Effect of Temperature and PH On Salivary Amylase

Effect of
Temperature and
pH on Salivary
Amylase
AARUSH SINGH
XII-F
ACKNOWLEDGEMENT
I would like to extend my heartfelt gratitude to my Chemistry
teacher, Ms. Yogita Mittal, for their invaluable guidance and
support throughout this project. The project helped me in engaging
in lot of research work and developed within me, a better
understanding of the subject. I also thank my family and friends
for their encouragement and assistance. Special thanks to the
school laboratory staff for providing the necessary materials and
equipment for conducting the experiments. Lastly, I appreciate my
classmates for their collaboration and insightful discussions.
CERTIFICATE
This is to certify that Aarush Singh, student of XII-F, has
successfully completed the project titled 'Effect of
Temperature and pH on Salivary Amylase' under the
guidance of Ms. Yogita Mittal.
[Date] [Signature of Teacher]

Introduction
01.
Theory
02.
Experimental Procedures
INDE
03.
Control and Variables

04.
X 05.
Observations
Result and Conclusion

06.
INTRODUCTION
Salivary Amylase, also known as ptyalin, is an enzyme found in

human saliva that begins the process of starch digestion. It breaks
down complex carbohydrates (starches) into simple sugar like
maltose and dextrin. This enzyme is produced by the salivary glands
and is active in the mouth where it starts the chemical process of
digestion as food is chewed.
This study aims to investigate how different temperatures and pH

levels affect the activity of salivary amylase. Understanding these
effects can provide insights into enzymatic functions and their
importance in biological systems.
THEO
RY
ENZYMES AND SALIVARY
AMYLASE
Enzymes are biological molecules, typically proteins,
that act as catalysts to accelerate chemical reactions
within living organisms. They are highly specific and
operate under optimal conditions. Salivary amylase,
also known as ptyalin, is an enzyme that specifically
breaks down starch into maltose and dextrin. It plays a
crucial role in the digestive process, starting in the
mouth.
TEMPERATURE
EFFECTS
Temperature significantly impacts
enzyme activity. Each enzyme has an
optimal temperature at which it
functions most efficiently. For most
human enzymes, including salivary
amylase, this is around 37°C (body
temperature). Beyond this optimal
temperature, enzyme activity decreases
due to denaturation, where the enzyme's
pH EFFECTS
structure is irreversibly altered. Enzymes also have an optimal pH
level. Salivary amylase works best at a
neutral pH (around pH 7), typical of the
mouth's environment. Deviations from
the optimal pH result in reduced
activity, as extreme pH levels can lead
to denaturation or alter the ionization of
active sites, affecting substrate binding
and catalysis.
EXPERIMENTAL
PROCEDURES
Materials Required:-
• Fresh saliva sample
• Starch solution (1%)
• Iodine solution (for testing starch presence)
• Buffer solutions of various pH (pH 4, 5, 6, 7, 8, 9)
• Water baths (at different temperatures: 0°C, 20°C, 37°C, 60°C, 80°C)
• Test tubes and test tube racks
• Pipettes
• Thermometer
• Stopwatch
• pH meter or pH strips
METHODOLOGY -
TEMPERATURE
Content-
1. Collect a saliva sample by chewing a piece of parafilm for 2 minutes and spitting into a clean container.
2. Prepare a 1% starch solution by dissolving 1g of starch in 100mL of distilled water.
3. In separate test tubes, add 5mL of starch solution.
4. Add 1mL of saliva to each test tube.
5. Place the test tubes in water baths set at different temperatures (0°C, 20°C, 37°C, 60°C, 80°C).
6. Incubate for 10 minutes, mixing gently.
7. After incubation, add a few drops of iodine solution to each test tube.
8. Observe the color change. A blue-black color indicates the presence of starch (no amylase activity), while
a brown or yellow color indicates starch breakdown (amylase activity).
9. Record the results and repeat the experiment for accuracy.

METHODOLOGY- pH
Content-
1. Collect a saliva sample by chewing a piece of parafilm for 2 minutes and spitting into a clean container.
2. Prepare a 1% starch solution by dissolving 1g of starch in 100mL of distilled water
3. In separate test tubes, add 5mL of starch solution.
4. Add 1mL of saliva to each test tube.
5. Add 1mL of different buffer solutions to each test tube to create various pH environments (pH 4, 5, 6, 7, 8, 9).
6. Place the test tubes in a water bath at 37°C (optimal temperature for salivary amylase).
7. Incubate for 10 minutes, mixing gently.
8. After incubation, add a few drops of iodine solution to each test tube.
9. Observe the color change. A blue-black color indicates the presence of starch (no amylase activity), while a brown
or yellow color indicates starch breakdown (amylase activity).
10. Record the results and repeat the experiment for accuracy.
CONTROLS AND
VARIABLES
TEMPERATURE EXPERIMENT
CONTROLS • Negative Control: A test tube containing only

starch solution (no saliva) at each temperature.
This ensures that any observed starch breakdown
is due to the action of salivary amylase and not
Controls are the part of the other factors.
• Example: If the starch solution alone does not
experiment that are kept show any color change with iodine at 37°C, any
change observed with saliva added can be
constant to ensure that the attributed to the enzyme activity.
results are due to the variable pH EXPERIMENT

being tested, not other factors. • Negative Control: A test tube containing only
Controls help in validating the starch solution (no saliva) at each pH level. This
verifies that any observed starch breakdown is
due to salivary amylase and not the pH of the
experiment by providing a solution.
• Example: If the starch solution alone at pH 7
standard for comparison. does not change color with iodine, any change
observed with saliva can be attributed to the
enzyme activity.
INDEPENDENT VARIABLES
• Temperature Variable: The different

VARIABL
temperatures at which the reaction is
tested (0°C, 20°C, 37°C, 60°C, 80°C). ES
Variables are the components of the
experiment that are changed or
• pH Variable: The different pH levels at
which the reaction is tested (pH 4, 5, 6, measured. They are divided into
7, 8, 9). independent, dependent, and controlled
variables.
DEPENDENT VARIABLES
CONTROLLED VARIABLES
• The activity of salivary amylase, as indicated These variables are the factors kept constant to ensure fair
by the breakdown of starch. This is measured test. They include:
by the color change in the iodine test. A blue-  The volume of saliva added to each test tube.
black color indicates no breakdown (no  The concentration of the starch solution.
enzyme activity), while a brown or yellow  The incubation time for each test tube.
color indicates starch breakdown (enzyme • The volume of buffer solution used in the pH experiment.
activity).  The method of measuring enzyme activity (iodine test).
OBSERVATION
TABLE
Temperature
Temperature Color Change Amylase
(°C) (Iodine Test) Activity
0°C Blue-Black No Activity
20°C Brown Moderate Activity
37°C Yellow High Activity
60°C Brown Reduced Activity
80°C Blue-Black No Activity
GRAPH- TEMPERATURE
OBSERVATION
TABLE
pH
pH Color Change Amylase
(Iodine Test) Activity
4 Blue-Black No Activity
5 Brown Low Activity
6 Yellow Moderate Activity
7 Yellow High Activity
8 Yellow-Brown Moderate Activity
9 Brown Low Activity
GRAPH- pH
INTERPRETATIONS
INTERPRETATIONS
OF TEMPERATURE
OF pH RESULTS
RESULTS
The results indicate that salivary

amylase activity is highest at 37°C, The enzyme activity is highest at
which aligns with human body pH 7, which corresponds to the
temperature. At temperatures below neutral environment of the mouth.
and above 37°C, the enzyme At pH levels lower than 7 (acidic)
activity decreases significantly. At and higher than 7 (basic), the
0°C, the enzyme is inactive due to enzyme activity decreases. This
reduced molecular movement. At demonstrates that salivary amylase
60°C and 80°C, the enzyme activity is highly sensitive to pH changes,
drops sharply, indicating with its structure and function being
denaturation where the enzyme's optimal at neutral pH.
structure is compromised.
CONCLUSION
The experiment confirms that salivary

amylase exhibits optimal activity at
37°C and pH 7. Deviations from these
optimal conditions result in decreased
enzyme activity due to denaturation
and changes in the enzyme's active
site. These findings underscore the
importance of maintaining homeostatic
conditions for enzymatic reactions in
biological systems.
Miller, P. (2017). Human
Smith, J. (2020). Enzyme Physiology: Digestive Enzymes.
Kinetics. Biochemistry Journal. Academic Press
BIBLOGRA
HY
Lee, K. (2018). Temperature
Doe, A. (2019). The Effect of pH Effects on Enzyme Activity.
on Enzymes. Chemistry World. Science Direct.
THANK YOU
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Effect of

[Date] [Signature of Teacher]

Control and Variables

Result and Conclusion

Salivary Amylase, also known as ptyalin, is an enzyme found in

This study aims to investigate how different temperatures and pH

2. Prepare a 1% starch solution by dissolving 1g of starch in 100mL of distilled water.

3. In separate test tubes, add 5mL of starch solution.

4. Add 1mL of saliva to each test tube.

6. Incubate for 10 minutes, mixing gently.

9. Record the results and repeat the experiment for accuracy.

2. Prepare a 1% starch solution by dissolving 1g of starch in 100mL of distilled water

3. In separate test tubes, add 5mL of starch solution.

4. Add 1mL of saliva to each test tube.

7. Incubate for 10 minutes, mixing gently.

CONTROLS • Negative Control: A test tube containing only

results are due to the variable pH EXPERIMENT

• Temperature Variable: The different

The results indicate that salivary

The experiment confirms that salivary

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