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Catalase
By: Morgan Haines, Grace Barton, Brett Cowan, Rachel Urban
Abstract:
The purpose of the report is to state the effect pH has on the enzyme, Catalase. The
enzyme Catalases purpose is to breaks down hydrogen peroxide that can harm the body. To test
the effect pH has on the enzyme Catalase is by measuring the pressure that is produced by the
chemical reaction when the enzyme is placed in a Hydrogen peroxide solution. The way of
comparing results is taking the slope of the pressure line through a 200 second test. The pH of
the hydrogen peroxide solution is changed each trial using pH buffers. As the pH went up during
the experiment the higher the slope was with the exception of one result that could be excluded
or redone for the best results. Using the data that was gathered during the experiment it was
shown the higher the pH was then more efficient the enzyme catalase was at breaking down the
Background:
The purpose of the experiment that we conducted was to find an answer to this question.
The question is this: How does changing the pH of the hydrogen peroxide solution affect the
efficiency of the catalase enzyme. Before a reliable experiment could be formulated/ conducted,
much research was done to obtain optimal information to help create the hypothesis and
experimental design.
First, the catalase chemical equation was researched and deciphered, as the reactants and
products were studied as well. Next, the meaning of a substrate and a product was studied to then
formulate a solid definition. Last, we ran a standard test of the reaction between catalase and
H2O2 to precurse later experiments. Also, keep in mind, that all information obtained was
written down in the PLTW lab journal. In the standard experiment, we tested the efficiency of the
catalase enzyme in these standard conditions: the enzyme was added to 50 mL of 1.5% H2O2
with a pH of 6.5. Through lots of research it was discovered that the optimal pH for catalase to
operate is 7. So, the pH of the H2O2 in the standard reaction was brought as close to 7 as
possible. The standard experiment showed how effectively catalase operated at its optimal pH
with no pH buffer.
Hypothesis:
If the pH of the hydrogen peroxide solution is changed, then the closer the pH is to seven,
the more efficient the catalase enzyme will be, because the optimal pH for the catalase enzyme is
7.
Materials:
fasten the flask to the ring stand. The flask was then positioned at the center of the
magnetic stirrer
6. The plastic tubing with two Luer-lock connectors was used to connect the two-hole
rubber stopper assembly to the Gas Pressure Sensor and the valve connected to the
micropipette
8. The flask was tightly sealed by twisting in the two-hole stopper connected to the Gas
Pressure Sensor
9. The stirrer was turned on at a speed of 4
10. Data collection was started
11. After 200 seconds had passed, data collection was stopped
12. The stopper was removed from the flask and the contents of the flask were
the 25mL graduated cylinder and 25mL of the 3% H2O2 solution was measure out in the
used to fasten the flask to the ring stand. The flask was then positioned at the center of the
magnetic stirrer
17. The plastic tubing with two Luer-lock connectors was used to connect the two-
hole rubber stopper assembly to the Gas Pressure Sensor and the valve connected to the
micropipette
19. The flask was tightly sealed by twisting in the two-hole stopper connected to the
Results:
Each test was ran with a 1.5% H2O2 solution and was measured by noting the pressure
(kPa) per second (s) and comparing the slopes that was produced in each line. The first test result
that was obtained was the standard. This was performed with a 1.5% H2O2 solution and slope
that was made was .001737 kPa/s. The next tests were all performed by making a new solution
that changes the pH with pH buffers and still contains a 1.5% H2O2 concentration. The first test
had a 3.3 pH made with a 3 pH buffer and the slope that was made was a .01689 kPa/s. The
second solution that was tested had a 5.2 pH made with a 5 pH buffer and ended with a slope of .
02507 kPa/s. The third solution has a 7.8 pH made with a 9 pH buffer then produced a slope that
was .02890 kPa/s. The final solution consists of 8.1 pH made with a 11 buffer and had a slope
Discussion:
Referring back to the background section, the optimal ph for catalase to operate in was 7.
The results refute to the information in the background, because it discussed how the optimal ph
The results showed a steady increase from the phs 3.3 and 5.2, but then had a sudden
drop at the ph of 6.5, then went right back up to a gradual increase from the phs 7.8 and 8.1. The
solution in which the catalase was the most efficient was the ph level 8.1 with a slope of .03380.
The solution in which the catalase was the least efficient was the ph level 6.5 with a slope of .
001737. The graph showed the sudden decrease of the ph 6.5, this indicates that an error
occurred, such as the ph buffer did not change the 3% hydrogen peroxide back to 1.5% hydrogen
peroxide. If the ph 6.5 would have had normal results the graph would have a steady increased
line and ph level 3.3 would be the least efficient. Other possible errors in the experiment could be
that the two-hole rubber stopper was not completely covering the Erlenmeyer flask and the
maximum pressure was not reached. Another possible mistake that could have occurred is that
the dilutions were not correct or the lab group but too little of an amount of catalase enzyme.
For improvement and elimination of any errors that occurred measure both parts of the
solution separately and double check the amount, to make sure the dilution will be effective. Also
test covering the Erlenmeyer flask with the two-hole stopper to make sure it fits and covers the
hole completely. Another improvement would be make sure to have the most updated version of
logger pro and a reliable device, also make sure to release ALL the catalase enzyme into the
flask. For further experiments, test more phs between, higher, and lower than 3,5,9, and 11, to
receive the most accurate results, then experiment with different dilutions of hydrogen peroxide
Conclusion:
In conclusion the hypothesis was not supported because generally the higher the ph
Citations:
BBC - Standard Grade Bitesize Biology - Enzymes and aerobic respiration : Revision, Page 5.
(n.d.). Retrieved January 12, 2017, from
http://www.bbc.co.uk/bitesize/standard/biology/investigating_cells/enzymes_and_aerobic
_respiration/revision/5/
Introduction to Enzymes. (n.d.). Retrieved January 12, 2017, from http://www.worthington-
Biochem.com/introbiochem/effectsph.html
Williams, J. (1928, March 20). THE DECOMPOSITION OF HYDROGEN PEROXIDE BY
LIVER CATALASE. Retrieved January 12, 2017, from
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2140981/