The Effect of the Enzyme Catalase on Hydrogen Peroxide When the Temperature is Changed

The Effect of the Enzyme Catalase on Hydrogen Peroxide When the Temperature is Changed

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The Effect of the Enzyme Catalase on Hydrogen Peroxide When the Temperature is Changed

Plan:

Chemical reactions take place in all living cells. All of these
reactions are enzyme-driven. Some of these reactions produce
by-products. One by-product is Hydrogen Peroxide (H2O2). It is a
poison and must be removed from the cells. The cell produces an enzyme
called Catalase to break down the Hydrogen Peroxide into water and
oxygen. This experiment will investigate the effect of temperature on
the rate of the breaking down of Hydrogen Peroxide by Catalase. I will
do this by measuring how much oxygen is produced when Catalase and
Hydrogen Peroxide react.

I predict that as the temperature increases, the rate of the reaction
will increase. I think that when the temperature is doubled, so the
reaction rate will also double. This is due to the particle theory. As
the temperature rises, the particles move faster and collide more
often. This gives the "lock and key" reaction more chance to occur.
Therefore, as it is a temperature coefficient with a Q10 of 2, if the
temperature is raised by 10ºC, the particles will move doubly fast,
collide twice as often and the rate of reaction will double. There
will be a point when this relationship isn't true. This will be at
around 60ºC when the enzyme denatures.

To make this a fair test, we need to keep the same all variables that
could affect the outcome except for the one being tested, temperature.
So the pH, amount of solutions used, concentration of solutions,
apparatus etc. must all remain the same.

As my source of Catalase, I will use potato. This is easier to cut and
measure than other sources such as liver, which would be hard to cut
into equal pieces. The potato "chip" will be pre-warmed in a boiling
tube in a water bath of each temperature for 10 minutes, as will 20cm3
of H2O2. Once the desired temperature is reached, the chip and H2O2
will be mixed in the boiling tube, shook a couple of times and placed

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back in the water bath with a rubber bung on top with a tube coming
out. This tube is connected to a burette of water whose bottom is
submerged in a bowl of water. I will measure the amount of oxygen
produced by recording the starting level of the burette and the end
level, and finding the difference, making sure I read it level with
the markings to avoid parallax error. The time will run for 3 minutes
from when the first bubble is seen rising up the burette. This will
all be done twice.

I will keep this experiment safe by adhering to all normal laboratory
rules, such as safety glasses, not running in the lab and so on.


The apparatus will be set out as shown below:

Preliminary Work

I did a preliminary experiment to give me an idea as to which values
and range I should use for my temperatures, and to check if the
equipment and setup I had chosen was suitable for the experiment.

Here are the results of that experiment:

Temperature (°C)

Oxygen Produced (cm3)

30

2.7

40

2.4

50

4.5

60

2.8

70

1.4

These results are not ideal, as they have only been done once so no
averages can be found to improve accuracy, but they give me an idea as
to which values to use as my temperatures. It does show however that
Catalase seems to start denaturing at around 60°C, and that it appears
to work best at around 50°C.

Table Of Results For Main Experiment

Temperature

(in °C)

Start Level

on burette

End Level

on burette

Oxygen
Produced (cm3)

Average of Expt. 1+2 (cm3)

2.5 Expt.1

47.1

44.3

2.8

2.3

2.5 Expt.2

46.4

44.6

1.8

-------------

Room Expt.1

46.3

41.5

4.8

4.65

Room Expt.2

45.3

40.8

4.5

-------------

41 Expt.1

40.4

35.0

5.4

5.45

41 Expt.2

41.0

35.5

5.5

-------------

51 Expt.1

34.3

29.8

4.5

4.5

51 Expt.2

35.0

30.5

4.5

-------------

59 Expt.1

28.7

28.5

0.2

0.2

59 Expt.2

30.3

30.1

0.2

-------------

69 Expt.1

27.0

27.0

0.0

0.0

69 Expt.2

29.0

29.0

0.0

-------------


Analysis

The table and graph show that as temperature increases, so does the
speed of the reaction, as the amount of oxygen produced is getting
bigger. This is because as the temperature rises, the particles have
more energy and move faster, and therefore collide more often, causing
the reaction to take place more times (this is the particle theory.)
This relationship is true, except for when it goes from around 55°C
down to around 70°C, the rate gets slower. This is due to the fact
that the enzyme is denaturing during this period.

The graph does not appear to show any kind of quantitative result, but
it does show that temperature rise does increase rate of reaction, and
it also shows the area where the enzyme is denatured - the sharp drop
between around 50-60°C, slowly down to 0.0cm3 at 69°C.

Evaluation

I think that my results are quite accurate, as I took care when
recording the results, measured amounts carefully and carried out the
experiment as carefully as possible. I do not think there are any
anomalous results, as the points lie on or very near to the best-fit
line.

This experiment has some faults. Firstly, as soon as the Catalase
comes into contact with the H2O2, it starts to lose its concentration
as the Catalase is used up in the reaction and therefore becomes less
concentrated towards the end of the reaction. This cannot really be
helped unless there was a way of keeping the concentration the same
all the way through the whole of the experiment, maybe by adding more
potato as it starts to run out.

The equipment could be improved. A syringe with a needle may be used
to inject the H2O2 through the bung directly into the boiling so that
there is no lag time for the oxygen to escape while the bung is being
put on the tube. I think the evidence I have is strong enough to
support the conclusion "as the temperature rises, the rate of reaction
increases except where the enzyme is denatured," but not strong enough
to give a quantitative result. This could be achieved by using more
sophisticated equipment so that no oxygen was lost at all, and testing
it several times instead of just two or three times.

More evidence could be acquired by carrying out more experiments. This
could be to get a more accurate reading of points not tested here,
such as 55°C or 15°C (these may be read off the best fit line on the
graph, but would not be as accurate as actually testing it).
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