Enzyme Activity

Lab 6

3/10/17

Report by Yeimi Guerra, RaeJean Bahena, and Josh Bradley

I. Introduction
II. Materials & Methods
III. Results
IV. Conclusion & Discussion
 Results
Below is a graph and table of the data collected every 30 seconds on the effects of pH on the
hydrogen Peroxide/Catalase Reaction during the Enzyme Activity lab.
Table 2: Effects of pH on the Hydrogen Peroxide/ Catalase Reaction
(ml Oxygen produced over time)
Time (sec) pH 2 pH 7 pH 10
0 0 0 0
30 1 5 4
60 1 11 7.5
90 1 14 9
120 1 17 11.5
150 1 20 13
180 1 21 14
210 1 23 15
240 1 25 16
270 1 27 17
300 1 29 17
330 1 31 17.5
360 1 35 18
390 1 37 18.5
420 1 37 19
450 1 37 20
480 1 37 20
510 1 37 20.5
540 1 37 21
570 1 37 21
600 1 37 21.5
Graph 2: Effects of pH on hydrogen peroxide/catalase reaction

pH Diffrences
50
45
40
ml Oxygen Produced

35
30
25
20
15
10
5
0
0 100 200 300 400 500 600 700
Time (sec)

2 Ph 7 Ph 10 Ph Linear (2 Ph) Linear (7 Ph) Linear (10 Ph)

 Conclusions & Discussion

The date exhibiting production of oxygen allowed the null hypothesis to be accepted. Various
temperatures affect the rate and amount of product produced of hydrogen peroxide breakdown
by catalase. Table 2 shows the most production of oxygen was at a neutral pH level of 7. Acidic
levels of pH had little to no oxygen producing. Whereas pH 10 (most alkaline) had enzymatic
activity that helped production of oxygen. Table 2 shows that as the temperature increases, the
rate of the production of oxygen also increases. However at 80C certain we can see the extreme
heat can denature the protein and can no longer work, therefore produced little to no oxygen.
Graph 2 shows that the optimal level for enzymatic activity producing the most oxygen was at
37C the second most optimal level for the enzyme to work along with the catalyst was at 2C and
the least temperature that the enzyme worked at was at 80C producing little to no oxygen.

1. What does the rate of oxygen production and/or amount of oxygen produced tell us
about the chemical reaction?
What the amount of oxygen production tells us about our chemical reaction is that the more
acidic the pH is, the less oxygen it will release, oxygen production is favorable In both a pH of 7
and 10. As shown is table 1 the temperature at 2C had a steady production of oxygen. 37C had
similarities of oxygen production as 2C and continued a steady rate of oxygen production.
Temperatures at 80C had a minor production of oxygen.

2. Given the conditions tested, which are the best for catalase activity?
Each enzyme has an optimal pH & Temperature in which it can function optimally. Optimal
conditions favor the most active shape for the enzyme molecule. The rate of oxygen produced
tell us that a neutral pH is where the enzyme thrives. The enzymes favors conditions in neutral
temperatures (not being too hot or cold). A favorable temperature of 37C produced the most
amount of oxygen.
3. How does 80C treatment affect the catalase? What experimental data supports your
answer?
80 C affects the treatment of the catalase by producing oxygen at a much slower rate when
compared to 37C and 4C. As we can see in graph 1, 80C had the least production of oxygen. The
first 60 seconds had no reaction. At 90 seconds we come to a slight oxygen production of 5ml.
The oxygen production levels of at 150 seconds producing no more oxygen.
4. How does 4C treatment affect the catalase? What experimental data supports your
answer?
4C affects the treatment of the catalase by producing oxygen at a minimum rate when compared
to 37C. As we can see in graph 1, 4C had a constant production of oxygen. At 30 seconds 3 ml
of oxygen was produced and at 600 seconds 11 ml of oxygen was produced.

5. How do various pH treatments affect the catalase? What experimental data supports
your answer?
Various pH can have several different effects on the activity and the structure of an enzyme. The
pH of an enzyme can charge properties of the substrate so that either the substrate cannot bind
to the active site or it cannot undergo catalyst. As seen in graph 2, the neutral level of pH 7
produced the most amount of oxygen. On the more alkaline level pH 10 there was a good amount
of oxygen produced as well, however not compared to the production of oxygen from pH 7. The
more acidic the pH is such as pH 2 had little to no production of oxygen.
6. Why does the production of oxygen slow down over time?
The production of oxygen slows down because different temperature can denature the enzyme
protein. Some high temperatures can cause the enzyme to denature, which permanently changes
the shape of the enzyme, causing the enzyme not to function correctly and cease the production
of the oxygen.
7. If you added more catalase to the 37C reaction that had stopped producing oxygen.
What do you expect would happen?
If you added more catalase to a 37C reaction since temperature is favorable for enzymatic
activity to take place the adding of the catalase would speed up the reaction or cause a reaction to
proceed at a lower temperature without it being consumed in the reaction itself. The catalyst
molecules would be able to be reused again.
8. If you added more hydrogen peroxide to the 37C reaction that had stopped producing
oxygen, what do you expect would happen?
If you added more peroxide to a 37C reaction the breakdown of oxygen gas and water will occur
with the hydrogen peroxide, at a much higher rate because you have optimal temperature for
enzymatic activity to take place. With the addition of more hydrogen peroxide the catalase will
be present and help facilitate the breakdown with less energy.
pH 2: Results
pH 2 Average (ml) : 1
Max Rate (ml): 1
Slope pH 2: 1
Total amount of O2 produced pH 2: 20 ml
Time of reaction until oxygen production levels off: 30 seconds
pH 7: Results
pH 7 Average (ml) : 24.62
Max Rate (ml): 37
Slope pH 7: 0.054245
Total amount of 02 produced pH 7: 554 ml
Time of reaction until oxygen production levels off: 390 seconds
pH 10: Results
pH 10 Average (ml) : 16.05
Max Rate (ml): 21.5
Slope pH 10: 0.025172
Total amount of 02 produced pH 10: 321 ml
Time of reaction until oxygen production levels off: still consistently producing oxygen
 Reference
Front Range Community College. (2017). Biology 111 Lab Manual. Fort Collins, CO: Front
Range Community College
"Introduction to Enzymes." Effects of PH (Introduction to Enzymes). N.p., n.d. Web. 09 Mar. 2017.

"PH and enzymes." PH and enzymes. N.p., n.d. Web. 09 Mar. 2017.