ENZYME PRACTICAL III

Sec 3 Biology

Name:……………………………………….( ) Class:…… Date:……………

Aim: To investigate show the effect pH on enzyme action.

This experiment is based on the fact that egg albumen (egg white) contains proteins.
Pepsin is a protein-digesting enzyme produced by the stomach.
Trypsin is a protein-digesting enzyme produced by the pancreas

Protein Polypeptide (shorter chains of proteins)

Pepsin/ Trypsin

Material and Methods

a. To each of the four test tubes, A, B, C and D, add 3cm3 of cooled egg
white suspension. Place the tubes in 37oC.
b. Make up the contents of the 4 tubes as shown in the table below.
c. Use dilute hydrochloric acid (0.1M), pepsin (5%) and sodium carbonate
solution (2%)
d. Record your observations at the beginning of the experiment.
e. Record your observations after 20 minutes.
f. Record down your conclusions for each tube.
[Note: If the egg white suspension becomes clear, it means that protein has been
digested or hydrolysed]

Results

Test tube A B C D
Contents 3cm3 egg white 3cm3 egg white 3cm3 egg white 3cm3 egg white
+ 10 drops dilute + 10 drops dilute + 10 drops dilute + 10 drops
HCL HCL Na2CO3 distilled water
+ 3cm3 pepsin + 3cm3 distilled + 3cm3 pepsin + 3cm3 pepsin
water
Observation

Conclusion

Adapted from Lam Peng Kwan ‘Practical Biology A Course for O-levels’ 1
Created by Daniel Lim, Dunman Secondary School
Pepsin is one of the enzymes found in the stomach’s gastric juice, which is a dilute
solution of hydrochloric acid of pH = 2.

The function of the hydrochloric acid

- stops the action of salivary amylase
- changes the inactive forms of enzymes in the gastric juice to the active forms
- provides a slightly acidic medium suitable for the action of gastric enzymes
- kills germs and potential parasites that enters the digestive system.

In the space below, sketch a graph showing the effect of pH on the rate of reaction
catalysed by pepsin. Indicate on the graph, the optimum pH as well as the pH of sodium
carbonate which is about 8.

In your own words, explain the nature of the curve in your graph in terms of the activity
of the effect of pH on pepsin.

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Adapted from Lam Peng Kwan ‘Practical Biology A Course for O-levels’ 2
Created by Daniel Lim, Dunman Secondary School
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Adapted from Lam Peng Kwan ‘Practical Biology A Course for O-levels’ 3
Created by Daniel Lim, Dunman Secondary School