectrophotometer
25. b a. Multiplex ligation-Dependent probe amplification c. pH meter
26. a (MLPA) d. Sphygmomanometer
27. b b. Chromosome painting 40. Test used for protein is/are: (PGI May 2014)
28. a c. Cytogenomic array technology a. Western blot b. Southern blot
29. a d. Southern blotting c. ELISA d. Dot blotting
30. b 30. Find the false statement about mi-RNAs: 41. Non coding RNA is/are:
31. a  (PGMEE 2015) a. miRNA b. tRNA
32. c a. Inhibit gene expression c. RNA d. si RNA
33. d b. Can encode proteins e. mRNA
34. b c. Gene silencing RNAs 42. Nucleic acid amplification techniques are:
35. d d. 21–30 nucleotides in length a. PCR
36. c 31. ChIP is used for : b. Real time PCR
37. a a. Protein DNA interactions and histone modifications c. DNA cloning
38. b b. Study mutations d. Next generation DNA sequencing
39. b c. Amplify DNA e. ELISA
40. a,c,d d. Movement of proteins 43. Gel used in RNA electrophoresis:
41. a,b, 32. In gene studies, the specific site to which the enzyme a. Agarose gel
c,d CRE Recombinase binds is : b. Polyacrylamide plain gel
42. a,b,c a. RE site b. INT site c. Polyacrylamide SDS impregnated gel
43. a c. Lox P site d. FRT site d. Acrylamide gel
44. Components/genes involved in RISC complex: 51. The recognition sequence for restriction endonucle-
 (PGI Nov 2017) ase, Hind III is:
a. Drosha a. AAGCTT b. AAGAAG 367
b. Pasha c. AAGTTC d. AAGAGA
c. Dicer 52. Commonly used prokaryotic host in rDNA technology
d. miRNA is:
e. rRNA a. Haemophilus aegypticus
45. Which of the following vector carry larger DNA b. Norcardia otitidis
segments? c. Escherichia coli
a. Plasmids d. Haemophilus influenza
b. Cosmids 53. Enzyme that produces single strand nicks in DNA:
c. Bacterial Artificial Chromosomes a. DNA polymerase I
d. Bacteriophage b. DNase I
46. By which method foreign DNA is introduced into a c. Polynucleotide kinase
cell by a virus or viral vector? d. λ – exonuclease
a. Transduction  (Recent Question Jan 2018) 54. Plasmids contain:
b. Transcription a. A circular double stranded DNA
c. Lysogenic conversion b. A circular single stranded DNA
d. Transformation c. A linear double stranded DNA
47. Genetic material is transferred from one bacteria to d. A linear single stranded DNA.
another by all EXCEPT :  (FMGE Nov 2018) 55. Transgenic animals are
a. Conjugation a. Genetically modified organisms with a new heritable
b. Transduction character
c. Transformation b. Serve as models for understanding the human
d. Transfection diseases
48. Which has more gene content X or Y?  c. Proteins produced by them are used as therapeutic
 (Recent Question June 2018) agents
a. X contents 2.8% more than Y d. All the above
b. Y contents 2.8% more than X 56. The most commonly used animal model in trans-
c. X contents 0.28% more than y genesis to represent humans:
d. X contents 0.28% more than Y a. Rabbit b. Mouse M
49. Hybridoma is used in:  (Recent Question June 2018) c. Monkey d. Dog C
a. In situ hybridization 57. The application(s) of rDNA technology: Qs
b. Sequencing DNA a. Understanding the molecular basis of diseases Ans.
c. Production of monoclonal antibodies b. Production of human proteins in large quantities 44. a,b
d. Production of continuous cell lines c. Gene therapy c,d
50. Which strategy in transcription factor research d. All the above 45. c
allows for the simultaneous identification of all the 58. Which of the following is TRUE about restriction 46. a
genomic sites bound by a given transcription factor enzymes? 47. d
under a given set of physiological conditions? a. Produced by viruses 48. a
a. Flourescence Energy Transfer (FRET) b. Cut only single stranded DNA 49. c
b. DNAase I sensitivity c. Can cut DNA anywhere 50. c
c. Chromatin Immuno Precipitation-Sequencing d. Restrict the expression of Viral DNA 51. a
d. FISH 52. c
53. b
54. a
55. d
56. b
57. d
58. d
 Answers with Explanations
368

1. Ans. (c) Radio-labelled DNA probe •• Edman’s reagent is PITC (Phenyl Isothiocynate),
CRO BIOCHEMISTRY

used for polypeptide sequencing.

[Ref: Harper 30th/e pg. 455]
6. Ans. (a) PCR
•• PCR (Polymerase Chain Reaction) is a method to
amplify DNA . [Ref: Harper 30th/e pg. 667]
•• Components of PCR are: DNA template, Primers (one
for each strand), DNA polymerase (Taq polymerase), •• Prenatal diagnosis of hemophilia is best done by
Deoxy-ribo-nucleoside tri-phosphates (d-NTPs), direct mutation analysis using PCR technique.
Buffer solution, Divalent cations- Mg++ or Mn++ Sample can be amniotic fluid, chronic villus sampling
(Generally Mg++is used), Monovalent cation- K+. or fetal blood sample.
•• No probe is used in PCR . Dideoxy nucleotide is also 7. Ans. (c) PCR


not used in PCR. If dideoxy nucleotide is used in PCR,

then DNA synthesis will stop,as it will be incorporated [Ref: Harper 30th/e pg. 455]
in the growing DNA chain and it will not allow the
•• In real time PCR, either SYBR Green dye or TaqMan
next normal nucleotide to be added.
dye can be used.
2. Ans. (a) HPLC
8. Ans. (d) FISH
[Ref: Robbins 10 /e pg 174,175]
[Ref: Harper 30th/e pg. 374]
•• Tests done in epigenetics are: Na bisulfite method,
•• Methods of gene transfer are Electrophoresis, Trans-
ChIP (Chromatin Immuno Precipitation), ChIP-
formation, Transfection, Lipofection, Conjugation,
on-chip, Fluorescent In Situ Hybridization (FISH),
and Transduction.
Methylation-sensitive restriction enzymes, PCR and
•• Site directed recombination is also called Conserva-
Microarray.
tive site specific recombination. This technique can
3. Ans. (a) Northern blot transfer specialized nucleotide sequences between
non-homologous sites within a genome. It can be
[Ref: Harper 30th/e pg. 457; Fig. 39.4] used as a method of gene transfer.
•• Blot transfer techniques: Northern blot detects •• Whereas FISH (Fluorescent In Situ Hybridization) is
RNA. Southern blot detects DNA. Western Blot a technique which uses fluorescently labelled DNA
detects Protein. South Western blot detects Protein- probes to detect gene or chromosome abnormalities
A DNA interactions. (cannot transfer gene).
N
S 4. Ans. (b); (e) 9. Ans. (a) Chromosome Painting
W [Ref: Harper 30th/e pg. 374]
E [Ref: Harper 30th/e pg. 362]
R •• Due to the presence of phosphates, DNA is negatively •• FISH (Fluorescent In Situ Hybridization) sometimes
charged and has tendency to move towards positive termed Chromosomal Painting is used to tell the
S exact location of gene on a chromosome.
side of the electrode during electrophoresis. Heavier
WITH fragment will move slower than lighter fragment •• Whereas Western blot detect proteins and FACS
which will move fast. So lighter fragment will be (Fluorescent Activated Cell Sorting) is a special type
E of flow cytometry which separate population of live
X towards the positive side and heavier fragment will
be towards the negative side. cella into groups of cells based on antibody binding.
P
L •• Out of the 5 options, heaviest fragment is 50 Kbp = 10. Ans. (b) FISH
A 50,000bp [1Kbp = 1000 bp]. Option b & e is same.
N [Ref: Harper 30th/e pg. 374]
5. Ans. (a) Dinitrobenzene
A •• For rapid chromosome identification, FISH is used.
T [Ref: Harper 30th/e pg. 458] FISH (Fluorescent in situ hybridization) can detect
I aneuploidy and it is a very rapid technique. Results
•• Sanger’s reagent or Di-Nitro-Fluoro-Benzene or
O can be obtained within 24 hours. Usually it is done in
DNFB - used for DNA sequencing and polypeptide
N intersex people.
sequencing.
S
 •• Karyotyping is a lengthy procedure (slow) but it is template sequence) and then Extension at 72°C
best method to detect aneuploidy. This question (polymerization or synthesis) occurs finally. These
mentions ‘Rapid’. three steps are repeated again and again to synthesize 369
•• Single-Strand Conformational Polymorphism (SSCP) large amount of DNA.
is a screening method which can identify genomic
variations in a large number of samples and in a 16. Ans. (d) 94°C

CHAPTER 14  TECHNIQUES IN MOLECULAR BIOLOGY

broad range of organisms.
[Ref: Harper 30th/e pg. 455]
11. Ans. (a) Analysis of DNA fragments •• High temperature (94°C) is required for denaturation
in PCR.
[Ref: Harper 30th/e pg. 463]
•• Restriction Fragment Length Polymorphism or RFLP 17. Ans. (c) Taq Polymerase
is used to identify a change in the genetic sequence
[Ref: Harper 30th/e pg. 454]
that occurs at a site where a restriction enzyme cuts.
•• RFLPs can be used to trace inheritance patterns, •• Taq Polymerase is a polymerase which is heat stable
identify specific mutations, and for other molecular and it is derived from a bacteria; Thermophillus
genetic-techniques. aquaticus or Thermus aquaticus. This bacteria
survives in boiling water so any enzyme of this
12. Ans. (c) Gene knockout animals bacteria can withstand high temperature.
[Ref: Harper 30th/e pg. 661] 18. Ans. (b) FRAP
•• Method which is most suited to assess function of a
[Ref: Harper 30th/e pg. 456]
gene include transgenic animals and gene knockout
animals. •• FRAP is Fluorescent Recovery After Photo bleaching.
•• Knock out means the particular gene of interest is This technique is used to detect movement of proteins
deleted from the organism’s genome and then the from one compartment of cell to another and lateral
effects of this gene’s absence can be used to study the diffusion of lipids and proteins in the membrane.
function of the gene.
•• Knockout mice have become widely used models for 19. Ans. (d) Restriction endonuclease
experimental study of human disease and provide [Ref: Harper 30th/e pg. 452]
essential information about gene function in vivo.
•• Restriction endonuclease enzymes cut DNA at
13. Ans. (d) Microarray specific sites known as palindromes and these
are also known as molecular scissors. They have
[Ref: Robbins 9th/e pg. 172-173] revolutionized genetic engineering and is the most
•• Microarray can detect variations in DNA sequences, common enzyme used in molecular techniques. A
and enables description of gene expression. N
•• Microarray technique is the most powerful tool for 20. Ans. (a) FISH
S
screening global pattern of gene expression. [Ref: Harper 30th/e pg. 464] W
14. Ans. (d) Microarray •• Gene therapy means when a gene or DNA is E
introduced into a cell for the treatment of a disease. R
[Ref: Robbins 9th/e pg. 172-173] •• FISH is Fluoresecent In Situ Hybridization, which is S
•• Aneuploidy (monosomy or trisomy) can be detected used for diagnosis, not for gene therapy or treatment.
WITH
by FISH (Fluorescent in Situ Hybridization), RT-PCR •• Other methods given in option are methods of gene
(Reverse Transcriptase PCR), QF-PCR (Quantitative transfer. E
Fluorescence PCR or Real Time PCR) and Multiplex X
21. Ans. (b) G-banding
ligation-dependent probe amplification (MLPA). P
•• These methods avoid the generation of cultured cells [Ref: Robbins 9th/e pg. 158; Internet] L
and can rapidly detect (within 1 or 2 days) aneuploidy. A
•• Microarray cannot detect aneuploidy. •• G-banding, or Giemsa banding (G stands for N
Giemsa) is used in cytogenetics to produce a visible A
15. Ans. (c) Denature DNA, Anneal Primers, Extend karyotype by staining condensed chromosomes T
DNA during metaphase. The metaphase chromosomes I
are treated with trypsin (to partially digest the O
[Ref: Harper 30th/e pg. 454] chromosome) and stained with Giemsa stain. It N
•• Three steps involved in PCR are Denaturation at yields a series of light and dark bands. Dark regions S
94°C (separation of DNA strands), then Annealing are heterochromatin and AT rich. The light regions
at 60°C (primer added and they get attached to are euchromatin and GC rich.
 •• R-banding is the reverse of G-banding (R stands for •• Single Nucleotide Polymorphisms (SNPs) are the
“reverse”). Dark regions are euchromatin and the common polymorphisms. Around 10 million SNPs
370 bright regions are heterochromatin. are found in humans.
•• Q-Banding (Q stands for Quinacrine Fluoroescent
Dye): Fluoroescent staining of chromosomes with 29. Ans. (a) Multiplex ligation-Dependent probe
Quinacrine gives bands that fluoresce on exposure to amplification (MLPA)
CRO BIOCHEMISTRY

UV light.
[Ref: Robbins 9th/e pg. 177]
Most common is G-banding.
•• Multiplex ligation-dependent probe amplification
22. Ans. (b) Metaphase (MLPA) can detect deletion or duplication of any
size (small or large). It is a kind of multiplex PCR
[Ref: Robbins 9th/e pg. 158-159]
which can simultaneously analyze multiple genomic
•• Karyotyping can be only done in metaphase. It regions.
cannot be done in any other phase of cell cycle.
30. Ans. (b) Can encode proteins
23. Ans. (d) Karyotyping


[Ref: Robbins 9th/e pg. 141]

[Ref: Ghai 8th/e/ ch 22, Nelson 20th/e/p 622]
•• mi-RNAs (micro-RNAs) are small non coding RNAs.
•• In Klinefelter’s syndrome, there are two or more They are 21–30 nucleotides in length. They do not
X-chromosome in a male. It is a trisomy (aneuploidy), code for any protein. These are transcribed but not
which is best detected by karyotyping. translated. They take part in gene silencing.

24. Ans. (b) Metaphase 31. Ans. (a) Protein DNA interactions and histone
modifications
[Ref: Robbins 9th/e pg. 158]
[Ref: Harper 30th/e pg. 465]
•• Colchicine causes the cell to arrest in metaphase, as
chromosomes are most condensed here and can be •• ChIP (Chromatin Immuno Precipitation) is used for
studied. interactions between DNA and proteins and histone
modifications.
25. Ans. (b) Lymphocytes
32. Ans. (c) Lox P site
[Ref: Robbins 9th/e pg. 161]
[Ref: Robbins 9th/e pg.172]
•• Peripheral blood lymphocytes are best cells for
chromosomal study as they do not undergo sub- •• CRE Recombinase binds to Lox P site. CRE Recom-
sequent cell divisions as they are terminally differen- binase causes site specific incorporation and is used
A tiated cells. in recombinant DNA technology.
N
26. Ans. (a) Study of multiple genes 33. Ans. (d) For monitoring the amplification target
S DNA
W [Ref: Robbins 9th/e pg. 174, 175]
E •• Microarray or chip can detect multiple mutations and
[Ref: Harper 30th/e pg. 455]
R multiple gene expression analysis simultaneously. •• Real-time PCR is used for monitoring the ampli-
S Thousands of oligonucleotide probes, corresponding fication of target DNA. In this PCR, we use SYBR-
to human genes are taken on a solid surface (gene green dye, which has a property of giving fluorescence
WITH
chip), allowing detection of multiple mutations at a when bound to dsDNA. Real time PCR is also known
E single go or multiple patient samples to be assessed as Quantitative PCR.
X for a single molecular marker in one experiment.
P 34. Ans. (b) RT PCR
L 27. Ans. (b) 1.5%
[Ref: Harper 30th/e pg. 29, 457]
A [Ref: Robbins 9th/e pg. 1]
N •• RT PCR is Reverse Transcriptase PCR, where starting
A •• Exons or coding genes comprise 1-2 % of the total material is RNA. It is used to make cDNA, which is
T genome. amplified. Thus RNA can be amplified by this method
I and quantification of mRNA can also be done.
28. Ans. (a) Single nucleotide polymorphism •• Western blot is to detect protein.
O
N •• Next generation sequencing & Sanger’s technique
[Ref: Robbins 9th/e pg. 3]
S are for DNA sequencing.
35. Ans. (d) GA TA TC CT AT AG 40. Ans. (a); (c); (d)
[Ref: Harper 30th/e pg. 463] [Ref: Wilson & Walker 7th/e pg 419-421] 371
•• RFLP is restriction fragment length polymorphism. •• Western blot, ELISA, Dot blotting all these techniques
This enzyme cut at a specific site known as can be used for protein detection.
palindromes. Palindrome is sequence same on both •• Dot blot assay is same like blotting but nucleic acid

CHAPTER 14  TECHNIQUES IN MOLECULAR BIOLOGY

strands. Direction is always from 5’ to 3’. is not separated by electrophoresis, instead sample is
directly applied on the membrane as a small dot.

41. Ans. (a); (b); (c); (d)

•• Non coding RNA means RNA that is transcribed from
DNA but is not translated.
•• Coding RNA is only mRNA, which is transcribed as
well as translated.
36. Ans. (c) Knock down 42. Ans. (a); (b); (c)
[Ref: Robbins 9th/e pg. 319] [Ref: Harper 30th/e pg. 455]
•• RNAi (RNA Interference) is gene knock down. This •• PCR: Polymerase chain reaction is for amplification
means that gene is present but its function is supp- of DNA.
ressed. •• DNA cloning: Technique used to produce multiple
copies of a gene or other piece of DNA.
37. Ans. (a) 3’ untranslated region •• ELISA: Enzyme linked Immuno Sorbent Assay →
[Ref: Harper 30th/e pg 368] used to detect and measure antigen or antibody in
blood
•• Gene knock down is when function of the gene •• New generation DNA sequencing: Describes a
present is suppressed by inhibiting the target mRNA number of different sequencing techniques which
so that it cannot form its protein. tells the base sequence of DNA.
•• miRNA is formed from RNA Polymerase II which
selectively inhibits the mRNA. It binds at 3’ end of 43. Ans. (a) Agarose gel
mRNA.
[Ref: Wilson & Walker 7th/e pg 419-421]
38. Ans. (b) Comparative genomic hybridization •• Separation of RNA is done by Agarose gel electro-
•• Comparative Genomic Hybridization is a cytogenetic phoresis, which is used in Northern blotting,
method for analyzing DNA copy number variations checking RNA integrity and size selection of RNA
in diseases like cancer. Frequently occurring regions cloning experiments. A
of DNA amplification usually harbour oncogenes, •• Agarose gel is usually used for Nucleic Acid separation N
whereas regions of recurrent deletion harbour tumour and it separate large molecules. S
suppressor genes. Such copy number changes can be •• Whereas Polyacrylamide gel is used for DNA or W
detected in tumours by this technique. It can compare Protein separation and it seperates small molecules. E
test sample with a reference sample, without the R
need for culturing cells. 44. Ans. (a); (b); (c); (d)
S
39. Ans. (b) Spectrophotometer [Ref: Robbins 9th/e pg. 319]
WITH
th •• RISC is RNA induced Silencing complex, which is
[Ref: Harper 30 /e pg 26] E
formed when single stranded miRNA is attached to
•• Spectrophotometry is based on the principle that every proteins. X
compound absorbs, transmits, or reflects light over •• Components/genes involved in RISC complex are : P
a certain range of wavelength. Spectrophotometry is ƒƒ Drosha (an endonuclease present in nucleus)- cut L
a measurement of how much a chemical substance primary mi RNA A
absorbs or transmits. So this measurement can also ƒƒ Drosha works with DGCR8, also known as Pasha N
be used to measure the amount of a known chemical (to rhyme with Drosha) A
substance. Spectrophotometry can be used in both ƒƒ Dicer (an endonuclease present in cytoplasm) – T
qualitative and quantitative analysis of DNA, RNA, cut pre mi RNA I
and proteins. ƒƒ mi RNA (micro RNA) O
ƒƒ si RNA (small interfering or silencing). N
S
 45. Ans. (c) Bacterial Artificial Chromosomes •• DNase I hypersensitive sites are regions of Chromatin
that are sensitive to cleavage by DNase I enzyme.
372 [Ref: Harper 30th/e p455] These regions contain the actively trans-cribed genes.
•• Yeast Artificial Chromosomes (YAC) carry the largest •• FISH (Fluorescence In situ Hybridization) uses fluor-
DNA segments (500-3000kb). But from the options escent DNA probes that binds to specific region of
given, Bacterial Artificial Chromosomes (BAC) carry Chromatin. It is used in detection of Aneuploidy and
CRO BIOCHEMISTRY

larger DNA segments (50-250kb). Refer Table 14.2. detection of gene location on chromosome.

46. Ans. (a) Transduction 51. Ans. (a) AAGCTT

[Ref: Satyanarayana 5th/e pg. 575] [Ref: Harper 30th/e pg. 451]

•• Transduction method can be used to introduce •• Do not get confused by the word ‘Hind III’ given. It
foreign DNA into a cell by a virus or viral vector. is just a restriction endonuclease enzyme, which will
•• Whereas Transcription, Lysogenic conversion, digest a restriction sequence or Palindrome
Transformation cannot introduce foreign DNA into •• Out of the four choices, select the inverted repeat /
a cell. palindromic sequence. Only option a) is a palin-


drome.
47. Ans. (d) Transfection

[Ref: Satyanarayana 5th/e pg. 575]

•• Transfection is the transfer of non viral genetic
material into eukaryotic cells, not bacterial cells. All
other options are methods of gene transfer from one
bacteria to another.

48. Ans. (a) X contents 2.8% more than Y 52. Ans. (c) Escherichia coli
[Ref: Harper 30th/e pg. 374] [Ref: Harper 30th/e pg. 451]
•• X chromosome is larger and always has higher •• The commonly used prokaryotic host in rDNA
content of DNA than Y chromosome. This percentage technology is bacteria, Escherichia coli (E. Coli).
is 2.8%.
•• Sperm Sorting or Microsort Method separates X- 53. Ans. (b) DNase I
bearing & Y- bearing sperms due to this difference
between these two sex chromosomes. [Ref: Harper 30th/e pg.373]
Deoxyribonuclease I (DNase I) acts on single-stranded
A 49. Ans. (c) Production of monoclonal antibodies DNA, double-stranded DNA, and chromatin and makes
N single stranded cuts in nearly any segment of DNA
S [Ref: Harper 30th/e pg 684-685]
i.e. has low sequence specificity. It will digest DNA
W •• In Hybridoma technique, two cells i.e. myeloma that is not protected, or not bound by protein, into its
E cells and spleen cells (from immunized animals), are component deoxynucleotides. It is used in DNA Foot
R fused and is used for the production of large numbers Printing.
S of identical antibodies known as monoclonal E.g. Pancreatic Deoxyribonuclease (DNase I) nick the
antibodies. single strands of double-stranded DNA. Two nicks
WITH
sufficiently close on opposite strands will lead to
50. Ans. (c) Chromatin Immuno Precipitation- breakage of the DNA molecule.
E Sequencing
X
54. Ans. (a) A circular double stranded DNA
P [Ref: Harper 30th/e p466 fig 39-11]
L [Ref: Harper 30th/e pg.455]
•• Chromatin Immuno Precipitation (ChIP) is used
A
N to find the regions of DNA to which DNA binding •• A plasmid is a small, circular ds DNA molecule within
proteins (like Transcription Factors) bind. It detects a cell that is physically separated from chromosomal
A
T PTMs (Post Translational Modifications) of histones DNA and can replicate independently. Plasmids
I and protein-DNA interactions. confer increased resistance to antibiotics.
O •• Fluorescence Resonance Energy Transfer (FRET) •• It is mostly found in bacteria. However, plasmids
N is used to detect the distance between two macro- are sometimes present in archaea and eukaryotic
S molecules e.g. Protein-Protein Interaction. organisms also.
 57. Ans. (d) All the above

[Ref: Harper 30th/e pg. 459] 373

•• Applications of recombinant DNA technology are
distant hybridization, development of transgenic
plants, development of root nodules in cereal crops,

CHAPTER 14  TECHNIQUES IN MOLECULAR BIOLOGY

development of C4 plants, production of antibiotics,
production of human proteins in large quantities,
production of hormone insulin, production of
vaccines, production of interferon, production
of enzymes, Gene Therapy solution of disputed
parentage, molecular analysis of disease.

Plasmid 58. Ans. (d) Restrict the expression of Viral DNA

[Ref: Harper 30th/e p452]

55. Ans. (d) All the above
•• Restriction enzymes are protective endonucleases
[Ref: Harper 30th/e pg. 464] present in bacteria that cleaves the infecting
•• A transgenic animal is one whose genome has been bacteriophage DNA
altered by the transfer of a gene or genes from another •• Each restriction enzyme recognize a short, double
species or breed. stranded sequence (4 to 6 bp) which is usually
palindromic.
56. Ans. (b) Mouse •• When the cleavage is directly proportional to each
[Ref: Harper 30th/e pg. 464] other, blunt ends are produced
•• When the cleavage is not opposite to one another, the
•• Transgenesis refers to the phenomenon of intro- resulting fragments can circularize. These ends are
duction of exogenous DNA into the host genome to known as sticky or cohesive ends.
create and maintain a stable heritable character. The
Uses of Restriction enzymes
foreign DNA that is introduced is called Transgene.
•• Recombinant DNA technology: DNA fragments
The animal whose genome is altered by adding one
from different species are cut with same restriction
or more transgenes is said to be a Transgenic animal.
enzymes.
•• The most commonly used animal model in
•• Detection of Restriction Fragment Length Poly-mor-
transgenesis to represent humans is Mouse, not
phism (RFLP)
only because their genomes are so similar to that of
•• Restriction Enzymes can be used to generate a
humans, but also because of their availability, ease of
restriction map of a particular organism.
handling, high reproductive rates, and relatively low A
cost of use. N
S
W
E
R
S
WITH

E
X
P
L
A
N
A
T
I
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UNIT
VI

MISCELLANEOUS
Miscellaneous 15
Overview of Chapter
H igh R eturn
•• Vitamins zz Vitamins synthesized by intestinal flora: B2, B5, B7, K
•• Minerals zz Royal jelly (bees) – rich in Vitamin B5, B6, B7
•• Xenobiotics
Vitamins are divided into two groups: Water soluble and Fat
•• Free radicals soluble.
•• RQ All water soluble vitamins act as coenzyme but vitamin
•• Alcohol K is the only fat soluble vitamin which acts as coenzyme. All
fat soluble vitamins can be stored in body but vitamin B12 is
the only water soluble vitamin which can be stored in body.
Vitamins differ from hormones in not being produced within
Micronutrients
the organism.

Water Soluble Vitamins

These include Vitamin B complex and Vitamin C.
Typical deficiency symptoms of this group are: Derma-
titis, Glossitis, Cheilosis, Diarrhoea and some neurological
symptoms, e.g. peripheral neuropathy.

Vitamin B Complex
Vitamins in this group are not related to each other in structure
but these are grouped together because all acts as coenzyme.
The larger the caloric intake, larger is the requirement of
B-complex vitamins. These are B1 (Thiamine), B2 (Riboflavin),
B3 (Niacin), B5 (Pantothenic acid), B6 (Pyridoxine), B7 (Biotin),
B9 (Folic acid), B12 (Cobalamine).

H igh R eturn
Choline, Inositol, Lipoic acid, PABA (Para-Amino Benzoic acid) →
are not vitamins as they are easily available in body.
VITAMINS
Vitamins: These are the organic essential compounds of low
Vitamin B1 (Thiamine)
molecular mass, which are required in minute quantities in
diet, as they cannot be synthesized in body. They must be
obtained from some exogenous source e.g. diet or bacterial H igh R eturn
flora in gut. zz Active moiety: TPP (Thiamine Pyro Phosphate)
zz Sources: Grains (Aleurone layer), unpolished rice, yeast.
Exception : Two vitamins can be synthesized in body, they
Richest source is rice polishings
are vitamin B3 (Niacin) and vitamin D, so these are called zz Main biochemical role: Oxidative decarboxylation
atypical vitamins. These vitamins are synthesized by human zz Biochemical assessment of deficiency: Erythrocyte Transketo-
enzymes during metabolism, not by intestinal bacterial lase activity reduced
enzymes. Vitamin B3 is synthesized from tryptophan amino zz Deficiency leads to Beri-Beri (most common in alcoholics)
acid and vitamin D is synthesized from 7-dehydro cholesterol
by UV light in skin. This vitamin has a pyrimidine like ring in its structure
that’s why the name ‘Thiamine’ is matching with the
pyrimidine ‘Thymine’.
 Biochemical Roles of Thiamine Beri-Beri is usually of mixed type (Wet and Dry): Wet
zz Required in oxidative decarboxylation reactions (for beriberi is called ‘Wet’ because it is associated with Oedema.
378 multienzyme dehydrogenase complexes e.g. pyruvate Infantile Beri-Beri – occurs in infants born to mothers
dehydrogenase, alpha-keto glutarate dehydrogenase, deficient in Vitamin B1. Sign and symptoms include restless-
branched chain alpha keto acid dehydrogenase) ness and sleeplessness. Symptoms appear suddenly and it
zz Transketolase (in HMP) can lead to cardiac failure and cyanosis.
CRO BIOCHEMISTRY

zz Transketolase is also the marker enzyme for B1 deficiency

zz Also, B1 required in phosphorylation of a chloride Vitamin B2 (Riboflavin)
channel in brain
H igh R eturn
H igh R eturn zz Active moiety: FAD (Flavin Adenine Dinucleotide), FMN (Flavin
NOTE: Main role of this vitamin is in carbohydrate metabolism. Mono Nucleotide)
So, vitamin for which RDA is based on carbohydrate intake is B1 zz Sources: Milk, egg, liver, fish, whole cereals, legumes, green
leafy vegetables. Flavanoids are colored, so more in broccoli
B1 Deficiency leads to Beri-Beri. As Vitamin B1 is mainly (than in Cauliflower), in colored bell peppers
involved in oxidative decarboxylation reactions, so in B1 zz Endogenously synthesized by bacterial flora


deficiency, highly aerobic tissues e.g. heart and brain fail first. zz Main biochemical role: Oxidation – Reduction reactions
Lactic Acidosis occurs in Beri-Beri because Pyruvate zz Biochemical assessment of deficiency: Erythrocyte Glutathi-
Dehydrogenase (enzyme which converts Pyruvate to Lactate) one Reductase activity reduced
requires Vitamin B1. In B1 deficiency, Pyruvate accumulates zz Deficiency: Riboflavinosis (Oro-Oculo Genital Syndrome)
leading to lactic acidosis.  Also known as Warburg Yellow Enzyme

Biochemical Role
zz FAD requiring enzymes are → Pyruvate Dehydrogenase
(link reaction ), Succinate dehydrogenase (TCA), alpha-
ketoglutarate dehydrogenase (TCA), branched chain
alpha keto acid dehydrogenase, complex II (ETC),
glutathione reductase (RBC ), D-amino acid Oxidase,
acyl CoA dehydrogenase, xanthine oxidase, and glycerol-
3-phosphate dehydrogenase -mitochondrial form
(involved in glycerol phosphate shuttle)
zz FMN requiring enzymes are → L-amino acid Oxidase,
Complex I (ETC) and Monoamine oxidase

B2 Deficiency: Riboflavinosis → It is characterized by

zz Corneal vascularization (earliest sign of deficiency).
zz Glossitis – inflammation of tongue (magenta tongue-
characteristic)
zz Cheilosis – redness and shiny appearance of lips
zz Angular stomatitis (lesions at mucocutaneous junction
at corners of mouth), leading to painful fissures
zz Seborrhoic dermatitis – rough scaly skin because of des-
quamation.
Fig. 15.1: Clinical features and types of Beri-Beri

T
H
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Fig. 15.2: Clinical features in B2 deficiency

Vitamin B3 (Niacin) Causes of Deficiency of Niacin
This is not strictly a vitamin as it can be synthesized from
tryptophan (60 mg of tryptophan can synthesize 1 mg of
Dietary deficiency of Tryptophan, Hartnup’s disease (Trypto- 379
phan is excreted in urine), Carcinoid syndrome (Tryptophan
Niacin). Rate limiting step in niacin synthesis is QPRTase
is used for the synthesis of serotonin), Vitamin B2 and B6
(Quinolinate Phospho Ribosyl Transferase). This synthesis
deficiency (affects conversion of tryptophan to niacin, which
also requires vitamin B2 and B6. So, B2 and B6 deficiency leads

CHAPTER 15  MISCELLANEOUS
requires these vitamins), Isoniazid therapy – anti TB drug
to B3 deficiency.
(inhibits PLP formation) so Isoniazid causes deficiency of
In high corn diet, requirement of dietary Niacin increases
both B3 and B6and High corn/maize diet.
because maize protein (zein) lacks Tryptophan. So pellagra is
more common when maize is the staple diet.
A dditional E dge
Incidence of Pellagra is more common in females as compared to
males as oestrogen metabolites inhibits Tryptophan metabolism.

Toxicity of Niacin
Cutaneous flushing causes itching and burning (this is
because of histamine release, leading to transient vaso-
H igh R eturn dilatation), gastric irritation, hepatic toxicity (most serious
Active moiety: NAD (Nicotinamide adenine dinucleotide), NADP toxic reaction).
(Nicotinamide adenine dinucleotide phosphate)
Sources: Meat, liver, fish, whole cereals, legumes, peanut Vitamin B5 (Pantothenic acid) – Meaning ‘Everywhere’
Main biochemical role: oxidation – reduction reactions
zz Contains beta alanine in its structure
Deficiency: Pellagra (meaning rough skin)
zz Present in Coenzyme A and Acyl Carrier Protein (ACP)

H igh R eturn
Active moiety: Coenzyme A and ACP (Acyl carrier protein)
Sources: Widely distributed in plants and animals. Yeast, liver
honey and egg are good sources
Synthesized by bacterial flora
zz Many enzymes use NAD+ (covered during metabolism).
NADPH is used in reductive biosynthesis. Main biochemical role: Acyl group transfer
zz Enzyme which can use both NAD and NADP → Glutamate Deficiency: Rare (Burning feet syndrome) i.e. pain sensation in
dehydrogenase (converts glutamate to a-ketoglutarate) lower extremities.
B3 Deficiency: Pellagra → characterized by:
zz Dermatitis (Photosensitive Biochemical Role of Pantothenic Acid
Dermatitis)
zz ACP: Role in Fatty Acid Synthesis
zz Diarrhoea 4 Ds of
Pellagra zz CoA: has role in TCA, haem synthesis, lipogenesis, fatty
zz Dementia acid Oxidation
zz Death zz Active group of CoA is a free SH group or Thiol group to
Photosensitive Dermatitis means patients have red, thick which acyl groups can be covalently linked to form thio
and scaly skin on exposure to sunlight. Patients have casal’s ester bond. e.g. Acetyl CoA
necklace.

Vitamin B6 (Pyridoxine) T
H
H igh R eturn E
O
Active moiety: PLP (Pyridoxal phosphate) R
Sources: Pork, fish, poultry (chicken or turkey), potatoes Y
vegetables, wholegrain cereals such as oatmeal, grain and brown
rice.
Fig. 15.3: Clinical features of pellagra (B3 deficiency)
Contd…
 Main biochemical role: Transamination, simple decarboxylation H igh R eturn
Deficiency: Neurological manifestations such as Peripheral Neuro-
380 Biotin is a prosthetic group for all carboxylases. But there are few
pathy, Mental Confusion, Depression, Carpel Tunnel Syndrome,
Microcytic Anemia, Convulsions, Kidney Stones, Pellagra (as B6 carboxylation reactions which do not require Biotin. They are:
zz CPS-I and II (Carbamoyl phosphate synthetase )
required for conversion of Tryptophan to Niacin)
zz CO2 addition in C6 of purines
CRO BIOCHEMISTRY

zz Pyruvate (3C) to malate (4C) conversion by malic enzyme

Biochemical Role of Pyridoxine zz Gamma carboxylation of glutamate (Vitamin K required, not
zz Transamination (first step in the catabolism of amino Biotin).
acids), Trans-sulfuration, Deamination
zz Simple decarboxylation (mainly histidine, gluatamate, Causes of Deficiency of Biotin
tryptophan)
zz ALA synthase (Haem synthesis) zz Excess consumption of raw eggs (contains a protein
zz Cystathionine synthase (synthesis of Cysteine) ‘Avidin’ – a Biotin binding protein.) When cooked, Avidin
is partially denatured and binding to Biotin is reduced.
zz Alkaline phosphatase
So cooked egg whites are safe to consume. So vitamin B7
zz Glycogen phosphorylase (RLE in glycogen breakdown)


is also known as Anti-egg white injury factor.

zz Tryptophan metabolism, synthesis of Niacin zz Multiple Carboxylase deficiency- in which there is defect
(vitamin B3) in association of biotin and Carboxylases.
H igh R eturn Treatment: Biotin supplementation.

NOTE: Here that main role of this vitamin is in amino acid and Vitamin B9 (Folic acid)
protein metabolism.
Vitamin for which RDA is based on protein intake –B6 H igh R eturn
Active moiety: THF (Tetra Hydro Folate)
Q. Vitamin B6 deficiency may lead to sleep distur- Sources: Green leafy vegetables, whole grains, fruits, eggs, dairy
bance because: products
T • Vitamin B helps regulate and synthesize GABA Main biochemical role: Carrier of one carbon units
H 6
& Serotonin. Even a mild deficiency in Vitamin Deficiency: Megaloblastic anemia, Neural tube defects, Homo-
I
N B6 may result in a down-regulation of GABA and cystenemia, Homocystinuria.
K serotonin synthesis. This deficiency would result
in GABA activity disruption, leading to obstructed Folic acid/Folate acts as carrier of one carbon units. Biotin
sleep as GABA is the central nervous system’s chief (carboxylation) and SAM (S-adenosyl methionine for
inhibitory neurotransmitter. methylation) are also involved in addition of one carbon unit
to a metabolic precursor. But CO2 or HCO3– (bicarbonate) is
not considered a member of one carbon pool.
Biotin (Vitamin B7)
Activation
H igh R eturn A single NADPH dependent enzyme DHFR (Dihydro Folate
Active moiety: Biocytin (Biotin + Lysine) Reductase) catalyzes both steps
Sources: Soyabean, whole grains, yeast, egg yolk and organ meat
Endogenously synthesized by bacterial flora
Main biochemical role: Carboxylation reactions
Deficiency: Alopecia, bowel inflammation, muscle pain

Biochemical Role of Biotin

zz Carboxylation, e.g. Pyruvate Carboxylase, Acetyl CoA
Carboxylase, Propionyl CoA Carboxylase, Methyl
Crotonyl CoA Carboxylase.
T zz Regulation of cell cycle
H
E
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Coenzyme Functions of THF

381

CHAPTER 15  MISCELLANEOUS
Fig. 15.4: One carbon pool
zz Major form of Folic acid for transfer of one carbon is –Methylene THF
zz Main form of Folic acid in blood is - Methyl THF
zz Active form of Folic acid is – Tetra Hydro Folate

T
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O
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 Folate Deficiency zz Ribonucleotide Reductase
DNA synthesis indirectly requires Folic acid (folate required zz Leucine Amino Mutase
382 for conversion of dUMP to dTMP, leading to decreased
thymidine, which is required for DNA synthesis). In Folic acid Deficiency of Vitamin B12
deficiency DNA replication and thus cell division are affected. Causes of Deficiency
Pernicious Anemia → an autoimmune disease in which
CRO BIOCHEMISTRY

The cytoplasm of folate deficient cell grows at a normal rate, zz

but cell division is delayed. So abnormally large cells are antibodies formed against intrinsic factor. So absorption
formed. This leads to megaloblastic anemia. Rapidly dividing of vitamin B12 is decreased.
cells e.g. bone marrow, intestinal mucosa are affected. zz Dietary deficiency of vitamin or in pure vegetarian diet
Clinical manifestations of deficiency:
Diagnosis of Folate Deficiency
zz Haematological: Megaloblastic Anemia, Hypersegmen-
tation of neutrophils
zz Neurological: Demyelination in peripheral nerves and
spinal cord
Megaloblastic Anemia is due to secondary deficiency of


folate (folate trap in B12 deficiency). Peripheral Neuropathy is

due to accumulation of L-methyl malonic acid.

H igh R eturn
zz To distinguish between folate and B12 deficiency → neurological
symptoms do not occur in Folate deficiency.
Vitamin B12 (Cobalamin)
Cobalamin is chemically the most complex vitamin. It
contains a cobalt atom in the structure with four pyrrole
rings (this is the only known role of cobalt in mammals).
This tetrapyrrole (called Corrin ring) structure is similar to
porphyrin.

H igh R eturn
zz Active moiety: Methyl Cobalamin and Deoxyadenosyl Coba-
lamin
zz Sources: milk, curd, cheese, egg, liver, meat, fish (only found
in animal sources)
zz Main biochemical role: Synthesis of Methionine, isomerization
of Methyl Malonyl CoA Fig. 15.5: Folate trap occurs in B12 deficiency. All the folate of
zz Deficiency: Megaloblastic Anemia, Subacute Combined De- body is present in the form of methyl THF, which is required only in
generation, Demyelination this reaction. But THF is required for so many reactions in the body,
which is now not available. So in cobalamin deficiency, functional
zz Cyanocobalamin → synthetic form of vitamin B12. It is Folate deficiency occurs
cheapest supplement option and it also gets stored in
liver. A dditional E dge
zz Methyl Cobalamin → Transport form There are five transmethylation reactions occurring in body. They
zz Hepatocorrin → a protein synthesized by liver, transports are:
B12 in plasma

Absorption
A glycoprotein called intrinsic factor, produced by parietal
T cells of stomach is required. The vitamin – intrinsic factor
H complex reaches ileum from where it is absorbed. Vitamin
1. Norepinephrine → Epinephrine
E B12 is then converted to methyl cobalamin and released into
2. Cephalin (Phosphatidyl Ethanolamine) → lecithin (Phos-
O bloodstream. Methyl cobalamin is the major circulating form.
phatidyl Choline)
R 3. Guanidine Acetate → Creatine
Biochemical Role
Y 4. Acetyl Serotonin → Melatonin
zz Homocysteine to Methionine by Methyl Cobalamin 5. Polynucleotide → Methylated Polynucleotide (e.g. methyl
zz Methyl Malonyl CoA to Succinyl CoA (enzyme Mutase) group on the 5’ cap on mRNA).
by deoxy adenosyl cobalamin
 Hydroxylase (Bile Acid ynthesis), Tryptophan Hydro-
xylase (Tryptophan to Serotonin conversion), formation
of catecholamines (Dopamine Beta Hydroxylase for 383
conversion of dopamine to norepinephrine), Carnitine
synthesis, alpha oxidation of fatty acids, hydroxylation of

CHAPTER 15  MISCELLANEOUS
steroid hormones and in Tyrosine catabolism.
zz Role in wound healing
zz Iron absorption (by reducing ferric to ferrous) and
Ferritin formation, so deficiency leads to anemia
zz It can convert metHb to Hb, so prevents hemolysis due
to metHb
Fig. 15.6: Megaloblastic anemia in Folate and B12 deficiency zz Antioxidant
zz Lowers blood cholesterol
Urinary Metabolites zz There is increased requirement in stress.
Vitamin deficiency Common Special
metabolite metabolite
Vitamin B6 deficiency Homocysteine Xanthurenic acid
Vitamin B9 deficiency Homocysteine FIGLU
Vitamin B12 deficiency Homocysteine L-methyl malonic
acid Fig. 15.7: Vitamin C role in Tyrosine catabolism

Vitamin C (Ascorbic Acid)

H igh R eturn
Active moiety: Ascorbic acid itself
zz Resembles monosaccharide in its structure.
Sources: Citrus fresh fruits, raw or minimally cooked green leafy
Fig. 15.8: Vitamin C role as antioxidant
vegetables. Amla is the best source. This is easily destroyed by
heat. zz Vitamin C can cross placenta and is secreted in milk if
Main biochemical role: Hydroxylation reactions (Vitamin C acts mother has good intake (but milk does not contain iron)
as reducing agent), antioxidant
Deficiency: Scurvy Vitamin C deficiency: Scurvy
This vitamin can be synthesized in non-primates due zz Characterized by bleeding gums, poor wound healing,
to presence of enzyme L-Gulono Lactone Oxidase. This anemia. (Fig. 15.9)
enzyme is not present in primates, thus humans cannot zz Infantile scurvy is known as Barlow’s disease
synthesize this vitamin.
Vitamin C toxicity
Biochemical Role:
Widely distributed in body mainly in glandular tissues zz Gastric irritation, flatulence, diarrhoea, oxalate
zz Hydroxylation reactions e.g. Prolyl Hydroxylase stones (as most of the oxalate in urine is derived from
and Lysyl Hydroxylase (collagen synthesis), 7-Alpha Vitamin C).

T
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384
CRO BIOCHEMISTRY


Fig. 15.9: Vitamin C deficiency (Scurvy)

Table 15.1: Water Soluble Vitamins

Vitamin Name Active Moiety Biochemical role Deficiency

B1 Thiamine TPP Oxidative decarboxylation, Beri-Beri
transketolase
B2 Riboflavin FAD, FMN Redox reactions Riboflavinosis (Oro-oculo genital syndrome)
B3 Niacin NAD, NADP Redox reactions Pellagra (Diarrhoea, Dermatitis, Demertia, Death)
B5 Pantothenic aid CoA, ACP Acyl carrier Burning feet syndrome
B6 Pyridoxine PLP Simple decarboxylation, Neurological manifestations, carpal tunnel syndrome
transamination
B7 Biotin Biocytin Carboxylation Rare
B9 Folic acid THF One carbon transfer Megaloblastic anemia, Neural tube defects
B12 Cobalamin Methyl Methylmalonyl CoA mutase, Megaloblastic anemia + Neurological symptoms
Cobalamin homocysteine to methionine
C Ascorbic acid Ascorbic acid Hydroxylation reactions Scurvy

T Fat Soluble Vitamins

H
E General Properties of Fat Soluble Vitamins
O zz Assembled from isoprenoid units
R zz Absorbed into intestinal lymphatics along with dietary lipids, then they reach liver and then other organs
Y zz They are non-polar, so they cannot be excreted via kidneys. So they tend to be stored in body
zz Toxicity can ccur
zz Coenzyme function is not known (Except Vitamin K).
Vitamin A (Carotene) zz Bitot’s spots- superficial deposition of keratin in conjunc-
tiva (foamy appearance on conjuction)
zz Mainly vitamin A is retinol
zz Keratomalacia: Corneal ulceration and scarring 385
zz Retinoids refer to all other compounds related to retinol
zz Follicular hyperkeratosis (gooseflesh) is an important
H igh R eturn early sign
Skin lesions, atrophy of epithelium leading to respiratory

CHAPTER 15  MISCELLANEOUS
zz
Active moiety: Retinol, Retinal, Retinoic acid and genitourinary infections.
Sources: Leafy vegetables, carrots, whole milk, fish liver oil, egg, zz Dry scaly skin, Immunosuppression.
butter, meat (richest plant source is carrot )
Main biochemical role: Antioxidant, Vision
Deficiency: Night blindness (Nyctalopia), Xerophthalmia, Bitot’s
spots, Keratomalacia

There are three biologically active forms:

zz Retinol: necessary for reproductive system
zz Retinal: involved in vision
zz Retinoic acid: involved in growth and cellular differen-
tiation
Carotenoids are present in plants, which is a provitamin.
It contains two molecules of retinal.
Retinoids are all natural/ synthetic compounds with
Vitamin A like activity. Vitamin A Toxicity
Characterized by dry, pruritic skin, hepatomegaly, arthralgia
A dditional E dge raised intracranial pressure which sometimes mimics brain
Transport of Retinol in blood: Retinol binds to Retinol Binding tumor. Organelle affected is lysosomes.
Protein (RBP), which has low MW. So RBP can be excreted via
kidneys. Therefore retinol – RBP complex reversibly binds with
Vitamin D
plasma protein, Transthyretin. RBP has single binding site for
Retinol. Retinoic acid binds with albumin. H igh R eturn
Active moiety: calcitriol
Functions of Vitamin A Sources: fish, egg yolk, liver, butter, cheese,
Main biochemical role: calcium and phosphate absorption from
zz Role in vision: Vitamin A is a prosthetic group of intestine and their reabsorption from kidneys
Rhodopsin (light sensing protein in retinal rod cells). Deficiency: rickets in children, osteomalacia in adults, osteopo-
dark adaptation is the time taken for the regeneration of rosis in elderly
rhodopsin which is required for vision in dark light. This
time is prolonged in Vitamin A deficiency Calcitriol is a hormone because:
zz Regulation of gene expression zz It is synthesized in body, released in circulation and has
zz Growth and differentiation distant target organs
zz Glycoprotein synthesis zz Mechanism of action resembles group II hormones i.e. it
zz Role in reproduction binds with nuclear receptors
zz Antioxidant role. Synthesis of Vitamin D
zz Prevent squamous metaplasia. zz Activation of 7-dehydro cholesterol by UV light in skin
produces cholecalciferol
zz All-trans Retinoic acid is used to treat measles and Acute
zz 25-hydroxylation in liver to form 25-hydroxy cholecalcif-
Pro-Myelocytic Leukemia (APL)
erol
zz Used topically for wrinkles and acne (oral Isotretinoin zz 1-alpha- hydroxylation in kidneys to form 1,25-dihydroxy
to treat severe cystic acne). Isotretinoin is teratogenic so cholecalciferol
prescribe it only when preg-nancy test is negative. zz PTH activates 1-a-hydroxylase of kidneys
Deficiency of Vitamin A Deficiency of Vitamin D
T
zz Most common cause of preventable blindness zz Rickets in children, osteomalacia in adults, osteoporosis H
zz Night blindness (Nyctalopia), in elderly
E
zz Xerophthalmia/Dry eyes
O
R
Y
 Functions of Vitamin E
Antioxidant role
386
zz
zz Antiatherogenic role: It converts oxidized LDL back to
normal LDL. Oxidized LDLs are strongly associated with
atherosclerosis and CAD.
CRO BIOCHEMISTRY

Vitamin K

H igh R eturn
Active moiety: Hydroquinone
Sources: Green leafy vegetables
Endogenously synthesized by bacterial flora
Main biochemical role: Coenzyme role in gamma carboxylation
of glutamate in prothrombin
Deficiency: Leads to easy bruising, Bleeding or fatal hemorrhagic


disease

zz K1 – in vegetables - Phylloquinone
zz K2- from intestinal bacteria - Menaquinone
zz K3- synthetic water soluble Vitamin K – Menadione

Biochemical Role of Vitamin K

zz Maintenance of normal levels of clotting factors (II, VII,
IX and X) and protein C and S, bone protein – Osteocalcin,
kidney protein – nephrocalcin. (a heterogenous group of
proteins).
zz This is done by coenzyme role of Vitamin K in the
gamma- carboxylation of glutamate and this is a post
translational modification in these proteins, specially
Vitamin E (α- Tocopherol) prothrombin- factor II. This is a critical step in the
activation of these proteins. Carboxy Glutamate has
H igh R eturn negative charge, which is attracted to the positive
charge of calcium, helping in coagulation. This way
Sources: Vegetable edible oils vitamin K introduces Ca2+ binding sites on these calcium
Main biochemical role: Antioxidant (most potent lipid phase dependent proteins. (Fig. 15.10)
antioxidant)
Deficiency: Haemolytic Anemia in premature infants. In adults,
degenerative changes in muscles and sterility.

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Fig. 15.10: Coenzyme role of vitamin K

Reasons of Deficiency of Vitamin K zz Maintainance of neuromuscular excitability
Fat malabsorption, Prolonged antibiotic exposure (as it zz Present in alkaline digestive juices e.g. pancreatic juice
zz
destroys intestinal flora), drugs like Warfarin, Dicumarol and bile 387
and Orlistat zz Glycogen and protein storage requires potassium
zz Cofactor for enzymes e.g. Pyruvate kinase

CHAPTER 15  MISCELLANEOUS
In Newborns, this vitamin is injected during birth
zz
Calcium
Deficiency is common in newborns so leads to Haemorrhagic
syndrome. This is because: It is found in highest concentration amongst all minerals, out
zz Intestine of newborns is sterile
of which 99% is present in bones.
zz Low fat stores
zz Limited storage of vitamin K in neonatal liver There are three forms of calcium present:
zz Breast milk has poor content of vitamin K 1. Ionized Calcium/ Diffusible Calcium: This form is freely
zz Liver immaturity
exchangeable between extracellular fluid, soft tissues
zz Poor placental transport
and blood. About 45-50% is in this form. It performs most
of the functions of calcium (metabolically active form of
Vitamin K Toxicity
Ca).
Vitamin K toxicity can occur due to excess use of synthetic 2. Protein bound Calcium: around 45-50% of serum
Vitamin K Menadione. It reacts with sulfhydryl groups of calcium is bound with proteins
many enzymes and can lead to brain damage, haemolytic 3. Calcium salts: Around 5-10 % is present in the form of
anemia and hyperbilirubinemia. This should be kept in mind calcium salts e.g. calcium phosphate, calcium citrate
while treating premature infants.
Biological Functions
zz Calcification of bones and teeth (Ca forms hydroxy
apetite crystals along with phosphate)
zz Activator of enzymes e.g. adenylate Cyclase, ATPase,
Protein Kinase
zz Blood clotting
zz Muscle contraction, nerve impulse conduction.
zz Capillary permeability and nerve excitability
zz Decrease membrane fluidity
zz Acts as second messenger for the secretion of hormones
MINERALS zz Regulates TCA cycle
Macroelements: Requirement is > 100 mg/day. These are
zz Activate glycogen phosphorylase kinase by binding to
sodium, potassium, calcium, phosphorus, magnesium,
Calmodulin and increase glycogenolysis.
chloride and sulfur.
zz Ca+Calmodulin → regulate kinases of various met-abolic
Sodium pathways.
Sodium is the main extracellular cation. It is Na+/ K+ ATPase
pump which maintains high extracellular concentration of Regulation of Serum Calcium
Na+. This is known as calcium homeostasis. This is regulated by
Biological Functions parathyroid hormone (PTH), Vitamin D and calcitonin.
zz Regulation of osmotic pressure PTH increases serum calcium. Calcitonin decreases
zz Acid base balance serum calcium.
zz Absorption of monosaccharides and amino acids (like zz PTH: increases serum calcium by increasing bone
sodium-glucose symport) resorption. It also increases Ca reabsorption by kidneys.
zz Nerve transmission It stimulates 1-alpha hydroxylase activity in kidneys,
zz Maintenance of blood viscosity which synthesize the active form of vitamin D, i.e.
zz Electrolyte and water balance calcitriol. Vitamin D active form increases intestinal T
absorption of calcium. H
Potassium zz Calcitonin: lowers serum calcium by doing calcium E
This is the main cation of intracellular fluid. deposition in bones. O
zz Factors which increase calcium absorption: Vitamin D, PTH,
R
Biological Functions Y
Lysine, Arginine.
zz Electrolyte and water balance
zz Factors which decrease calcium absorption: Phytates
zz Regulation of pH of body fluids
 Phosphorus zz Iron toxicity: Haemosiderosis/Haemochromatosis
 Peroxidase contains iron required for Phagocytosis.
388 It is widely distributed in body. Around 75% of phosphorus
is present in bones and teeth, in combination with calcium.
It is present in two forms: Copper
zz Inorganic phosphorus
CRO BIOCHEMISTRY

Functions
zz Organic phosphorus
zz Iron absorption and its incorporation in heme
Biological Functions zz Is required for all oxidases
zz Mineralization of bones and teeth zz Two oxidases do not require copper. They require
zz Acid base balance and regulation of pH molybdenum. They are xanthine oxidase and sulphite
zz Present in phospholipids (required in lipoproteins and oxidase.
membranes) zz Cytoplasmic SOD also requires copper.
zz Constituent of nucleic acids zz Copper has mainly biliary excretion (minimally excreted
zz Constituent of high energy phosphates like ATP via kidneys). So a disease which hampers biliary
zz Metabolism of carbohydrates e.g. Glucose-6-phosphate. excretion will lead to accumulation of copper in liver.


zz Regulation by phosphorylation and dephosphorylation. This will cause increased blood levels of copper and
Phosphorus Deficiency: Occurs in premature infants increase urinary excretion.
exclusively fed on breast milk and in alcoholics fed with high
C linical B ox
carbohydrate diet.

Magnesium Wilson’s disease: (Condition of Cu excess in body)

zz Defect in Cu binding protein – P-type ATPase in liver cells.
Required for all Carboxylases, Phosphorylases and Kinases
zz It causes decreased excretion of copper in bile and reduction
(Exception: Pyruvate kinase uses K+ More than Mg)
in incorporation of copper in ceruloplasmin, leading to
accumulation of copper in liver, brain, kidney and RBCs.
Sulfur
Menke’s Disease: (X linked , a condition of Cu deficiency)
It is not present in body as inorganic Sulfur. It is present in zz Deficient production of Cu binding P- type ATPase in intestine.
vitamin B1, B5, Biotin and Lipoic acid, sulfur containing Cu increased in intestinal cell as it is not released into
amino acids, glycosaminoglycans. circulation.
zz Cu decreased in blood and urine.

Trace Elements zz Clinical features → Mental retardation, Grey kinky hair.

Requirement is < 100 mg/day. These are iron, iodine, copper,

chromium, manganese, zinc, molybdenum, selenium, Selenium
fluoride.
H igh R eturn
Iron zz Selenium is a constituent of 21st amino acid- Selenocysteine.
zz Haem iron: Haemoglobin, Myoglobin, Catalase, Trypto- zz Sources of Se: mainly sea foods and animal food like pork,
phan Pyrrolase, Peroxidase, Cytochromes beef, meat
zz Vegetable source include: nuts (Brazil nuts) and soya products.
zz Non-haem iron: Ferritin Hemosiderin, Transferrin, Fe-S
protein, Ribonucleotide Reductase.
Deficiency of Keshan’s Disease
H igh R eturn zz It was mainly observed in province of China- Keshan
zz Iron storage: Ferritin and Haemosiderin (Fe 3+) zz It mainly affects children and women of child bearing
zz Haemosiderin has higher iron content than Ferritin age.
zz Iron transport: Transferrin (Fe 3+)
zz Clinical manifestations include cardiomyopathy, muscle
zz Hepcidin: Regulate iron transport in circulation
weakness, hypothyroidism, eczema, increased risk of
 Haem (Fe2+)
stroke, hypertension and cancer
T  For absorption of Iron, Fe2+ required (Vitamin C plays role)
H Zinc
E Biological Functions
O zz Transport and storage of O2 (Hb and Mb) and CO2 zz Zinc, a divalent cation, is important for normal growth
and development of children both with and without
R zz In ETC as component of cytochromes
diarrhoea.
Y zz Cofactor for many enzymes e.g. catalase, peroxidase,
tryptophan pyrrolase zz Zinc is not synthesized within the human body and
therefore requires intake to maintain adequate levels.
zz Role in phagocytosis
zz Zinc benefits children with diarrhoea because it is a structure and function of the gastrointestinal tract, and
vital micronutrient essential for protein synthesis, cell impaired immune function
growth and differentiation, enhances immune response, zz Administration of zinc along with new low osmolarity 389
intestinal transport/absorption of water and electrolytes, oral rehydration solutions/salts (ORS), can reduce the
improves regeneration of the intestinal epithelium, risk, duration and severity of diarrheal episodes for up to
increases the levels of brush border enzymes, critical three months.

CHAPTER 15  MISCELLANEOUS
role in metallo-enzymes, polyribosomes, and the cell zz Acquired Zn deficiency can occur from:
membrane and cellular function  Decreased intake
zz Zinc is required for around 100 enzymes e.g. Carbonic  Malabsorption
Anhydrase, Alkaline Phosphatase, Carboxy Peptidase ,  Inability to absorb the micronutrient
Lactate Dehydrogenase etc.
 Increased metabolic demand
zz Zinc deficiency is associated with an increased risk
 Excessive loss
of gastrointestinal infections, adverse effects on the

C linical B ox
Acrodermatitis Enteropathica
zz Inherited deficiency of Zinc (not absorbed)
zz Rare disease
zz It is characterized by simultaneous occurrence of skin inflammation (Dermatitis) and Diarrhoea. Skin on cheeks, elbows and knees and
tssue around mouth and anus are inflamed.
zz Absorption of Zinc from intestine is affected in both acquired form and genetic form.
zz If inherited then it is autosomal recessive.

A dditional E dge
Metallothioneins
These are proteins which are induced at the time of excess intake of certain minerals (like Zinc, Copper, Cadmium, mercury). They help
to regulate the concentration of minerals at tissue level. These proteins are rich in cysteine.

Table 15.2: Minerals

Mineral Major Functions Deficiency

Sodium • Na /K ATPase pump
+ +
• Hyponatremia- T
• Major Extracellular Cation ƒƒ Excess Sodium loss H
• Acid Base Balance ƒƒ Muscle cramps, Headache, seizures can
• Regulation of osmotic pressure occur
E
• Nerve transmission O
• Maintenance of blood viscosity R
• Electrolyte and Water Balance Y

Contd…
 Mineral Major Functions Deficiency
390 Potassium • Na+/K+ ATPase pump • Hypokalemia-
• Major Intracellular Cation ƒƒ Muscular weakness
• Electrolyte and Water Balance ƒƒ Irregular heart beat
• Regulation of pH of body fluids ƒƒ Tachycardia
CRO BIOCHEMISTRY

• Maintenance of Neuromuscular excitability ƒƒ Altered ECG pattern

• Cofactor for enzymes e.g. Pyruvate Kinase
Chloride • Formation of HCl in stomach • Hypochloraemia
• Regulation of osmotic pressure of extracellular fluids. ƒƒ Severe vomiting
ƒƒ Addison’s Disease
Calcium • Calcification of bones and teeth • Rickets in Children
• Blood Clotting • Osteomalacia in Adults
• Decrease membrane Fluidity • Osteoporosis in elderly
• Capillary permeability and nerve excitability
Phosphorus • Mineralization of bones and teeth • Rickets


• Constituent of High energy phosphates like ATP • Osteomalacia

• Constituent of Nucleic Acids
Magnesium • Constituent of bones and teeth • Neuromuscular weakness
• Cofactor for Kinases, Phosphorylases, Carboxylases
• Improves Glucose Tolerance
Iron • Constituent of haem • Microcytic, hypochromic anaemia
• Component of oxido-reductase enzymes
• Electron transfer reactions
Copper • Cofactor for Oxidases • Microcytic, hypochromic anaemia
• Fe absorption and its incorporation in heme • Menke’s Disease
• Component of several enzymes or proteins involved in redox
reactions
Selenium • Antioxidant Function • Keshan’s Disease
• Its action is complementary to Vitamin E • (Endemic cardiomyopathy)
• Cofactor for Deiodinase
Zinc • Zinc fingers-Important constituent of regulatory proteins that • Poor Wound Healing
control transcription • Growth Retardation
• Cofactor for 100 + enzymes e.g. Carbonic Anhydrase, Alcohol • Infertility
Dehydrogenase, Lactate Dehydrogenase, Carboxy peptidase, • Hypogonadism
Alkaline Phosphatase • Acrodermatitis enteropathica
• Role in spermatogenesis.
Chromium • Component of ‘Glucose Tolerance Factor’ • Hyperglycemia
• Facilitates action of Insulin • Impaired Glucose Tolerance
• Regulation of Gene Expression
Molybdenum • Cofactor for Xanthine oxidase, Sulfite Oxidase, Aldehyde oxidase, • Severe Neurological Abnormalities
Lipoprotein Lipase, Nitrite Reductase
Cobalt • Constituent of Vitamin B12 • Anaemia, low WBC count
• Maturation of RBCs
Manganese • Required for Glycoprotein and Proteoglycan Synthesis • Growth Retardation
• Cofactor for mitochondrial SOD, Pyruvate Carboxylase, Glutamine • Bone Deformity
Synthesis • Sterility
• Fatty infilteration in Hepatocytes
T Iodine • Constituent of Thyroid hormones • Cretinism
H • Goiter
E • Myxedema
O Flouride • Formation of Bones and teeth • Dental caries
R • Provides resistance to tooth enamel to sustain acid attacks
Y
XENOBIOTICS Cytochrome P 450 Enzymes

zz Xeno’ means foreign, ‘Biotics’ means metabolism. zz Most important enzyme of xenobiotic metabolism 391
zz Xenobiotics are any substances which should not be zz Catalyse hydroxylation of phase I, using NADPH and O2
present in human body. It can be a drug or a chemical zz Hydroxylases are also known as mono-oxygenases or
or a substance which is present in excess of normal limit. mixed function oxidases. Here, one oxygen atom is

CHAPTER 15  MISCELLANEOUS
zz Detoxification: Xenobiotics & drugs are inactivated incorporated into the substrate & other atom of oxygen
by a series of enzymatic reactions & they are excreted is used to reduce water.
by kidneys. This process is known as detoxification. zz It is most versatile biocatalyst.
Basically this metabolism of drugs makes them more
zz It is a haem containing enzyme
soluble so that they can be easily excreted out of body.
zz It is membrane bound enzyme, present in microsomes
zz Xenobiotic metabolism – also known as Biotransforma-
tion reactions. & IMM
zz Major organ – Liver zz There are various isoforms but CYP 3A 4 is used for the
zz Metabolism of xenobiotics involves Phase-I and Phase-II metabolism of most drugs
reactions. zz They absorb light at 450 nm
zz Lipophilic drugs first enter phase I reactions and are zz These enzymes are highly inducible by their own
made polar. Then they enter Phase II reactions and are substrate by increasing the rate of transcription
made hydrophilic by bioinactivation conjugation and zz Advantage of high inducibility – efficient detoxification
thus can be easily excreted out in urine. zz Disadvantage of high inducibility – rapid development
zz Not all polar molecules are hydrophilic. of tolerance against drugs e.g. in case of epileptic drug
zz Those drugs or compounds which are already polar, phenobarbital, the dose has to be increased 3-4 fold
directly goes to Phase II reactions in which they are within a week of starting
converted to hydrophilic and are then excreted. E.g.
Paracetamol, Isoniazid.
zz Paracetamol undergoes Glucuronidation, also Sulfation.

T
H
E
O
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Fig. 15.11: Cytochrome P450 (CYP450) catalytic cycle. The enzyme cytochrome P450 oxidizes the drugs using a reactive heme ring, with
an iron atom as the ultimate electron acceptor and NADPH as a necessary co-factor
 Conjugating Agents

392 zz Glutathione
zz Glycine (e.g. benzoic acid conjugated with glycine to
form hippuric acid)
zz Glucuronic acid (e.g. bilirubin)
CRO BIOCHEMISTRY

zz Sulfation (by PAPS- Phospho Adenosyl Phospho Sulfate)

zz Acetylation (by Acetyl CoA)
zz Methylation (by SAM- S-Adenosyl Methionine)

FREE RADICALS (FR)

How OFRs are Generated in Body?
It is a molecule or molecular fragment that has one or more
than one unpaired electron in its outer orbit and has an zz Electron leakage from ETC
independent existence. They are extremely short lived but zz Normal oxidation- reduction reactions
they readily extract electron from other molecules, converting zz Exogenous agents: Carbon tetrachloride, ionizing radia-


tions, cigarette smoke

them to free radicals and thus start a chain of reaction.
zz Transition metals like iron, copper generate free radicals.
A free radical is represented by a superscript dot.
Transition metals can react with superoxide radical to
form hydroxyl radical. Ferrous and Cuprous ions are
more reactive than their oxidized counterparts i.e. ferric
and cupric (Fe+3, Cu+2). Therefore these metals are never
found in free state in body.

Fenton Reaction
Fe2+ +H2O2 Fe3+ + OH–+.OH
By Fenton’s reaction, H2O2 is converted to highly dangerous hyd-
roxyl radical. This reaction is mediated by transition metal – iron.
Fig. 15.12: Generation of OFR (Oxygen Free Radicals) or ROS
(Reactive Oxygen Species) Uses of OFR in Body
zz Phagocytes generate free radicals with a purpose of
F undamental B ox destroying engulfed bacteria. As a result, their oxygen
consumption rises, which is known as Respiratory burst.
zz Some enzymes generate FR at their active sites which
help in catalysis e.g. Ribonucleotide Reductase

Damage caused by OFRs

One of the most sensitive sites for FR damage is cell mem-
brane, which is rich in PUFAs (Polyunsaturated Fatty acids).

zz OFR with highest activity which is most powerful is OH Damage by FR is because of their extreme high reactivity
(hydroxy). It is most reactive & thus most dangerous. Its zz Lipids are most susceptible (which is known as LP-
half-life is in nanoseconds Lipid Perox-idation). PUFAs are more prone to lipid
peroxidation. LP is harmful as it is an amplifying chain
zz H2O2 is not a free radical but it can generate free radicals.
reaction providing a continuous supply of free radicals
So it is considered a reactive oxygen species but not a free that initiate further peroxidation. The compounds
radical. formed by lipid peroxidation reacts with proteins to form
zz Precursor of all ROS is superoxide radical. adducts & cross links known as advanced lipoxidation
T zz Other products of peroxidation are: oxidized LDL, MDA end products (ALE).
H (Malon Dialdehyde), oxysterols, prostanoids. MDA is zz Nucleic acids e.g. DNA (chain breaks or alterations in
E also considered the marker of lipid peroxidation. nitrogenous bases which may lead to mutations, cell
O zz Nitric oxide (endothelium derived relaxation factor) is death or cancers)
R also a free radical. It can yield peroxynitrite, which can zz Haemoglobin: It gets converted to metHb.
Y then form hydroxyl radical.
zz Proteins: Alteration of conformation of proteins or SOD enzyme has three forms:
enzymes because of oxidation of sulfhydryl group or zz Cytoplasmic form – requires Copper
modification of certain amino acid residues. zz Mitochondrial form – requires manganese 393
zz ROS also react with carbohydrates and form compounds zz Extra cellular form – requires Cu and Zn
that react with proteins to form adducts & cross links,
known as glycoxidation products or Advanced glycation Reaction by Catalase

CHAPTER 15  MISCELLANEOUS
end products (AGE).
Due to these kind of damages done by free radical,
they are often involved in the etiology of diseases like par-
kinsonism, Alzheimer’s disease, cataract, rheumatoid
arthritis, cancer, diabetes, atherosclerosis, ageing, infertility,
autoimmune diseases etc.
Fig. 15.14: Catalase

Reaction by Glutathione Peroxidase

zz Vitamins with antioxidant activity : Vit A, C , E , D

zz Vitamin E is the most powerful lipid phase antioxidant. There
is a strong negative association between plasma alpha-
tocopherol levels & coronary artery disease.
zz Vitamin D is also a powerful natural membrane antioxidant.
It inhibits iron-dependent liposomal lipid peroxidation.

Artificial Antioxidants
zz Propyl Gallate
zz Butylated Hydroxy Toluene (BHT)
zz Butylated Hydroxy Anisole (BHA)

Measuring FR
zz FOX assay: (Ferrous oxidation in Xylenol)
zz Estimation of dialdehydes (e.g. MDA- MalonDialdehyde)
zz Pentane and methane measurement in exhaled air.

RQ – RESPIRATORY QUOTIENT
RQ is equal to amount of CO2 produced divided by amount
of O2 utilized

Reaction by SOD (Superoxide Dismutase)

zz For carbohydrates, RQ is 1
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Fig. 15.13: SOD

 zz For Fats, RQ is 0.7 (calculated indirectly) ALCOHOL METABOLISM
zz For Proteins, RQ is 0.8 (calculated indirectly)
394 zz For Mixed diet, RQ is 0.85 H igh R eturn
zz RQ for Alcohol = 0.66
zz Organ – Liver
zz Exclusive Carbohydrate diet = 1
zz Organelle – Cytoplasm and Mitochondria
CRO BIOCHEMISTRY

zz But for carbohydrate rich diet, RQ is > 1

zz Energy – 7 Kcal/gm
zz Because in case of carbohydrate rich diet, Carbohydrates zz Site of absorption – mainly small intestine (to a lesser extent
are converted to Fats. in stomach)
zz Carbohydrates have high O2 content as compared zz Zero order kinetics - means rate of elimination is fixed (does
to fats. So because more O2 containing compound not depend on drug concentration in body)
(carbohydrates) is converted to less O2 containing zz Pleasurable effect is because of increase dopamine
compound (fat), so O2 will be released in this process.
This O2 released will be used for oxidation. So less O2 is Ingestion of alcohol (CH3CH2OH i.e. ethyl alcohol) gives
required from outside. So RQ rises energy i.e. 7 kcal/gm but these are empty calories because they
So RQ tells us about the: are not associated with any nutrients (vitamins or minerals).
Type of macromolecule used in body Food interferes with absorption in stomach. So alcohol


zz
zz Conversion of one macromolecule to another ingestion on empty stomach leads to faster absorption.
There is no negative feedback control for alcohol
Acidosis Alkalosis metabolism. So alcohol oxidation is preferred over other
RQ increases because CO2 RQ decreased because
macromolecules.
output increases respiration is depressed
CO2 output is greater than O2 CO2 retained in boby
consumption So less CO2 produced

zz Fasting/ Starvation - RQ decreases

zz Fever: RQ increases because of increase in breathing.
There is wash out of CO2 occurring and CO2 production
increases
zz Exercise- lactic acidosis occurs, so, RQ increases
zz During Recovery from Exercise- Less CO2 is produced.
So RQ decreases. Gradually it goes back to normal

H igh R eturn
RQ in Diabetes Fig. 15.15: Alcohol metabolism leads to NADH increase
In a normal person, main fuel for body is carbohydrates. But in a Enzyme Alcohol dehydrogenase- ADH
diabetic patient, main fuel for body is fats as carbohydrate/glucose
zz There are many isoenzymes
is not entering the cells due to relative or absolute deficiency of
insulin. So RQ decreases in Diabetes as RQ of fats is less than the zz Most abundant – ADH-1A
RQ for carbohydrates. zz Present in cytoplasm of liver and adrenal glands
But on giving Insulin, it again rises as glucose starts entering zz Has NAD containing domain known as Rossman fold
cells. Now cells will again start using glucose. Enzyme Aldehyde dehydrogenase-(ALDH)
zz Present in Mitochondria
zz 2 Isoenzymes:
1. ALDH-1 → Cytoplasm (Minor role)
2. ALDH-2 → Mitochondria (Major)

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 395

CHAPTER 15  MISCELLANEOUS
Microsomal Ethanol Oxidizing System (MEOS)
zz In ER/ microsomes
zz Inducible system
zz Induced after ingestion of lots of alcohol
zz CYP-2E1 → has high Km
zz NADPH involved
zz So protective in chronic alcoholics as this does not further leads to increase NADH
zz But too much use of this system will produce ROS, which can damage DNA, Proteins and Lipids
There is one Accessory pathway in peroxisomes which produces H2O2. Catalase is needed to detoxify this.
Table: Biochemical changes in Alcoholism

Increased NADH (due to Alcohol Dehydrogenase and Aldehyde Dehydrogenase)

• Oxaloacetate to Malate conversion occurs as these reactions
• Pyruvate to Lactate conversion occurs use excess NADH

Pathways which are decreased in alcoholism:

• Gluconeogenesis (as decreased Pyruvate and Oxaloacetate)
• β- oxidation (as it produces NADH)
• TCA (due to decreased Oxaloacetate)
Pathways which are increased in alcoholism in liver:
• Lipogenesis (because excess Acetyl CoA present as Oxaloacetate depleted and not for TCA)
Leads to Alcoholic
• Increased Fatty Acid Synthesis (as β-oxidation is decreased)
Fatty Liver
• Cholesterol synthesis due to excess
• Ketosis Acetyl CoA

The reasons of alcoholic fatty liver are: zz Decreased beta oxidation, so this excess fatty acid is not
zz Increased TG synthesis (by reciprocal regulation as beta broken down.
oxidation is inhibited)
zz No NAD+ available in cells, so there is increased H igh R eturn
glyceraldehyde-3-P which cannot be taken by enzyme NOTE: Alcoholic fatty liver is not because of fatty acids derived
glyceraldehyde-3-P dehydrogenase, which uses NAD+. from adipose tissue. Rather it is because of endogenous synthesis T
So glyceraldehyde-3-P gets converted to DHAP (Di of TG in liver. So production of TG increases and breakdown H
hydroxy acetone phosphate), which gets converted to inhibited. E
glycerol-3-P, which is used for the formation of TGs in zz Alcoholic fatty liver leads to Alcoholic hepatitis which can lead O
liver. to alcoholic cirrhosis R
zz Impaired formation or release of VLDL (because acetal-
Y
dehyde inhibits microtubule formation responsible for
the movement of TG into VLDL)
 Wernicke-Korsakoff Syndrome Treatment of Wernicke-Korsakoff Syndrome
396 zz Occurs in thiamine (Vitamin B1) deficiency, which zz Thiamine supplementation before giving glucose for
occurs in alcoholics hypoglycemia
zz 2 reasons of thiamine deficiency in alcoholism: zz But there is incomplete recovery of memory
 Patients mostly not eating food
A dditional E dge
CRO BIOCHEMISTRY

 Alcohol interferes with absorption of vitamin B1

zz Wernicke peripheral neuropathy + Korsakoff psychosis, Antabuse – DISULFURAM
characterized by ataxia, memory loss, confabulations, zz Inhibits Aldehyde dehydrogenase
ophthalmoplegia, nystagmus and cerebral haem- zz Also, genetic variation occurs in this enzyme specially in people
orrhage. Highly aerobic tissue e.g. brain and heart of china and korean origin, known as Asian flush syndrome.
are affected here as vitamin B1 is mainly required for zz This occurs due to increased acetaldehyde which causes

oxidative decarboxylation nausea, vomiting, flushing, sweating, tachycardia and hyper-

ventilation
zz Acetaldehyde is toxic because it forms adducts with proteins
and amino acids. It binds glutathione, damages mitochondria
and inhibits microtubules.


Methanol Poisoning
zz Methanol is metabolized by alcohol dehydrogenase, forming toxic product formic acid
zz Ethanol used in methanol poisoning as ethanol acts as competitive inhibitor of enzyme alcohol dehydrogenase. As
ethanol has low km for alcohol dehydrogenase, thus it inhibits the binding of methanol with the enzyme. So alcohol
dehydrogenase metabolize ethanol to form acetate, which is non-toxic.
zz Also fomepizole can be used in methanol poisoning but it is costly.

Fig. 15.16: Methanol Poisoning

Antifreeze Compound – Ethylene Glycol defects. All the effects caused by alcohol are irreversible in
the growing baby. Child has low birth weight, low IQ, short
Ethylene glycol is metabolized by same alcohol height, small head size.
dehydrogenase, producing toxic products which can cause Both alcohol and acetaldehyde crosses placenta. It causes
severe acidosis and renal damage. For the treatment, ethanol brain damage of the fetus as alcohol crosses blood brain
is used as antidote in ethylene glycol poisoning. Ethanol barrier. Also inhibition of enzyme alcohol dehydrogenase
has low Km for alcohol dehydrogenase, thus it inhibits the (due to high ethanol) of fetus occurs. This enzyme is required
binding of ethylene glycol with the enzyme. Ethylene glycol is to produce retinoic acid in fetus for growth and development.
present in automobile antifreeze.

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Fetal Alcohol Syndrome Markers for Chronic Alcoholism:
R
CDT-Carbohydrate deficient transferrin
Y If mother taking alcohol during pregnancy, then fetal alcohol
syndrome occurs, leading to fetal alcohol spectrum disorders zz A plasma protein which is a marker for chronic alcoho-
which are a range of disorders including physical and mental lism.
zz Alcohol inhibits glycosylation of Transferrin. So an altered transferrin is found which is deficient in 4-5 sialic acid residues.
γ-GGT- Gamma Glutamyl Transpeptidase
zz Also a marker for chronic alcoholism. 397
zz This is a microsomal enzyme widely distributed in body. This enzyme is responsible for hydrolysis of gamma glutamyl
peptide bonds.

CHAPTER 15  MISCELLANEOUS
Pearls of the Chapter
zz Selenium deficiency can lead to hypothyroidism
zz Three hormones involved in homeostasis of blood calcium levels are vitamin D, Parathyroid hormone & Calcitonin.
zz Keshan’s disease occurs due to selenium deficiency

zz Fluorosis occurs when concentration of fluorine is > 20 ppm

zz Vitamins which can cause dementia: Vitamin B1, B3, and B12

zz Nutritional causes of cardiomyopathy are deficiency of Thiamine, Selenium, Ca, Mg and excess of Iron

zz Micronutrients in glycolysis: Vitamin B1, B2, B3, Mg (for Kinases)

zz Minerals required in energy – Mg, Zn, Chromium

zz Vitamins those are necessary for neurological function are Thiamine, Pyridoxine and Cobalamin
Micronutrients required for bones:
zz Vitamin D – for absorption of calcium.

zz Vitamin C – for formation of bone matrix.

zz Vitamin K, Mg, P, copper, zinc and manganese.

zz OFR with highest activity which is most powerful is OH (hydroxy)

zz H2O2 is not a free radical but it can generate free radicals. Precursor of all ROS is superoxide radical

zz MDA is also considered the marker of lipid peroxidation

zz Nitric oxide (endothelium derived relaxation factor) is also a free radical

zz FOX assay – (Ferrous oxidation in Xylenol) is for the measurement of FR

zz Chain breaking anti-oxidants are Alpha-Tocopherol, Beta- carotene and Super oxide dismutase (SOD)

zz Vitamins with antioxidant activity: Vitamin A, C, E

zz Vitamin E is the most powerful lipid phase antioxidant

zz Lipids are most susceptible (known as LP- Lipid Peroxidation)

zz Enzymatic antioxidants are Superoxide dismutase, Catalase and Glutathione Peroxidase.

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 398 Multiple Choice Questions
Vitamins and Minerals 11. Which of the following cannot be synthesized in
1. True statement about Vitamin K is: humans: (PGI)
 (Recent Question Jan 2018) a. Vitamin C b. Riboflavin
a. Vitamin K is water soluble c. Vitamin B3 d. Cobalamin
b. Vitamin K is needed for action of clotting factor 8 e. Thiamine
c. Vitamin K deficiency leads to DVT 12. Deficiency of thiamine leads to nerve weakness
d. Vitamin K affects bone health by activating proteins because of: (PGMEE 2015)
that bind calcium a. Difficulty to produce ACh molecules
2. Vitamin which is excreted in urine is:  (PGMEE 2013) b. Inactivation of Chloride Channel
a. Vitamin A b. Vitamin C c. Hypocalcemia
c. Vitamin D d. Vitamin K d. Hypomagnesima
3. In Vitamin C deficiency, post translational modification 13. Excess of Avidin causes deficiency of: (PGMEE 2015)
of which amino is defective: a. Folate b. Choline
 (PGMEE 2013, 2015) c. Vitamin B12 d. Biotin
a. Arginine b. Glycine 14. What is the other name of Hallervorden-Spatz syn-
c. Lysine d. Alanine drome? (PGMEE 2013)
4. Boy who refuses to eat fruits, comes with knee a. Mitochondrial encephalomyopathy; Lactic acidosis;
swelling and ecchymoses, deficiency of which Stroke
vitamin is suspected: (PGMEE 2012) b. Pantothenate kinase – associated neuro
a. Vitamin E b. Vitamin D degeneration
c. Vitamin B1 d. Vitamin C c. Thiamine-responsive megaloblastic anemia
5. The vitamins those are necessary for neurological syndrome
function: (PGMEE 2009) d. Riboflavin sensitive myopathy
a. Thiamine, Riboflavin, Cyanocobalamin 15. Which of the following vitamin causes mental
M disorder ?
b. Thiamine, Riboflavin, Pyridoxine
C c. Thiamine, Pyridoxine, Cyanocobalamin a. Thiamine b. Pyridoxine
Qs d. Thiamine, Folic acid, Cyanocobalamin c. Niacin d. Biotin
Ans. 6. Thiamine deficiency is known to occur in all of the 16. Vitamin deficiency causing dementia: (PGMEE 2015)
1. d following EXCEPT: (PGMEE 2003) a. Vitamin B12 b. Pyridoxine
2. b a. Food Faddist c. Thiamine d. Biotin
3. c b. Homocystinemia 17. The form of THFA used in treatment is:
4. d c. Chronic alcoholic  (PGMEE 2015)
5. c d. Chronic heart failure patient on diuretics a. N5 Formyl THFA b. N5 Methyl THFA
6. b 7. A young lady presented with tingling sensation in legs c. N10 Formyl THFA d. N5 Formimino THFA
7. a and hands. On examination she had fissure tongue and
18. Seborrheic dermatitis is produced by deficiency of:
8. d lesions in angle of mouth. On investigation, she had
 (PGMEE 2015)
9. a low RBC glutathione reductase activity. Diagnosis is
a. Vitamin C b. Vitamin B1
10. d deficiency of: (MAY AIIMS 2017)
c. Vitamin B2 d. Vitamin A
11. a, a. Vitamin B2 b. Vitamin B6 19. Riboflavin is a constituent of: (PGMEE 2013)
d,e c. Vitamin B12 d. Vitamin B1 a. FMN b. NAD
12. b 8. Restless leg syndrome can occur due to: c. PLP d. THF
13. d  (PGMEE 2013)
20. Co-factor for L-amino acid oxidase is- (PGMEE 2015)
14. b a. Folic acid b. Iron a. Niacin b. Folic acid
15. a c. Manganese d. All of the above c. Thiamine d. Riboflavin
16. a 9. Which of the following is an Atypical Vitamin? 21. Glutamate Dehydrogenase requires cofactor-
17. a  (PGMEE 2015)
a. Vitamin B3 (Niacin)
18. c a. NAD+ b. NADP
b. Vitamin B12 (Cobalamin)
19. a c. Both a and b d. None
c. Vitamin C (Ascorbate)
20. d 22. Beta alanine is a component of: (PGMEE 2013)
d. Vitamin B9 (Folate)
21. c a. Biotin
10. Which of the following deficiency can cause genera-
22. c b. Pyridoxal phosphate
lized oedema ?
c. Pantothenic acid
a. Vitamin B2 b. Vitamin B6
d. Folic acid
c. Vitamin B12 d. Vitamin B1
23. Pantothenic acid containing coenzyme is involved 36. What type of anemia is seen in B12 deficiency:-
in: (PGMEE 2015)  (PGMEE 2016-17)
a. Carboxylation b. Acetylation a. Normocytic b. Macrocytic 399
c. Decarboxylation d. Dehydrogenation c. Microcytic d. Dimorphic
24. Vitamin synthesized from tryptophan- 37. Which does not requires Biotin?
 (PGMEE 2015) a. Acetyl CoA to Malonyl CoA
a. Riboflavin (Vitamin B2) b. Pyruvate to Oxaloacetate
b. Thiamine (Vitamin B1) c. Glutamate to GABA
c. Niacin (Vitamin B3) d. Propionyl CoA to Methyl Malonyl CoA
d. Pyridoxine (Vitamin B6) 38. THIAMINE is a cofactor for all EXCEPT:
25. Niacin deficiency causes all EXCEPT- a. Alpha ketoglutarate dehydrogenase
 (PGMEE 2013-14) b. Succinate dehydrogenase
a. Diarrhoea b. Dementia c. Pyruvate dehydrogenase
c. Dactylitis d. Dermatitis d. Branched chain alpha keto acid dehydrogenase
26. Pellagra, parkinsonism, convulsions, anemia and 39. Thiamine is NOT used in which of the following
kidney stones are seen in deficiency of: reactions
 (PGMEE 2013) a. Non oxidative phase of HMP
a. FADH b. Niacin b. Lactate to Pyruvate
c. Pyridoxal phosphate d. Coenzyme A c. Oxidative decarboxylation of Alpha-keto amino
27. Vitamin B6 is used in treatment of- (PGMEE 2013-14) acids
a. Homocystinuria b. Cystathionuria d. Alpha-KetoGlutarate to Succinyl Co-A
c. Xanthourenic aciduria d. All 40. Hypervitaminosis A causes:  (PGMEE 2013-14)
28. Vitamin responsible for decarboxylation of amino a. Alopecia
acid: (PGMEE 2013) b. Benign intracranial hypertension (Pseudo-
a. Vitamin A b. Vitamin D Tumourcerebri)
c. Vitamin B6 d. Vitamin E c. Liver damage
29. Major form of folic acid for transfer of one carbon is: d. All
 (PGMEE 2013) 41. Vitamin A is present in all EXCEPT:
a. Methylene THF b. Methyl THF a. Sunflower seeds b. Egg
c. Formyl THF d. All c. Milk d. Tomato
42. Vitamin which is more in cow’s milk than breast milk M
30. In one carbon metabolism when serine is converted C
to Glycine, which carbon atom is added to THF: is: (PGMEE 2011)
a. Vitamin A b. Vitamin C Qs
 (PGMEE 2015)
c. Vitamin D d. None of the above Ans.
a. Alpha Carbon b. Gamma Carbon
43. Vitamin acting on intranuclear receptors- 23. b
c. Beta Carbon d. Delta Carbon
 (PGMEE 2015) 24. c
31. Which amino acid does not take part in one carbon
a. Vitamin K b. Vitamin D 25. c
transfer reactions: (PGMEE 2007)
c. Vitamin B1 d. Vitamin E 26. c
a. Glycine b. Serine
44. Richest source of Vitamin D is: (PGMEE 2011) 27. d
c. Threonine d. Tyrosine
a. Milk b. Fish liver oils 28. c
32. Vitamin given in pregnant women to prevent neural
c. Sunlight d. Carrots 29. a
tube defect: (PGMEE 2015)
45. 1, 25 (OH)2 cholecalciferol is formed in- 30. c
a. Vitamin A b. Vitamin C
 (PGMEE 2015) 31. d
c. Folic acid d. Vitamin B12
a. Skin b. Liver 32. c
33. FIGLU excretion is seen in urine in which vitamin
c. Spleen d. Kidney 33. b
deficiency: (PGMEE 2016-17)
46. Lipid soluble plasma membrane associated anti-oxidant 34. c
a. Tryptophan b. Folic acid
is - (PGMEE 2015) 35. b
c. Vitamin B12 d. Vitamin C
a. Glutathione b. Vitamin E 36. b
34. Neurological worsening with anemia what is the
c. Ubiquitin d. Vitamin C 37. c
treatment to be given: (PGMEE 2015)
47. Vitamin E deficiency causes all EXCEPT: 38. b
a. Iron
 (PGMEE 2015) 39. b
b. Folic acid alone 40. d
a. Ataxia b. Neuropathy
c. Folic acid along with hydroxycobalamin 41. a
c. Ophthalmoplegia d. Areflexia
d. Pyridoxine 42. c
48. Which of the following trace element has Vitamin E
35. Which vitamin is essential for metabolism of sulphur 43. b
sparing effect? (PGMEE 2013)
containing amino acids? 44. b
a. Copper
 (PGMEE 2011, 2014, 2015) 45. d
b. Iron
a. Thiamine b. Folic acid c. Magnesium 46. b
c. Vitamin C d. Biotin d. Selenium 47. c
48. d
 49. The vitamin synthesized by bacteria in the intestine 63. Which of the following mineral has antioxidant
is: (PGMEE 2004) property- (PGMEE 2015)
400 a. Vitamin K b. Vitamin B1 a. Calcium b. Magnesium
c. Vitamin D d. Vitamin C c. Zinc d. Iron
50. The water soluble form of vitamin K is which one of 64. Number of iron in transferrin: (PGMEE 2015)
the following: (PGMEE 2010) a. 1 b. 2
a. Phylloquinone b. Primidone c. 3 d. 4
c. Menadione d. Menaquinone 65. Which of the following is used for the transport of
51. Vitamin K is required for: (PGMEE 2009) Retinol Binding Protein:
a. Hydroxylation b. Chelation a. Transferrin b. Transthyretin
c. Transamination d. Carboxylation c. Transretinol d. Transalbumin
52. In carboxylation of clotting factors by Vitamin K, 66. Which is not involved in iron metabolism?
which amino acid is carboxylated: (PGMEE 2008)  (AIIMS Nov 2017)
a. Glutamate b. Aspartate a. Transthyretin b. Ceruloplasmin
c. Histamine d. Histidine c. Hepcidin d. Ferritin
53. Which of the following is true about vitamin K? 67. Which vitamin is required as cofactor for enzyme
 (PGMEE 2015) glycogen phosphorylase?  (AIIMS Nov 2017)
a. Vitamin K dependent factors undergo post- a. Biotin
transcriptional modification b. Niacin
b. Menadione is a natural water insoluble vitamin K c. Thiamine
used in clinical practice d. Pyridoxal Phosphate
b. Stuart-Prower factor is not vitamin K dependent 68. Vitamin-B12 is required for all EXCEPT:
d. Prothrombin is a vitamin K dependent factor  (AIIMS Nov 2015)
54. Vitamin required for production of thrombin is: a. Glycogen phosphorylase
 (PGMEE 2013) b. Methionine synthase
a. Vitamin A b. Vitamin D c. Leucine amino mutase
c. Vitamin K d. Vitamin E d. Methyl Malonyl CoAmutase
55. In a well fed person, Vitamin A stores last for: 69. Thiamine deficiency cause (s):  (PGI Nov 2014)
 (PGMEE 2013) a. Glossitis b. Polyneuropathy
a. > 6 months b. <1 months c. Pellagra d. Angular stomatitis
M c. 3 months d. 3-6 months e. Cardiomegaly
C 56. Which is non-essential mineral: (PGMEE 2013) 70. Pyridoxine is required for: (PGI May 2018)
Qs a. Lead b. Iron a. Decarboxylation
Ans. c. Manganese d. Sodium b. Carboxylation
49. a 57. Coenzyme required for activation of sulfite oxidase is: c. Transmination
50. c  (PGMEE 2011) d. Transsulfuration
51. d a. Copper b. Zinc
52. a e. Oxidative deamination
53. d
c. Iron d. Molybdenum 71. Vitamin E deficiency in adult causes: (PGI May 2018)
54. c 58. Zinc is present in: (PGMEE 2015) a. Hemolysis
55. a a. Glutathione Reductase b. Posterior column involvement
56. a b. Glutathione Synthetase c. Peripheral neuropathy
57. d c. Carbonic Anhydrase d. Hair loss
58. c d. Xanthine Oxidase
59. d
e. Impaired immunity
59. Which one among the following is not an anti- 72. Vitamin C deficiency is associated with:
60. b
61. d oxidant? (PGMEE 2013-14)  (PGI May 2018)
62. d a. Zinc b. Selenium a. Decreased immunity
63. c c. Copper d. Iron b. Improper wound healing
64. b 60. The co-enzyme for the enzyme Phosphofructokinase c. Epistaxis
65. b is: (PGMEE 2010)
66. a
d. Seizures
a. Manganese b. Magnesium e. Anemia
67. d
68. a c. Copper d. Zinc 73. Vitamin A is stored in: (Recent Question Jan 2019 )
69. b,e 61. Menke’s Kinky Hair syndrome is characterized by a. Kupffer cells b. Hepatocytes
70. a,c, congenital deficiency of: (PGMEE 2010) c. Ito cells d. Endothelial cells of liver
d,e a. Serum copper b. Serum ceruloplasmin 74. In Wilson disease there is less urinary excretion of:
71. a,b, c. Ferrochelatase d. Copper binding ATPase
c,d,e
 (Recent Question Jan 2019 )
62. Zinc deficiency can cause which one of the following: a. Phosphorus
72. a,b,
c,d
 (PGMEE 2009) b. Methyl-Histidine
73. c a. Myopathy b. Multiple sclerosis c. Phosphotyrosine
74. b c. Goitre d. Infertility d. Serine
75. A 20 year old alcoholic malnourished patient Xenobiotics and Free Radicals
presented to hospital with respiratory distress. 85. Which of the following enzyme is free radical scaven-
His pulse was 112/minute. Patient had edema,
ger? (Recent Question June 2018)
401
hypertension, systolic murmur along the left sternal
a. Glutathione peroxidase
edge. Bilateral crepitations were felt in the lungs. A
b. NADH oxidase
diagnosis of congestive high output cardiac failure
c. Hydrogen peroxidase
was made. Which vitamin is deficient?
d. Hypochlorous
 (Recent Question Jan 2019)
86. Which of the following is not a free radical?
a. Vitamin B1 b. Vitamin C
a. Hydrogen peroxide b. Hydroxyl
c. Vitamin B2 d. Vitamin B6
c. Superoxide d. Hydroperoxyl
76. Keshan’s disease is due to deficiency of:
87. Most powerful chain breaking antioxidant:
 (Recent Question June 2018)
a. Selenium b. Iron  (Recent Question 2018)
c. Copper d. Zinc a. Vitamin C
77. Dysesthesia is caused by deficiency of: b. β – Tocopherol
 (Recent Question June 2018) c. Catalase
a. Zinc b. Selenium d. Glutathione peroxidase
c. Iron d. Copper 88. Free radical with highest activity: 
78. Which of the following vitamin increases the  (Recent Question 2018)
absorption of iron? (AIIMS May 2018) a. Hydrogen peroxide
a. Vitamin A b. Vitamin C b. Hydroxyl radical
c. Thiamine d. Riboflavin c. Superoxide radical
79. Vitamin K in its coenzyme form is regenerated by d. Hydroperoxyl radical
which enzyme: (AIIMS May 2018) 89. Enzyme which carries dismutation reaction is:
a. Glutathione Reductase  (Recent Question 2018)
b. Pyruvate Carboxylase a. Catalase
c. Dihydrofolate Reductase b. Glutathione peroxidase
d. Epoxide Reductase c. SOD
80. Use of Zinc in diarrhoea is because it: d. Glutathione reductase
 (FMGE Nov 2018) 90. Enzyme which carries the reaction H2O2 gives H2O
a. Reduces the risk, duration and severity of diarrheal +O2: (Recent Question 2017) M
episodes a. Catalase C
b. Enhances immune response b. Glutathione peroxidase Qs
c. Regulates intestinal transport & absorption of water c. SOD Ans.
and electrolytes d. Glutathione reductase 75. a
d. All 91. Assay for lipid peroxidation is: 76. a
81. Fishy odour can be due to deficiency of which  (Recent Question 2017) 77. b
vitamin? (Recent Question Jan 2018) a. MTT Assay 78. b
a. Biotin b. Thiamine b. FOX assay 79. d
c. Riboflavin d. Vitamin A c. Ame’s test 80. d
82. Most abundant form of pro-vitamin A is: d. Guthrie’s test 81. c
 (Recent Question Jan 2018) 92. Which is the marker for lipid peroxidation: 82. a
a. Beta carotene a. Catalase 83. d
b. Alpha- carotene b. Glutathione reductase 84. d
c. Retinol c. Maltase 85. a
d. Retinaldehyde d. Malondialdehyde 86. a
83. False statement regarding Schilling test is: 93. O2– + H2O → OH– + H2O2 + O2 catalyzed by: 87. b
 (JIPMER May 2018) a. Iron (PGMEE 2013) 88. b
a. Unlabelled B12 is given IM b. Superoxide dismutase 89. c
b. Labelled B12 given orally c. Catalase 90. a
c. Vitamin B12 excreted in urine d. Glutathione peroxidase 91. b
d. Abnormal test rules out primary intestinal 94. Which of following is not a free radical-  92. d
malabsorption  (PGMEE 2015) 93. a
84. Which of the following is/are applications of RIA a. OH– b. Superoxide amino 94. c
(Radio Immuno Assay)? (JIPMER May 2018) c. HOCl– d. H2O2 95. d
a. Barbiturates assay 95. Phase-II reaction is- (PGMEE 2015)
b. Hormone analysis a. Hydroxylation b. Cyclization
c. Analysis of anti-DNA antibodies c. Oxidation d. Conjugation
d. All
 96. True regarding Cytochrome P450 are all EXCEPT- 107. Respiratory quotient after an exclusive carbohydrate
 (PGMEE 2013) meal is: (AIIMS November 2016)
402 a. Present in endoplasmic reticulum a. 0.7 b. 0.8
b. Absent in liver c. 1 d. 1.2
c. Involved in phase I metabolism 108. Respiratory quotient of carbohydrate:
d. All are hemoproteins  (PGMEE 2015)
97. True regarding glucuronidation: (PGMEE 2013) a. 0.5 b. 0.7
a. Phase I reaction c. 0.8 d. 1
b. Water solubility is decreased 109. Respiratory Quotient 0.7 is seen in: (PGMEE 2015)
c. Phase II reaction a. Carbohydrates
d. Done by CYP enzymes b. Alcohol
98. Phase II reactions include: (PGI Nov 2017) c. Protein
a. Acetylation d. Fat
b. Glycine conjugation 110. RQ is least in – (PGMEE 2012-13)
c. Methylation a. Brain
d. Reduction b. Adipose tissues
e. Glucuronidation c. RBC
99. Tocopheroxyl radical is converted to Tocopherol by d. Heart
which vitamin: (Recent Question 2016)
a. Vitamin D b. Vitamin C Alcohol
c. Vitamin K d. Vitamin B 111. Fatty liver caused due to excess alcohol consumption
100. In crystalline lens, level of tocopherol and ascorbate is because of the excess ratio of:
is maintained by:
a. NAD/NADH (Recent Question June 2018)
a. Glutathione b. Lipoic acid
b. NADP/NADPH
c. Vitamin D d. Fatty acid
c. NADH/NAD
101. Which of the following has antioxidant property?
d. NADPH/NADP
 (PGI Nov 2012)
112. Biomarker of Alcoholic Hepatitis: (AIIMS Nov 2018)
a. Tocopherol b. Citrulline
a. ALP b. AST
c. Vitamin K d. Lycopene
c. LDH d. GGT
e. Reduced Glutathione
M 102. Minerals which can generate free radical are all
113. 4 year old baby boy landed in emergency with rapid
C EXCEPT: breathing. He was cold and clammy, confused,
Qs a. Copper b. Selenium lethargic. His mother gave a history of accidental
Ans. c. Cobalt d. Nickel ingestion of automobile antifreeze. Ethanol is used
96. b 103. The Fenton reaction leads to free radical generation as treatment in this poisoning because:
97. c when: (PGMEE 2007) a. It conjugate with ethylene glycol
98. a,b, a. Radiant energy is absorbed by water b. Inhibit enzyme alcohol dehydrogenase
c,e b. Hydrogen peroxide is formed by myeloperoxidase c. Inhibit binding of ethylene glycol to alcohol dehy-
99. b c. Ferrous ions are converted to ferric ions drogenase
100. a d. Nitric oxide is converted to peroxynitrite anion d. Stimulate the excretion of ethylene glycol
101. a,d,e 104. Cytochrome P450 is/are involved in: (PGI May 2016) 114. Asians and native americans may flush and feel ill
102. b a. Hydroxylation of xenobiotics after drinking small amount of ethanol. This reaction
103. c b. Methylation of xenobiotics is due to genetic variation in which enzyme?
104. a,c, c. Deamination reaction a. Alcohol Dehydrogenase
d,e d. Involved in hydroxylation of steroids b. Aldehyde Dehydrogenase
105. a,b, e. Drug metabolism c. Isocitrate Dehydrogenase
c,d,e 105. Which of the following act as antioxidants: d. Alpha-Ketoglutarate Dehydrogenase
106. b  (PGI Nov 2009) 115. Alcohol ingestion leads to production of acetaldehyde
107. c a. Vitamin D and NADH. This excess NADH promotes which of the
108. d b. Vitamin C following reaction:
109. d c. Selenium a. Malate to Oxaloacetate
110. d d. Glutathione peroxidase b. Oxaloacetate to Malate
111. c e. Vitamin E c. Acetyl CoA to Citrate
112. d d. Lactate to Pyruvate
113. c Respiratory Quotient (RQ) 116. Feature (s) of Korsakoff psychosis: (PGI Nov 2014)
114. b 106. Respiratory quotient after heavy carbohydrate meal a. Confabulation
115. b is: (AIIMS Nov 2018) b. Retrograde amnesia
116. a,b,c a. 1 b. 1.2 c. Ophthalmoplegia
c. 0.8 d. 0.7 d. Delirium
 Answers with Explanations
403
1. Ans. (d) Vitamin K affects bone health by activating Transketolase. Clinical features of B2 deficiency i.e.
proteins that bind calcium Riboflavinosis are corneal vascularization, cheilosis,

CHAPTER 15  MISCELLANEOUS
glossitis, angular stomatitis.
[Ref: Harper 30th/e pg. 554]
Vitamin K is fat soluble. It is need for the activation of 8. Ans. (d) All of the above
clotting factors II, VII, IX and X and also for Protein
[Ref: Harper 30th/e pg. 550]
C and S. It also activates proteins – Nephrocalcin &
Osteocalcin, which binds calcium. Vitamin K deficiency Restless leg syndrome is a disorder of nervous system
will lead to bleeding, not deep vein thrombosis. and a sleep disorder where patient has an urge to move
legs. Factors associated with this disorder are deficiency
2. Ans. (b) Vitamin C of iron, folic acid, biotin, manganese, kidney failure,
[Ref: Harper 30th/e pg. 561] Diabetes and Peripheral Neuropathy.

Water soluble vitamins are excreted in urine like B – 9. Ans. (a) Vitamin B3 (Niacin)
complex vitamins and vitamin C.
[Ref: Harper 30th/e pg. 557]
3. Ans. (c) Lysine
Vitamin B3 (Niacin) and Vitamin D are considered
[Ref: Harper 30th/e pg. 562] Atypical Vitamins as they can be synthesized in body.
Do not consider those Vitamins which are synthesized
Vitamin C is required for hydroxylation reactions.
by intestinal flora. (Vitamin B3 is synthesized from
Proline to hydroxyl proline conversion occurs by
amino acid - Tryptophan).
enzyme prolyl hydroxylase. Lysine to hydroxyl lysine
conversion occurs by lysyl hydroxylase. These reactions 10. Ans. (d) Vitamin B1
are required in collagen post translational modification.
[Ref: Harper 30th/e pg. 555]
4. Ans. (d) Vitamin C
Vitamin B1 deficiency is Beri-Beri, which can cause
[Ref: Nelson 20th/e/p 329, 330, Ghai 8th/e/p 120] oedema in wet beri-beri which affects CVS, leading to
cardiac failure, pulmonary and peripheral oedema.
The source of Vitamin C is fresh citrus fruits and
deficiency of vitamin C presents with bleeding gums
11. Ans. (a); (d); (e)
and petechiae. Collagen post translational modification
is affected here, wound healing is impaired. [Ref: Harper 30th/e pg. 557] A
Human enzymes can synthesize vitamin B3 and Vitamin N
5. Ans. (c) Thiamine, Pyridoxine, Cyanocobalamin
D (atypical vitamins). Intestinal flora can synthesize S
[Ref: Harper 30th/e pg. 62] vitamin B2, B5, B7 and Vitamin K. W
B1 deficiency leads to Dry Beri–Beri where nervous E
system is affected. Pyridoxine and B12 deficiency also 12. Ans. (b) Inactivation of Chloride Channel R
leads to neurological manifestations. S
[Ref: Harper 30th/e pg. 62]
6. Ans. (b) Homocystinemia WITH
Thiamine normally activates a chloride channel in
the membrane of nervous system. So deficiency leads E
[Ref: Harrison 18th/p 594, 595, 3218]
to inactivation of this chloride channel and nerve X
Homocystinemia occurs due to deficiency of vitamins weakness. P
– B6, B9 and B12. Alcohol interferes with the absorption L
of Thiamine. Patients using diuretics have increased 13. Ans. (d) Biotin A
excretion of Thiamine in urine. N
[Ref: Harper 30th/e pg. 556]
A
7. Ans. (a) Vitamin B2 Excess consumption of raw eggs, which contains a T
protein ‘Avidin’ – a biotin binding protein leads to biotin I
[Ref: Harper 30th/e pg. 556]
deficiency. When cooked, avidin is partially denatured O
The marker enzyme of B2 deficiency is Glutathione and binding to biotin is reduced. So cooked egg whites N
Reductase and the marker for B1 deficiency is are safe to consume. S
 14. Ans. (b) Pantothenate kinase – associated neurode- 21. Ans. (c) Both a and b
generation
404 [Ref: Harper 30th/e pg. 556]
[Ref: Harper 30th/e pg. 559]
Glutamate dehydrogenase is the only enzyme which
Hallervorden-Spatz syndrome: can use both NAD+ and NADP.
•• Pantothenate Kinase – associated neurodegeneration
CRO BIOCHEMISTRY

•• Rare, autosomal recessive, excessive accumulation of 22. Ans. (c) Pantothenic acid
iron containing pigments. In brain accumulation of
N-pantothenoyl-cysteine and pantetheine, leading to [Ref: Harper 30th/e pg. 162]
free radical cell damage characterized by parkinson Beta Alanine is component of Pantothenic acid i.e.
like features and mental retardation. Vitamin B5.
15. Ans. (a) Thiamine 23. Ans. (b) Acetylation
[Ref: Harper 30th/e pg. 567] [Ref: Harper 30th/e pg. 561]
Both Thiamine and Niacin causes psychosis but Pantothenic acid containing coenzyme is Coenzyme A,


Thiamine is best option to be marked as Niacin is an which is involved in acetylation reactions.

atypical vitamin.
24. Ans. (c) Niacin (Vitamin B3)
16. Ans. (a) Vitamin B12
[Ref: Harper 30th/e pg. 557]
[Ref: Harper 30th/e pg. 560]
Dementia can occur due to deficiency of Vitamins B1, This amino acid tryptophan can synthesize a vitamin
B3 and B12. i.e. Niacin (Vitamin B3).

17. Ans. (a) N5 Formyl THFA 25. Ans. (c) Dactylitis

[Ref: Harper 30th/e pg. 559] [Ref: Ghai 8th/e pg. 118, Nelson 20th/e/p 323 table 49-1]
N5 Formyl THFA/ Folinic acid has high stability so it is Three Ds of Pellagra (Niacin deficiency) are Diarrhoea,
used therapeutically. dermatitis and dementia.
Dactylitis /Sausage digit is the inflammation of an entire
18. Ans. (c) Vitamin B2 digit (a finger or toe). This does not occur in pellagra.
[Ref: Harper 30th/e pg. 556]
26. Ans. (c) Pyridoxal phosphate
B2 Deficiency: Riboflavinosis is characterized by
Corneal vascularization (earliest sign of deficiency), [Ref: Harper 30th/e pg. 558]
A Glossitis – inflammation of tongue (Magenta tongue- In Vitamin B6/Pyridoxine deficiency: Microcytic
N characteristic), Cheilosis – (redness and shiny anemia, kidney stones, convulsions, mental confusion,
S appearance of lip), Angular Stomatitis (lesions at muco- depression, carpel tunnel syndrome, neuropathy
W cutaneous junction at corners of mouth), leading to occurs. Pellagra occurs as vitamin B6 is required for the
E painful fissures, Seborrhoic Dermatitis – (rough scaly
conversion of tryptophan to niacin.
R skin because of desquamation).
S 27. Ans. (d) All
19. Ans. (a) FMN
WITH [Ref: Nelson 20th/e p 326]
[Ref: Harper 30th/e pg. 62]
E Riboflavin i.e. vitamin B2 is a constituent of FMN Deficiency of Vitmain B6 results in accumulation of
X (Flavin Mono Nucleotide) and FAD (Flavin Adenine Homocysteine and Cystathionine (in the metabolism of
P Dinucleotide). Niacin i.e. Vitamin B3 is a constituent of methionine & formation of cysteine).
L NAD and NADP. Also, in Tryptophan metabolism, 3-Hydroxy
A Kynurenine is one of the intermediate, which gets
N 20. Ans. (d) Riboflavin converted to next intermediate and this conversion
A requires vitamin B6 (Pyridoxine). In vitamin B6
[Ref: Harper 30th/e pg. 556]
T deficiency, this reaction does not occur and 3-Hydroxy
I Riboflavin i.e. Vitamin B2 is a constituent of FMN Kynurenine is diverted to form alternate metabolite,
O and FAD. FMN requiring enzymes are L-amino acid Xanthurenic acid, which is excreted in urine.
N Oxidase, Complex I (ETC), Monoamine Oxidase.
S
28. Ans. (c) Vitamin B6 34. Ans. (c) Folic acid along with hydroxycobalamin

[Ref: Harper 30th/e pg. 554] [Ref: Harper 30th/e pg. 560] 405
Vitamin B6 is responsible for simple decarboxylation. In folate deficiency, megaloblastic anemia occurs.
And vitamin B1 is responsible for oxidative decar- In B12 deficiency, both megaloblastic anemia and
neurological manifestations occur.

CHAPTER 15  MISCELLANEOUS
boxylation.

29. Ans. (a) Methylene THF 35. Ans. (b) Folic acid

[Ref: Harper 30th/e pg. 560] [Ref: Harper 30th/e pg. 500]

Major form of Folic Acid for transfer of one carbon Methionine (S-containing) metabolism requires three
is –Methylene THF. Main form of Folic Acid in vitamins: folate, cobalamin and pyridoxine.
blood is –Methyl THF. Active form of Folic Acid is - 36. Ans. (b) Macrocytic
Tetrahydrofolate.
[Ref: Wintrobes Hematology 12th/e p. 794, 795]
30. Ans. (c) Beta Carbon
Vitamin B12 deficiency causes Macrocytic anemia.
[Ref: Harper 30th/e pg. 559]
37. Ans. (c) Glutamate to GABA
Beta carbon is added to THF when serine is converted
to glycine. [Ref: Harper 30th/e pg. 556]
•• Biotin, Vitamin B7 is required mainly in carboxylation
reactions.
•• Acetyl CoA to Malonyl CoA is done by enzyme Acetyl
CoA Carboxylase
•• Pyruvate to Oxaloacetate is done by enzyme Pyruvate
Carboxylase
•• Propionyl CoA to Methyl Malonyl CoA is done by
enzyme Propionyl CoA Carboxylase
•• Glutamate to GABA conversion is a simple
decarboxylation reaction, which requires vitamin B6
i.e. Pyridoxine
31. Ans. (d) Tyrosine
38. Ans. (b) Succinate dehydrogenase
[Ref: Harper 30th/e pg. 599]
[Ref: Harrison 18th/e pg. 594, 595]
Amino acids which take part in one carbon transfer A
Thiamine i.e. Vitamin B1 is needed mainly in oxidative N
reactions are: glycine, serine, threonine, methionine.
decarboxylation reactions and for enzyme Transketolase S
Vitamins which take part in one carbon transfer which is an enzyme in HMP pathway.
reactions are folate and cobalamin. W
Alpha Keto-Glutarate Dehydrogenase, Pyruvate Dehy-
E
drogenase and Branched Chain Alpha Keto Acid
32. Ans. (c) Folic acid R
Dehydrogenase are oxidative decarboxylation steps
[Ref: Harper 30th/e pg. 558] requiring Thiamine. S
But Succinate Dehydrogenase, enzyme of TCA is doing WITH
Supplements of folate given in appropriate dose before oxidation, not oxidative decarboxylation.
conception can result in a significant reduction in the E
incidence of spina bifida, and other neural tube defects. 39. Ans. (b) Lactate to Pyruvate X
P
33. Ans. (b) Folic acid [Ref: Harrison 18th/e pg.594, 595, 3218]
L
Thiamine acts as a co-enzyme for multienzyme A
[Ref: Harper 30th/e pg. 299]
complexes doing Oxidative Decarboxylation e.g. N
Folate is required for the conversion of FIGLU (Form Pyruvate Dehydrogenase complex, Alpha Keto- A
Imino Glutamate) to Glutamate. This is the basis of Glutarate Dehydrogenase complex, Branched Chain T
FIGLU excretion test in folic acid deficiency. Amino Acid Dehydrogenase complex, Transketolase, I
(used in non-oxidative phase of HMP), Lactate Dehy- O
drogenase converts Lactate to Pyruvate. Here NAD is N
involved so Vitamin B3 is used, not B1. S
 40. Ans. (d) All 48. Ans. (d) Selenium

406 [Ref: Nelson’s 20th/e pg. 320] [Ref: Harper 30th/e pg. 567]
Vitamin A toxicity leads to alopecia, dry pruritic skin, Selenium is required for glutathione peroxidase, which
hepatomegaly, liver damage, raised intracranial is required for the antioxidant action of Vitamin E.
pressure which sometimes mimic brain tumor. This is
CRO BIOCHEMISTRY

known as benign intracranial hypertension (Pseudo- 49. Ans. (a) Vitamin K

tumour cerebri). Organelle affected is lysosomes. [Ref: Harper 30th/e pg. 554]
41. Ans. (a) Sunflower seeds Vitamins which are synthesized by intestinal flora are
vitamin B2, B5, B7 and Vitamin K.
[Ref: Harrison’s 18th ed chapter 74]
Vitamin A is present in leafy vegetables, carrots (being 50. Ans. (c) Menadione
the richest source in plants), whole milk, fish liver oil, [Ref: Harper 30th/e pg. 554]
egg, butter, meat, tomato.
Menadione is the synthetic water soluble form of


42. Ans. (c) Vitamin D Vitamin K.

[Ref: Harper 30th/e pg. 508] 51. Ans. (d) Carboxylation

Cow’s milk has more Vitamin D than breast milk. [Ref: Harper 30th/e pg. 554]
43. Ans. (b) Vitamin D Refer to Ans. 52

[Ref: Harper 30th/e pg. 508] 52. Ans. (a) Glutamate

Vitamin D is considered as a hormone as it is synthesized [Ref: Harper 30th/e pg. 554]
in body, released in circulation and acts by binding to
the nuclear receptors. Q. 51 & 52 Vitamin K is required in the gamma-
carboxylation of glutamate and this is a post transla-
44. Ans. (b) Fish liver oils tional modification in proteins, specially prothrombin-
factor II.
[Ref: Harper 30th/e pg. 508]
53. Ans. (d) Prothrombin is a vitamin K dependent
Fish liver oils are the richest source of Vitamin D. Other
factor
sources of Vitamin D are egg yolk, liver, butter, cheese.
[Ref: Harper 30th/e pg. 554]
45. Ans. (d) Kidney
Vitamin K dependent factors undergo post translational
A [Ref: Harper 30th/e pg. 551] (not post-transcriptional) modifications. Menadione
N
•• 25-hydroxy cholecalciferol is formed in liver is a synthetic water soluble vitamin K. Stuart-Prower
S
•• 1, 25 (OH)2 cholecalciferol is formed in kidneys and it factor is Factor X, which is vitamin K dependent.
W
E is the most potent form of Vitamin D.
54. Ans. (c) Vitamin K
R
46. Ans. (b) Vitamin E [Ref: Harper 30th/e pg. 554]
S
[Ref: Harper 30th/e pg. 553] Vitamin K brings about post translational modification
WITH
Vitamin E is a fat soluble and anti-oxidant vitamin. of certain blood clotting factors. The clotting factor II
E (Prothrombin), VII, IX and X are synthesized as inactive
X 47. Ans. (c) Ophthalmoplegia precursors (Zymogens) in the liver.
P
[Ref: Harper 30th/e pg. 567] 55. Ans. (a) > 6 months
L
A Deficiency of vitamin E can lead to neurological [Ref: Harper 30th/e pg. 547]
N problems, such as difficulty in coordinating movements
A (Ataxia) and speech (Dysarthria), loss of reflexes in the Vitamin A stores last for more than 6 months in a well
T legs (lower limb Areflexia), and a loss of sensation in the fed person.
I extremities (peripheral neuropathy).
O Thiamine deficiency can cause Ophthalmoplegia in
N susceptible persons.
S
56. Ans. (a) Lead 63. Ans. (c) Zinc

[Ref: Harper 30th/e pg.562] [Ref: Harper 30th/e pg. 549] 407
Lead is a non-essential mineral which is not required Selenium is the best answer to be marked. If selenium
in human nutrition instead it is considered as toxic not given then mark zinc, as zinc is required for enzyme
chemical while sodium, manganese, iron are essential Superoxide Dismutase.

CHAPTER 15  MISCELLANEOUS
elements.
64. Ans. (b) 2
57. Ans. (d) Molybdenum
[Ref: Harper 30th/e pg. 674, 541]
[Ref: Text book of biochemistry by G.P talwar and LM Number of iron in transferrin is 2. Transferrin is the
srivastava page – 585, Shinde and chaatterjee 6th ed p.
transport form of iron which has two binding sites for
555]
iron.
All oxidases require copper but two oxidases require Ferritin is the storage form of iron. It can carry 4000-
molybdenum. They are xanthine oxidase and sulphite 4500 iron atoms. It is a measure of body iron stores.
oxidase.
65. Ans. (b) Transthyretin
58. Ans. (c) Carbonic anhydrase
[Ref: Harper 30th/e pg. 551]
[Ref: Harper 30th/e pg. 643]
Transthyretin (TTR) is a transport protein in plasma
Zinc requiring enzymes are carbonic anhydrase, alcohol and CSF that carries T4 and retinol-binding protein
dehydrogenase, aldehyde dehydrogenase, carboxy bound to retinol.
peptidase, alkaline phosphatase. Liver secretes Transthyretin into blood and choroid
plexus secretes TTR into CSF. TTR was originally
59. Ans. (d) Iron called prealbumin (or Thyroxine-Binding Prealbumin)
[Ref: Harper 30th/e pg. 564] because it moves faster than albumin in electrophoresis.

Iron is not an anti-oxidant, rest all are anti-oxidants. 66. Ans. (a) Transthyretin
Zn play a role as anti-oxidant as it binds with SH group
[Ref: Harper 30th/e pg. 551]
of biomolecules protecting them from oxidation. Also
it increases the activation of anti-oxidant properties. Transthyretin: Transports thyroxine and retinol
Action of Se is complementary to Vitamin E, constituent binding protein. Proteins for iron storage are Ferritin
of glutathione peroxidase and selenocysteine. Cu has and haemosiderin. Haemosiderin has higher iron
role as anti-oxidant as it is present as a constitutent in content than ferritin. Transferrin is for iron transport.
Cu/Zn SOD and ceruloplamin. Ceruloplasmin is a Cu containing enzyme which is also A
having ferroxidase activity. Ferroxidase converts Fe+2 to N
60. Ans. (b) Magnesium Fe+3 (i.e. Ferrous to ferric). Ferric form of iron is required S
for transport by transferrin in plasma. Hepcidin W
[Ref: Harper 30th/e pg. 120]
regulates iron transport in circulation. E
All kinases require magnesium. But pyruvate kinase Heparin is an anticoagulant used for the measurement R
requires K >> Mg. of serum electrolyte.
S
61. Ans. (d) Copper binding ATPase 67. Ans. (d) Pyridoxal phosphate WITH
[Ref: Harper 30th/e pg. 47,281] [Ref: Harper 30th/e pg. 557] E
In Menke’s syndrome, there is dietary deficiency of Vitamin B6 i.e. Pyridoxal phosphate is required as a X
copper. Defect occurs in the production of Cu binding cofactor for enzyme glycogen phosphorylase. P
P-type ATPase in intestinal mucosa, leads to copper L
deficiency. 68. Ans. (a) Glycogen phosphorylase A
N
62. Ans. (d) Infertility [Ref: Harper 30th/e pg. 557] A
Glycogen phosphorylase requires Vitamin B6 – Pyri- T
[Ref: Harper 30th/e pg. 549] I
doxine.
Zinc deficiency leads to hypogonadism, infertility, O
dermatitis, diarrhoea, poor wound healing and growth N
retardation, alopecia immunodeficiency. S
 69. Ans. (b); (e) 74. Ans. (b) Methyl-Histidine

408 [Ref: Harrison 18th/e pg.594, 595, 3218] [Ref: Harper 30th/e pg. 667]
Thiamine deficiency cause polyneuropathy and cardio- Wilson disease is a condition of excess copper in body.
megaly. Also, there exists a renal lesion in Wilson’s disease.
Glosssitis and angular stomatitis are characteristics of Plasma amino acids are normal both qualitatively &
CRO BIOCHEMISTRY

vitamin B2 deficiency whereas pellagra is characteristic quantitatively, both in fed & fasting state. Aminoaciduria
of Vitamin B3 deficiency. (especially lysine, threonine, cystine) occurs. But
taurine & 1 Methyl-Histidine & 3 Methyl-Histidine are
70. Ans. (a); (c); (d); (e) less in urine.
This is a journal based very rare question, just cram it.
[Ref: Harper 30th/e pg. 550]
Pyridoxine is Vitamin B6, active form of which is PLP. It is 75. Ans. (a) Vitamin B1
mainly required for proteins and amino acid metabolism. [Ref: Harper 30th/e pg. 554]
Examples: Haem synthesis, Transamination, Trans- This is a case of Wet/Cardiac Beri-Beri, i.e. Vitamin B1
sulfuration and simple decarboxylation & catabolism of


deficiency.
tryptophan to niacin.
76. Ans. (a) Selenium
71. Ans. (a); (b); (c); (d); (e)
[Ref: Harper 30th/e pg 286]
[Ref: Harper 30th/e pg. 553] Keshan’s disease is due to the deficiency of Selenium.
Vitamin E deficiency causes hemolysis, posterior
column involvement, peripheral neuropathy, hair loss, 77. Ans. (b) Selenium
Impaired immunity. Haemolytic anaemia is associated [Ref: Harper 30th/e pg. 380]
with deficiency of vitamin E. Dysesthesia is caused by deficiency of Zn. Also known as
It is associated with Lichtheim's disease. Lichtheim's burning mouth syndrome. Other causes of this syndrome
disease is subacute combined degeneration of spinal are deficiency ofvitamin B1, vitamin B2, vitamin B6,
cord and involves the degeneration of posterior and vitamin B12 and folate deficiency.
lateral column of spinal cord.
(Deficiency of Vitamin C and Vitamin K is associated 78. Ans. (b) Vitamin C
with bleeding.)
[Ref: Harper’s illustrated biochemistry, 30th ed. Pg. 541]
72. Ans. (a); (b); (c); (d) Factors which increase iron absorption are Vitamin C/
Ascorbic acid, Cysteine (SH containing amino acid)-
[Ref: Lippincott 4th/e pg377]
A they convert ferric to ferrous form, which is required
Deficiency of vitamin C leads to scurvy. Features for absorption. Factors which decrease iron absorption
N
are: Decreased immunity, improper wound healing, are antacids, alkalies, phosphates (present in egg yolk),
S
epistaxis, bleeding, haemorrhages, microcytic hypo- Phytates (present in wheat, maize), drug- tetracycline.
W
chromic anaemia because absorption of Fe requires presence of food in stomach also decreases absorption
E
vitamin C. Low grade fever, irritabililty, digestive distur- of iron.
R
bances, rachitic rosary - prominant ribs with disrupted
S costochondral junction occurs. 79. Ans. (d) Epoxide Reductase
WITH [Ref: Harper’s illustrated biochemistry, 30th ed., pg. 555]
73. Ans. (c) Ito cells
E Vitamin K in its coenzyme form is regenerated by
[Ref: Harper 30th/e pg. 545] epoxide reductase
X
P HSCs (Hepatic Stellate Cells) (also called vitamin
L 80. Ans. (d) All
A-storing cells, lipocytes, interstitial cells, fat-storing
A cells or Ito cells, perisinusoidal cells) are pericytes [Ref: Harper 30th/e pg. 62]
N which exist in the space between parenchymal cells and Zinc, a divalent cation , is important for normal growth
A liver sinusoidal endothelial cells of the hepatic lobule and development of children both with and without
T and store 50-80% of vitamin A in the whole body as diarrhoea. It regulates intestinal electrolytes. It also
I retinyl palmitate. enhances immune function.
O
N
S
81. Ans. (c) Riboflavin 86. Ans. (a) Hydrogen peroxide
[Ref :https://www.ncbi.nlm.nih.gov/pmc/articles/ [Ref: Harer30th/e pg. 586] 409
PMC3848652/]
Hydrogen peroxide is not a free radical but it can
•• Riboflavin-Responsive Trimethylaminuria (TMAU):
generate free radicals. Precursor of all ROS is superoxide
Riboflavin (vitamin B2) administration has been
radical

CHAPTER 15  MISCELLANEOUS
associated with reduction of trimethylamine
excretion in some TMAU patients presumed to be 87. Ans. (b) a– Tocopherol
due to increased FMO3 activity with riboflavin acting
as a co-factor. [Ref: harper 30th/e pg. 125]
•• Trimethylamine gives fishy odour
Chain breaking anti-oxidants are a- Tocopherol, β-
82. Ans. (a) Beta carotene Carotene and Super oxide dismutase (SOD), out of
which a- Tocopherol is most powerful.
[Ref: Harper 30th/e pg.219]
88. Ans. (b) Hydroxyl radical
Carotenoids are provitamins of vitamin A which are
present in plants. Most important carotenoid is beta [Ref: N.V. Bhagavan 4th/e p. 925]
carotene , which consists of two molecules of vitamin A
(retinal) joined end to end. OFR with highest activity which is most powerful is OH
(hydroxy) radical.
83. Ans. (d) Abnormal test rules out primary intestinal
malabsorption 89. Ans. (c) SOD

[Ref:https://www.ncbi.nlm.nih.gov/books/NBK507784/] [Ref: Harper 30th/e pg. 124]

•• Schilling test is a test for B12 deficiency, named after Enzyme which carries dismutation reaction is Superox-
the scientist who discovered. It uses radioactive ide Dismutase (SOD). Refer Fig. 15.3
cobalt 60.
•• Shilling test – administration of oral (radioactive) 90. Ans. (a) Catalase
supplement is followed by IM administration. If
[Ref: Harper 30th/e pg. 556}
Radioactive B12 is excreted in urine, it suggests that it
was absorbed (no pernicious anemia). Catalase is an enzymatic antioxidant which destroys
•• If there is no radioactive B12 in urine - then give H2O2. Refer Fig. 15.4
oral radioactive vitamin along with intrinsic factor
(followed by IM administration). If Radioactive B12 is 91. Ans. (b) FOX assay
excreted in urine, it suggests pernicious anemia.
[Ref: Harper 30th/e pg. 563-565]
•• IM administration is supposed to saturate liver rece-
A
ptors, but not enough to completely replenish the Free radical measurement can be done by:
N
body needs. •• FOX assay – (Ferrous oxidation in Xylenol)
S
•• Estimation of Dialdehydes (e.g. MDA- Malondia-
84. Ans. (d) All W
ldehyde)
E
[Ref: Harper 30th/e pg.596] •• Pentane and methane measurement in exhaled air
R
•• Applications of RIA (Radio Immuno Assay): 92. Ans. (d) Malondialdehyde S
ƒƒ In drug assays e.g. Barbiturates, Morphine,
[Ref: Harper 30th/e pg. 220] WITH
Digitoxin, Amphetamine etc.
ƒƒ Vitamin analysis e.g. Folic acid, Riboflavin MDA (Malondialdehyde) is also considered the marker E
ƒƒ Hormone analysis e.g. Aldosterone, Insulin, of lipid peroxidation. X
Growth Hormone, Thyroxine P
ƒƒ Analysis of anti-DNA antibodies like in SLE 93. Ans. (a) Iron L
(Systemic Lupus Erythematosus) A
[Ref: Harper 30th/e p. 566]
N
85. Ans. (a) Glutathione peroxidase
Iron is a transition metal which can take part in the A
[Ref: Harper 30th/e pg. 200] generation of free radicals. T
I
Glutathione peroxidase is a Se containing protein O
having anti oxidation or radical scavenging property. N
S
 94. Ans. (c) HOCl– in ER, liver, intestine and also found in mitochondria,
in some tissues. It is involved in both phase I & phase
410 [Ref: Principles in medical pathology p. 391] II reactions.
Though both HOCl– & H2O2 are both not free radicals,
but if both given in options then HOCl– is better option 97. Ans. (c) Phase II reaction
as HOCl– is a precursor for many free radicals in the
CRO BIOCHEMISTRY

[Ref: Harper30th/e pg. 586]

human body. (HOCl– >>> H2O2).
Glucuronidation reactions is phase II reaction. Phase II
95. Ans. (d) Conjugation reactions make the molecules more water soluble and
eventually they are excreted in urine or bile.
[Ref: Harper 28th/e p. 609-612; Essentials of biochemistry
p. 412] 98. Ans. (a); (b); (c); (e)

•• Phase II reactions include conjugation and [Ref: Harper30th/e pg. 586]

methylation. Conjugation is the most common •• Phase I reactions are – hydroxylation, oxidation,
xenobiotic reaction. reduction, hydrolysis.


•• Conjugating agents are: glutathione, glycine (e.g. •• Phase II reactions include conjugation and
benzoic acid conjugated with glycine to form methylation. Conjugation is the most common
hippuric acid), glucuronic acid (e.g. bilirubin), xenobiotic reaction. Conjugating agents are:
sulfation (by PAPS- Phospho Adenosyl Phospho glutathione, glycine (e.g. benzoic acid conjugated
Sulfate), acetylation (by Acetyl CoA) and methylation with glycine to form hippuric acid), glucuronic
(by SAM- S-Adenosyl Methionine). acid (e.g. bilirubin), sulfation (by PAPS-
phosphoadenosylphospho sulfate), acetylation (by
96. Ans. (b) Absent in liver Acetyl CoA) and methylation (by SAM- S-Adenosyl
[Ref: Harper30th/e pg. 584] methionine).

Cytochrome P450 are important superfamily of heme

containing monooxygenases. They are located mainly

99. Ans. (b) Vitamin C

[Ref: Harper 30th/e pg. 553]
When vitamin E or Tocopherol scavenges lipid peroxide radicals, then it is converted to Tocopheroxyl radical. This is
A reduced back to Tocopherol with the help of vitamin C. Glutathione helps in regenerating ascorbate.
N
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100. Ans. (a) Glutathione 108. Ans. (d) 1

[Ref: Harper 30th/e pg. 546] [Ref: Dinesh puri 3rd/e p. 712] 411
When Vitamin E or Tocopherol scavenges lipid peroxide For carbohydrates, RQ is 1
radicals, then it is converted to Tocopheroxyl radical.
This is reduced back to tocopherol with the help of 109. Ans. (d) Fat

CHAPTER 15  MISCELLANEOUS
Vitamin C. Glutathione helps in regenerating ascorbate.
[Ref: Harper 30th/e pg. 119]
101. Ans. (a); (d); (e) For Fats, RQ is 0.7 (calculated indirectly)
[Ref: Harper 30th/e pg. 549] 110. Ans. (d) Heart
Tocopherol, Lycopene and Reduced glutathione all act
as antioxidants. [Ref: Harper 30th/e pg. 119]
Heart uses fats as the main fuel even in fed state. RQ for
102. Ans. (b) Selenium fats is 0.7.
[Ref: Harper 30th/e pg. 556] 111. Ans. (c) NADH/NAD
Selenium helps in the scavenging of free radical as it is
[Ref: Harper 30th/e pg 261]
required for enzyme glutathione peroxidase.
Transition metals like iron, copper, cobalt, nickel •• The above reaction takes place in alcohol metabolism
generate free radicals. and there is increase in NADH. So ratio of NADH/
NAD increases.
103. Ans. (c) Ferrous ions are converted to ferric ions •• In liver cells when there is increase in NADH.
β oxidation does not occur because cell has enough
[Ref: Harper 30th/e pg. 674] NADH and it does not have the need to generate it.
In Fenton reaction, hydroxy free radical is produced Fatty acid synthesis increases leading to fatty liver.
when ferrous is converted to ferric form. To use the excess NADH in the cell, oxaloacetate
gets converted to malate, and oxaloacetate is no
104. Ans. (a); (c) ; (d); (e) longer available for TCA. So TCA is inhibited, as a
result acetyl CoA accumulates, which is diverted to
[Ref: Harper 30th/e pg. 584]
fatty acid synthesis. These fatty acids form lots of fats
Cytochrome P450 plays role in drug detoxification leading to fatty liver.
and steroid synthesis, hydroxylation of xenobiotics
and steroids. Cytochrome P450 belongs to phase I 112. Ans. (d) GGT
metabolism catalysing reactions such as hydroxylation, [Ref: Harper 30th/e pg. 674]
deamination, desulfuration, epioxidation, peroxy- A
genation and reduction. Whereas methylation is a Markers for alcoholism: N
phase II reaction. (a) CDT- Carbohydrate deficient transferrin S
(b) g- GGT- Gamma Glutamyl Transpeptidase
W
105. Ans. (a); (b); (c); (d); (e)
113. Ans. (c) Inhibit binding of Ethylene Glycol to Alcohol E
[Ref: Harper 30th/e pg. 582] Dehydrogenase R
All these vitamins D, C, E act as antioxidants. Glutathione
S
[Ref: Harper 30th/e pg. 261]
peroxidase has selenium in it, so act as antioxidant. WITH
Ethylene Glycol is present in automobile antifreeze.
106. Ans. (b) 1.2 Ethanol is used as antidote in Ethylene Glycol poisoning. E
Ethanol has low km for Alcohol Dehydrogenase, thus it X
Refer to ans 107. inhibits the binding of Ethylene Glycol with the enzyme. P
107. Ans. (c) 1 L
114. Ans. (b) Aldehyde Dehydrogenase
A
[Ref: Harper 30th/e pg. 119; Harper 30th /e p541 [Ref: Harper 30th/e pg. 261] N
Q106, 107 If exclusive carbohydrates given in Q, then A
Genetic variation occurs in enzyme aldehyde dehy-
RQ is 1. If excess carbohydrates given in Q, then RQ is T
drogenase, specially in people of china, korea and
more than 1. Because if excess carbohydrates taken, I
native america, known as Asian flush syndrome.
then carbohydrates get converted to fats in body. O
This occurs due to increased acetaldehyde which causes
N
nausea, vomiting, flushing, sweating, tachycardia and
S
hyperventilation.
 115. Ans. (b) Oxaloacetate to malate 116. Ans. (a); (b); (c)

412 [Ref: Harper 30th/e pg. 184, 261] [Ref: Harper 30th/e pg. 594]
In alcoholism, NADH excess occurs. So these following Wernicke-Korsakoff Syndrome occurs in Thiamine
reactions start occurring. (Vitamin B1) deficiency, which occurs in alcoholics. It
is characterized by ataxia, memory loss, confabulations,
CRO BIOCHEMISTRY

Ophthalmoplegia, nystagmus and Cerebral haem-

orrhage.


A
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P
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Extra Edge Tables

BIOCHEMICAL TESTS
Tests Specific for

Hay’s Sulphur test Bile salts in urine

Fouchet’s test Bilirubin in urine
Ehrlich Aldehyde test Urobilinogen
Vandenberg test Differentiation of conjugated and unconjugated Bilirubin (Quantitative)
Rothera’s test Acetoacetate and Acetone (Ketone Bodies)
Gerhardt’s test Acetoacetate
Sulkowitch test Urinary Calcium

Schilling test Absorption of Vitamin B12

HOMOPHONES IN BIOCHEMISTRY
• D and L • Structural Isomers (Enantiomers)
• d and l • Optical Isomers (Dextro-rotatory and Levo-rotatory)
• Racemic Mixture • Equal d + l present (optically inactive)
• Racemase Enzyme • Enzyme which converts D and L (name Racemase is misnomer)
• Dextrin • Hydrolytic Product of Starch
• Dextran • Branched, HMW Homopolysaccharide made up of α-Glucose (used as Plasma Volume Expander)
• Dextrose • Solution of Glucose
• Lactose • Disaccharide, made up of Galactose + Glucose (Present in Milk), broken by enzyme Lactase
• Lactulose • Disaccharide, made up of Galactose + Fructose (used as Laxative), fermented by intestinal
bacteria
• Lactate • Dead end of Glycolysis, Produced in Anaerobic Glycolysis
• Lactase • Enzyme which breakdown Lactose (Disaccharide)
• Lectin • Carbohydrate Binding Proteins, which play a role in Recognition and Attachment E.g. Mannose
binding Lectin which is specific for Mannose containing Polysaccharide
• Pectin • Heteropolysaccharide, made up of Galacturonic Acid, acting as a Soluble Dietary Fibre
• Lignin • A complex organic polymer, which is the major Insoluble Dietary Fibre. It is not a Carbohydrate.
It is a Poly phenolic compound, a polymer of Phenyl propane
• Insulin • Hormone, secreted from β-Islet cells of Pancreas
• Inulin • A Homopolysaccharide, made up of β-Fructose, also Ideal substance for GFR
• Fructose 1,6 Bisphosphate • Involved in Glycolysis
• Fructose 1,6 Bisphosphatase • Involved in Gluconeogenesis
• Fructose 2,6 Bisphosphate • Reciprocal regulator which increases the rate of Glycolysis and decrease the rate of
Gluconeogenesis
• Fructose 2,6 Bisphosphatase • Present in TIGAR (TP-53 Induced Glycolysis and Apoptosis Regulator)

Contd…
 • Transferrin • Transports Iron
• Transcortin • Transports Cortisol, Aldosterone and Progesterone
414 • Transthyretin • Transports Thyroxine and Retinol Binding Protein
• Hemopexin • Transports Free Heme released from Haemoglobin
• Haemosiderin • Are storage forms of Iron
• •
CRO BIOCHEMISTRY

Hepcidin Regulates Iron transport in circulation.

COGNOMEN
Name Other Name
Sarcosine N-Methyl Glycine
Betaine Trimethyl Glycine
Choline Trimethyl Ethanolamine
Carnosine β-Alanyl Histidine


Anserine Carnosine on methylation

Homo Carnosine GABA + Histidine
Melatonin Acetyl Methyl Serotonin
Serotonin 5-OH Tryptamine
Ethanolamine Serine on Decarboxylation
β-Mercapto Ethanolamine Cysteine on Decarboxylation

MOST COMMON
1st most common Human Enzyme Deficiency Glucose 6- Phosphate Dehydrogenase (HMP pathway)
2nd most common Human Enzyme Deficiency Pyruvate Kinase (Glycolysis Pathway)
Most common SCID X- linked
Most common Gene Disorder worldwide Thalassemia
Most common Hemoglobinopathy Sickle Cell Anemia
Most common Chromosomal Disorder Down Syndrome
Most common Urea Cycle Defect OTC (Ornithine Trans-Carbamoylase) Deficiency or
Hyperammonemia Type II
Most common cause of Inherited Non-Spherocytic Haemolytic Anemia Pyruvate Kinase Deficiency
Most common Saturated Fatty Acid of cells Palmitic Acid (16-C)
Most common Plasma Membrane Receptor G-Protein Coupled Receptor (GPCR)
Most common Covalent Modification Phosphorylation and Dephosphorylation
Most common Chronic Liver Disease Non- Alcoholic Fatty Liver Disease
Most common Prosthetic group Metal
Most common Glycogen Storage Disease Von-Gierke’s Disease
Most common Clotting Factor Deficiency Factor VIII
Most common Lysosomal Storage Disease Gaucher’s disease
Most common Gene Mutated in Cancer p53
Most common Secondary Structure of Protein α-helix
Most common form of DNA B-DNA
Most common type of Mutation Transition
Most common Oncogene involved in the Development of Cancer Ras
Most common defect of Fatty Acid Oxidation MCAD deficiency
Contd…
Most common cause of preventable Blindness in children. Vitamin A deficiency
Most common DNA Binding Motif Helix turn helix 415
Most common Insoluble Dietary Fibre Cellulose
Most common Amino Acid Transport Defect Cystinuria

EXTRA EDGE TABLES

Most common Porphyria PCT (Porphyria CutaneaTarda)
Most common Hepatic Porphyria AIP (Acute Intermittent Porphyria )
Most common Porphyria in children Erythropoeitic Porphyria
Most common Enzyme Defect of Homocystinuria Cystathionine β Synthase
Most common Acceptor in Transamination α Ketoglutarate
Most common Trinucleotide Repeat CAG
Most common RNA Editing Adenosine to Inosine
Most common point of Gene Regulation in Eukaryotes Initiation of Transcription
Most common cause of Insulin Resistance Obesity
Most common cause of cobalamin deficiency Pernicious Anemia
Most common O-linked Glycoprotein in body Mucin
Most common Renal Stones in children Cystine Stones

MOST ABUNDANT
Most abundant Collagen in Bone and White Fibro Cartilage Type I
Most abundant Collagen Type I
Most abundant Collagen in Cartilage Type II
Most abundant Collagen in Basement Membrane Type IV
Most abundant Nucleoprotein Histones
Most abundant Amino Acid in Plasma Glutamine
Most abundant Free Nucleotide in Mammalian Cells ATP
Most abundant Heteropolysaccharide GAG ( Glycosaminoglycan)
Most abundant GAG Chondroitin sulfate
Most abundant Fatty Acid synthesized in the body Palmitic acid (16C)
Most abundant Saturated Fatty Acid in circulation Palmitic acid
Most abundant Ketone Body during Ketosis β-OH-Butyrate
Most abundant Membrane Proteins of RBC Glycophorin and Band 3 Anioninc Transporter
Most abundant Peripheral Membrane Protein of RBC Spectrin
Most abundant Platelet Receptors GP II b – GP III a complex
Most abundant DNA Polymerase of Prokaryotes DNA polymerase I
Most abundant of all human proteins Collagen
Most abundant form of Vitamin A α-Tocopherol
Most abundant Polysaccharide in nature Cellulose
Second Most abundant Polysaccharide in nature Chitin
Most abundant organic molecule in nature Carbohydrates
Most abundant Monosaccharide in nature D-Glucose
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Image–Based Questions
1. Which glucose transport present at points X and 3. The following graph represents effect of
Y? substrate concentration on the initial velocity
of an enzyme- catalyzed reaction. WRONG
statement about this graph is:

a. The curve is hyperbolic in shape.

b. “?” here represents Km of the enzyme.
c. At point C, only a small amount of the enzyme
is present as the Enzyme-Substrate complex.
a. X- SGLT-2, Y- GLUT-2 d. At point C, Vi is independent of [S]
b. X- GLUT-3, Y- SGLT-1
c. X- SGLT-1, Y- GLUT-2
d. X- GLUT-2, Y- SGLT-2
4. Which of the following is CORRECT about the
enzyme which has sigmoidal graph?

2. Number of isomers possible for the carbohydrate Ans.

given are: 1. c
2. c
3. c
4. b

a. 32 b. 64
c. 16 d. 8
a. Allosteric modifier binds in a concentration
dependent manner
b. Modifier can affect the catalytic site by
binding to the allosteric site
c. Adding more substrate to the enzyme can
displace the allosteric modifier
d. Allosteric modifiers change the binding
constant of the enzyme but not the velocity of
reaction
 5. This pathway is occurring in which cell of the 8. A 48 year old female presented with bony pain
body? and hepatosplenomegaly. On examination
of biopsy from spleen, crumpled tissue paper
418 appearance is seen. Which of the following
product is likely to have accumulated?
CRO BIOCHEMISTRY


a. RBCs b. Liver a. Ganglioside

c. Muscles d. Brain b. Sulfatides
c. Sphingomyelin
d. Glucocerebroside

6. This serum of patient shows increased:

9. Products of complete hydrolysis of cardiolipin

are:

a. 3 glycerol, 4 fatty acids, 2 phosphates

Ans. b. 3 glycerol, 4 fatty acids, 1 phosphates
c. 3 glycerol, 3 fatty acids, 2 phosphates
5. a
d. 5 glycerol, 4 fatty acids, 2 phosphates
6. d
7. a
a. Cholesterol b. LDL
8. d
c. VLDL d. Chylomicrons
9. a
10. b 7. Which of the following is NOT found in lipid
profile after an overnight fast? 10. In the structure of Acetyl CoA, which is thioester
bond?

a. X
b. Y
c. Z
d. All

a. Chylomicrons b. VLDL
c. LDL d. HDL
11. Which group is removed during this conversion: 14. The given structure is:
419

 IMAGE–BASED QUESTIONS
a. Phosphocholine
b. Sphingosine
c. Fatty acid
d. Phosphoethanolamine

a. Epimer of glucose at C-4

b. Enantiomer of glucose at C-4
c. Epimer of glucose at C-2
12. A fatty acid is shown. This is: d. D-Enantiomer of glucose at C-2 and C-3

a. Omega-3 category
b. A saturated fatty acid
c. Omega-6 category 15. A child with pellagra like symptoms, amino
d. Omega 9 category acids in urine, family history of two siblings
affected and two normal. Parents are normal.
What is the diagnosis?

Ans.
11. a
13. The given structure is: 12. a
13. b
14. c
15. d

a. Phenylketonuria
b. Alkaptonuria
a. Cephalin b. Lecithin
c. Maple syrup urine disease
c. Sphingomyelin d. Glucocerebroside
d. Hartnup’s disease
 16. A 12 year old girl is brought to emergency by her 18. A 3-month-old infant presents with hepa-
420 parents with complaints of severe polyuria and tospleno- megaly and failure to thrive. A liver
polydipsia. Laboratory examination reveals a biopsy reveals glycogen with an abnormal,
purple ring when a test was done in her urine. amylopectin like structure with long outer
Which of the following is the most likely source chains and missing branches. Which of the
CRO BIOCHEMISTRY

of this compound which is positive in this following enzymes would most likely be
patient? deficient?


a. Alpha Amylase
a. Protein breakdown
b. Branching enzyme
b. Fatty acid breakdown
c. Debranching enzyme
c. Gluconeogenesis
d. Glucose-6-phosphatase
d. Side chain of cholesterol

19. Identify the given structure:

17. A breast-fed infant began to vomit frequently

and lose weight. Several days later she
Ans. developed jaundice, hepatomegaly. Bilateral
cataract found in patient is shown. What is the
16. b
possible cause for these symptoms?
17. a
18. b a. Glycerol
19. b b. D-Glyceraldehyde
20. a c. L- Glyceraldehyde
d. Dihydroxyacetone

20. Dextrose is:

a. Galactosemia
b. Juvenile Diabetes Mellitus
c. Hereditary Fructose Intolerance
d. Gaucher disease
a. D + glucose b. D – glucose
c. L + glucose d. L – glucose
21. In which phase of cell cycle, proofreading 24. The given structure of protein is:
occurs?

421

 IMAGE–BASED QUESTIONS
a. Primary
b. Secondary
c. Tertiary
a. G1 b. S d. Quaternary
c. G2 d. M

22. Which group is removed in the following 25. Enzyme of transcription is:
conversion?

a. Amino group b. Carboxy group

c. Sulphate group d. Ester group a. DNA dependent DNA Polymerase
b. Reverse transcriptase
c. DNA polymerase Ans.
d. DNA dependent RNA Polymerase 21. b
22. a
23. d
23. What is the name of the given cycle? 24. d
25. d
26. b
26. Tell the bond in the given structure of a disac-
charide:

a. Alpha 1 → 4
b. Beta 1 → 4
c. Beta 1 → 2
a. Cori’s cycle b. Carnitine shuttle d. Alpha 1 → 2
c. Alanine cycle d. Cahill cycle
 27. Which enzyme synthesize Okazaki fragments?

422
CRO BIOCHEMISTRY


a. DNA Polymerase III

b. DNA polymerase I
c. DNA primase
d. DNA Ligase

28. How many ATPs are used when glucose enters 29. Which isoenzyme has maximum electrophoretic
in this diagram? mobility?

Ans.
27. a
28. d a. LDH-1 b. LDH-5
29. a c. LDH-2 d. LDH-3
30. d

30. These osazones belong to:

a. 1 b. 2
c. 3 d. Zero

a. Glucose b. Fructose
c. Mannose d. All
31. A 38 year old female patient was on isoniazid 32. The following picture show which mechanism of
therapy for tuberculosis. She developed rashes transport across the membrane?
on the exposed parts of her body. She has
disoriented memory. Family members gives 423
history of diarrhoea also. What is the diagnosis?

 IMAGE–BASED QUESTIONS
a. Ping pong b. Symport
c. Uniport d. Antiport

33. What is the enzyme involved in the following

a. Tuberculosis skin lesions conversion?
b. Niacin deficiency
c. Isoniazid neuropathy
d. Some other drug has caused this

a. Sorbitol dehydrogenase
b. Glucose reductase
c. Aldose reductase
d. Glucose oxidase Ans.
31. b
32. a
34. Which is coding strand in the given figure?
33. c
34. a

a. DNA Sense strand b. DNA antisense strand

c. RNA strand d. Both DNA sense and antisense strand
 35. Structure of DNA is shown. What are the bonds 37. Which is the parent nucleotide which synthesize
424 between 2 nitrogenous bases? both AMP and GMP?
 (AIIMS Nov 2017)
CRO BIOCHEMISTRY


a. Hypoxanthine
a. 3’ 5’ phosphodiester bonds b. Inosine
b. 2’5’ phosphodiester bonds c. IMP (Inosine mono phosphate)
c. Hydrogen bonds d. OMP (orotidine mono phosphate)
d. Covalent bonds

Ans.
35. c 36. Following are the tests done for proteins, sugar 38. Sample from the area given in picture can be
36. c and ketones. Which of the following will be used to know:
37. c positive in starvation state in urine?
38. b  (AIIMS Nov 2017)

1.   2.   3.

a. Blood sugar
b. Inborn errors of metabolism
a. 1 and 2 b. Only 2
c. Hepatitis
c. Only 3 d. 2 and 3
d. Cataract
39. Which amino acid present at this point of a 42. The given is the structure of protein. Which
protein? bond is responsible for this structure?

425

 IMAGE–BASED QUESTIONS
a. Glycine b. Lysine
c. Methionine d. Glutamate

40. What is your diagnosis from the following

picture: a. Disulfide bond
b. Hydrophobic bond
c. Hydrogen bond
d. Covalent bond

43. Best method to detect the given structure of

protein is:

Ans.
a. Magnesium deficiency 39. a
b. Selenium deficiency 40. b
c. Iron deficiency 41. c
d. Beta oxidation defect 42. d
43. c

41. Given is the structure of which amino acid ?

a. Mass spectrometry
b. Edman’s technique
c. X-ray crystallography
a. Tryptophan b. Threonine d. NMR spectrometry
c. Tyrosine d. Phenylalanine
 44. A patient presented with muscle weakness, 46. Bond shown in the picture:
426 sleep disturbance, constipation and fatigue.
On examination, blue lines were seen in gums
(shown in picture). By occupation, he is a
battery recycling worker. Which enzyme is
CRO BIOCHEMISTRY

increased in this patient?



a. Has partial double bond character

a. ALA synthase b. It is a non-covalent bond
b. Porphobilinogen deaminase c. Occurs in cis configuration
c. Ferrochelatase d. Is a single bond
d. ALA dehydratase

45. Which protein has this primary sequence? 47. Which type of column chromatography is this?

Ans.
44. a
45. c
46. a
47. b

a. Elastin a. Affinity chromatography

b. Haem b. Size exclusion chromatography
c. Collagen c. HPLC
d. Albumin d. Ion exchange chromatography
48. Identify the given compound?
427

 IMAGE–BASED QUESTIONS
a. Uric acid
b. Guanine
c. Thymine
d. Uracil

49. In the given chromatography, the negative charged compound used is:

Ans.
48. c
49. b

a. Sephadex
b. CMC (Carboxy methyl Cellulose)
c. DEAE cellulose (Di Ethyl Amino Ethyl cellulose)
d. Methyl cellulose media
 50. Which bond is present in the given structure of a 52. Low purine diet is:
protein?

428
CRO BIOCHEMISTRY

a. b.

c. d.


a. Hydrogen
b. Disulphide bond
c. Electrostatic
d. All 53. When this kind of nucleotide is added in PCR,
what will happen?

51. End product of purine catabolism is:

a. DNA synthesis stops
b. Fluorescence occurs
c. DNA synthesis is faster
Ans. d. DNA synthesis is slower
50. d
51. a
52. c (a)    (b) 54. All of the following can occur in the given
53. a organelle EXCEPT:
54. b

(c)    (d)

a. Protein synthesis
b. Fatty acid synthesis
c. Beta oxidation of fatty acids
d. DNA synthesis
55. A girl presented with tingling sensation in legs 57. Type of isomers shown are:
and hands. On examination, she had fissure
tongue and lesions in angle of mouth. On
investigation, she had low RBC glutathione 429
reductase activity. Diagnosis is deficiency of?

 IMAGE–BASED QUESTIONS
a. Anomerism b. Enantiomerism
c. Epimerism d. Diastereomers

58. What is the structure indicated below?

a. Vitamin B2
b. Vitamin B6
c. Vitamin B12
d. Vitamin B1

56. This image shows the results of which technique

of molecular biology?
a. Guanosine b. Adenosine
c. Pyrimidine nucleoside d. Purine nucleoside
Ans.
55. a
59. Given is the structure of glycogen. What is the 56. b
linkage at point ‘a’, ‘b’ and ‘c’ respectively? 57. b
58. b
59. c

a. Microarray a. α1 → 6, β 1 → 2 and α1 → 4
b. Karyotyping b. α1 → 6, β 1 → 6 and α1 → 6
c. RFLP c. α1 → 4, α 1 → 6 and α1 → 4
d. ChIP d. α1 → 4, α 1 → 6 and α1 → 6
 60. Which of the following is correct regarding the location of codon and anticodon?

430
CRO BIOCHEMISTRY

a. b.


c. d.
Ans.
60. c
61. a

61. Which of the following is CG site in DNA?

62. The given picture shows which technique of molecular biology?

431

 IMAGE–BASED QUESTIONS
a. ChIP b. Microarray
c. RNA interference d. RFLP

63. Which of the following is correct regarding the Lactose Operon in E.coli?

a. b.

c. d.
Ans.
62. b
63. d
64. c
65. a

64. Which mutation is shown: 65. Which mutation is shown:

a. Missense mutation
a. Missense mutation b. Non sense mutation
b. Non sense mutation c. Silent mutation
c. Silent mutation d. Frameshift mutation
d. Frameshift mutation
 66. Tell the enzymes involved in these two reaction: 68. Which is the enzyme involved in the following
432 reaction:
CRO BIOCHEMISTRY

a. LCAT (Lecithin cholesterol acyl transferase)

b. ACAT (Acyl CoA cholesterol acyl transferase)
a. X is Pyruvate carboxylase, Y is Lactate c. Lipoprotein lipase
dehydrogenase d. HDL acyl transferase
b. X is Pyruvate Dehydrogenase, Y is Lactate
dehydrogenase


c. X is Lactate dehydrogenase, Y is Pyruvate

69. What is the ligand for the following receptor?
Dehydrogenase
d. X is phospho-enol-Pyruvate carboxy kinase, Y
is Lactate dehydrogenase

67. Tell the bonds (X,Y,Z) in the structure of DNA?

a. Apo B 100 b. Apo E

c. Both a and b d. Apo B 48

Ans.
66. b
67. a 70. Identify the isomerism:
68. a
69. c
70. b

a. X- Hydrogen bond, Y- beta N glycosidic bond,

Z- 3’ 5’ phospho diester bond
b. X- Hydrogen bond, Y- beta O glycosidic bond,
Z- 2’ 5’ phospho diester bond
c. X- Hydrogen bond, Y- beta N glycosidic bond, a. Enantiomerism
Z- 2’ 3’ phospho diester bond b. Optical isomerism
d. X- Hydrogen bond, Y- phospho diester bond, c. Epimerism
Z- 3’ 5’ beta N glycosidic bond d. Mirror images
71. See the image and tell which complexes are 74. Type of collagen present in the following organ:
involved when FADH2 enters ETC:

433

 IMAGE–BASED QUESTIONS
a. Complex I, II, III, IV
b. Complex II and III a. Type I b. Type II
c. Complex II, III and IV c. Type VII d. Type V
d. Complex I, III and IV

72. Which form of glucose transport is present in 75. Recognize the protein shown in the picture. This
the given organ? is also the most abundant of all human proteins.
It has many types. Which type of this protein is
most abundant?

a. GLUT 1 b. GLUT 2 Ans.

c. GLUT 3 d. GLUT 5 a. Type I b. Type II
c. Type VII d. Type V 71. c
72. b
73. c
73. Which of the following enzyme is present inside 74. a
the given organelle? 75. a
76. Identify the transport mechanism marked with
76. b
arrow:

a. Glucokinase
b. Isocitrate dehydrogenase
c. Glucose-6- phosphatase a. Antiport b. Symport
d. Fructose 2, 6 bisphosphatase c. Uniport d. Endocytosis
 77. Identify the following transport mechanism: 79. Identify the given disaccharide. It is present in:
434
CRO BIOCHEMISTRY


a. Spinach b. Milk
c. Table sugar d. Germinating grain
a. Endocytosis
b. Exocytosis
c. Phagocytosis
d. Symport 80. Identify the given disaccharide. It is present in:

78. Identify the kind of channel. This channel is

associated with which of the following transport:

a. Spinach b. Milk
c. Table sugar d. honey
Ans.
77. b
78. d
79. b 81. What is the rate limiting step in the biosynthesis
80. d of following compound?
81. d

a. GLUT-1
b. GLUT-4
c. GLUT- 7
d. SGLT-1
a. HMG CoA synthase
b. Thiolase
c. Thiophorase
d. HMG CoA Reductase
82. What is the rate limiting step in the synthesis of 85. Identify the given structure. The portion marked
the following compound? in box is synthesized by liver so it is: 435

 IMAGE–BASED QUESTIONS
a. HMG CoA synthase
b. Thiolase
c. Thiophorase
d. HMG CoA Reductase

a. Apo D
b. Apo B 100
c. Apo B 48
d. Apo B 50
83. These are five lipoproteins shown. What is ‘A’?

86. Which of the following is involved in the

conversion of these amino acids

a. HDL
b. LDL Ans.
c. VLDL
82. a
d. Chylomicron a. Ammonia 83. d
b. THF 84. a
c. Vit B12 85. b
d. Vit B1 86. b
87. a
84. These are five lipoproteins shown. Which has
maximum percentage of proteins?
87. Which group is added in the following
conversion?

a. 5 a. Methyl
b. 2 b. Amino
c. 3 c. Carboxy
d. 4 d. Phosphate
 88. Which group is added in the following 91. Identify the condition. This occurs in which
436 conversion? vitamin deficiency?
CRO BIOCHEMISTRY

a. Methyl
b. Amino
c. Carboxy
d. Phosphate

a. Vitamin A b. Vitamin C


c. Vitamin D d. Vitamin E
89. Transport shown in picture is done by which
amino acid:
92. Identify the condition. This occurs in which
vitamin deficiency?

a. Glutamate
b. Alanine
c. Glutamine
d. Glycine

a. Vitamin A b. Vitamin B2
c. Vitamin D d. Vitamin B12
Ans.
88. b 90. Identify the given condition. This is due to
89. c deficiency of which vitamin?
90. a 93. Parents gives history with these (shown in
91. c image) self inflicted injuries in the child. The
92. b child also has joint pains. Which of the following
93. a enzyme is most likely to be deficient?

a. Vitamin A
b. Vitamin C
c. Vitamin D a. HGPRT b. APRT
d. Vitamin E c. LCAT d. ACAT
94. Which of the following test tube shows the 97. Amino acid ‘X’ shown in the Reaction (Photo-
presence of sugar in the sample: graph) is 437

 IMAGE–BASED QUESTIONS
a. Alanine
b. Aspartate
c. Phenylalanine
d. Arginine

a. 1 b. 2
c. Both d. None
98. Enzyme catalyzing reaction shown in the Photo-
graph is:
95. A patient’s urine turned black on exposure to
air. Identify the enzyme which is deficient.

a. Phenylalanine hydroxylase
b. Tyrosine hydroxylase
c. DOPA decarboxylase
d. Transaminase

Ans.
a. Homogentisate oxidase 94. b
b. Homogentisate dioxygenase 95. b
c. Phenyl alanine hydroxylase 99. This picture shows an intermediate of Histidine
96. b
d. Phenyl alanine oxidase metabolism which is excreted in deficiency of
97. d
98. a
99. d
96. Enzyme involved in this conversion is

a. Melanin dehydrogenase a. Thiamine

b. Tyrosinase b. Riboflavin
c. Phenyl alanine hydroxylase c. Biotin
d. Tyrosine dehydrogenase d. Folic acid
 100. A person suffers from obstructive jaundice. 102. The given figure demonstrates the structure of
Hay's Sulphur test was performed. The test was an amino acid which has Pyrrolidine ring and is
positive as sulphur powder sinked to the bottom not found in a-helix. Identify the amino acid?
438 as shown in figure. This test is positive for:
CRO BIOCHEMISTRY

a. Histidine b. Proline
c. Tryptophan d. Phenylalanine


103. Identify the following structure ?

a. Cholesterol
b. Bile Salts
c. Bile pigments
d. Sulfur containing amino acids

a. Adenine
101. The given figure represents tRNA. Base triplet at b. Guanine
3'OH end required for the attachment of amino c. Xanthine
acid on the tRNA is: d. Hypoxanthine

Ans.
104. The enzymes in the given figure (marked with
100. b
red arrows) uses:
101. a
102. b
103. b
104. a

a. NADPH
b. NADH
c. FADH2
a. CCA b. CAC
d. NADP
c. ACA d. AAC
105. The following picture depicts epidermolysis 107. The amino acid in the given figure has maximum
bullosa. Which type of collagen is defective in buffering capacity. Identify the amino acid
this disease?
439

 IMAGE–BASED QUESTIONS
a. Histidine b. Arginine
c. Proline d. Tryptophan

108. The figure below shows the base sequence on

template & non-template strand of DNA. What
is the base sequence of RNA?

a. Type I
b. Type VI
c. Type VII
d. Type IV

106. The figure represents disease due to absence

of pigment Melanin from skin, hair and eyes.
Identify the disease.
a. CGGUAAAGCUUA b. AUUCGAAAUGGC
c. GCCATTTCGAAT d. UAAGCUUUACCG
Ans.
105. c
109. The synthesis of creatinine involves following 106. b
reactions. The site of reactions 1, 2, 3 are? 107. a
108. a
109. b

a. Phenylketonuria a. Liver, Kidney, Muscle

b. Albinism b. Kidney, Liver, Muscle
c. Alkaptonuria c. Liver, Liver, Muscle
d. Vitilgo d. Muscle, Liver, Kidney
 110. A 9 year old patient has bluish discoloration of 111. A 20 year old boy with severe mental retar-
both scleras. Careful inquiry reveals that they dation, mousy odour in body fluids, hypo-
have occasionally noticed very dark urine on pigmentation. Patient has frequent episodes of
440 the periphery of their asian style toilet. Which seizures and aggressive behaviour. What is your
amino acid pathway is implicated in this diagnosis?
disease?
CRO BIOCHEMISTRY


a. Tyrosine
b. Tryptophan
c. Histidine
d. Glycine
a. Phenylketonuria
b. Albinism
c. MSUD
d. Untreated PKU (Phenylketonuria)

112. Which abnormality is shown in the figure by FISH ?

Ans.
110. a
111. d
112. c

a. Deletion
b. Inversion
c. Translocation
d. Insertion
113. Identify the following techniques of molecular biology:

441

 IMAGE–BASED QUESTIONS
(I) (II)

(III) (IV)

a. (I) Microarray, (II) FISH showing microdeletion, (III) PCR, (IV) RFLP
b. (I) FISH showing microdeletion, II) Microarray, (III) RFLP, (IV) PCR
c. (I) RFLP, (II) Microarray, (III) FISH showing microdeletion, (IV) PCR
Ans.
d. (I) Microarray, (II) RFLP, (III) PCR, (IV) FISH showing microdeletion
113. a
114. c

114. A tall and slim girl got operated for dislocated lens. After careful history from ophthalmologist, it came out that
her sister also had similar operation. Cyanide nitroprusside test was positive in her urine sample. What is the
diagnosis?

a. Albinism
b. Pheochromocytoma
c. Homocystinuria
d. Rickets
 115. A 4 year old girl presented to hospital with 116. A 48 year old male presented to clinic with chest
failure to thrive. Her PBF showed megaloblastic pain. His ECG was normal. Careful examination
anemia. Vitamin B12 & folate were given but showed tendon xanthomas on his lower limbs.
442 anemia did not improve. Enzyme measurement His father & elder brother died of heart attack.
in leucocytes showed deficiency of OPRT His blood cholesterol was 350 mg/dl & TGs were
(Orotate Phospho Ribosyl Transferase). What is normal. What is the diagnosis?
the diagnosis ?
CRO BIOCHEMISTRY


a. Familial hypercholesterolemia
b. Familial hyperchylomicronemia
a. Orotate deficiency b. Gout
c. Wolman’s disease
c. Orotic aciduria d. SCID
d. Dysbeta-lipoproteinemia

117. A 12 year old child was brought to OPD with defective vision. He had constant dribbling thick mucous from
mouth, coarse facial features, mental retardation. Skeletal deformities appeared after 1 year and child was in
vegetative state by age 15. Complete urine analysis showed heparan sulfate and dermatan sulfate. What is the
diagnosis?

Ans.
115. c
116. b
117. a

a. Hurler disease b. Hunter disaese

c. MCAD deficiency d. Wolman’s disease
 Answers with Explanations
443
1. Ans. (c) X- SGLT-1, Y- GLUT-2 This is Michaelis Menton graph (rectangular hyperbola).
[Ref: Harper 30th/e p. 191] Km is Michaelis Menton constant. Km is that substrate

 IMAGE–BASED QUESTIONS
concentration at which velocity of reaction is half of
In intestine, both facilitative transport (GLUT) and
Vmax. Here '?' point represents Km (on X-axis). At
secondary active (SGLT) transport is present. On apical
point C, maximum amount of enzyme is present as
side of intestinal cells (towards the lumen), SGLT-1 is
E-S complex. Initial portion of this graph has first order
present i.e. sodium dependent glucose transport, and
kinetics i.e. velocity is directly proportional to substrate
at the basolateral membrane, facilitative transport i.e.
concentration and later portion of the graph has zero
GLUT-2 is present.
order kinetics i.e. velocity is independent of substrate
concentration.

4. Ans. (b) Modifier can affect the catalytic site by

binding to the allosteric site
[Ref: Harper 30th/e p. 80]
Sigmoidal graph is for allosteric enzymes. They have
active site where substrate binds. But they also have
allosteric site, where allosteric modulator binds.
Allosteric modulators can be activators or inhibitors.
They bind at allosteric/regulatory site and they induce
changes in the active site, where substrate binds and they
modulate the binding of substrate. Allosteric modifiers
can change the binding constant of the enzyme and
thus the velocity of reaction. Allosteric modifier binding
is not concentration dependent. Adding more substrate
to the enzyme cannot displace the allosteric modifier.

2. Ans. (c) 16 5. Ans. (a) RBCs

[Ref: Harper 30th/e p. 153] [Ref: Harper 30th/e p. 172]
The structure shown here is linear structure of glucose. This is RL shunt i.e. Rapoport Leubering shunt/ cycle.
C1 and C6 of glucose are symmetric carbons. C2 to C5 This occurs only in RBCs, for the production of 2,3
are asymmetric carbons. Number of isomers possible bisphosphoglycerate. This compound is required in
RBCs to release oxygen from HbA at tissue level. In RL A
for any compound is given by the formula where 2n,
shunt, substrate level phosphorylation step by phospho N
n = number of asymmetric carbons. In case of glucose,
number of asymmetric carbons are 4. So total number glycerate kinase enzyme does not occur. Net gain of ATP S
of isomers possible are 16. in RL shunt is zero. W
E
6. Ans. (d) Chylomicrons R
[Ref: Harper 30th/e p. 257] S
The sample taken from patient shows milky plasma, WITH
which is characteristic of increased chylomicrons in
blood. Chylomicrons contains TG (triglycerides). E
X
7. Ans. (a) Chylomicrons P
[Ref: Harper 30th/e p. 257] L
Chylomicrons are formed by intestinal cells after A
taking a lipid rich meal. Chylomicrons are removed N
from circulation within 2 hours of food intake. So after A
3. Ans. (c) At point C only a small amount of the enzyme overnight fast, chylomicrons are not present in blood. T
is present as the Enzyme-Substrate complex I
O
[Ref: Harper 30th/e p. 79]
N
S
 8. Ans. (d) Glucocerebroside 13. Ans. (b) Lecithin

444 [Ref: Harper 30th/e p. 245] [Ref: Harper 30th/e p. 245]

This is Gaucher’s disease (most common lysosomal This structure is lecithin (Glycerol + 2 Fatty acids +
storage disease). Glucocerebrosides accumulate Phosphocholine). Lecithin is a phospholipid which acts
in reticuloendothelial system. Macrophages have as a surfactant in the lungs.
CRO BIOCHEMISTRY

typical crumpled tissue paper appearance/ wrinkled

appearance. These patients have bony pain, patho- 14. Ans. (c) Epimer of glucose at C-2
logical fractures, hepatosplenomegaly, but no mental [Ref: Harper 30th/e p. 154]
retardation. This structure shown is Mannose.
Epimerism is different H and OH orientation around
9. Ans. (a) 3 glycerol, 4 fatty acids, 2 phosphates
only one carbon, other than the penultimate carbon.
[Ref: Harper 30th/e p. 217] Mannose is epimer of glucose at C2 and Galactose is
Cardiolipin is a complex phospholipid, which can be epimer of glucose at C4.
antigenic. The structure is glycerol attached to two
phosphatidic acid. Each phosphatidic acid contains one


glycerol, 2 FA and one phosphate. So in total 3 glycerol,

4 fatty acids, 2 phosphates are present in cardiolipin.

10. Ans. (b) Y

[Ref: Harper 30th/e p. 162]
The bond between C and S is known as thioester bond.
Ester bond is – COOR and Thioester is –COSR.

15. Ans. (d) Hartnup’s disease

[Ref: Harper 30th/e p. 304]
The diagnosis is Hartnup's disease. It is autosomal
recessive. There is failure to reabsorb tryptophan from
urine. So tryp-tophan comes in urine (Aminoaciduria).
Patient has Pellagra like symptoms (as Tryptophan
A 11. Ans. (a) Phosphocholine forms Niacin – Vitamin B3 in body).
N
[Ref: Harper 30th/e p. 251] 16. Ans. (b) Fatty acid break down
S
W Sphingomyelin is made up of alcohol – sphingosine,
[Ref: Harper 30th/e p. 288]
one fatty acid and phosphocholine. Sphingosine and
E The test shown in figure is Rothera’s test in which a
fatty acid together join to form ceramide. Enzyme
R purple ring is formed at the top, if ketone bodies are
sphingomyelinase removes phosphocholine from
S sphingomyelin to form ceramide. positive.
Fatty acid breakdown provides Acetyl CoA that serves
WITH
12. Ans. (a) Omega – 3 category as a precursor for ketone bodies. In Diabetes Mellitus,
E glucose utilization is impaired due to absolute or
[Ref: Harper 30th/e p. 242]
X relative insulin deficiency.
Starting from methyl end of fatty acid, count the double Fatty acid breakdown occurs to provide energy and the
P bond is at which number. It is at omega 3. So this fatty
L resultant excessive Acetyl CoA enters the pathway of
acid belongs to omega-3 category. ketogenesis.
A
N
A 17. Ans. (a) Galactosemia
T [Ref: Harper 30th/e p. 205]
I The cataract shown is oil drop cataract, which is
O characteristic of galactosemia.
N
S
 These clinical manifestations of liver affected when 22. Ans. (a) amino group
milk is the diet, are typical of classical galactosemia.
Bilateral cataract rules out the possibility of Von Gierke’s
[Ref: Harper 30th/e p. 165] 445
disease and hereditary fructose intolerance although Alpha-keto glutarate forms glutamate on addition
other symptoms are there in both these diseases. In of amino group. Glutamate further forms glutamine
juvenile diabetes mellitus, jaundice and hepatomegaly on addition of amino group. So if we have to convert

 IMAGE–BASED QUESTIONS
are not observed. In Gaucher disease, hepatomegaly is glutamine to glutamate, then amino group is removed.
observed but cataract is never there
23. Ans. (d) Cahill cycle
18. Ans. (b) Branching enzyme [Ref: Harper 30th/e p. 383]
[Ref: Harper 30th/e p. 177] •• This is glucose-alanine cycle shown, also known
During glycogenesis, branching enzyme creates as Cahill cycle. Alanine is formed from glucose in
branch points and further elongation is carried out muscles during fasting conditions. This alanine
by Glycogen synthase. In the deficiency of branching comes out in blood, then taken up by liver and in liver
enzyme stored glycogen is abnormal, in the form of it is converted back to glucose by a pathway known as
long polysaccharide chains with few branch points, gluconeogenesis. Then this glucose is again supplied
resembling the structure of Amylopectin, thus this to the fasting muscle. So, a cycle is formed known as
defect is also called Amylopectinosis. Cahill cycle.
Alpha Amylase is an enzyme for digestion of starch and •• Also alanine is the most glucogenic amino acid
glycogen. (Option a) •• Cori’s cycle is glucose lactate cycle, which occurs in
In Debranching enzyme deficiency, Limit dextrins are exercising muscles.
accumulated and thus it is also called Limit dextrinosis.
24. Ans. (d) Quaternary
(Option c)
Glucose-6-phosphatase deficiency is observed in Von- [Ref: Harper 30th/e p. 39]
Gierke’s diseases, (type 1 glycogen storage disease), In the given structure, you can see four different fully
the stored glycogen is always normal in chemistry. folded polypeptide chains are present. So it is quaternary
(Option d) structure. If single fully folded chain is present, then it is
tertiary structure.
19. Ans. (b) D-Glyceraldehyde
[Ref: Harper 30th/e p. 154] 25. Ans. (d) DNA dependent RNA Polymerase
In this structure, there is aldehyde present at C1. At C2 [Ref: Harper 30th/e p. 395]
(second last carbon here), H is on left side and -OH is •• Enzyme of transcription is DNA dependent RNA
on right side. OH on right means it is D –form. So it is Polymerase as the template is DNA and RNA
D-Glyceraldehyde. synthesis occurs on it.
In L-Glyceraldehyde, -OH is on left side of C2. Dihydroxy •• Enzyme of replication is DNA dependent DNA A
acetone is a 3C keto form. Glycerol has –OH group Polymerase as the template is also DNA and DNA N
present at all carbons. synthesis occurs on it. S
•• Enzyme of reverse transcription is RNA dependent W
20. Ans. (a) D + glucose DNA Polymerase or reverse transcriptase as the E
[Ref: Harper 30th/e p. 786] template is RNA and DNA synthesis occurs on it. R
Solution of glucose is known as dextrose. ‘D’ means S
26. Ans. (b) Beta 1→ 4
structural isomer i.e. Enantiomer. Abundant form of a
carbohydrate is always D- form. (+) means dextrorotatory [Ref: Harper 30th/e p. 157] WITH
(d). Glucose is always dextrorotatory (d)(+). The given structure is of lactose. Lactose is made up of
E
galactose + glucose. In galactose, at C4 –OH is upwards
21. Ans. (b) S Phase X
but H and –OH orientation at C4 is opposite in glucose.
Rest all carbons are same for galactose and glucose. Now P
[Ref: Harper 30th/e p. 381] L
look at the bond- C1 of galactose is making a bond with
Proofreading occurs in S phase of cell cycle. Most A
C4 of glucose. At C1 of galactose, H is downward (OH
repairs occur in G1 phase of cell cycle. But mismatch N
upwards), so it is beta position. So the bond is beta 1→ 4.
repair occurs in G2 phase of cell cycle. A
T
I
O
N
S
 Always number ‘1’ is having maximum electrophoretic
mobility. LDH-1 and CK-1 have maximum electropho-
446 retic mobility
CRO BIOCHEMISTRY

27. Ans. (a) DNA Polymerase III

[Ref: Harper 30th/e p. 383]
DNA Polymerase III synthesizes both leading and


lagging strands (Okazaki fragments). DNA polymerase

I removes RNA primers and replace it with DNA. DNA
Primase synthesize RNA primers. DNA ligase join the
fragments.
28. Ans. (d) Zero
[Ref: Harper 30th/e p. 538]
This is Na-glucose symport shown. SGLT-1 (Sodium
dependent glucose transort) is present in intestine.
This transport of glucose is against the concentration
gradient. But when glucose enters then ATPs used are
zero here. Because ATPs are not used directly here. ATPs
are used indirectly by Na-K ATPase pump present in the 30. Ans. (d) All
basolateral membrane.
[Ref: Harper 30th/e p. 157]
All these three monosaccharides i.e. glucose, fructose
and mannose have same shape of their osazone crystals
i.e. needle shaped or broom stick appearance. Non
reducing sugars do not show any osazones.
A 31. Ans. (b) Niacin deficiency
N
[Ref: Harper 30th/e p. 556]
S
W Isoniazid inhibits the endogenous synthesis of niacin.
E So the patient has developed pellagra i.e. dermatitis,
dementia and diarrhoea.
R
Prolonged treatment with isoniazid leads to pyridoxine
S deficiency, which is required for endogenous synthesis
WITH of niacin.

E 32. Ans. (a) Ping pong mechanism

X [Ref: Harper 30th/e p. 191]
P
This is ping pong mechanism of transport shown.
L
In Ping state, carrier is loaded from the extracellular
A
environment. Then conformation change occur. In
N
Pong state, carrier is unloaded towards other side of the
A
membrane. This way glucose is transported down the
T 29. Ans. (a) LDH-1
concentration gradient. This occurs in case of facilitative
I [Ref: Harper 30th/e p. 69] transport for glucose i.e. GLUTs.
O
N
S
 447

 IMAGE–BASED QUESTIONS
Rate at which glucose (solute) enters a cell via GLUT (Facilitative transport) depends on:
1. Number of GLUTs (most important)
2. Concentration of glucose
3. Affinity of GLUT for glucose

33. Ans. (c) Aldose reductase Test tube number 1 is showing protein precipitation
test. Test tube number 2 is showing benedict’s test
[Ref: Harper 30th/e p. 380] which is performed for sugar. Test tube number 3 shows
Monosaccharides gets converted to their respective Rothera’s test (purple ring at the top), which is a test for
alcohols on reduction. Glucose gets converted to its ketone bodies.
alcohol i.e. Sorbitol. Galactose gets converted to its
alcohol i.e. galactitol. In both cases, a common enzyme 37. Ans. (c) IMP (Inosine mono phosphate)
is used i.e. Aldose reductase. [Ref: Harper 30th/e p. 248]
34. Ans. (a) DNA Sense strand IMP i.e. Inosine mono phosphate is the parent
nucleotide which synthesize both AMP (Adenosine
[Ref: Harper 30th/e p. 205] A
mono phosphate) and GMP (Guanosine mono
N
In case of transcription, sense strand is known as coding phospahte). Also, IMP is the first purine nucleotide to
strand or non-template strand. The strand where gene S
be formed.
is present is always the template / antisense or non W
coding strand. The direction as well as codons of new 38. Ans. (b) Inborn errors of metabolism E
RNA are matching with sense strand. R
[Ref: Harper 30th/e p. 596]
S
35. Ans. (c) Hydrogen bonds The sample in this picture is taken from heel of the
newborn. This sample can be dried and can be used WITH
[Ref: Harper 30th/e p. 361] for the measurement of more than 50 analytes to detect
In case of DNA, the two nitrogenous bases are joined Inborn errors of metabolism. Method used is mass E
by hydrogen bonds. In case of A and T, two hydrogen spectrometry. X
bonds are present. In case of G and C, three hydrogen P
bonds are present. 39. Ans. (a) Glycine L
[Ref: Harper 30th/e p. 165]
A
36. Ans. (c) Only 3 N
This is a beta bend/turn shown in the picture. Two A
[Ref: Rafi’s Practical pg.4] amino acids are frequently found in beta turn, these are T
In case of starvation, adipose tissue is broken down to glycine and proline. I
release fatty acids. These fatty acids release Acetyl CoA,
40. Ans. (b) Selenium deficiency O
which is used to synthesize ketone bodies. So, ketone
N
bodies are raised during starvation. [Ref: Harper 30th/e p. 286] S
 This is Keshan cardiomyopathy. This is an endemic 46. Ans. (a) Has partial double bond character
disease found in areas where the soil is deficient in
448 selenium. It was named after a place named ‘keshan’
[Ref: Harper 30th/e p. 37]
in China. This leads to cardiomyopathy and pulmonary This is amide/ peptide bond shown in between two
edema. amino acids. General name of this bond is amide bond.
If amide bond is present in proteins, then it is known as
CRO BIOCHEMISTRY

41. Ans. (c) Tyrosine peptide bond. This amide or peptide is a covalent bond.
[Ref: Harper 30th/e p. 326] It has partial double bond character and the double
bond is in trans configuration.
The given amino acid is tyrosine (–OH group attached
to phenylalanine). 47. Ans. (b) Size exclusion chromatography
[Ref: Harper 30th/e p. 26-27]
This is size exclusion or gel filtration chromatography.
In this method, a gel is taken in the column. The size
of gel is such that smaller protein can enter the gel but
bigger protein cannot. The mixture of proteins to be


separated is poured on the top of the column. Both

proteins start coming down with gravity. But smaller
protein takes time to come down as it enters the gel.
But bigger protein is eluted first. This way, separation of
proteins can be done.

42. Ans. (d) Covalent bond

[Ref: Harper 30th/e p. 22]
This is primary structure of protein shown i.e. sequence
of amino acids. In primary structure, bond is present
between the amino acids. This bond is amide/ peptide
bond. General name of this bond is amide bond. If
amide bond is present in proteins, then it is known as
peptide bond. This amide or peptide is a covalent bond.

43. Ans. (c) X-ray crystallography

[Ref: Harper 30th/e p. 41]
This is alpha helix shown i.e. secondary structure of
A protein. Best method to detect secondary, tertiary and
N quaternary structure of protein is X-ray crystallography.
S In this technique, protein is first crystallized and then 48. Ans. (c) Thymine
W X-ray beam is thrown over it and then 3 D structure can
[Ref: Harper 30th/e p. 342]
E be determined.
This is a nitrogenous base. Only one ring present means
R
44. Ans. (a) ALA synthase it is a pyrimidine, not purine. Methyl group present,
S which is characteristic of thymine.
[Ref: Harper 30th/e p. 326]
WITH This is a case of lead poisoning. During battery recycling, 49. Ans. (b) CMC (Carboxy Methyl Cellulose)
E workers have to use molten lead. These blue lines (also
[Ref: Harper 30th/e p. 435]
X known as burton’s lines) in gums are characteristic of
lead poisoning. Lead poisoning is the most common This is ion exchange chromatography shown. In this
P either positive charge can be used on the column or
L cause of acquired porphyria. In porphyria, the rate
limiting enzyme – ALA Synthase is increased. (lead negative charge. Carboxy always gives negative charge
A and amino group (NH2) always gives positive charge.
N affects two enzymes in haem synthesis pathway- ALA
Because carboxy -COOH ionize to give COO– and
A dehydratase and Ferrochelatase).
NH2 ionize to give NH3+. So CMC i.e. Carboxy Methyl
T Cellulose is negatively charged. DEAE cellulose (Di
45. Ans. (c) Collagen
I Ethyl Amino Ethyl cellulose) is positively charged.
O [Ref: Harper 30th/e p. 47]
N In this picture shown, you can observe that every 3rd 50. Ans. (d) All
S amino acid is a glycine. This is very typical of collagen. [Ref: Harper 30th/e p. 47]
Collagen is most abundant of all human proteins.
 This is a fully folded protein shown and there is only 53. Ans. (a) DNA synthesis stops
single polypeptide chain present. So it is tertiary
structure of protein. If more than one polypeptide chain
[Ref: Harper 30th/e p. 458]
449
is present, then it is known as quaternary structure. This is a dideoxy nucleotide. If this is added in a growing
In tertiary structure, bonds present are disulphide, DNA chain or during PCR (polymerase chain reaction),
hydrogen, hydrophobic and ionic (electrostatic). So then DNA synthesis stops.

 IMAGE–BASED QUESTIONS
answer here is all. A 2’ deoxy nucleotide means oxygen is removed
from number 2 position. But in case of a 2’ 3’ dideoxy
51. Ans. (a) nucleotide, oxygen is removed from both number 2 and
[Ref: Harper 30th/e p. 354] 3 positions. This kind of nucleotide if incorporated in
End product of purine catabolism is uric acid. In the DNA, then it will not allow the next normal nucleotide
structure of uric acid, two rings are present (like the to be incorporated. So, DNA synthesis will stop.
structure of purines) (Option a). (Option b) is urea.
(Option c) is thymine (pyrimidine). (Option d) is beta
alanine, which is one of the end product of pyrimidines.

(a) (b)

(c) (d)

52. Ans. (c)

54. Ans. (b) Fatty acid synthesis
[Ref: Harper 30th/e p. 348]
[Ref: Harper 30th/e p. 162]
Low purine diet is milk (cow’s milk), yoghurt, cheese A
and vitamin C. These help in the excretion of uric acid This picture shown is of mitochondria. In mitochondria,
all the vital pathways occur i.e. TCA and ETC. All N
from body.
catabolic pathways occur in mitochondria (except
S
High purine diet is meat, liver, tuna fish, mushroom,
glycogenolysis). Also replication, transcription
W
cauliflower, peas and spinach.
and translation for mitochondrial genes occur in
E
(a) (b) mitochondria. But anabolic pathways occur in
R
cytoplasm e.g. glycogenesis, fatty acid synthesis. S
WITH
55. Ans. (a) Vitamin B2
[Ref: Harper 30th/e p. 198] E
X
Fissure tongue, lesions at the corners of mouth are
P
characteristic of riboflavin deficiency. Low RBC
L
(c) (d) glutathione reductase activity is confiramtory for ribo- A
flavin deficiency because this is the marker enzyme for N
B2 / riboflavin deficiency. Transketolase activity is the A
marker for Vitamin B1 deficiency. T
I
O
N
S
 56. Ans. (b) Karyotyping •• ChIP is Chromatin Immuno Precipitation. This
technique is used to study protein DNA interactions
450 [Ref: Harper 30th/e p. 375]
and histone modifications.
This is a karyogram shown where number and type
•• RFLP is restriction fragment length polymorphism.
of all chromosomes of a eukaryotic cell are shown.
This technique uses restriction endonulclease
The chromosomes of an organism are arranged into
enzyme which cut at the palindromic sites.
CRO BIOCHEMISTRY

homologous pairs according to size (with sex chromo-

•• RNA interference, also known as silencing technique.
somes shown last).
This can inhibit protein synthesis at the level of
This is best method to detect monosomy, trisomy. E.g.
translation, by using micro RNA, which inhibits the
Down syndrome (Trisomy 21)
target mRNA.
57. Ans. (b) Enantiomerism
63. Ans. (d)
[Ref: Harper 30th/e p. 416]
[Ref: Harper 30th/e p. 430]
D and L isomerism is also known as Enantiomerism or
Repressor tetramer binds at operator site. RNA Poly-
mirror images of each other. In this type of isomerism,
merase binds at promotor. CAP-cAMP complex binds at
there is different –H and -OH orientation at the


CAP site. CAP is Catabolic Activator Protein.

penultimate carbon. In case of glucose, at C5 if –OH is
on right side, then it is D –glucose. If at C5, -OH is on left 64. Ans. (c) Silent mutation
side then it is L-glucose.
[Ref: Lehninger 7th /e g.241]
58. Ans. (b) Adenosine In silent mutation, a codon is replaced by another
[Ref: Harper 30th/e p. 341] codon which codes for same amino acid. So, primary
structure of the protein is not changed. In this case, a
This is adenosine. Look at the nitrogenous base. It
stop codon (UAA) is replaced by stop codon (UAG). So
is having two rings, so it is a purine. Amino group is
this is silent mutation.
present so, it is adenine. But sugar is also attached so
In nonsense mutation, a codon which is coding for
it is a nucleoside (nitrogenous base + sugar). In case of
some amino acid is replaced by a stop codon. so primary
adenine, nucleoside is known as adenosine.
structure of the protein is changed.
Note: Nucleotide = nitrogenous base + sugar + phosphate
65. Ans. (a) Missense mutation
59. Ans. (c) α1 → 4, α1 → 6 and α1 → 4
[Ref: Harper 30th/e p. 417]
[Ref: Harper 30th/e p. 153]
This is missense mutation. AUG is the initiation codon,
In the structure of glycogen, linear chains have alpha
which codes for methionine. This amino acid does
(1 → 4) bonds. Only the branch point is alpha (1 → 6)
not show degeneracy, means it has only one codon.
bond. Point ‘a’ and ‘c’ lies in linear chain. Point ‘b’ lies at
So when codon for methionine is changed by some
A the branch point.
another codon, then amino acid will be changed. So
N 60. Ans. (c) this is missense mutation. The new codon is not a stop
S codon (UAA, UAG, UGA), so it is not nonsense mutation.
W [Ref: Harper 30th/e p. 365]
E Codon is present on mRNA. Anticodon is present on 66. Ans. (b) X is Pyruvate Dehydrogenase, Y is Lactate
R tRNA. Amino acid is attached to the 3’ end of the tRNA. dehydrogenase
First amino acid is methionine. [Ref: Harper 30th/e p. 172]
S
61. Ans. (a) Pyruvate to Acetyl CoA conversion is Link reaction, by
WITH
enzyme Pyruvate Dehydrogenase complex. Pyruvate to
[Robbins 10 th/e pg. 174-175] Lactate conversion occurs in anaerobic conditions, by
E
X CG site means C andG present on same strand of DNA. enzyme Lactate Dehydrogenase (LDH).
P The cytosine of CG site can be methylated to inhibit
L the gene. This mechanism is the most commonly used 67. Ans. (a) X- Hydrogen bond, Y- beta N glycosidic
A method in Genomic Imprinting. bond, Z- 3’ 5’ phospho diester bond
N [Ref: Harper 30th/e p. 71]
62. Ans. (b) Microarray
A X is hydrogen bond i.e. in between the two nitrogenous
T [Ref: Harper 30th/e p. 465] bases. There is double bond between A and T, triple
I •• Microarray is also known as ‘chip’. It’s a small chip on bond between G and C.
O which lots of probes can be placed and thus multiple Y is the bond between nitrogenous base and sugar.
N mutations can be detected. Its like a chip in computer This is beta-N glycosidic bond, which is a covalent bond.
S where lots of information present on a small ‘chip’.
 This bond is between C1 of sugar and N9 of purines and glucose-6- phosphate to free glucose. This enzyme is
N1 of pyrimidines. common to two pathways i.e. gluconeogenesis and
Z is the bond between sugar and phosphate. This is 3’ glycogenolysis. This enzyme is present in the membrane 451
5’ phosphodiester bond. of endoplasmic reticulum.

68. Ans. (a) LCAT (Lecithin cholesterol acyl transferase)

 IMAGE–BASED QUESTIONS
[Ref: Harper 30th/e p. 253]
LCAT i.e. Lecithin Cholesterol Acyl Transferase is
involved in the conversion of cholesterol to cholesterol
ester. In this reaction, fatty acid is added. This fatty acid
is taken from Lecithin, which is a phospholipid present
within the structure of HDL.

69. Ans. (c) Both a and b

[Ref: Harper 30th/e p. 275]
LDL receptors are present on liver which can recognize
LDL circulating in blood and can uptake LDL into
hepatocytes. The main ligand for LDL receptors is Apo
B 100 but also Apo E acts as a minor ligand. So answer
is both A and B 74. Ans. (a) Type I

70. Ans. (b) Optical isomerism [Ref: Harper 30th/e p. 631]

Type of collagen in skin is type I, which is the most
[Ref: Harper 30th/e p. 153] common type. Around 28 types of collagen are known.
In this picture you can see that a plane polarized light Type II is in connective tissues, type III in vascular tissue
is passed through the solution. After passing through and type IV is in basement membrane.
the solution, light gets rotated. If light is rotated towards
right side, then the sugar present in solution is known 75. Ans. (a) Type I
as dextrorotatory (d is +). If light is rotated towards left [Ref: Harper 30th/e p. 426]
side, then the sugar present in solution is known as This picture shows collagen in which every 3rd amino
levorotatory (l is –). These dextro and levo are optical acid is glycine. Collagen is the most abundant of all
isomers. human proteins. Around 28 types of collagen are
Enantiomers are D and L- isomers or mirror images. known. Type I is most abundant.
Epimerism is different –H and -OH orientation around
only one carbon, other than the penultimate carbon. 76. Ans. (b) Symport
A
71. Ans. (c) Complex II, III and IV [Ref: Harper 30th/e p. 486] N
The transport mechanism in the diagram marked by S
[Ref: Harper 30th/e p. 132]
arrow is symport i.e. both molecules going in same W
When NADH enters ETC, then the complexes involved direction. The first one is uniport. The third one is E
are complex I, III and IV. But if FADH2 enters ETC, then antiport. R
the complexes involved are complex II, III and IV.
S
72. Ans. (b) GLUT 2
WITH
[Ref: Harper 30th/e p. 191]
E
GLUT-2 is present in liver, intestine and pancreas.
X
Refer Table 2.13
P
73. Ans. (c) Glucose-6- phosphatase L
A
[Ref: Harper 30th/e p. 178] N
This cell organelle shown is endoplasmic reticulum. A
Glucose-6-phosphatase is an enzyme which converts T
I
O
N
S
 77. Ans. (b) Exocytosis phagocytosis, pinocytosis and receptor mediated
endocytosis (RME).
452 [Ref: Harper 30th/e p. 476]
In Phagocytosis, cell engulf large particles. Pinocytosis
Exocytosis is a kind of transport by which cell or fluid endocytosis cell engulf liquid or small particles.
transports molecules out of the cell, with the help of a In RME, a ligand is present on the extracellular side of
membrane vesicle. If molecule is taken inside the cell the membrane and a specific molecule is taken in by
CRO BIOCHEMISTRY

using a membrane bound vesicle, then it is known making a membrane bound vesicle in which ligand
as pinocytosis. Endocytosis can be of three types: is also taken inside. Ligand can later return to the
membrane.


78. Ans. (d) SGLT-1 82. Ans. (a) HMG CoA synthase
[Ref: Harper 30th/e p. 492] [Ref: Harper 30th/e p. 222]
This is sodium potassium pump shown in the picture. It This is acetoacetate – a ketone body. Rate limiting step
is associated with SGLT i.e. sodium dependent glucose is HMG CoA synthase.
transport. It is not associated with GLUTs. GLUTs are
facilitative transport and are sodium independent. 83. Ans. (d) Chylomicron
[Ref: Harper 30th/e p. 252]
79. Ans. (b) Milk
•• Chylomicron is largest in size. HDL is smallest in size.
A [Lippincott 4th /e pg.84 •• So the one which is largest in size is chylomicron. The
N •• This is lactose in which there is beta (1→ 4) glycosidic one which is smallest in size is HDL.
S bond between galactose and glucose. Lactose is
present in milk. 84. Ans. (a) 5
W
E •• Sucrose is present in table sugar, sugarcane juice [Ref: Harper 30th/e p. 845]
R and sugarbeet. Maltose/malt sugar is present in The one which is largest in size is chylomicron. The one
germinating grain. Fructose is present in fruits, honey which is smallest in size is HDL. HDL has maximum
S and corn syrup. density and chylomicron has least density. Density is
WITH directly proportional to the percentage of protein. So
80. Ans. (d) Honey
HDL has maximum protein content. In this picture
E [Ref: Harper 30th/e p. 155] number 5 (smallest) represent HDL, which has
X
This is fructose structure. Fructose is present in fruits, maximum density and thus maximum proteins.
P
honey and corn syrup.
L 85. Ans. (b) Apo B 100
A 81. Ans. (d) HMG CoA Reductase
N [Ref: Harper 30th/e p. 410]
A [Ref: Harper 30th/e p. 266] This is the structure of a lipoprotein. The portion
T This is cholesterol shown. Rate limiting step is HMG marked in box is the apoprotein 4. Liver synthesize Apo
I CoA Reductase. B 100 and intestine synthesize Apo B 48.
O
N
S
86. Ans. (b) THF 89. Ans. (c) Glutamine
[Ref: Harper 30th/e p. 283] [Ref: Harper 30th/e p. 289] 453
Serine to glycine conversion is reversible. THF (tetra Ammonia is transported from brain to liver via
hydro folate) is required. glutamine. In liver this ammonia is used for the
production of urea.

 IMAGE–BASED QUESTIONS
90. Ans. (a) Vitamin A
[Ref: Satyanarayana 5 th /e pg. 126]
These are Bitot’s spots shown in the picture. In vitamin
A deficiency, there is build up of keratin located
superficially in conjunctiva. This can be oval, triangular
or irregular in shape. This is also associated with
conjunctival xerosis.

91. Ans. (c) Vitamin D

[Ref: Harper 30th/e p. 553]
This is rachitic rosary, which occurs in rickets in vitamin
D deficiency. There is nodularity at the costochondral
junction due to expansion of the anterior rib ends.

92. Ans. (b) Vitamin B2

87. Ans. (a) Methyl [Ref: Harper 30th/e p. 198]
[Ref: Harper 30th/e p. 353] This is circumcorneal vascularization, which occurs
A methyl group is added to make deoxy thymidine in Vitamin B2 deficiency. Other clinical features of
Vitamin B2 deficiency are: redness in lips, cheilosis,
mono phosphate from deoxy uridine mono phosphate.
glossitis, seborrheic dermatitis.
This conversion can occur only at the level of mono
phosphate. The source of this methyl group is THF. 93. Ans. (a) HGPRT
[Ref: Harper 30th/e p. 356]
HGPRT (hypoxanthine guanine phospho ribosyl
transferase) is enzyme of salvage pathway of purine
synthesis. The complete deficiency of this enzyme leads
to Lesch nyhan syndrome, in which patient has gout
and self mutilation behaviour.
A
N
94. Ans. (b) 2 S
W
[Ref: Rafi’s Practical pg.4]
E
This is benedict’s test shown. In this test benedict’s
R
88. Ans. (b) Amino reagent is taken and then the sample added. Then test
tube is heated. The color of benedict’s reagent is blue. S
[Ref: Harper 30th/e p. 118]
So after heating blue color shows negative results. But WITH
This conversion from UTP to CTP occurs at the level of after heating if color changes to green, orange or brick
triphosphate. ATP is used and amino group added. This red then it shows a positive test for reducing sugar. E
amino group is taken from glutamine. X
95. Ans. (b) Homogentisate dioxygenase P
[Ref: Harper 30th/e p. 304] L
Diagnosis is alkaptonuria. Urine turns black on
A
exposure to air due to oxidation of homogentisic acid.
N
Enzyme is homogentisate dioxygenase.
A
T
I
O
N
S
 96. Ans. (b) Tyrosinase 101. Ans. (a) CCA

454 [Ref: Harper 30th/e p. 317] [Ref: Harper 30th/e pg. 365]
The enzyme involved in the conversion of tyrosine to
melanin is tyrosinase. This enzyme is an oxidase. It
requires copper. It is deficient in Albinism.
CRO BIOCHEMISTRY

97. Ans. (d) Arginine

[Ref: Harper 30th/e. p. 661]
Nitric Oxide Synthase Reaction
•• Nitric oxide (NO) is an endothelium derived relaxing


factor
•• NO is formed by the action of the enzyme NO
synthase, which is cytosolic
•• The endothelial and neuronal forms of NO synthase
are activated by Ca2+
•• The substrate is arginine, and the products are
Citrulline and NO

98. Ans. (a) Phenylalanine hydroxylase 102. Ans. (b) Proline

[Ref: Harper 30th/e p. 304] [Ref: Harper 30th/e pg. 45]
•• Phenylalanine hydroxylase is an enzyme that Proline is referred to as Imino Acid. The a-amino
catalyzes the hydroxylation of the aromatic side- nitrogen form a rigid five membered Pyrrolidine ring.
chain of phenylalanine to generate tyrosine Therefore, this amino group is called secondary amino
group.
99. Ans. (d) Folic acid
[Ref: Harper, 30/e p 299, 302] 103. Ans. (b) Guanine
•• Catabolism of histidine proceeds through urocanate, [Ref: Harper 30th/e pg. 342]
4-imidazolone-5- propionate, and N form-imino- The given structure is Guanine (a purine). You have
A glutamate (FIGLU) to learn recognizing the structures of purines and
N •• Formimino group is transferred to tetrahydrofolate pyrimidines.
S forming glutamate, then alpha-ketoglutarate
W •• In folic acid deficiency, group transfer is impaired
E and FIGLU is excreted:
R
100. Ans. (b) Bile Salts
S
[Ref: Practical pg. 32]
WITH
•• Hay’ Sulphur test is a test for bile salts. In the
E control, sulphur powder remains immiscible with
X the underlying liquid whereas in the positive test, the
P sulphur powder sinks to the bottom.
L •• Foucher’s test is a test for bile pigments.
A •• Sulfur test is for Sulfur containing amino acids. But
N only cysteine and cystine gives positive sulphur test.
A Methionine does not give positive sulphur test due to
T sulphur in thioether linkage in it.
I
O
N
S
104. Ans. (a) NADPH 108. Ans. (a) CGGUAAAGCUUA
[Ref: Harper 30th/e pg. 672] [Ref: Harper 30th/e pg. 362] 455
•• Both heme oxygenase and biliverdin reductase use •• Base sequence and direction of new RNA is always
NADPH. same as the non-template strand. But there is uracil
•• These both enzymes are involved in haem cata- in place of thymine.

 IMAGE–BASED QUESTIONS
bolism. •• Whenever sequence is written without mentioning
the direction, then it is always 5’ on left side & 3’ on
right side.

109. Ans. ( b) Kidney, Liver, Muscle

[Ref: Harper 30th/e pg. 320]
Synthesis of Guanidinoacetate occurs in kidney.
Synthesis of creatine occurs in liver and synthesis of
105. Ans. (c) Type VII Phosphocreatine or Creatinine Phosphate occurs in
[Ref: Harper 30th/e pg. 631] muscle.
In epidermolysis bullosa, Type VII collagen is 110. Ans. (a) Tyrosine
defective, which anchors dermis- epidermis junction.
Glycosylation of collagen is impaired. Patient has skin [Ref: Harper 30th/e pg. 303]
blisters •• This is a case of alkaptonuria, where patient has
bluish-black pigmentation of connective tissue. Fresh
106. Ans. (b) Albinism urine is normal in color, but on standing it turns black
due to oxidation of excess homogentisic acid. A
[Ref: Harper 30th/e pg. 269]
•• Alkaptonuria is a defect in catabolic pathway of N
•• Albinism (milky white skin, white hair, red eye color) S
is absence of pigment Melanin from skin, hair and phenylalanine and tyrosine
W
eyes. Enzyme deficient is Tyrosinase (an Oxidase and
111. Ans. (d) Untreated PKU (Phenylketonuria) E
Cu required).
R
•• Oculo Cutaneous Albinism is associated with defects [Ref: Harper 30th/e pg.304]
in Vision and Photophobia. •• Untreated PKU (Phenylketonuria) >> Phenylketonuria S
•• In Vitiligo, enzyme Tyrosinase is normal but there is •• This is clinical picture of PKU (Phenylketonuria) WITH
lack of Melanoblasts in regional areas. with mental retardation. These patients have mousy
•• Alkaptonuria is associated with defects in Catabolism or musty odour due to excess phenyl-acetate. Hypo- E
of Tyrosine and Phenylalanine. pigmentation occurs as tyrosine not formed, which is X
•• Phenylketonuria (PKU) is due to defect in enzyme responsible for making pigment Melanin. PKU can be P
Phenylalanine Hydroxylase. treated if diagnosed early. L
A
107. Ans. (a) Histidine 112. Ans. (c) Translocation N
[Ref: Harper 30th/e pg. 17] A
[Ref: Harper 30th/e pg.459]
Histidine has maximum buffering capacity because T
•• Here, chromosomes 1, 2, and 4 are labeled yellow I
Histidine is the only amino acid which can ionize within with FISH and the other chromosomes are stained
the physiological pH range. O
red. Translocations between yellow and red chromo- N
S
 somes are found. The left picture represents a skeletal abnomalities resembling marfan syndrome
normal cell (the numbers in the figure indicate the i.e. they are usually tall and thin. Progressive mental
456 number of chromosome) and the right picture is an retardation is common.
example of reciprocal translocation with two bi-color •• Cyanide Nitroprusside test is positive as homocystine
chromosomes (indicated by two arrows). is excreted in urine.
CRO BIOCHEMISTRY

115. Ans. (c) Orotic aciduria

[Ref: Harper 30th/e pg. 346]
Orotic aciduria is a rare, autosomal recessive disorder.
It is the most common metabolic error in pyrimidine
synthesis. OPRT (Orotate Phospho Ribosyl Transferase)
& Decarboxylase are bifunctional enzymes which are
deficient in orotic aciduria. Treatment is administration
of uridine (results in the improvement of anemia and
decreased excretion of orotate). All pyrimidines are


not getting formed in these patients but only uridine is

given as other pyrimidines (C & T) will be synthesized
from uridine because those enzymes are not deficient.

113. Ans. (a) (I) Microarray, (II) FISH showing 116. Ans. (a) Familial hypercholesterolemia
microdeletion, (III) PCR, (IV) RHP [Ref: Harper 30th/e pg.275]
[Ref: Harper 30th/e pg.462] •• This is a case of Familial hypercholesterolemia which
•• I- Microarray is AD (Autosomal Dominant).
•• II- FISH showing microdeletion •• There is history of coronary artery disease in the
•• III- PCR family. Patient has high cholesterol and normal TGs
•• IV- RFLP
117. Ans. (a) Hurler disease
114. Ans. (c) Homocystinuria
[Ref: Harper 30th/e pg.179]
[Ref: Harper 30th/e pg.302] •• This is a case of Hurler disease.
•• Homocystinuria has autosomal recessive inheri- •• In hunter disease, vision is clear but this patient has
tance. Infants with disorder are normal at birth. They defective vision. Heparan sulfate & Dermatan sulfate
have sub-luxated lens, thromboembolic episodes, are excreted in both hurler’s & hunter’s disease

A
N
S
W
E
R
S
WITH

E
X
P
L
A
N
A
T
I
O
N
S