Properties of Enzymes

By: Kevin Curran

Honors Biology
11/20/15
Period 3

Introduction:
Enzymes have a multitude of functions that we can use for a variety of uses. They are
specific in their function, and they can be used more than once. Enzymes work by binding to a
reactant, known as a substrate, from a location on the enzyme called the active site. They act as a
catalyst, which is a substance that speeds the rate at which a chemical reaction takes place. They
do this by weakening the bonds between molecules, so that the activation energy required for the
chemical reaction takes place is lowered. There are three factors that go into the effectiveness of
an enzyme, the environmental conditions, cofactors and coenzymes, and enzyme inhibitors. The
environmental conditions that are best for the enzyme depends entirely on which enzyme it is.
Each enzyme has their own optimal temperature and optimal pH level. Any environmental
conditions that are not favorable to the enzyme may denature it, which means that it unfolds and
becomes ineffective. Some examples of cofactors and coenzymes are inorganic substances such
as iron or zinc. Enzyme inhibitors are chemicals that closely resemble the enzymes natural
substrate, and it competes with the regular substrate for the active site one the enzyme. Another
example of an inhibitor is something that binds somewhere on the enzyme other than the active
site.
In this lab, I hope to calculate the rate of the reaction between hydrogen peroxide and
peroxidase. Doing this, I will also find the optimum pH at which peroxidase works best at. The
rate at which chemical reactions take place can be measured by a Spec 20. It can do this by
measuring the absorption of the liquid put into the test tube. When hydrogen peroxide is
combined with peroxidase, it results in water and oxygen. Guaiacol is an indicator, which means
that it reacts with oxygen, and turns brown. If Guaiacol is mixed in with the hydrogen peroxide
and the peroxidase, then the oxygen from the hydrogen peroxide and peroxidase mix will react

with the Guiaicol, which should turn the substance brown. Absorption is the amount of light that
is being absorbed by the substance. If the substance is turning brown, the absorption will raise,
and the Spec 20 can measure that. If we test a pH of 3, 5, 8, and 9 with the hydrogen peroxide
and peroxidase, then the pH of 5 will be the optimal pH of the peroxidase.

Materials:

Spec 20
Hydrogen Peroxide
Turnip
Guiaicol
Test Tubes

Procedures:
1.
2.
3.
4.
5.
6.

Insert test tube filled with water into the Spec 20

Calibrate the Spec 20 so that the absorbance is 0
Mix the test tubes filled with the turnip and substance with a pH of 9
Invert it twice and then insert it into the Spec 20
Observe and record the absorbance readings at the given times
Repeat steps 1-5 with the substances with a pH of 8, 5, and 3

Results
2.5
2
1.5

Absorbance

1
0.5
0

20 40 60 90 120 150 180 210 240 270 300 330 360 390420450480510540570600630 660

Time (Seconds)
control

pH 9

pH: 8

pH: 5

pH: 3

Results

This graph shows the rate at which the Spec 20 measured the absorbance of the mixtures. In the
graph, the control reaches its max of about 2 at around 180 seconds into the experiment, while
the pH of 5 substance reached the same max at about 510 seconds. The pH of 8 substance
steadily increased throughout the entire experiment never reaching a maximum. The pH of 9
reached its max of about 1.3 at around 570 seconds. The substance with a pH of 3 reached its
max of about 0.01 at around 570 seconds.

Discussion
After the experiment I conclude that my original hypothesis of the optimal pH being 5 as
incorrect. After running the tests, I came to the conclusion that the optimum pH that peroxidase
works at was 7. I came to this because in the control, the absorbance reached the highest in the

least amount of time. The next best pH was 5. It was able to reach the same absorbance as the
control, but it took about 230 more seconds to get there. The pH of 9 was able to reach its
maximum of about 1.3 at around 570 seconds. Because it was not able to get to as high of an
absorbance as the control or the pH of 5, 9 is not the optimal pH of the peroxidase. The pH of 3
never really had any absorbance, which leads me to believe that the substance was too acidic for
the peroxidase, which caused it to denature. An error might have been made with the testing and
recording of the pH of 8. This is because its graph looks different from the other ones.

Works Cited

Biggs, Alton. Biology. Columbus, OH: McGraw-Hill Education, 2012. Print.