Experiment 6

ENZYME ACTIVITY

OBJECTIVE

To determine how enzymatic activity is affected by changes in pH and temperature.

MATERIALS

9 test tubes water bath Reagents:

test tube rack wire gauze 1% starch solution
10-mL graduated cylinder tripod 0.2 M NaCl
medicine dropper thermometer 0.01 M Iodine solution
2 100-mL beakers test tube brush buffer solutions (pH 3, 7
bunsen burner test tube holder (and 10)
lukewarm water
ice

PROCEDURES

A. Preparation of Amylase Solution

1. Rinse the mouth with clean drinking water.

2. Gargle with lukewarm water for 1 minute to induce the release of enzyme in the
mouth.
3. Collect the washing in a beaker. This serves as the diluted solution of amylase which
will be used for procedures B and C.

B. Influence of pH on Enzyme Activity

1. Prepare a mixture of 5 mL 1% starch solution, 1 mL 0.2 M NaCl and 1 drop of iodine

solution in 3 separate test tubes.
2. Add 2 mL of pH 7 buffer solution in test tube no. 1. In test tube no. 2, add 2 mL of pH
3 buffer solution and 2 mL of pH 10 buffer solution to test tube no. 3.
3. Place the 3 test tubes in a water bath whose temperature is maintained at 37oC.
4. Immediately add 3 mL of amylase solution to each test tube. Mix well. Let the
mixtures stand for a while.
5. Compare the intensity of the blue color produced in each test tube.
6. Record all observations in the report sheet.

C. Influence of Temperature on Enzymatic Activity

1. Prepare a mixture of 5 mL 1% starch solution, 1 mL 0.2 M NaCl and 1 drop of Iodine

solution in three separate test tubes.

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 2. Measure and place 5 mL of amylase solution in 3 other test tubes. Immerse the first
test tube in an ice bath. Maintain the temperature of the second test tube at 37 oC

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 using the water bath. Heat the third test tube directly in an open flame until the
temperature reaches 80oC. Use the thermometer to monitor the increase in the
temperature.
3. Simultaneously add the solutions prepared in procedure C1 to each test tube
prepared in procedure C2.
4. Compare the intensity of the blue color produced in the three test tubes.
5. Record all observations in the report sheet.

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 REPORT SHEET
Group No: 1 Score:
Name: Sean Jodi M. Cosepe Date Performed:
C/Y/S: BSN 1A Date Submitted:

Experiment 6
ENZYME ACTIVITY

DATA and RESULTS

A. Influence of pH on Enzymatic Activity

pH Intensity of the Blue Color

Test Tube No. 1 pH 7 (2 drops) Dark Blue
Test Tube No. 2 pH 4 (2 drops) Dark Blue
Test tube No. 3 pH10 (2 drops) Dark Blue

B. Influence of Temperature on Enzyme Activity

Temperature Intensity of the Blue Color

Test Tube No. 1 37oC Regular Color Violet
Test Tube No. 2 37oC Darker
Test tube No. 3 More than 37oC Colorless

POST LABORATORY DISCUSSION

1. Why is there a need to place the test tubes in water heated to 37oC ?

There is a need to place the test tubes in water heated to 37 oC to simulate body temperature.

2. How is enzymatic activity related to the differences in the intensity of the colors obtained?

Under different circumstances, such as pH and temperature, enzymatic activity can have varied
degrees of activity, which can impact how effectively an enzyme function variations in this situation
can result in variation in color intensity when the enzyme's activity is adjusted.

3. Based on the activity, what is the best pH and temperature for optimum enzymatic activity?

Based on the activity, the best pH is pH7 and best temperature is 37oC for optimum enzymatic
activity.

4. Give examples of other enzymes.

Example of enzymes includes digestive enzymes like amylase, proteases, lipases, and nucleases.

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 CONCLUSION

Identifying enzyme activity and its impacts on pH and temperature is critical for expanding
scientific knowledge, generating practical applications, and solving real-world problems in
sectors ranging from healthcare to biotechnology and environmental research. It helps
researchers and businesses to optimize operations, enhance products, and create new
solutions.

CLINICAL APPLICATIONS

1. What is the importance of enzymes in body processes such as digestion and metabolism?

Enzymes are biological catalysts that help and control a variety of chemical processes in the body.
Enzymes break down complex food molecules into simpler, more absorbable ones during digestion.
Amylase is an enzyme that breaks down carbohydrates (starches) into simpler sugars, proteases are
responsible for breaking down proteins into amino acids, lipases are responsible for breaking down
dietary fats (lipids) into fatty acids and glycerol, and nucleases are responsible for breaking down
nucleic acids (DNA and RNA) in our food into their constituent nucleotides. Enzymes are required
for energy generation, biomolecule synthesis, detoxification, homeostasis maintenance, and
cellular respiration.

2. Can a patient's disease/illness be diagnosed by means of enzymatic activity? Give examples of

diseases resulting from abnormal enzymatic activities.

Yes, enzyme activity in the body can be used to detect or identify specific diseases or disorders.
Lysosomal storage disorders (LSDs), phenylketonuria (PKU), hemolytic anemias, Wilson's disease,
acid-base problems, liver illnesses, and inborn errors of metabolism (IEMs) are examples of
diseases caused by aberrant enzyme activity.

3. How does fever affect enzymatic activity?

Fever, or an increase in body temperature, can have a major impact on enzyme activity. Enzymes
are extremely temperature sensitive, and their activity is frequently controlled by the body's
temperature. Prolonged, extremely high fevers can be harmful to the body, potentially deactivating
vital enzymes and causing other issues.