PowerPoint® Lecture

Presentations prepared by
John Zamora
Middle Tennessee State
University

CHAPTER 3
Microbial
Metabolism

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 I. Laboratory Culture of Microorganisms

• 3.1 Cell Chemistry and Nutrition

• 3.2 Culture Media

• 3.3 Laboratory Culture

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 3.1 Cell Chemistry and Nutrition

• Nutrients
• Supply of monomers (or precursors of) required by cells
for growth

• Macronutrients
• Nutrients required in large amounts

• Micronutrients
• Nutrients required in trace amounts

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© Pearson Education Limited 2015
 3.1 Cell Chemistry and Nutrition

• Carbon
• Required by ALL cells

• Typical bacterial cell is ~50% carbon (by dry weight)

• Major element in ALL classes of macromolecules

• Heterotrophs use organic carbon

• Autotrophs use carbon dioxide (CO2)

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 3.1 Cell Chemistry and Nutrition

• Nitrogen
• Typical bacterial cell is ~13% nitrogen
(by dry weight)

• Key element in proteins, nucleic acids, and many more

cell constituents

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 3.1 Cell Chemistry and Nutrition

• Other macronutrients
• Phosphorus (P)
• Synthesis of nucleic acids and phospholipids

• Sulfur (S)
• Sulfur-containing amino acids (cysteine and methionine)

• Vitamins (e.g., thiamine, biotin, lipoic acid) and coenzyme

A

• Potassium (K)
• Required by enzymes for activity
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 3.1 Cell Chemistry and Nutrition

• Other macronutrients (cont'd)

• Magnesium (Mg)
• Stabilizes ribosomes, membranes, and nucleic acids

• Also required for many enzymes

• Calcium (Ca)
• Helps stabilize cell walls in microbes

• Plays key role in heat stability of endospores

• Sodium (Na)
• Required by some microbes (e.g., marine microbes)
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 3.1 Cell Chemistry and Nutrition

• Iron
• Key component of cytochromes and FeS proteins
involved in electron transport

• Growth factors
• Organic compounds required in small amounts by
certain organisms
• Examples: vitamins, amino acids, purines, pyrimidines
• Vitamins
• Most commonly required growth factors
• Most function as coenyzmes
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 3.2 Media and Laboratory Culture

• Culture media
• Nutrient solutions used to grow microbes in the
laboratory

• Two broad classes

• Defined media: precise chemical composition is known

• Complex media: composed of digests of chemically

undefined substances (e.g., yeast and meat extracts)

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 3.2 Media and Laboratory Culture

• Enriched media
• Contain complex media plus additional nutrients

• Selective media
• Contain compounds that selectively inhibit growth of
some microbes but not others

• Differential media
• Contain an indicator, usually a dye, that detects
particular chemical reactions occurring during growth

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 3.2 Media and Laboratory Culture

• For successful cultivation of a microbe, it is

important to know the nutritional requirements and
supply them in proper form and proportions in a
culture medium

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 3.2 Media and Laboratory Culture

• Pure culture: culture containing only a single kind

of microbe

• Contaminants: unwanted organisms in a culture

• Cells can be grown in liquid or solid culture media

• Solid media are prepared by addition of a gelling agent
(agar or gelatin)

• When grown on solid media, cells form isolated masses

(colonies)

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 3.2 Media and Laboratory Culture

• Microbes are everywhere

• Sterilization of media is critical

• Aseptic technique should be followed

(Figure 3.3)

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© Pearson Education Limited 2015 Figure 3.3
 Aseptic Transfer and the Streak Plate Method

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 3.2 Media and Laboratory Culture

• Pure culture technique

• Streak plate (Figure 3.4)

• Pour plate

• Spread plate

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 Isolated colonies
Subsequent streaks
at end of streak
are at angles to
the first streak. Confluent growth at
1 2
5 beginning of streak
3
4

1. Loop is sterilized and a 2. Initial streak is worked in 3. Appearance of well-streaked

loopful of inoculum is well in one corner of the plate after incubation shows
removed from tube. agar plate. colonies of the bacterium
Micrococcus luteus on a
blood agar plate.

© Pearson Education Limited 2015 Figure 3.4

© Pearson Education Limited 2015
 II. Energetics, Enzymes and Redox

• 3.3 Energy Classes of Microorganisms

• 3.4 Bioenergetics

• 3.5 Catalysis and Enzymes

• 3.6 Electron Donors and Electron Acceptors

• 3.7 Energy-Rich Compounds

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 Metabolism: Overview

[insert Metabolism_Overview.jpg]

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 3.3 Energy Classes of Microorganisms

• Metabolism
• The sum total of all of the chemical reactions that occur
in a cell
• Catabolic reactions (catabolism)
• Energy-releasing metabolic reactions
• Microorganisms grouped into energy classes
(Figure 3.5)
• Chemorganotrophs
• Chemolithotrophs
• Phototrophs
• Heterotrophs
• Autotrophs
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© Pearson Education Limited 2015 Figure 3.5
 3.4 Bioenergetics

• Energy is defined in units of kilojoules (kJ), a

measure of heat energy

• In any chemical reaction, some energy is lost

as heat

• Free energy (G): energy released that is available

to do work

• The change in free energy during a reaction is

referred to as ΔG0′
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 3.4 Bioenergetics

• Reactions with a negative ΔG0′ release free

energy (exergonic)

• Reactions with a positive ΔG0′ require energy

(endergonic)

• To calculate free-energy yield of a reaction, we

need to know the free energy of formation (Gf0; the
energy released or required during formation of a
given molecule from the elements)
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 3.4 Bioenergetics

• For the reaction A + B C + D,

• ΔG0′ = Gf0 [C+D] - Gf0[A+B]

• ΔG0′ is not always a good estimate of actual free-

energy changes
• ΔG: free energy that occurs under actual
conditions
• ΔG = ΔG0′ + RT lnK
• where R and T are physical constants and K is the
equilibrium constant for the reaction in question
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 3.5 Catalysis and Enzymes

• Free-energy calculations do not provide

information on reaction rates

• Activation energy: energy required to bring all

molecules in a chemical reaction into the reactive
state (Figure 3.6)
• Catalysis is usually required to breach activation energy
barrier

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© Pearson Education Limited 2015 Figure 3.6
 3.5 Catalysis and Enzymes

• Catalyst: substance that

• Lowers the activation energy of a reaction

• Increases reaction rate

• Does not affect energetics or equilibrium of a reaction

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 3.5 Catalysis and Enzymes

• Enzymes
• Biological catalysts

• Typically proteins (some RNAs)

• Highly specific

• Generally larger than substrate

• Typically rely on weak bonds

• Examples: hydrogen bonds, van der Waals
forces, hydrophobic interactions

• Active site: region of enzyme that binds substrate

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 3.5 Catalysis and Enzymes

• Enzymes (cont'd)
• Increase the rate of chemical reactions by 108 to 1020
times the spontaneous rate

• Enzyme catalysis: E + S E—S E+P

• Catalysis dependent on
• Substrate binding

• Position of substrate relative to catalytically active amino

acids in active site

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 Products
Substrate

1. Substrate is Active site

bound to 4. Products
enzyme are released.
2. Enzyme–
active site.
substrat 3. Strain is
e placed
complex on bond.
forms.

5. Enzyme is ready
to begin new
catalytic cycle.

© Pearson Education Limited 2015 Figure 3.7

 3.5 Catalysis and Enzymes

• Many enzymes contain small nonprotein molecules that

participate in catalysis but are not substrates
• Prosthetic groups
• Bind tightly to enzymes
• Usually bind covalently and permanently
(e.g., heme group in cytochromes)
• Coenzymes
• Loosely bound to enzymes
• Most are derivatives of vitamins
(e.g., NAD+/NADH)

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 3.6 Electron Donors and Electron Acceptors

• Energy from oxidation–reduction (redox) reactions

is used in synthesis of energy-rich compounds
(e.g., ATP)
• Redox reactions occur in pairs (two half reactions;
Figure 3.8)
• Electron donor: the substance oxidized in a redox
reaction
• Electron acceptor: the substance reduced in a
redox reaction
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 Half reaction
donating e– Electron Electron
donor acceptor

Formation Net reaction

Half reaction of water
accepting e–

© Pearson Education Limited 2015 Figure 3.8

 3.6 Electron Donors and Electron Acceptors

• Reduction potential (E0′): tendency to donate

electrons
• Expressed as volts (V)

• Substances can be either electron donors or

electron acceptors under different circumstances
(redox couple)
• Reduced substance of a redox couple with a more
negative E0′ donates electrons to the oxidized
substance of a redox couple with a more positive E0′
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 3.6 Electron Donors and Electron Acceptors

• The redox tower represents the

range of possible reduction
potentials
• The reduced substance at the
top of the tower donates
electrons
• The oxidized substance at the
bottom of the tower accepts
electrons
• The farther the electrons
"drop," the greater the amount
of energy released
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 3.6 Electron Donors and Electron Acceptors

• Redox reactions usually involve reactions between

intermediates (carriers)

• Electron carriers are divided into two classes

• Prosthetic groups (attached to enzymes)

• Coenzymes (diffusible)
• Examples: NAD+, NADP (Figure 3.10)

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 3.6 Electron Donors and Electron Acceptors

NAD+ reduction

NAD+ Active Enzyme–substrate

binding site complex
1. Enzyme I reacts with e– donor
site
and oxidized form of
coenzyme, NAD+.
Enzyme I

2. NADH and
reaction
Substrate
+ product are
NAD+ + NADH
(e– donor) formed.
+
Product
4. NAD+ is
released. Product NADH Activ
binding e
site site
Enzyme Il Substrate
3. Enzyme II reacts with e– (e– acceptor)
acceptor and reduced Enzyme–substrate
NADH oxidation form of coenzyme, NADH. complex

• NAD+ and NADH facilitate redox reactions without

being consumed; they are recycled
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 3.7 Energy-Rich Compounds

• Chemical energy released in redox reactions is

primarily stored in certain phosphorylated
compounds (Figure 3.12)
• ATP; the prime energy currency

• Phosphoenolpyruvate

• Glucose 6-phosphate

• Chemical energy also stored in coenzyme A

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 Anhydride bonds Ester bond
Ester bond

Anhydride bond

Phosphoenolpyruvate Adenosine triphosphate (ATP) Glucose 6-phosphate

Compound G0′kJ/mol
Thioester
Anhydride bond
bond ΔG0′< 30kJ
Phosphoenolpyruvate –51.6
1,3-Bisphosphoglycerate –52.0
Acetyl phosphate –44.8
ATP –31.8
ADP –31.8
Acetyl Coenzyme A Acetyl phosphate Acetyl-CoA –35.7
Acetyl-CoA ΔG0′< 30kJ
AMP –14.2
Glucose 6-phosphate –13.8

© Pearson Education Limited 2015 Figure 3.12

 3.7 Energy-Rich Compounds

• Long-term energy storage involves insoluble

polymers that can be oxidized to generate ATP
• Examples in prokaryotes
• Glycogen
• Poly-β-hydroxybutyrate and other
polyhydroxyalkanoates
• Elemental sulfur
• Examples in eukaryotes
• Starch
• Lipids (simple fats)
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 III. Fermentation and Respiration

• 3.8 Glycolysis
• 3.9 Fermentative Diversity and the Respiratory
Option
• 3.10 Respiration: Electron Carriers
• 3.11 Respiration: The Proton Motive Force
• 3.12 Respiration: Citric Acid and Glyoxylate Cycle
• 3.13 Catabolic Diversity

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 3.8 Glycolysis

• Two reaction series are linked to energy

conservation in chemoorganotrophs: fermentation
and respiration (Figure 3.13)
• Differ in mechanism of ATP synthesis
• Fermentation: substrate-level phosphorylation; ATP is
directly synthesized from an energy-rich intermediate
• Respiration: oxidative phosphorylation; ATP is produced
from proton motive force formed by transport of
electrons
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 Intermediates
Energy-rich
Pi intermediates ADP ATP

A B B~P C~P D
Substrate-level phosphorylation

Energized
membrane
Dissipation of proton ADP + Pi
motive force coupled
to ATP synthesis
ATP

Less energized
membrane

Oxidative phosphorylation
© Pearson Education Limited 2015 Figure 3.13
 3.8 Glycolysis

• Fermented substance is both an electron donor

and an electron acceptor

• Glycolysis (Embden–Meyerhof pathway): a

common pathway for catabolism of glucose
(Figure 3.14)
• Anaerobic process

• Three stages

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© Pearson Education Limited 2015 Figure 3.14
 3.8 Glycolysis

• Glycolysis
• Glucose is consumed

• Two ATPs are produced

• Fermentation products are generated

• Some harnessed by humans for consumption

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 3.9 Fermentative Diversity and the Respiratory
Option
• Fermentations classified by products formed
• Ethanol

• Lactic acid

• Propionic acid

• Mixed acids

• Butyric acid

• Butanol

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 3.9 Fermentative Diversity and the Respiratory
Option
• Fermentations classified by substrate fermented
• Usually NOT glucose

• Amino acids

• Purines and pyrimidines

• Aromatic compounds

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 3.9 Fermentative Diversity and the Respiratory
Option
• Saccharomyces cerevisiae can carry out
fermentation or respiration
• Carries out the one most beneficial
• Respiration generates more ATP

• Fermentation occurs when conditions are anoxic

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 3.10 Respiration: Electron Carriers

• Aerobic respiration
• Oxidation using O2 as the terminal electron acceptor

• Higher ATP yield than fermentations

• ATP is produced at the expense of the proton motive
force, which is generated by electron transport

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 3.10 Respiration: Electron Carriers

• Electron transport systems

• Membrane-associated

• Mediate transfer of electrons

• Conserve some of the energy released during transfer

and use it to synthesize ATP

• Many oxidation–reduction enzymes are involved in

electron transport (e.g., NADH dehydrogenases,
flavoproteins, iron–sulfur proteins, cytochromes,
quinones)
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 3.11 Respiration: The Proton Motive Force

• Electron transport system oriented in cytoplasmic

membrane so that electrons are separated from
protons (Figure 3.20)

• Electron carriers arranged in membrane in order of

their reduction potential

• The final carrier in the chain donates the electrons

and protons to the terminal electron acceptor

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 During electron transfer, several
protons are released on outside
of the membrane
Protons originate from NADH and
the dissociation of water
Results in generation of pH
gradient and an electrochemical
potential across the membrane
(the proton motive force)
The inside becomes electrically
negative and alkaline
The outside becomes electrically
positive and acidic
© Pearson Education Limited 2015 Figure 3.20
 3.11 Respiration: The Proton Motive Force

• During electron transfer, several protons are

released on outside of the membrane
• Protons originate from NADH and the dissociation of
water

• Results in generation of pH gradient and an

electrochemical potential across the membrane
(the proton motive force)
• The inside becomes electrically negative and alkaline
• The outside becomes electrically positive and acidic
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 3.11 Respiration: The Proton Motive Force

• ATP synthase (ATPase): complex that converts

proton motive force into ATP; two components
(Figure 3.21)
• F1: multiprotein extramembrane complex; faces
cytoplasm

• Fo: proton-conducting intramembrane channel

• Reversible; dissipates proton motive force

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 δ
α δ
β
ADP + Pi α
α α
β β F1 F1

ATP In In
b2 b2
γ γ
ε
ε

F0
Membrane F0
c12 Out Out

© Pearson Education Limited 2015 Figure 3.21

 3.12 Respiration: Citric Acid and Glyoxylate
Cycle
• Citric acid cycle (CAC): pathway through which
pyruvate is completely oxidized to CO2 (Figure
3.22a)
• Initial steps (glucose to pyruvate) same as glycolysis
• Per glucose molecule, 6 CO2 molecules released and
NADH and FADH generated
• Plays a key role in catabolism AND biosynthesis

• Energetics advantage to aerobic respiration

(Figure 3.22b)
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© Pearson Education Limited 2015 Figure 3.22a
© Pearson Education Limited 2015 Figure 3.22b
 3.12 Respiration: Citric Acid and Glyoxylate
Cycle
• The citric acid cycle generates many compounds
available for biosynthetic purposes
• - α-Ketoglutarate and oxaloacetate (OAA): precursors of
several amino acids; OAA also converted to
phosphoenolpyruvate, a precursor
of glucose

• Succinyl-CoA: required for synthesis of cytochromes,

chlorophyll, and other tetrapyrrole compounds

• Acetyl-CoA: necessary for fatty acid biosynthesis

© Pearson Education Limited 2015
 3.12 Respiration: Citric Acid and Glyoxylate
Cycle
• Organic acids can be metabolized as electron
donors and carbon sources by many microbes

• C4-C6 citric acid cycle intermediates (e.g., citrate,

malate, fumarate, and succinate) are common
natural plant and fermentation products and can
be readily catabolized through the citric acid cycle
alone

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 3.12 Respiration: Citric Acid and Glyoxylate
Cycle
• Glyoxylate cycle
• Catabolism of C2-C3 organic acids typically involves
production of oxaloacetate through the glyoxylate cycle
(Figure 3.23)

• A variation of the citric acid cycle

• Glyoxylate is a key intermediate

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© Pearson Education Limited 2015 Figure 3.23
 3.13 Catabolic Diversity

• Microorganisms demonstrate a wide range of

mechanisms for generating energy (Figure 3.24)
• Fermentation

• Aerobic respiration

• Anaerobic respiration

• Chemolithotrophy

• Phototrophy

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 Electron donor
Fermentation
(organic compound)
Electron transport/
generation of pmf

Aerobic
Electron Organic e– respiration
acceptors acceptors
Chemotrophs

Anaerobic respiration
Chemoorganotrophy
Photoheterotrophy Photoautotrophy
Light

Phototrophs
Organic
compound
Electrons from
Electron transport/ Electron H2O (oxygenic)
generation of pmf transport H2S (anoxygenic)

Electron Aerobic respiration Generation of pmf

acceptors
Cell material Cell material
Anaerobic respiration
Chemolithotrophy Phototrophy

© Pearson Education Limited 2015 Figure 3.24

 3.13 Catabolic Diversity

• Anaerobic respiration
• The use of electron acceptors other than oxygen
• Examples include nitrate (NO3–), ferric iron (Fe3+), sulfate
(SO42–), carbonate (CO32–), certain organic compounds

• Less energy released compared to aerobic respiration

• Dependent on electron transport, generation of a proton

motive force, and ATPase activity

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 3.13 Catabolic Diversity

• Chemolithotrophy
• Uses inorganic chemicals as electron donors
• Examples include hydrogen sulfide (H2S), hydrogen gas
(H2), ferrous iron (Fe2+), ammonia (NH3)

• Typically aerobic

• Begins with oxidation of inorganic electron donor

• Uses electron transport chain and proton motive force

• Autotrophic; uses CO2 as carbon source

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 3.13 Catabolic Diversity

• Phototrophy: uses light as energy source

• Photophosphorylation: light-mediated ATP synthesis

• Photoautotrophs: use ATP for assimilation of CO2 for

biosynthesis

• Photoheterotrophs: use ATP for assimilation of organic

carbon for biosynthesis

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 IV. Biosyntheses

• 3.14 Sugars and Polysaccharides

• 3.15 Amino Acids and Nucleotides

• 3.16 Fatty Acids and Lipids

• Regulation: biofeedback

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 3.14 Sugars and Polysaccharides

• Prokaryotic polysaccharides are synthesized from

activated glucose (Figure 3.25a)
• Adenosine diphosphoglucose (ADPG)
• Precursor for glycogen biosynthesis

• Uridine diphosphoglucose (UDPG)

• Precursor of some glucose derivatives needed for
biosynthesis of important polysaccharides

• Examples: N-acetylglucosamine,
N-acetylmuramic acid
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 Glucose

Uridine diphosphoglucose (UDPG)

© Pearson Education Limited 2015 Figure 3.25a

 3.14 Sugars and Polysaccharides

• Gluconeogenesis (Figure 3.25b)

• Synthesis of glucose from phosphoenolpyruvate
• Phosphoenolpyruvate can be synthesized from
oxaloacetate

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© Pearson Education Limited 2015 Figure 3.25b
 3.14 Sugars and Polysaccharides

• Pentoses are formed by the removal of one

carbon atom from a hexose (Figure 3.25c)

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 Glucose-6-P
Pentose
phosphate
Ribulose-5-P + CO2
pathway, see
Figure 3.26
Ribose-5-P

Ribonucleotides Ribonucleotides
NADPH
NADPH-dependent
ribonucleotide reductase
forms deoxyribonucleotides.

RNA Deoxyribonucleotides DNA

© Pearson Education Limited 2015 Figure 3.25c

 3.14 Sugars and Polysaccharides

• Pentoses are required for the synthesis of nucleic

acids

• If pentoses are not readily available from the

environment, organisms must synthesize them

• The major pathway for pentose production is the

pentose phosphate pathway (Figure 3.26)

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 From glycolysis

Glucose 6- 6-Phosphogluconate
phosphate
Production of
NADPH and CO2 Ribulose 5-
phosphate

To nucleic acid synthesis

(see Figure 3.25)

Ribose 5- Transaldolase
phosphate (C5) Transketolase
Ribulose 5-
Isomerase
phosphate
Xylulose 5-
phosphate (C5)
Gluconeogenesis
Other pentose
sugars feed in here.

© Pearson Education Limited 2015 Figure 3.26

 3.15 Amino Acids and Nucleotides

• Biosynthesis of amino acids and nucleotides often

involves long, multistep pathways
• Amino acid biosynthesis
• Carbon skeletons come from intermediates of glycolysis
or citric acid cycle (Figure 3.27)
• Ammonia is incorporated by glutamine dehydrogenase
or glutamine synthetase
(Figure 3.28)
• Amino group transferred by transaminase and synthase
(Figure 3.28)
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 Amino acid biosynthesis
Carbon skeletons come from intermediates of glycolysis or
citric acid cycle (Figure 3.27)

© Pearson Education Limited 2015 Figure 3.27

 Ammonia is incorporated by glutamine dehydrogenase or glutamine synthetase
(Figure 3.28)
Amino group transferred by transaminase and synthase (Figure 3.28)

© Pearson Education Limited 2015 Figure 3.28

 3.15 Amino Acids and Nucleotides

• Purine and pyrimidine biosyntheses are complex

• Purines (Figure 3.29a and b)

• Adenine and guanine

• Inosinic acid intermediate

• Pyrimidines (Figure 3.29c and d)

• Thymine, cytosine, and uracil

• Orotic acid intermediate

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 CO2
Amino group Glycine
of aspartate

Formyl
group
(from folic
acid)
Amide nitrogen
of glutamine Ribose-5-P
Purine skeleton Inosinic acid

Purine biosynthesis

NH3 Aspartic acid

CO2
Orotic acid Uridylate

Pyrimidine biosynthesis

© Pearson Education Limited 2015 Figure 3.29

 3.16 Fatty Acids and Lipids

• Fatty acids are biosynthesized two carbons at a

time (Figure 3.30)
• Acyl carrier protein (ACP) involved

• ACP holds the growing fatty acid as it is being

synthesized

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© Pearson Education Limited 2015 Figure 3.30
 3.16 Fatty Acids and Lipids

• In Bacteria and Eukarya, the final assembly of

lipids involves addition of fatty acids to glycerol

• In Archaea, lipids contain phytanyl side chains

instead of fatty acids
• Phytanyl biosynthesis is distinct from that of fatty acids

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 Regulating the Activity of Biosynthetic Enzymes

• Two major modes of enzyme regulation

• Amount
• Regulation at the gene level

• Activity
• Temporary inactivation of the protein through
changes in enzyme structure

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© 2012 Pearson Education, Inc.
 Regulating the Activity of Biosynthetic Enzymes

• Feedback Inhibition: mechanism for turning off the

reactions in a biosynthetic pathway
• End product of the pathway binds to the first enzyme
in the pathway, thus inhibiting its activity

• The inhibited enzyme is an allosteric enzyme (Figure

4.29)
• Two binding sites: active and allosteric

• Reversible reaction
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 Figure 4.28

Starting substrate
The allosteric
enzyme
Enzyme A

Intermediate I
Enzyme B
Intermediate II Feedback
inhibition
Enzyme C
Intermediate III
Enzyme D

End product
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 End product
(allosteric effector)

Allosteric site Active site

Enzyme
Substrate
INHIBITION: ACTIVITY:
Substrate cannot Enzyme reaction
bind; enzyme proceeds
reaction inhibited

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 Regulating the Activity of Biosynthetic Enzymes

• Some pathways controlled by feedback

inhibition use isoenzymes

• Isoenzymes
• Different enzymes that catalyze the same
reaction but are subject to different regulatory
controls

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© 2012 Pearson Education, Inc.
 Figure 4.30

Phosphoenol-  Erythrose Initial

pyruvate 4-phosphate substrates

DAHP synthases
(isoenzymes 1, 2, 3)
DAHP

Chorismate

Tyrosine Tryptophan Final

Phenylalanine products
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© 2012 Pearson Education, Inc.
 Regulating the Activity of Biosynthetic Enzymes

• Biosynthetic enzymes can also be regulated by

covalent modifications
• Regulation involves a small molecule attached to or
removed from the protein

• Results in conformational change that inhibits activity

• Common modifiers include adenosine monophosphate

(AMP), adenosine diphosphate (ADP), inorganic
phosphate (PO42), methyl groups (CH3)
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