MODERN HIGHER SECONDARY SCHOOL

MALAYNAGAR, DATTAPALLI
AGARTALA, WEST TRIPURA

“ ”
Study of digestion of starch by salivary amylase
and effect of pH and temperature on it

SUBMITTED BY
NAME : Parinita Choudhury
CLASS : XII [SCIENCE]
ROLL NO. : 55
 CERTIFICATE

This is to certify that PARINITA CHOUDHURY, a student of

Modern Higher Secondary School of class XII (Science) has
successfully completed the research on the Chemistry project
"Study of digestion of Starch by salivary amylase and effect of
pH and temperature on it" under the guidance of Miss Prakriti
Roy, during the academic session of 2024-2025 as per the
guidelines of Central Board of Secondary Education (CBSE).

___________________________
Mr. Sujit Dasgupta (Principal)
SIGNATURE OF THE PRINCIPAL

_______________________ _______________________
SIGNATURE OF EXTERNAL SIGNATURE OF INTERNAL
EXAMINER EXAMINER
It gives me great pleasure to express my gratitude towards our
Chemistry Teacher Miss Prakriti Roy for his guidance, support and
encouragement throughout the duration of the project. Without his
motivation and help the successful completion of this project would
not have been possible.
I would also like to thank Mr. Sujit Dasgupta, Principal of Modern
High Secondary School for his support and encouragement.
Last but not the least; i want to thank my Family members and
friends for their unlimited support.

XII (Science)
Modern High Secondary School
 INDEX
Sl No. Content Page no.
1. Aim 05
2. Objective 06
3. Introduction 07 - 09
4. Structure and function of starch 10 - 12
5. Mechanism of Starch Digestion 13 – 14
6. Experimental Study 15 – 16
7. Precaution 17 – 26
8. Graphical Representations 27 – 28
9. Summary of Findings 29 – 31
10. Conclusion 32
11. Bibliography 33
 – Aim –
To study of digestion of starch by salivary amylase
and effect of pH and temperature on it

5
 – Objective –
The goal of the project is to study about digestion
of starch by salivary amylase and effect of pH and
temperature on it.

6
 Introduction
What is Digestion?

Digestion is one among many life

processes observed in nearly all living
organisms. The process involves
breaking down complex molecules
into simpler molecules that can be
readily absorbed by body.

Importance of Digestion in Breaking Down Complex

Molecules

1. Energy Production: Starch is a polysaccharide that, when broken down,

releases glucose, the body's main source of energy. Without digestion, the
energy stored in starch cannot be accessed.
2. Facilitating Absorption: The small intestine can only absorb small
molecules. Enzymes like salivary amylase break down starch into glucose
and maltose, making them absorbable.
3. Blood Glucose Regulation: Digestion ensures a steady release of glucose
into the bloodstream, maintaining stable blood sugar levels and providing
energy for immediate use or storage.

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4. Supporting Growth and Repair: Glucose fuels cells and provides raw
materials for growth and tissue repair.
5. Preventing Digestive Issues: Proper breakdown of starch reduces the
risk of bloating and discomfort caused by undigested food.

Role of enzymes in chemical process

Enzymes are biological catalysts that speed up chemical reactions in living
organisms without being consumed in the process. They lower the
activation energy required for reactions, making processes like digestion,
metabolism, and cellular repair more efficient. Enzymes are highly specific,
meaning each enzyme typically acts on a particular substrate, converting it
into a product. Their activity can be influenced by factors like temperature,
pH, and substrate concentration. Without enzymes, many vital chemical
reactions in the body would occur too slowly to sustain life.

Salivary Amylase : The Key Enzyme

Salivary amylase is an enzyme that catalyzes the hydrolysis of starch, a

polysaccharide, into simpler sugars like maltose and dextrins. Chemically,
it is a calcium metalloenzyme, meaning it requires calcium ions for stability
and activity. Salivary amylase is classified as a glycoside hydrolase,
specifically acting on α-1,4 glycosidic bonds in starch. It operates optimally
at a neutral pH (around 7), which is typical of the oral cavity.

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Its activity begins the chemical digestion of carbohydrates in the mouth,
breaking down complex molecules into simpler forms that are more easily
processed during digestion.

Function:
Catalyzes the hydrolysis of starch (polysaccharide) into simpler sugars like
maltose and dextrins.

(C₆H₁₀O₅)ₙ + nH₂O → nC₁₂H₂₂O₁₁

where:

 (C₆H₁₀O₅)ₙ represents starch (a polysaccharide made of glucose units).

 H₂O is water, which is required for the hydrolysis reaction.
 C₁₂H₂₂O₁₁ is maltose (a disaccharide).

NOTE
Salivary amylase is produced and secreted by the
salivary glands in the mouth. The three main
types of salivary glands responsible for its
production are:

1. Parotid Glands: Located near the ears,

these are the largest salivary glands and
produce a watery secretion rich in salivary
amylase.
2. Submandibular Glands: Located beneath
the lower jaw, these glands produce both
watery and mucous secretions containing
amylase.
3. Sublingual Glands: Located beneath the
tongue, these glands mainly produce
Fig : 3D structure of salivary amylase mucous secretions but also contribute a
small amount of amylase.

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 Structure and
Function of Starch

Starch consists of glucose molecules synthesized by the green leaves of

plants during photosynthesis and found in the form of granules in plants.

Sources of Starch

Starch is present in the leaves of

green plants, stems (sago), roots of
the cassava plant, all vegetables,
fruits like banana and plantain,
tubers like potatoes, cereals such as
wheat, corn, maize, sorghum and
rice, and some algae.

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 Structure of Starch

Starch or Amylum is a homopolymer (each yields only one type of

monosaccharide (glucose) after complete hydrolysis) composed of D-
glucose units linked by 1 -> 4) glycoside bonds. The (1 -> 4)
glycoside linkage between the glucose units is formed by starch
synthases. It is also called glucosan or glucan.,–amylase specifically
act on catalyzing the degradation of 1,4 linkage. Starch hydrolyzes to
liberate dextrins and then maltose and glucose units with the help of an
amylase enzyme.
It constitutes two polysaccharide components: water-soluble amylose
(20-30%) and water-insoluble amylopectin (70-80%).
Both are present in starch granules

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5
 Difference

Amylose Amylopectin
Amylose has Linear, unbranched Amylopectin has Highly branched
structure. structure.
Amylose only contains α-1,4 Amylopectin contains both α-1,4
glycosidic bonds. glycosidic bonds (in the linear chains)
and α-1,6 glycosidic bonds (at the
branch points).
Amylose typically makes up 20- Amylopectin constitutes the majority
30% of starch. of starch, around 70-80%.
Amylose is less soluble in water Amylopectin is more soluble in water
due to its linear structure, which due to its branched structure, which
allows the chains to pack tightly. prevents tight packing.
Amylose is more resistant to Amylopectin is easier to digest due to
digestion because of its compact, its highly branched, open structure.
linear structure.

Q. Why starch needs to be broken down into simpler sugars for

absorption?

Ans. Starch, a large polysaccharide, needs to be broken down into simpler

sugars like glucose because its complex structure is too big for direct
absorption through the small intestine. Enzymes like amylase catalyze the
hydrolysis of glycosidic bonds, converting starch into smaller, water-soluble
molecules like glucose. These simpler sugars can then be absorbed into the
bloodstream and used by the body for energy production via cellular
respiration.

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 Mechanism of
Starch Digestion
Breakdown of Starch

Starch hydrolyzed by Amylase into maltose units, with water acting as a

reactant in the process can be represented by the following chemical
equation:
𝒂𝒎𝒚𝒍𝒂𝒔𝒆
(C6H10O5)n + nH2O nC12H22O11
Where:

 (C₆H₁₀O₅)n represents starch, a polysaccharide made of repeating

glucose units.
 H₂O represents water, which is involved in the hydrolysis reaction.
 C₁₂H₂₂O₁₁ is maltose, a disaccharide produced from starch breakdown.

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 Optimal Conditions for Amylase

Temperature: Optimal activity

occurs at 37°C, which is normal
body temperature. Activity
decreases at higher or lower
temperatures.

pH: Amylase works best at a neutral

pH (~7), which is the pH of saliva in
the mouth.

Substrate Concentration: Higher

concentrations of starch increase
amylase activity, up to a saturation
point.

Enzyme Concentration: Increased

amounts of amylase boost the
Fig: 3D Molecular
reaction rate, provided there is
structure of amylase
enough starch available.

Cofactors: Amylase requires calcium

ions (Ca²⁺) for stability and activity.

14
 Experimental
study
This experiment explores how salivary amylase breaks down starch
into simpler sugars and examines the effects of different variables
such as temperature and pH on enzyme activity.

Aim

To study the digestion of starch by salivary amylase and determine the

effect of temperature and pH on the enzymatic activity.

Materials Required

G Starch solution (1%).

G Fresh saliva (source of salivary amylase).
G Iodine solution (for starch detection).
G Test tubes.
G Droppers.
G pH buffer solutions (pH 4, 7, and 9).
G Thermometer.
G Water baths (for temperature control).
G Stopwatch.
G Spotting tile.
15
 ()

Starch solution Fresh Saliva Iodine solution

Test Tubes Droppers pH Buffer Solutions

Water Bath Thermometers Stopwatch

16
 Precautions

G Ensure saliva is fresh and not contaminated, as it may affect

enzyme activity.

G Maintain the correct temperature, especially for the test tubes

in the water bath. Too high or low temperatures can denature
amylase.

G Accurately adjust the pH using buffers to observe the effect of

different pH levels on enzyme activity.

G Use clean and dry test tubes to avoid contamination that could
alter results.

G Handle iodine solution carefully as it is a staining reagent and

can be harmful if ingested.

G Ensure consistent timing when adding saliva to each test tube

to allow for accurate comparison of results.

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 Experiment 1: Effect of Time on Starch
Breakdown
AIM
This experiment studies how the duration of exposure to salivary amylase
affects the digestion of starch into simpler sugars.

PROCEDURE

Prepare Starch Solution:

 Dissolve 1 g of starch in 100 mL of hot water and allow it to cool. This

will serve as the substrate for the reaction.

Collect Saliva:

 Collect fresh saliva as the source of salivary amylase.

Start the Reaction:

 Take a test tube and add 5 mL of starch solution.

 Add 1 mL of saliva to the test tube to initiate the enzymatic reaction.
 Start the stopwatch immediately.

Testing for Starch Breakdown:

 At regular intervals (e.g., every 2 minutes), use a dropper to take a

small sample from the test tube.
 Place a drop of the sample on a spotting tile that has a drop of iodine
solution.
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  Observe the color change:
o Blue-black indicates the presence of starch.

o The disappearance of the blue-black color indicates that starch

has been digested.

Repeat Sampling:

 Continue testing the sample every 2 minutes, recording the time at

which the blue-black color fades completely.

OBSERVATIONS:
 Initially, the iodine turns blue-black, showing the presence of starch.
 Over time, the intensity of the blue-black color fades, indicating the
progressive breakdown of starch into simpler sugars.
 Record the time at which the blue-black color disappears completely,
indicating full digestion of starch.

RESULTS:
 The longer the starch is exposed to salivary amylase, the more starch
is broken down into maltose.
 The complete disappearance of the blue-black color marks the time
at which the starch has been fully digested.

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CONCLUSION:
The breakdown of starch by salivary amylase is a time-dependent process. As
time progresses, more starch is digested, with complete digestion occurring
after a specific duration. The rate of starch breakdown can be observed
through the disappearance of the blue-black color in the iodine test,
indicating the enzymatic hydrolysis of starch into simpler sugars.

20
 Experiment 2: Effect of pH on Enzyme
Activity
AIM
To study the effect of pH on the activity of salivary amylase during starch
digestion.

PROCEDURE

Prepare Starch Solution:

o Dissolve 1 g of starch in 100 mL of hot water and allow it to

cool. This will serve as the substrate for the enzyme reaction.

Collect Saliva:

o Collect fresh saliva to use as a source of salivary amylase.

Set Up pH Conditions:

o Take three test tubes and label them with pH 4, pH 7, and pH 9.

o Add equal volumes of starch solution (e.g., 5 mL) and saliva
(e.g., 1 mL) to each test tube.

Add pH Buffers:

o Add a few drops of the pH 4 buffer to the first tube, pH 7 buffer

to the second tube, and pH 9 buffer to the third tube.

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Incubate at 37°C:

o Place all test tubes in a water bath set at 37°C (body temperature)
to simulate optimal conditions for enzyme activity.

Test for Starch Breakdown:

o At regular intervals (e.g., every 2 minutes), remove small samples

from each test tube and place them on a spotting tile with drops
of iodine solution.
o The iodine will turn blue-black if starch is still present. Once the
blue-black color fades, it indicates the starch has been broken
down.

Record Time:

o Record how long it takes for the starch to be completely digested

(when no blue-black color appears in the iodine test) in each pH
condition.

OBSERVATIONS:

 pH 4 (Acidic): The reaction proceeds slowly or may stop, indicating

that salivary amylase is less active in acidic conditions.
 pH 7 (Neutral): The reaction occurs the fastest, suggesting optimal
enzyme activity at neutral pH.
 pH 9 (Alkaline): The reaction is slower compared to neutral pH,
showing reduced activity in basic conditions.

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RESULTS:

 Salivary amylase is most effective at pH 7 (neutral), which is the pH of

saliva in the mouth.
 Enzyme activity decreases at acidic (pH 4) and alkaline (pH 9)
conditions, indicating that extreme pH levels denature or reduce the
efficiency of the enzyme.

CONCLUSION:
The activity of salivary amylase is highly dependent on pH. It works
optimally at a neutral pH (~7), while activity significantly decreases in both
acidic and basic environments.

23
 Experiment 3: Effect of Temperature on
Enzyme Activity

AIM:
To study the effect of temperature on the enzymatic activity of salivary
amylase during starch digestion

PROCEDURE:

Prepare Starch Solution:

o Dissolve 1 g of starch in 100 mL of hot water and allow it to cool.

Collect Saliva:

o Collect fresh saliva as a source of salivary amylase.

Divide the Reaction:

o Take three test tubes and label them according to the

temperatures: 0°C, 37°C, and 60°C.
o Add 5 mL of starch solution to each test tube.
o Add 1 mL of saliva to each test tube to initiate the reaction.

Set Up Water Baths:

o Place each test tube in a water bath set at the appropriate

temperature:

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  0°C (cold, ice bath).
 37°C (body temperature).
 60°C (higher temperature).

Test for Starch Breakdown:

 At regular intervals (e.g., every 2 minutes), take a small sample from

each test tube and place it on a spotting tile with a drop of iodine
solution.
 Observe the color change:
o Blue-black indicates the presence of starch.

o The disappearance of the blue-black color signifies the digestion

of starch.

Record Time:

 Record the time taken for the starch to be fully digested (when no
blue-black color appears) in each temperature condition.

OBSERVATIONS:

 0°C (Cold): Little or no starch breakdown, as enzyme activity slows

down at low temperatures.
 37°C (Body Temperature): Starch digestion occurs rapidly, indicating
optimal amylase activity at body temperature.
 60°C (High Temperature): Starch breakdown is slow or may stop
completely, as the enzyme may be denatured at high temperatures.

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RESULTS:
 0°C: Low enzyme activity; starch digestion is very slow.
 37°C: Optimal enzyme activity; starch is digested most quickly.
 60°C: Enzyme denaturation occurs, leading to little or no starch
breakdown.

CONCLUSION:
Salivary amylase functions most efficiently at 37°C, which is normal body
temperature. Lower temperatures (0°C) slow down enzyme activity, and
higher temperatures (60°C) can denature the enzyme, stopping the
digestion process. Temperature significantly affects the rate of starch
breakdown by salivary amylase.

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Graphical
Representation

Fig: pH v/s Amylase

27
Fig: Temperature v/s Amylase

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 Summary of
Findings

A. Summarizing the Optimal pH and Temperature for

Salivary Amylase Activity.

Optimal pH

G The optimal pH for salivary amylase

activity is around pH 7 (neutral).
G At this pH, enzyme activity is highest,
as it corresponds to the natural pH of
the mouth.
G Activity decreases in more acidic (pH
4) and alkaline (pH 9) environments,
where enzyme function is less
effective or inhibited.

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 Optimal Temperature

G Salivary amylase exhibits maximum

activity at a temperature of 37°C
(normal body temperature).
G Low temperatures (0°C) slow down
enzyme activity significantly, reducing
the rate of starch breakdown.
G At high temperatures (above 60°C),
the enzyme denatures, losing its
ability to function.

B. Summarizing how Extreme pH or Temperature

Affects Enzyme Efficiency.

Effect of Extreme pH:

Acidic (Low pH):

 At a very low pH (e.g., pH 4), salivary amylase undergoes structural

changes. This disrupts the enzyme’s active site, reducing its ability to
bind to the starch substrate.

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  The enzyme becomes less efficient or inactive because the ionic
bonds and hydrogen bonds maintaining its structure are disrupted.

Alkaline (High pH):

 At high pH levels (e.g., pH 9), the enzyme's shape also alters, leading
to a decrease in catalytic efficiency.
 In extreme alkaline conditions, the enzyme may denature,
permanently losing its functionality.

Effect of Extreme Temperature

Low Temperatures:

 At temperatures close to 0°C, enzyme activity slows down

significantly because molecular movements decrease.
 The enzyme remains structurally intact, but the rate of reaction is
low due to reduced collision frequency between enzyme and
substrate.

High Temperatures:

 Temperatures above 60°C can cause the enzyme to denature, where

the protein structure unfolds and the active site loses its shape.
 Denatured enzymes cannot catalyze reactions, so starch digestion
effectively stops.

31
 Conclusion
This project illustrates the critical role of salivary amylase, a
biological catalyst, in the hydrolysis of starch (a polysaccharide)
into maltose (a disaccharide), and ultimately glucose, which can
be absorbed by the body for energy production. The experiment
demonstrated that amylase exhibits optimal catalytic activity
under specific chemical conditions, namely at a neutral pH (~7)
and 37°C. Deviations from these conditions—such as lower or
higher pH or non-physiological temperatures—were shown to
significantly reduce the enzyme's efficiency by affecting its active
site, and thus, its ability to break the glycosidic bonds in starch.

The practical relevance of these findings is evident in daily life, as

changes in pH (e.g., consumption of acidic foods) or body
temperature (e.g., during fever) can impact the catalytic efficiency
of enzymes involved in digestion. Proper enzyme-substrate
interaction relies on maintaining optimal environmental
conditions for amylase to function efficiently.

In conclusion, the enzymatic breakdown of starch by salivary

amylase follows the principles of enzyme kinetics and
biochemical catalysis, providing a deeper understanding of
digestion at the molecular level. This study highlights the
importance of optimal chemical conditions in digestive enzyme
activity and has applications in nutritional science and healthcare,
especially for improving digestion and nutrient absorption through
proper diet and health management.

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 Bibliography

WEBSITES

1. LibreTexts Chemistry: 5.1: Starch and Cellulose - Chemistry LibreTexts

2. Byju’s: Starch - Definition, Formula, Uses, & Facts with Examples
(byjus.com)
3. Topper: Enzyme: Structure, Function and Classification with Videos,
Examples (toppr.com)
4. Microbe Notes: Starch: Structure, Composition, Properties, Uses,
Types (microbenotes.com)

BOOKS

1. NCERT (2020). Chemistry Textbook for Class 12. National Council of

Educational Research and Training (NCERT).
2. EZEE Notes Chemistry for class 12 (with new 2024-25 chemistry
syllabus)
3. BHARAT PANCHAL chemistry book class 12 | IIT-JEE | NEET | CUET.

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