Bio Lab Assignment

Research Question:
The effect of temperature on the enzyme activity of Catalase. The catalase will be
used on potato shavings. The amount of oxygen released will be measured. The
range of temperatures used will start from 10 to 80 with an interval at every
10. 8 pieces of potato measuring 2x2x2 mm will be used. All the samples will be
given 3 minutes for the reaction.

Background:
Enzymes are biological catalysts that speed up a chemical reaction within the body
without getting used up. They can anabolize (build up) or catabolize (break down) a
reaction within or outside the cells. Enzymes work best at a certain temperature
(optimum temperature) and certain pH (optimum pH). For instance: the optimum pH
for pepsin is 1.5. Pepsin is an enzyme found within the stomach which helps in
digestion. The stomach is acidic because of the presence of HCL. Therefore, the
acidic condition of the stomach is very conducive for this reaction. Similarly, the
optimum temperature for most of the enzymes found within our body is the body
temperature i.e. 37. This is also true for catalase.
Hydrogen Peroxide is produced within our cells during metabolism. It is very toxic
and it must be removed from our body. Catalase helps in breaking down Hydrogen
Peroxide into oxygen and water. This oxygen and water is then used up by our body.

Independent Variable:
The independent variable in this experiment is the temperature. A wide range of
temperatures from 10 to 80 will be used.

Dependent Variable:
The amount of oxygen released is the dependent variable.

Controlled Variable:

Controlled Variable
1) pH

2) Volume and
Concentration of
catalase

3) Volume of potato
pieces

4) Time

Why should it be
controlled?

How should it be
controlled?

The pH needs to be
controlled because this
reaction is pH sensitive.
The enzyme has an
optimum pH i.e. it works
the best at that pH. If there
is a slight discrepancy in
the ph of the samples, the
outcome will be different.
The optimum pH for
catalase is 7 (neutral)
The volume and the
concentration of catalase
should remain constant. A
slight difference in the
volume or the
concentration of the
samples can lead to a
wrong result.

It can be controlled by
adding a pH buffer to all
the samples. This will make
the pH of all the samples
constant.

The volume of the potato

pieces need to remain
constant. Large pieces and
the reaction will slow down
and small pieces will cause
the reaction to occur
quickly.
The time given will have to
remain constant for all the
samples. If a sample is
given too much of time in

Clean and dry the test tube

thoroughly. Carefully
measure the volume of the
catalase using a pipette
before putting it in the test
tube. Ensure that the same
concentration of catalase is
being used for all the
samples.
Use a ruler to accurately
measure the size of the
potato pieces.

Each reaction will collect

data for 3 minutes after the
H2O2 is poured into the
catalase solution. After 3

comparison to the rest,

then excess oxygen will be
collected for that sample
thus giving a wrong result.

minutes, I will measure the

amount of oxygen that is
released.

Hypothesis:
As the temperature increases from 10 to 40 the amount of oxygen released will
rise. This is because the reaction will receive more energy so the number of
successful collisions will increase. The reaction occurs when the Hydrogen Peroxide
fits in the active site of the Catalase. The Catalase then catalysis the reaction. The
Catalase changes its shape slightly to accommodate the Hydrogen Peroxide. This is
known as the induced fit hypothesis. The optimum temperature for Catalase is 37.
So after 40, the amount of oxygen released will fall. This is because the enzyme
will slowly get denatured. At around 70, the enzyme will get denatured
completely. The enzyme wont function; therefore no oxygen will be released. The
high temperature will cause a conformational change in the shape of the active site
of the Catalase. Therefore, the Hydrogen Peroxide (Substrate) will no longer fit in
the active site of the Catalase.

Result:
Sample

Temperature

10

2
3
4
5
6

20
30
40
50
60

Volume of oxygen
released
4.8 cm3
7 cm3
10 cm3
15 cm3
9 cm3
4 cm3

70

0 cm3

80

0 cm3

Apparatus:

Beaker
Thermometers
Bunsen Burners
8 Test Tubes
Pipette
Gas Syringe
Delivery Tube
Small Knife

Methodology:
I conducted each experiment one after the other. I started by cleaning the 8 test
tubes thoroughly. Then, with the help of a ruler and a knife, I cut 8 pieces measuring
2x2x2 mm from a potato. I connected the delivery tube to the gas syringe. I did the
first experiment at room temperature i.e. 20. I used a Pipette to pour 2 ml of
catalase into the first test tube. Then, I put the first potato piece into the test tube.
Immediately, I connected the delivery tube with the test tube and started the stop
clock. After 3 minutes, I measured the amount of oxygen that was released. Then, I
noted down the result. I repeated the same with all the temperatures. I used a hot
water bath, fridge and thermometer to bring each test tube to the correct
temperature.