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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:
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(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)
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