Kashyap Patel

EGHS AP Biology Lab Report

How pH affects catalases rate of reaction when breaking down hydrogen peroxide.
Section 1: Introduction/Hypothesis Research information about your enzyme that is
relevant to your procedure. Length should be about a paragraph.
Enzymes are organic catalysts found in all living organisms. They assist in all the living
functions of an organism by lowering the activation energy and facilitating a chemical reaction.
Their ability to perform reactions is affected by 3 environmental factors, temperature, pH, and
salinity. If the enzyme is exposed to a environment that it is not suited to, it will denature and
lost its folding which creates active sites for the reaction. Hydrogen peroxide is a highly acidic
solution found in almost all living organisms. It is found in the peroxisome in the cell after being
produced through various chemical reactions that metabolise food in the body. If a body has too
much hydrogen peroxide healthy cells begin to die. Peroxidase/catalase in the body breaks down
2 hydrogen peroxide molecules into 2 water molecules and 2 oxygen atoms. Yeasts optimum
pH is is between 4-4.5. If yeast is introduced to a more basic environment, the rate of reaction
will decrease because yeast will denature in the solution.
Section 2: Variables
Independent Variable The independent variable is the pH value of the yeast solution
Dependent Variable The dependent variable is the amount of O2 produced by the reaction.
Controlled Variables/Experimental Constants The experimental constants we maintain
throughout the experiment are temperature, salinity, and the amount of enzyme/substrate used.
Experimental Group The experimental groups are the the pH 5, 7, and 10 solutions that the
yeast are mixed into.
Control Group The control group is the pH 4 solution that the yeast is exposed to.
Section 3: Materials and Methods
Materials: (List in bullets)
250 mL Nalgene bottles
Pipettes for yeast, and different pH solutions

O2 Sensor
10 mL graduated cylinder per pH
100 L yeast of trial
Beakers (mix pH & yeast)
10 mL 3% H2O2 per trial
Labquest 2
3 mL pH solution per trial (pH 4, 5, 7, 10)
Safety goggles

Procedure:
1. Mix 100 L of yeast with 3 mL of pH solution being tested and allow it to sit for 3
minutes.
2. Test pH of solution to ensure it has changed to the amount being tested.
3. Use graduated cylinder to measure out 10 mL 3% H2O2 and pour into Nalgene bottle.
4. Set up labquest 2 and O2 sensor to measure the rate of reaction.
5. Mix hydrogen peroxide solution and yeast/pH solution in Nalgene bottle and cover with
O2 to measure the rate of the reaction.
6. Allow reaction to run for 200 seconds while observing the reaction.
7. Record the rate of reaction from the labquest analytics.
8. Pour solution down the sink, clean equipment that was exposed to solutions and reset for
next trial.
9. Repeat steps 1-8 for multiple trials as needed, at least 3 is required.
10. After the last trial is run, complete data table, summarize lab, and graph data for analysis.
11. Clean all lab equipment and prepare for next class.
Section 4: Results (must be computer generated)
Data Tables:
Catalases Rate of Reaction With Varying PH
PH

Trial 1 (Rate of
Reaction
%O2/sec)

Trial 2 (Rate of
Reaction
%O2/sec)

Trial 3 (Rate of
Reaction
%O2/sec)

Average (Rate
of Reaction
%O2/sec)

.0048

.0056

.0045

.0050

.0025

.0028

.0025

.0026

.0023

.0029

.0024

.0025

10

-.003

.0012

.0047

.0010

Graphs:

Section 5: Discussion/Conclusion: In CER format

Our hypothesis was correct in stating that yeast would be more effective in an acidic
environment rather than a neutral or basic environment. According to our data, pH 4 had the
highest rate of reaction, .00497, while pH 10 had .001, or the lowest of all the reactions. It is
evident that catalase performs optimally in pH 4 but denatures in more neutral and basic
environments. Because of this higher reaction rate, more Oxygen and water is produced through
the reaction which is proof as to why the reaction is better in an acidic environment.
Furthermore, the graph in our lab report further emphasizes the decrease in the overall rate of
reaction over multiple pHs. This perfectly illustrates the effect that pH has on the efficiency of
enzymes inside the yeast.
However, we had an outlier in out pH 10 trial 1 when we measured a negative %O2/sec
output. A source of error in our experiment may be the mixing of solutions that occurred during
the data collection phase. While the reaction occurred, a group member swirled the Nalgene
bottle to facilitate the reaction, however, sometimes a different group member did it or at a
different speed which may affect the amount of collisions that facilitated the reaction rate. To fix
this in a future experiment, I believe having a hotplate turned off and only on the swirl mode
may be able to keep the amount of mixing that occurs in the solution the same. Also, through our

observations, we noticed that there usually a slight decrease in Oxygen levels before the
chemical reaction was fully active. The actual activation of yeast may have been a difference in
the experiment. The solution was prepared multiple times and may have changed throughout the
experiment. We are also measuring in the thousandths place which may be inaccurate since the
labquest only measures to the hundredths place. Therefore, in a future lab we would like to test
why this happens and what factors other than pH affect the amount the Oxygen levels dip before
the reaction can occur.

Section 6: Works Cited (Use MLA format, Search on Purdue OWL for a template)
"Fermented and Vegetables. A Global Perspective. Chapter 3." Fermented
and Vegetables. A Global Perspective. Chapter 3. FAO, n.d. Web. 05 Oct.
2016.
"Yeast Are Greatly Affected by Temperature Too Cold and - ENGLISH - 1001.
"Yeast Are Greatly Affected by Temperature Too Cold and - ENGLISH 1001. FAO, n.d. Web. 05 Oct. 2016.
"FERMENTED FRUITS AND VEGETABLES. A GLOBAL PERSPECTIVE..
"CHAPTER 3 YEAST FERMENTATIONS. FAO, n.d. Web.