LAB #10

Title: ENZYMES
Aim: To investigate how pH, temperature, substrate concentration and enzyme concentration
affect the activity of the catalase enzyme.

List all apparatus and materials used.

Method:
1. Remove the skin of the potato and cut into minute pieces. Homogenize with an equal
volume of water in a blender.

2. Dice the liver and homogenize with distilled water in a blender.

3. Place 1cm3 of liver into boiling tube A (take the pH using pH paper) and add 5cm3 of
hydrogen peroxide.

4. Quickly place a rubber bung connected with tubes over the boiling tube and place the
open end of the tube into a tube of water. Ensure that the rubber bung or tube does not
come into contact with the testing solution.

5. Count the number of bubbles produced in a one minute.

6. Repeat the procedures 3-5 using the potato.

7. Record all observations and results.

8. Place 1cm3 of liver and potato into boiling tubes C and D then add 8 cm3 of hydrogen
peroxide. Place the rubber bung over the tube and count the number of bubbles
produced.

9. Place 1cm3 of liver and potato into 2 different labelled boiling tubes.

10. Expose boiling tube E and F to 65⁰c water bath for 2 minutes. Remove from heat;
allow cooling and adding 5cm3 of hydrogen peroxide.

11. Quickly place a rubber bung connected with tubes over the boiling tube and place the
open end of the tube into a tube of water.

12. Count the number of bubbles produced in a one minute.

13. Place 1cm3 of liver and potato into 2 different labelled boiling tubes.

14. Add 1cm3 of HCl to boiling tube G and H. Take the pH of each solution. Add 5cm3 of
hydrogen peroxide to each tube.

15. Quickly place a rubber bung connected with tubes over the boiling tube and place the
open end of the tube into a tube of water.

16. Count the number of bubbles produced in a one minute.

 17. Tabulate all results.

Diagram: set up of apparatus.

Description of the reaction which occurred as soon as the hydrogen peroxide was added to
the tissue and oxygen evolution under each condition for both liver and potato.
State the control used and the manipulated variables in each experiment.
Discussion:
 Give background information on enzymes e.g. what it is, how it works, factors which
affect its functioning.
 Say that catalase is found in both liver and potato.
 Reaction broke down hydrogen peroxide into water and oxygen (state in worded and
chemical equation form).
 Bubbles represented the bubbles formed which indicated the rate of production of
oxygen hence the breakdown of hydrogen peroxide.
 Were the results expected? Why?
 Theoretically what are the known concentrations of the enzyme in both potato and
liver. Relate to results (in experiment A, did the liver or potato produce the higher
number of bubbles? Why?)
 Use the first experiment (5cm3 of hydrogen peroxide at RT and pH 6) of liver and
potato for comparisons – experiment A.
 Compare results of potato and liver when temperature and pH were varied with
experiment A. For example, when temperature increased, the number of oxygen
evolved decreased from 127 in experiment A to 20.
 Why did results vary in varying conditions, that is, how did each condition affect the
activity of the enzymes. Mention activation energy, kinetic energy, active site, bonds
in enzyme structure.

Sources of Error, Precautions taken, One Limitation must be stated.

Criteria : A / I Marks Awarded

A Definition of related terms such as what is an enzyme and how do they 1

work.
B Briefly discussing relevant theory of experiment such as what type of 1
reaction occurs between catalase and hydrogen peroxide, what type of
gas is the bubbles, what factors affect enzyme activity.
C State the theoretical basis of the experiment. 1
D Describing appropriate trends 1
If only data / Tables / graphs stated award 0 mk only
Discuss fully what is the relationship between the results and the aim
Giving full details of what the results were in relation to background
theory.
Points should address comparison between experiments and why the
results were expected (relate to bonds affected within protein
structure of enzyme)
E Temperature 1
F pH 1
G Substrate and enzyme concentration 1

H Explanation of limitations / precautions / sources of error that may 2

have impacted the lab in some way.
(1mk each)
I Reflection stated- how can the experiment be changed and improved 1
J Conclusion should be a Concise summary statement 1
K Conclusion Must be related to the aim 1
TOTAL 12
POSSIBLE
MARKS