BIOLOGY LABORATORY REPORT

Enzyme Concentration and Rate of Reaction.

Name: Sharifah Nabaaha Binti Syed Yusof

IC Number: 971108-14-6522
S.I.D: 1511170006
Title: Enzyme Concentration and Rate of Reaction.
Date of Experiment:
Date of Submission:
Lecturer: Miss Nurul Nadia

Title: Enzyme Concentration and Rate of Reaction.

Objectives:
1) To investigate the effect of different enzyme concentration on the rate of reaction.
2) To highlight experimental and investigative skills.

Introduction

Enzymes are biological catalysts that catalyses biochemical reactions in living cells. In an
enzyme catalyzed reaction, the substrate binds to the active site and forms enzyme-substrate
complex with the enzyme. The enzyme then breaks the bonds in the substrate. The product of the
reaction then leaves the enzyme, which remains unchanged after the reaction. Catalase in an
enzyme produced by our liver to break down hydrogen peroxide a common end product of
metabolism, but highly toxic if accumulated in the body into water and oxygen.1 The equation
of the reaction is as follows:

1
http://www.bbc.co.uk/schools/gcsebitesize/science/add_aqa_pre_2011/enzymes/enzy
mes1.shtml

All living things possess catalysts, or substances within them that speed up chemical reactions
and processes. Enzymes are molecules that enable the chemical reactions that occur in all living
things on earth. In this catalase and hydrogen peroxide experiment, we will discover how
enzymes act as catalysts by causing chemical reactions to occur more quickly within living
things. Using a potato and hydrogen peroxide, we can observe how enzymes like catalase work
to perform decomposition, or the breaking down, of other substances. Catalase works to speed up
the decomposition of hydrogen peroxide into oxygen and water.

The protein has a certain 3D structure when it is active, which contains a channel into which the
hydrogen peroxide can diffuse. In the channel is a heme group which is a iron molecule bound to
the center of a ring-like structure called a porphyrin ring. The heme group in catalase is very
important to the reaction, because it can be oxidized from Fe(III) to the very oxidized and less
common Fe(IV) form. This is the first part of the reaction:
H2O2 + Fe(III)-Enzyme -- H2O +O=Fe(IV)-Enzyme (1)
Hydrogen peroxide has bound to the heme group and oxidized it to Fe(IV). Now for the second
part of the reaction: The enzyme has to go back to the Fe (III) form and reduce the second
molecule of hydrogen peroxide to water. 2
H2O2 + O=Fe(IV)-Enzyme -- H2O + Fe(III)-Enzyme (2)

2 http://scienceline.ucsb.edu/getkey.php?key=166

The highly-oxidizing Fe(IV) form now reacts with the second peroxide molecule, releasing water
and an oxygen molecule.3

Problem Statement: How does enzyme concentration affects the rate of reaction of
decomposition of hydrogen peroxide?

Hypothesis: As the enzyme concentration increases, the rate of reaction of decomposition of

hydrogen peroxide increases until it reaches optimum.

Variables:
Manipulated variable: The enzyme concentration (amount of scoop of potato)
Responding variable: The rate of reaction of decomposition of hydrogen peroxide
Constant variable: pH of the enzyme surrounding

Apparatus: Spatula, stopwatch, test tube, conical flask, graduated tube, beaker, delivery tube.

Materials: Hydrogen peroxide solution, blended potato.

3 http://www.ebi.ac.uk/interpro/potm/2004_9/Page2.htm

Procedure
1) Place 1 spatula of blended potato and add 5 cm3 of buffer solution of pH 6.5 into a test tube.
2) Gently swirl the test tube to mix the both.
3) Replace the bung in the flask. Fill the graduated tube with water and insert with care into the
beaker. Do not place over the end of the delivery tube yet.
4) Measure 2.5cm3 of H2O2 solution using a syringe and connect it to the flask.
5) Immediately place the graduated tube over the end of the delivery tube and measure the
volume of oxygen gas collected every 20 seconds for 5 minutes.
6) Record your results in a suitable table, showing the times volumes of oxygen, O2 collected.
7) Plot a graph to show how enzyme concentration affects rate of reaction of decomposition
H2O2.

H2O2 + catalase

Results
1 spatula of blended potato
Time/s

1st Reading/
cm3

2nd Reading/cm3

Average/cm3

Rate of
Reaction
(cm3/s)

0
30
60
90
120
150
180
210
240
270
300

0
2.0
2.0
3.0
3.0
3.5
4.0
4.0
4.0
4.0
4.0

0
1.0
1.0
1.0
2.0
2.0
2.0
2.0
3.0
3.0
3.0

0
1.5
1.5
2.0
2.5
2.75
3.0
3.0
3.5
3.5
3.5

0
0.050
0.050
0.067
0.083
0.092
0.100
0.100
0.117
0.117
0.117

2 spatulas of blended potato

Time/s

1st Reading/cm3

2nd reading/cm3

Average/cm3

Rate of
Reaction
(cm3/s)

0
30
60
90
120
150
180
210
240
270
300

0
1.5
2.0
2.0
3.0
4.5
4.5
5.0
5.0
5.0
5.0

0
1.5
1.5
1.5
2.5
3.0
3.0
4.5
4.5
4.5
4.5

0
1.5
1.75
1.75
2.75
3.75
3.75
4.75
4.75
4.75
4.75

0
0.050
0.058
0.058
0.092
0.125
0.125
0.158
0.158
0.158
0.158

3 spatulas of blended potato

Time/s

1st Reading/cm3

2nd reading/cm3

Average/cm3

Rate of
Reaction
(cm3/s)

0
30
60
90
120
150
180
210
240
270
300

0
0.5
1.0
2.0
3.0
4.0
4.5
5.0
5.5
6.0
6.0

0
1.0
2.0
3.0
4.0
4.5
5.0
5.0
5.0
6.0
6.0

0
0.75
1.5
2.5
3.5
4.25
4.75
5.0
5.25
6.0
6.0

0
0.025
0.050
0.083
0.117
0.142
0.158
0.167
0.175
0.20
0.20

Graph of Rate of Reaction of Catalase against Time.

Graph of Rate of Reaction(cm3s-1) against Time(s)

0.5
0.45
0.4
0.35
0.3

Three Scoop Potato

Two Scoop Potato

Rate of Reaction/cm3s-1

0.25
0.2
0.15
0.1
0.05
0

Discussion

One Scoop Potato

During the experiment, when catalase enzyme breaks down the hydrogen peroxide solution, it
releases a gas, oxygen gas. Thus, using a graduated beaker tilted under water, the volume of
oxygen gas was measured and recorded. This data is used to calculate the rate of reaction of
catalase enzyme, with a difference in 30 seconds. This experiment was carried out with different
enzyme concentration, defined by the different number of scoops of potato.
In overall, three scoops potato produced the highest rate of reaction due to the highest enzyme
concentration among the other two experiment. The highest data achieved was 0.2 cm3s-1,
compared to two scoops with the highest, 0.16cm3s-1 and one scoop with 0.12 cm3s-1. This
comfirms the hypothesis that stated the higher the enzyme catalase concentration, the higher rate
of reaction. This is because the three scoops contains the most amount of catalase enzyme, thus
the same amount of hydrogen peroxide used in both one scoop and two scoop, is able to be
broken down and catalyzed in a much shorter time and at a higher rate. The rate of gas bubbles
released also increased, in three scoops, compared to the one scoop and two scoops that took a
longer time.
In one scoop, the complete reaction was achieved at 240 seconds. In two scoops, the complete
reaction was already achieved at 210 seconds. In three scoops, the complete reaction was
achieved at approximately 270 seconds. For three scoops, the longer time taken might be due to
the errors that occurred during the experiment such as the enzymes being losing its ability to
catalyze, due to being exposed for so long, with non-optimal surroundings. Thus, this might have
caused the data to vary from usual, where it would take lesser than 210 seconds, lesser than the
time for two scoops.
Based on the graph, the rate of reaction for three scoops had the highest gradient that shows that
it has had the highest values that varied compared to the one scoop and two scoop. Thus, this
sums up that three scoops has the highest rate of reaction compared to the one scoop of potato
and two scoops of potato.

Precautions

1) Wear eye protection and cover clothing when handling hydrogen peroxide.
2) Wash splashes of pureed potato or peroxide off the skin immediately.
3) Be aware of pressure building up if reaction vessels become blocked.
4) Take care inserting the bung in the conical flask it needs to be a tight fit, so push and twist the bung
in with care.

Conclusion
The hypothesis is accepted. As the enzyme concentration increases, the rate of reaction of
decomposition of hydrogen peroxide also increases until it reaches optimum.