4.2 Studying the effects of pH on the activity of pepsin Problem statement: What are the effects of pH on the activity of pepsin? Hypothesis: The optimum pH for activity of pepsin is an acidic medium of pH 3. Variables: MV – pH of medium RV – Rate of enzymatic reaction CV – Volume and concentration of albumen suspension, volume and concentration of pepsin solution and temperature of medium Materials and apparatus: Albumen suspension, 1% pepsin solution, 0.1M hydrochloric acid, 0.1M sodium hydroxide solution, distilled water, beakers, droppers, thermometer, test tubes, 5 ml syringe, pH paper, wire gauze, a stopwatch and test tube rack Procedure:

1. 2. 3. 4.


(P1) Prepare an albumen suspension by mixing the egg white from an egg with 500 ml of distilled water. Boil and cool down the suspension. Remove the large particles using a glass wool. (P1) Prepare and label three test tubes P, Q and R respectively. (P1) Add 5 ml of albumen suspension to each test tube using a syringe. (P1, P2) Add the solution below to the test tube: P: 1 ml of 0.1M hydrochloric acid + 1 ml of 1% pepsin solution Q: 1 ml of distilled water + 1 ml of 1% pepsin solution R: 1 ml of 0.1M sodium hydroxide solution + 1 ml of 1% pepsin solution (P1, P2) Dip a piece of pH paper into each test tube. Determine and record the pH value. (P1, P4)

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6. Immerse all the test tube in the water bath which is maintained at 37 o C for 20 minutes. (P2, P5) 7. Observe the condition of the beginning of the experiment and again after 20 minutes. (P3) 8. Record the result in the table. (P3) Results: Test tube P Q R pH 3 7 8 Mixture At the beginning of the experiment After 20 minutes

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