Elena Knudsen

29 September 2016

Mrs. Handest

AP Biology

Peroxidase Lab

Part A:

Part A Measurements:

Substrate tube:

8 mL distilled water

0.5 mL peroxide

0.25 mL guaiacol

Enzyme Tube:

7 mL distilled water

1.7 mL peroxidase

Hypothesis: The color will rise to a ten within 5 minutes.

Procedure:

1. Label a tube with an ‘S’ for substrate. Add 8 mL water, 0.5 mL peroxide, and 0.25 mL

guaiacol to the tube and gently mix.

2. Label a tube with an ‘E’ for enzyme. Add 7 mL water and 1.7 mL peroxidase, then gently

mix.

3. Carefully pour the contents of both test tubes into a new test tube, then record the color

every 15 seconds. Graph results.

Part B:

Part B Measurements:

Substrate tube:

8 mL distilled water

0.5 mL peroxide

0.25 mL guaiacol

Enzyme Tube:

7 mL distilled water

2.5 mL peroxidase

Hypothesis: The altered measurement of peroxidase will increase the speed of the reaction as

compared to the results from Part A.

Procedure:

4. Label a tube with an ‘S’ for substrate. Add 8 mL water, 0.5 mL peroxide, and 0.25 mL

guaiacol to the tube and gently mix.

5. Label a tube with an ‘E’ for enzyme. Add 7 mL water and 2.5 mL peroxidase, then gently

mix.

6. Carefully pour the contents of both test tubes into a new test tube, then record the color

every 15 seconds. Graph results.

Part C:

Part C Measurements:

Substrate tube:

8 mL distilled water

0.5 mL peroxide
1 mL guaiacol

Enzyme Tube:

7 mL distilled water

1.7 mL peroxidase

Hypothesis: The altered measurement of peroxidase will increase the speed of the reaction as

compared to the results from Part A.

Procedure:

7. Label a tube with an ‘S’ for substrate. Add 8 mL water, 0.5 mL peroxide, and 1 mL

guaiacol to the tube and gently mix.

8. Label a tube with an ‘E’ for enzyme. Add 7 mL water and 1.7 mL peroxidase, then gently

mix.

9. Carefully pour the contents of both test tubes into a new test tube, then record the color

every 15 seconds. Graph results.

Data/Analysis:

Time(Seconds) Color(Indicated by Color(Indicated by Color(Indicated by

Guaiacol) Part A Guaiacol) Part B Guaiacol) Part C

0 0 8 7

15 0 9 8

30 1 9 9

45 2 10 10

60 2 10 10

75 2 10 10
 90 3 10 10

105 3 10 10

120 3 10 10

135 4 10 10

150 5 10 10

165 5 10 10

180 5 10 10

195 5 10 10

210 6 10 10

225 6 10 10

240 6 10 10

255 7 10 10

270 7 10 10

285 7 10 10

300 7 10 10
Fig. 1: The chart displays the results of Part A, Part B, and Part C experiments.
 Fig. 2: Part A data graph. It demonstrates the rise in the enzyme action. The relative rate of

enzyme action was determined by the guaiacol. The guaiacol reacts with the oxygen that was

released from the enzyme reaction, and the contents of the test tube turn brown. The intensity of

the brown determines the relative rate of enzyme reaction. It is not possible to find the absolute

rate of reaction in this lab with the given materials.

Fig. 3: Part B data graph. It demonstrates the rise in the enzyme activity. The relative rate of

enzyme action was determined by the guaiacol. The guaiacol reacts with the oxygen that was

released from the enzyme reaction, and the contents of the test tube turn brown. The intensity of

the brown determines the relative rate of enzyme action. The amount of peroxidase was altered

compared to the original. The increased amount of enzymes led to an increased reaction rate.
 Fig. 4: Part C data graph. It demonstrates the rise in the enzyme action. The relative rate of

enzyme action was determined by the guaiacol. The guaiacol reacts with the oxygen that was

released from the enzyme reaction, and the contents of the test tube turn brown. The intensity of

the brown determines the relative rate of enzyme action. The amount of guaiacol was altered

compared to the original. The increased amount of guaiacol indicator caused the oxygen reaction

to intensify in color faster, skewing the results compared to Part A and Part B, which both had

the same amount of indicator.

Conclusion:

The data from each experiment supported its hypothesis. For Part A, the relative rate of

enzyme reaction did reach 10 within five minutes. In Part B, the increased amount of peroxidase

sped the rate of enzyme activity. Finally, in Part C, the increased guaiacol changed the color

faster than Part A, even though nothing about the enzyme reaction was altered. Part A was the
control of the experiment, because it was the base of every other task. The amount of materials

added were the independent variables, and the relative rate of activity was the dependent

variable. Sources of error included the specificity of measurements, and also the timing. The

results were not recorded exactly by the second throughout the fifteen second intervals. The

reason why there aren’t bubbles in this version of the lab as compared to the celery pre-lab is

because guaiacol is present. The guaiacol reacts with the oxygen before it can be released, and

changes the color instead of emitting bubbles.

Enzyme reactivity can be slowed by denaturing and inhibitors. However, it can be slowed

without the use of unrelated obstacles. As evidenced in the Toothpickase lab, the enzyme can

pick up part of their previous substrates, or products, which block full substrates from entering

the active sites. This happens increasingly frequently the more substrates are broken down,

because when an enzyme is surrounded by more products than substrates, it becomes harder for

the enzyme to find an intact substrate, therefore slowing its process. That is why when there is an

overabundance of substrates, the reactivity becomes very high but then begins to decrease until

there is no activity. If there was a large amount of peroxide, the peroxidase would break it down

at a fast rate, but eventually would not be able to break down any more, just because it would be

difficult to discover a full substrate. There are ways for enzymes to dissolve, though. Proteolytic

enzymes are enzymes that break down proteins into amino acids. Therefore, an enzyme, which is

a protein, can be broken down by proteolytic enzymes. If active peroxidase were exposed to

trypsin, a proteolytic enzyme, the peroxide would no longer function. Peroxide is naturally

produced within the human body, but if it were able to accumulate, it would kill the body. An

enzyme, called catalase, breaks down the peroxide into water and oxygen. Without the use of

catalase, humans would die because of the rising levels of cell destroying peroxide.
 Works Cited

Activity, Factors Affecting Enzyme, and Purdue University Instrument Van Project.

"Factors Affecting Enzyme Activity." Factors Affecting Enzyme Activity: n. pag. Web. 29 Sept.

2016.

"Johnson Matthey Catalysts - Education - Enzymes." Johnson Matthey Catalysts -

Education - Enzymes. Web. 29 Sept. 2016.

"Proteolytic Enzyme." Encyclopedia Britannica Online. Encyclopedia Britannica. Web.

29 Sept. 2016.