FAKULTI SAINS GUNAAN

UNIVERSITI TEKNOLOGI MARA

CAWANGAN PAHANG, KAMPUS JENGKA

BIO150 – METABOLISM & CELL DIVISION

SCIENTIFIC LAB REPORT

EXPERIMENT TITLE: ENZYME I – THE FASTEST ENZYME: CATALASE

LECTURER: SIR MOHAMAD AZAM AKMAL BIN ABU BAKAR

DATE OF SUBMISSION: 7th MAY 2021

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Declaration of Academic Honesty

Academic honesty or academic integrity is a very important virtue that all students should uphold at all
times.

I declare that the lab report submitted is not plagiarised and is entirely my own work, and that no part of it
has been copied from any work produced by other person(s)/ source(s) or provided by any other
student(s).

I understand that issuing a false declaration can result in severe penalties and I am willing to be penalized
if any form of copying found valid.
INTRODUCTION

The enzyme catalase breaks down hydrogen peroxide into water and oxygen.

2 H 2 O2 2 H 2 O+ O2

Hydrogen peroxide is a highly active chemical formed continually as a by-product of chemical

reactions in living cells. It is poisonous and if it were not immediately broken down by the cells, it
would destroy them. Hence, the importance of the enzyme is known. It is one of the fastest known
enzymes and its turnover number is 6 million, which means the number of substrate molecules which
one molecule of the enzyme turns to products per minute.

Catalase is a known enzyme. Based on the knowledge of the effects of enzymes on reaction rates, it
is hypothesized that the smaller the size of the particles, the higher the rate of reaction of catalase.
Further, at very high temperatures the enzyme catalase will have become denatured, and this
changed shape, unable to catalyze the decomposition reaction.

So, how does the temperature and size of particle affect the presence of the oxygen bubble produced
when liver, kidney and potato catalase react with hydrogen peroxide?
 OBJECTIVES

1. To identify the factors that influence the reactions of the catalase enzyme.
2. To analyze the reaction of catalase on different samples.

APPARATUS & MATERIALS

1. 7 test tubes
2. Beaker 250 cm3
3. Pestle and mortar
4. Hydrogen peroxide, H 2 O 2
5. Bunsen Burner
6. Tweezer
7. Wooden Splinter
8. Liver
9. Kidney
10. Potato

PROCEDURE

1. 2 ml hydrogen peroxide solutions were poured into a clean test tube. Cut 1cm cube of liver and
dropped it into the test tube of hydrogen peroxide. Observed the changes and recorded them. Tested
the oxygen presence with a wooden splint.
2. A piece of liver in the same size as before was placed in a mortar. The liver has been ground and
transferred into a test tube that contains the fresh hydrogen peroxide. The activity of the ground liver
had been viewed and compared with the activity observed for the whole piece of the liver.

3. Another piece of liver was added into a test tube that has been placed in a beaker of boiling water.
Placed it for three minutes.

4. Then, dropped the piece of liver into fresh hydrogen peroxide and observed if the enzyme is still
capable to break down the hydrogen peroxide.
5. The experiment repeated using kidney and potato to find out if they contain the enzyme catalase.
6. Observation has been recorded in Table 1.
 RESULT

Table 1: Catalase reaction

Test tube containing Observations Presence of Oxygen

Cube of liver  Bubble overflowed the test tube at a Present

slower rate
 Wooden splinter lights up

Ground liver  Bubbles overflowed the test tube at a Present

fast rate
 Wooden splinter lights up

Boiled liver  No bubbles produced Absent

 Wooden splinter does not light up

Cube of kidney  Fewer bubbles produced Absent

 Wooden splinter does not light up

Ground kidney  More bubbles produced Present

 Wooden splinter lights up

Cube of potato  Less bubble produced Absent

 Wooden splinter does not light up

Ground potato  Less bubble produced Absent

 Wooden splinter does not light up
DISCUSSION

I carried out an experiment to observed the presence of oxygen and the reaction of the liver,
kidney and potato after added the same amount of hydrogen peroxide. Before I experimented, I know
that the enzyme that I am using in this experiment can break down hydrogen peroxide into water and
also oxygen. Catalase is an important enzyme because it protects the cell from oxidative damage by
reactive oxygen species.

The experiment was divided into three parts for liver while kidney and potato were divided into two
parts. In the first part, a cube of liver, heart, and potato was added into a clean tube filled with 2ml
hydrogen peroxide respectively and observed the reaction. Then, tested the presence of oxygen by
using a wooden splint. In the second part, each liver, heart and potato were ground but with the same
size as before. Then, observed what happened. In the third part, only for the liver, the same size of
liver was put in boiled water for three minutes. After that, put into a tube of hydrogen peroxide
respectively and observed.

In the first experiment, the cube of liver produced bubbles that overflowed the test tube in a few
seconds once it made contact with hydrogen peroxide. The foams are oxygen produced from the
catalyzation. For the presence of oxygen, a wooden splinter was inserted into the test tube. The
wooden splinter lights up, as I expected. For the ground liver result, the bubbles were produced at a
fast rate than the cube liver in the experiment. A wooden splinter was inserted and it lit up, which
means oxygen is present. Based on the previous research, a smaller size of particles increases the
rate of reaction. This is because the smaller size of particles used less energy to break down
molecules than the larger ones. The larger the size of particles, the rate of reaction decreased
because they required more energy to break down. This proves that there is no error had occurred
during this experiment.

For the third part which was the boiled liver, the experiment shows that it did not produce any
bubbles. There was no oxygen presence because the wooden splinter did not light up when inserted
into the test tube. The boiled sample did not show any reaction because the catalase enzyme in the
liver was denatured after being put into the water at a high temperature. In the previous research, the
reaction of the enzyme should be increased when the temperature increases but at 40℃ , the rate of
reaction decreases because the enzymes would start experience denaturation. When the
temperature reached 60℃ , the enzyme is denatured. They are damaged and would not be able to
break down hydrogen peroxide. Therefore, the result is what I expected.
In the second experiment, the cube of the kidney produced fewer bubbles. When the wooden splinter
was inserted into the test tube, it lights up shows that oxygen was present. This proves that since the
kidney was raw, the catalase in the enzyme of the kidney was not damaged at all. It same goes for
the enzyme of the liver. To continue the experiment, a cube of the liver was ground up and inserted
into a test tube. A more amount of bubble was produced than the cube of the kidney. Also, the
bubbles produced were faster than the cube of the kidney which proves that the size of particles
affects the rate of reaction. Next, a wooden splinter was inserted but it shows that there is no
presence of oxygen. Actually, the oxygen should present but because the oxygen gas was not
enough to ignite the wooden splinter, the wooden splinter did not light up.

In the last experiment, the cubed potato produced fewer and tiny bubbles. When a wooden splinter
was inserted and it did not light up. This means that there is no presence of oxygen gas. Same goes
with ground potato, less bubble was produced but the reaction was faster than the cube of potato.
Then, when a wooden splinter was inserted and it did not light up, it clear that there was no presence
of oxygen gas.

From the result, the difference from my expectation is most of my sample formed bubbles but does
not light up the splint such as a cube of potato, cube of kidney and ground potato. During this
experiment, I encountered one error that I found out that can be the reason why the splint did not lit
up because the split is wet and the tip of it dipped into the surface of the solution in the test tube.

To overcome all of this error, other factors that can affect the reaction such as temperature, pH, the
concentration of substrate and concentration of enzyme must be controlled to avoid error and the
result becomes clearer and persist. Most importantly, make sure all the samples are the same size,
so the result will be more accurate.
CONCLUSION

In conclusion, the size and temperature of the sample influence the reaction of catalase. The
smaller the size of the particles, the higher the rate of reaction of catalase. When the temperature is
over 60℃ the enzyme denatured. Therefore, the enzyme is unable to break down the harmful
hydrogen peroxide.

The ground-up liver, kidney and potato have a faster rate of reaction than the cube. All of this is
proof that the size of particles affects the rate of reaction. While, the boiled liver did not produce
bubbles nor oxygen gas, shown that when the temperature is over 60℃ the enzyme denatured.
 REFERENCES

1. N. A., Reece, J. B.(2005).Campbell Italic Biology. (7th ed.). Benjamin Cummings, San Francisco.
154-155pp.

2. Sarina Mohamad (2012). Laboratory Manual for Metabolism & Cell Division. Shah Alam, Selangor:
UiTM Press.

3. Aebi, H. (1974). Catalase. In Methods of enzymatic analysis. Academic press. 673-684pp.

Retrieved from https://www.sciencedirect.com/science/article/pii/B9780120913022500323

4. Bonnichsen, R. K. (1947). Catalase activity. 685-709pp. Retrieved from

http://actachemscand.org/pdf/acta_vol_01_p0685-0709.pdf

5. Beers, R. F., & Sizer, I. W. (1952). A spectrophotometric method for measuring the breakdown of
hydrogen peroxide by catalase. J Biol chem. 195(1), 133-140pp. Retrieved from
https://www.semanticscholar.org/paper/A-spectrophotometric-method-for-measuring-the-of-by-
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