Omlor 1

Gregory Omlor
Dr. Sadler
BIOL 101 02
October 16, 2014

Exercise 11 Enzymes: Factors Affecting the Rate of Activity

Qualitative and Quantitative Color Changes as Catechol Oxidase Activity Produces Brown Benzoquinone
Qualitative Color Change Results
Quantitative Absorbance Results
Tube
0 min
5 min
10 min
15 min
20 min
10 min
20 min
1
0
0
0
0
0
0
0
2
0
0
0
0
0
0
0
3
0
0
0
0
0
0.3
1.6
4
0.1
3
3
2.5
2.5
0.35
0.7
5
3
3
3
3
3
0.72
0.7
6
3
2.5
2.5
2.5
2.5
0.45
0.43
7
3
2
2
2
2
0.3
0.5
Question One
A. The Production of Benzoquinone from a variety of environmental conditions in potato juice extract.
In this study we investigated the effects of temperature on enzymatic activity with catechol oxidase, a
plant enzyme that converts catechol to benzoquinone. Catechol oxidase was combined with potato juice
extract and water. To provide more results the solutions and temperatures were changed and alterations
were made in pH. The solutions absorption was measured by a spectrophotometer, which determined
what conditions produced the most benzoquinone.
The materials used in this procedure consisted of 7 test tubes, a wax pencil, distilled water, a pH buffer,
potato extract, a 1% catechol solution, a spectrophotometer, and heating/cooling appliances.
We started by numbering the test tubes 1-7 and added the appropriate solutions as indicated in table 11.1.
We immediately recorded our initial readings of the color changes. We then placed the tubes in the
appropriate water-bath, refrigerator, etc. Every 5 minutes we observed and recorded the color changes of
each tube for a total of 20 minutes, returning the tubes to their original temperature locations
immediately after taking each reading. To further quantify our data we measured the absorbance of the
solution in each tube using the spectrophotometer once at 10 minutes and again at 20 minutes.
B. The enzyme was catechol oxidase, the substrate was potato juice, and the product was benzoquinone.
C. To allow for the substances to react.
D. There was little to no result in tubes 1-3. This was the control group. The reason for these tubes was to
prove that the results we were getting were accurate with the hypothesis. We altered these variables in a
known way to test the hypothesis. To make note that the potato extract and the catechol were required to
prove the results we were getting.

Omlor 2
E.

Qualitive Color Change Results

3.5
3
2.5
2
1.5
1
0.5
0
0 minutes

5 minute
Tube 4

10 minutes
Tube 5

15 minutes
Tube 6

20 minutes

Tube 7

Quantatitive Absorbance Results

0.8
0.7
0.6
0.5
0.4
0.3
0.2
0.1
0
10 minutes
Tube 4

20 minutes
Tube 5

Tube 6

Tube 7

F. Catechol oxidase functions equally and efficiently at various temperatures. Of the temperatures we
tested this statement appears to be true but if the temperatures were much higher I dont think we would
have gotten much of a reaction.
G. The temperature where the catechol oxidase appeared to be the greatest was a 4C (tube 5). Catechol
oxidase had its biggest reaction at the lowest temperature we tested. Is this correct? Strange, I wonder
what wouldve happened if we dropped the temperatures lower. Maybe the optimum temperature is
even lower than 4C thats what we should test.

Omlor 3
H. The temperature which the catechol oxidase activity was the greatest was at 4C. We should test
temperatures even lower than this to determine the optimum temperature.
I.

Catechol oxidase denatured at 80C because the reactions seemed to be slower.

J.

Denaturation of an enzyme by heat changes the shape of the molecule. The substrate will no longer fit
into the required location (lock and key mechanism). Therefore the enzyme can no longer catalyze the
reaction that it is supposed to.

K. With more than one enzyme to catalyze the same reaction, we would have a wider range of temperatures
and pH balances that are bodies could still catalyze the same reactions effectively. Making us more
adaptive to various environments.
L. The results supported our hypothesis. There is still more testing we could do to get optimum results.
Questions for Further Thought and Study
1. In an enzyme substrate reaction you need 1 enzyme for 1 substrate. Enzymes can only bind to one
substrate at a time. So if you add more substrate, it will take longer for the enzymes to complete the
reaction and attach to a new substrate.
2. Denaturation. The pH level, as well as different inhibitors, and activators can also influence a
proteins structure.
3. An enzymes function relies on its shape. When an enzyme is denatured its shape changes and it loses
its function. In other words, the enzyme becomes useless.
4. No because when it is active it is held together by hydrogen bonds. When you denature it and break
these hydrogen bonds and the enzyme unravels. The denaturing process is irreversible but the
remains are recycled to form new enzymes.
5. I suspect the group of organic compounds that these enzymes would react with would be protein.
To test this you could test the reactions of these enzymes with different forms of proteins,
carbohydrates, and lipids.