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Studying the effect of pH on the activity of pepsin

 oblem statement :
What are the effects of pH on the activity of pepsin?

Va iables
Manipulated variable : pH of medium
Responding variable : Rate of enzymatic reaction
Fixed variable : Volume and concentration of albumne suspension

Hypothesis
The optimum pH for the activity of pepsin is an acidic medium of pH 3

Mate ials
Albumen suspension, 1% pepsin solution, 0.1 M hydrochloric acid, 0.1 M sodium
hydroxide solution and distilled water.

Appa atus
Beakers, droppers, thermometers, test tubes, 5 ml syringes, pH paper, wire gauze,
a stopwatch and a test-tube rack.

Technique
Observe and record the condition of mixture before and after 20 minutes.

 ocedu e
1. An albumen suspension is prepared by mixing the egg white from an egg, with
500 ml of distilled water. The suspension is boiled and cooled down. Large
particles are removed by using glass wool.
2. Three test tubes are prepared and labeled P, Q and R respectively.
3. A syringe is used to add 5 ml of the albumen suspension to each test tube.
4. The solution below are added to the test tubes.

P: 1 ml of 0.1 M hydrochloric acid + 1 ml of 1% pepsin solution.

Q: 1 ml of distilled water + 1 ml of 1% epsin solution.

R: 1 ml of 0.1 M sodium hydroxide
solution +1 ml of 1% pepsin

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Figure 1

5. A piece of pH paper is dipped into each test tube and the pH value is
determined and recorded.
6. All the test tubes are immersed in a water bath maintained at 37 C for 20
minutes. (Figure 1)
7. The conditions of the mixtures are observed at the beginning of the
experiment and again after 30 minutes.
8. The result are recorded in a table

Result:

Mixture
Test tube pH At the beginning of the After 20
experiment minutes
P 3 Cloudy Cloudy
Q 7 Cloudy Cloudy
R 8 cloudy Cloudy

üiscussion:
1. The test tube are immersed in the water bath maintained at 37 C because
that is the optimum temperature for the action of pepsin
2. Pepsin, hydrolyses albumen (a protein) into polypeptides in an acidic medium.
This solution turns clear because polypeptides are soluble in water.
3. (a) The pH condition in test tube P (pH 3) is optimum for the function of
pepsin
(b) This is because the contents of test tube P become clear at the end of
the experiment.
(c) this shows that albumen has been digested or hydrolysed by pepsin.
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4. (a) The content of test tubes Q and R are still cloudy at the end of the
experiment
(b) This shows that a neutral (test tube Q) and an alkaline pH (test tube R) are
not suitable for the activity of pepsin

 onclusion :
The activity of pepsin is affected by the pH of its medium. An acidic medium
(pH 3) is the most suitable medium for pepsin to function effectively.
The hypothesis is accepted