Experiment 7

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.
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 37oC

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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.: _______________________________ Score: _______________

Name: __________________________________ Date Performed: _______
C/Y/S: ___________________________________ Date Submitted: _______

Experiment 7
ENZYME ACTIVITY

DATA and RESULTS

A. Influence of pH on Enzymatic Activity

pH Intensity of the Blue Color

Test Tube No. 1
Test Tube No. 2
Test tube No. 3

B. Influence of Temperature on Enzyme Activity

Temperature Intensity of the Blue Color

Test Tube No. 1
Test Tube No. 2
Test tube No. 3

POST LABORATORY DISCUSSION

1. Why is there a need to place the test tubes in water heated to 37oC ?
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2. How is enzymatic activity related to the differences in the intensity of the colors obtained?
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3. Based on the activity, what is the best pH and temperature for optimum enzymatic activity?
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4. Give examples of other enzymes.

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CONCLUSION

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CLINICAL APPLICATIONS

1. What is the importance of enzymes in body processes such as digestion and metabolism?
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2. Can a patient's disease/illness be diagnosed by means of enzymatic activity? Give examples of

diseases resulting from abnormal enzymatic activities.
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3. How does fever affect enzymatic activity?

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