Problem Statement

Farmer john place a fungicide called copper sulfate on the potato plant and a few
days later he noticed that the fungus was no longer there. Plan and design an
experiment to explain the reason why the fungus was no longer there
Hypothesis
An increase in copper sulfate (competitive inhibitor) concentration would result in
the decrease of the rate of reaction of the enzyme (catalase found in potato).
Aim
To determine how different concentration of the inhibitor (copper sulfate) causes the
rate of reaction of the enzyme (catalase) to decrease.
Apparatus/ Materials
 Mortar and pestle
 One (1) retort stands and clamp
 Analytical balance
 One (1) 25ml burette
 Three (3) 10 ml measuring cylinder
 Two (2) potatoes
 One (1) stopper
Reagents:
 Hydrogen peroxide
 Ph7 buffer solution
 0.5,1,1.5,2,2.5 M copper sulfate
Method:
1. Peel the potatoes and slice into dices after which crush the potato dices using
a mortar and pestle.
2. Weigh 20g of the crush potato with an analytical balance
3. Set up the retort stand and clamp with the burette.
4. To the 25ml burette, add the crush potatoes and label it A
5. Using a measuring cylinder measure 5 ml of the ph7 buffer and add it to the
burette contain the crush potatoes.
 6. Using another measuring cylinder measure 5ml 0.5M copper sulfate and add
it to the burette followed by 0.5 M hydrogen peroxide.
7. Using a rubber stopper, stopper the burette and remove from clamp and shake
to in cooperate the different solution.
8. Attach the burette back to the clamp and observe the reaction
9. In a suitable table, record the froth level every 20 seconds for 2 minutes
10. Repeat steps 1-9 using the other concentrations of copper sulfate
(1,1.5,2,2.5M respectively) with the burette label from B-E respectively.
11.In a burette label F repeat steps 1-9 without the use of copper sulfate.
12.Plot a graph with rate of enzyme activity(cm3/s) against concentration (M) of
copper sulfate solution.
Variables
Independent variable: Concentrations of copper sulfate
Dependent variable: rate of froth level produced every 20 seconds for 2 minutes
Controlled variable:
1. Mass and volume of sample used
2. Concentration of hydrogen peroxide (0.5M)
Treatment of data
Time (s) Volume of Froth produced with varying
Conc. Of Lead Nitrate (cm^3)
0.5M 1M 1.5M 2M 2.5M
0
20
40
60
80
100
120

TABLE SHOWING THE VOLUME OF FROTH PRODUCED FOR

EVERY 20S FOR 2 MINUTES
 Conc. Of Rate of Enzyme Activity
Lead Nitrate [volume(cm^3)/time(s)]
(M)
0.5
1
1.5
2
2.5

TABLE SHOWING THE RATE OF THE ENZYME ACTIVITIES

Expected Results:
The lock and key theory utilizes the concept of an "active site." The concept holds that one
particular portion of the enzyme surface has a strong affinity for the substrate. The substrate is
held in such a way that its conversion to the reaction products is more favorable. If we consider
the enzyme as the lock and the substrate the key. The key is inserted in the lock, is turned, and
the door is opened and the reaction proceeds hence the burette without any copper sulfate would
have a normal rate of reaction that is the hydrogen peroxide would bind with the enzyme catalase
until the saturation point is reached.
However, when an inhibitor which resembles the substrate is present, it will compete with the
substrate for the position in the enzyme lock. When the inhibitor wins, it gains the lock position
but is unable to open the lock. Hence, the observed reaction is slowed down because some of the
available enzyme sites are occupied by the inhibitor. Therefore, increasing the concentration of
copper sulfate solution from 0.5M to 2.5M would cause a decrease in the rate of reaction of the
enzyme hence causing a decrease in the volume of froth produced every 20 seconds for 2
minutes.
A possible source of error would be the stopper was not inserted properly causing oxygen to
escape which would cause inaccurate results. A way to correct this is to ensure the stopper is
inserted properly so that the oxygen produce does not escape.