NAME: MICHAEL TIMSON

DATE:
FORM: L6-4
SUBJECT: Biology
TITLE: THE EFFECT OF THE DECOMPOSITION OF HYDROGEN PEROXIDE
CONCENTRATIONS BY CATALASE
AIM: To determine the effects of the decomposition of hydrogen peroxide concentrations by
catalase.

INTRODUCTION:
Molecules that accelerate, or catalyze a chemical reactions are known as enzymes. In
reactions such as these, the molecules found at the beginning of the process are called substrates.
Enzymes converts these substrates into different molecules, called products. Most metabolic
reactions found in cells, require involution of enzymes catalyst to stimulate these processes at a
faster rate, to sustain life. The type of enzymes synthesized in a cell determines which metabolic
pathways occur in that cell. Enzymes are known to catalyze more than 5,000 biochemical
reaction types. Most enzymes are proteins with the exception of a few being catalytic RNA
molecules. Enzymes' specificity comes from their unique three-dimensional structures. Enzymes
are generally globular proteins, acting alone or in larger complexes. Like all proteins, enzymes
are linear chains of amino acids that fold to produce a three-dimensional structure. The sequence
of the amino acids specifies the structure which in turn determines the catalytic activity of the
enzyme.
Catalase is a common enzyme found in nearly all living organisms exposed to oxygen,
such as vegetables, fruit or animals. It catalyzes the decomposition of hydrogen peroxide to
water and oxygen. It is a very important enzyme in protecting the cell from oxidative damage by
reactive oxygen species. Likewise, catalase has one of the highest turnover numbers of all
enzymes; one catalase molecule can convert approximately 5 million molecules of hydrogen
peroxide to water and oxygen each second. Catalase is a tetramer of four polypeptide chains,

each over 500 amino acids long. It contains four porphyrin heme (iron) groups that allow the
enzyme to react with the hydrogen peroxide.
The reaction of catalase in the decomposition of hydrogen peroxide in living tissue:
2H2O2 2H2O + O2
The presence of catalase in a microbial or tissue sample can be tested by adding a volume of
hydrogen peroxide and observing the reaction. The formation of bubbles, oxygen, indicates a
positive result. This easy assay, which can be seen with the naked eye, without the aid of
instruments, is possible because catalase has a very high specific activity, which produces a
detectable response.
Concentration, being one of the factors affects the rate at which an enzyme catalyze a
reaction. In this experiment, the effects of substrate concentration on enzymatic activity will be
investigated using various concentration of hydrogen peroxide.

APPARATUS:
Boiling Tubes, H2O2 solution, 5 cm3 Syringe, Cork borer, Scalpel, Ruler, Potato, Glass
Rod, Forceps, Stopwatch

METHOD
Five boiling tubes were labelled A-D. Twelve potato cylinders were cut into 0.5cm each
using a cork borer, ruler and scalpel. Using a syringe, 5cm 3 of the H2O2 solution was added to a
boiling tube and stirred using a glass rod. Using a forceps, one potato cylinder was placed into
the boiling tube containing hydrogen peroxide while simultaneously starting the stopwatch. The
boiling tube was observed and the time taken for the potato cylinders to elevate from the bottom
of the boiling tube was recorded. The steps were repeated using another potato cylinder with the
same concentration of hydrogen peroxide concentration. The experiment was repeated using
different concentrations of hydrogen peroxide.

RESULTS
TABLE SHOWING THE INITIAL RATE OF DECOMPOSITION OF HYDROGEN
PEROXIDE FOR EACH SUBSTRATE CONCENTRATIONS BY ENZYME CATALASE
Substrate

Time taken

Initial Rate of

concentration (%)

T1

T2

Average time taken

Reaction (1/t)

3.0

91

69

80

0.0125

4.5

50

30

40

0.0250

6.0

30

28

29

0.0340

9.0

21

23

22

0.0450

DISCUSSION
From the results obtained in this experiment, it is seen, in the tabulated results, as the
concentration of substrate increased, the time taken to complete the reaction decreased and thus,
the initial rate of reaction had increased. A graph of initial rate of reaction versus substrate
concentration was derived. From the graph, it was seen that there was a significant increase from
an initial rate of reaction 0 at substrate concentration 1.5% to an initial rate of 0.034 at 6%.
However, the increases in the initial rate of reaction decreased. That is, addition of the substrate
began to display little effect beyond that point. Therefore, as the concentration of substrate
increases, the rate of reaction also increases until the point saturation occurs. It means that as
there is an increase in the concentration, the initial rate keeps increasing until the one point
where the maximum rate is achieved and there is no free enzyme to bind with substrate and all
the active sites of enzyme are bound to the substrate. Therefore, addition of substrate beyond this
point have no effect on the rate of reaction.
Within this experiment there were various sources of errors. There were contamination of
reagents to some extent which would have affected the time taken in the results and hence the
rate of decomposition of hydrogen peroxide by catalase. The factor of reaction time was also
involved in stopping the stopwatch and the experiment was conducted in an air conditioning
environment and thus temperature fluctuations occurred. Temperature, being one of the factors
affecting initial rate of reactions of the enzymatic activities, would thus cause a slight depletion
in the accuracy of the results. It is recommended that before conducting the experiment, that all
apparatus are washed thoroughly to ensure that there are no residue of chemicals on the
apparatus when conducting the experiment. Also, the experiment could be conducted using more
concentrations, hence acquiring a more accurate observations pertaining to the effects of the
increase of hydrogen peroxide concentration

CONCLUSION: Within the limit of experimental error, in increasing substrate concentration, the
time taken for the reaction to be complete decreased. Therefore, the initial rate of reaction was
increased.