KYAMBOGO UNIVERSITY

FACULTY OF SCIENCE

DEPARTMENT OF CHEMISTRY

BACHELOR OF SCIENCE IN CHEMICAL AND

PROCESS ENGINEERING

NAME; OKELLO DAVIS JOSHUA

REG NO ;19/U/CHE/1642/PE

YEAR OF STUDY ; THREE

COURSE UNIT; BIOCHEMICAL ENGNEERING

TASK; LABARATORY REPORT

EXPERIMENT TO FIND OUT THE EFFECTS OF TEMPERATURE ON ENZYME
ACTIVITY OF CATALASE FOUND IN POTATOES USING HYDROGEN PEROXIDE

Objectives

 To find out the effect of temperature on the enzyme activity of catalase in potatoes using
hydrogen peroxide as a substrate.
 To find out the optimum temperature at which catalase works well.

Abstract

This experiment was carried out to find out the effect of temperature on the rate of enzyme
activity of catalase found in potatoes using hydrogen peroxide.

The potatoes were pealed crushed and same amount of it was measured and a known amount of
hydrogen peroxide was measured for each temperature under investigation both placed in a hot
water bath being maintained at a certain temperature and then the hydrogen peroxide was pored
in the test tube containing the crushed potato it was then left for one minute the foam height was
measured.

For each temperature under investigation.

A graph was obtained using the data obtained.

It was found out that the optimum rate of reaction was at 250c which is the room temperature.

The experiment was successful since the aims of the experiment were obtained.

Introduction

An enzyme is a substance that acts as a catalyst in living organisms by speeding up the rates of
chemical reactions in that organism . Enzymes, which are proteins, are the most common
biological catalysts . Enzymes are able to increase the rate of chemical reactions by lowering the
activation energy needed for a reaction to occur. However, one thing that enzymes do not do is
supply free energy to a reaction or change the total energy of a reaction . Enzymes also follow a
rule of substrate specificity. This means that each enzyme catalyzes the reaction of a single
substrate or a group of closely related substrates, and is also the reason why a typical cell in the
human body needs over 4,000 enzymes to function . Enzymes are also reusable, because they are
not consumed in the reactions they catalyze . Temperature and pH are two main factors that affect
enzyme activity. When fluctuations in temperature and pH occur, an enzyme may denature and
lose its function . For most enzymes, extremely high temperature and an extremely low and acidic
pH can cause the protein to denature. In lab, the enzyme catalase reacted with the substrate
hydrogen peroxide to produce water and oxygen in the form of bubbles. The catalase was sourced
from potatoes, since they contain the enzyme naturally.

Theory

. Enzymes are large protein molecules (i.e. macromolecules) that act as catalysts in the
biochemical reactions that occur in living things. A catalyst is a specific type of protein that
increases the rate of a chemical reaction that would otherwise occur too slowly. How do
enzymes increase the rate of a reaction? They lower the amount of activation energy that is
needed to get the reaction started.

In an enzyme-catalyzed reaction, the substance that the enzyme acts upon is known as the
substrate. Each enzyme is highly specific for a particular substrate because the enzyme has a
special area the active site. The active site has a unique shape, and like pieces of a puzzle, the
enzyme’s active site can only bind to a substrate with a complimentary shape. When the correct
substrate binds, it forms a union known as the enzyme-substrate complex, in which the enzyme
begins to change the substrate from its original form. After a short period of time, the substrate
is turned into a new product; the enzyme will be recycled to find a new substrate. This process is
summarized below:

Enzyme + Sustrate  Enzyme-Substrate complex  Enzyme + Product

Thus, it is the highly specific three-dimensional structure of an enzyme that will determine its
ability to function properly. However, there are environmental conditions that can change the
shape of an enzyme. When this happens, we say that the enzyme has become denatured, and its
activity decreases or possibly completely stops. There are two factors that can alter the shape of
(i.e. denature) enzymes: pH and temperature. An enzyme’s optimal pH and temperature are
present when it works most efficiently. For most human enzymes, an optimal pH is 7.4 (i.e.
neutral conditions) and an optimal temperature is 37oC. Most human enzymes become denatured
at extremely low/high pH levels (i.e. acidic conditions, pH 0-3, and basic conditions, pH 12-14).
Temperatures around 40-50oC will denature some human enzymes, but others won’t denature
until they reach 70-80oC; almost all enzyme activity will cease at100oC (i.e. boiling).

The enzyme you will investigate is this lab is called catalase. Catalase is found in the tissues of
many organisms (both plants and animals) because it plays a very important role in protecting
cells from a harmful chemical called hydrogen peroxide (H2O2). Hydrogen peroxide is a normal
by-product of cellular metabolism; however, it quickly becomes a poisonous toxin if it builds up
in the cell. The enzyme catalase is responsible for speeding up the breakdown of toxic hydrogen
peroxide into two harmless substances, water and oxygen. This chemical reaction is represented
by the following chemical equation:

2 H2O2  2 H2O + O2

(catalase)

Materials And Requirements

 Test tubes
 Hot water bath
 Thermometer
 6 % H2O2
 Measuring cylinder
 Potatoes
 Crusher
 Knife
 Weighing balance
Procedure

i. Six test tubes were set up each containing 10mL of 6 % H2O2, measured with a measuring
cylinder.
ii. the thermometer was used to measure the temperatures in the test tubes.
iii. 5 test tubes were set up with one piece of potato in each. two drops of detergent was
added.
iv. Using hot water bath, one test tube of H2O2 and one tube of potato was brought to the
appropriate temperatures.
v. the H2O2 was added to the potato
vi. the amount of reaction that occurs in each test tube was measured (the amount of
reaction was indicated by the height of the bubbles produced in 1 min)
vii. the experiment was repeated two more times and calculate the average reaction rates

Results

Rate of reaction
Suggested Actual
Height of foam produced in volume/reaction
Tube Temperature Temperature
reaction time (mm) time
(o C) (o C)
(mm/min)
      1 2 3 4 Av  
1 RT 25 70 58 55 60 60.75 60.75
2 35 32 45 48 45 31 42.25 42.25
3 45 45 52 36 34 30 38 38
4 55 54 34 32 28 31 31.25 31.25
5 75 70 31 31 32 28 30.5 30.5
 graph of rate of reaction against temper-
ature
rate of rection(mm/min)

70
60
50
40
30
20
10
0
20 30 40 50 60 70 80
temperature(0c)

Observation

The rate of reaction decreases with increase in temperature were by at room temperature which is
250c the reaction is higher it then decreases rapidly to a temperature of about 320c.the rate of
reaction then decreases slowly which is almost a constant between 32 and 45 .between 45and
550c the rate of reaction decreases constantly and between 54 and 700c the rate of reaction is
almost a constant.

 The bubbling reaction seen is the metabolic process of decomposition, This reaction is caused
by catalase, an enzyme within the potato
Discussion

The rate of reaction is high at 250c showing that the catalase in the sweet potatoes work best at
that temperature and in our experiment the highest foam was formed at this temperature

The rate of reaction decrease between 32 and 450c because enzymes are being denatured

At 70 0c almost all the enzymes are denatured hence least foam is formed this because the
enzyme catalase found in the potatoes is composed of proteins which are highly affected by high
temperatures.

Sources of Errors

 Errors in thermometer reading.

 Errors in measuring the time taken.
 Errors in measuring the height of the foam.
 Errors due to improper maintaining of constant temperatures of the hot water bath.
 Errors in weighing the potatoes.
 Errors in measuring the amount of hydrogen peroxide.

Recommendations to the errors

 Accurate readings of the thermometer should be taken.

 A stop clock should be used to measure accurate time.
 Height of the foam should be measured twice to ensure accuracy.
 The hot water bath should be well maintained at the temperature under investigation to
reduce errors.
 The hydrogen peroxide should be well measured.
 The amount of potatoes to be used should well weighed.
Conculusion

Catalase acts as the catalyzing enzyme in the decomposition of hydrogen peroxide. Nearly all
living things possess catalase, including us! This enzyme, like many others, aids in the
decomposition of one substance into another. Catalase decomposes, or breaks down, hydrogen
peroxide into water and oxygen. The experiment was successfully done since the objectives for
the experiment were obtained.
REFERENCES

Lab Manual: Duquesne University. 2019. “Catalytic Activity of Enzymes Part 2,”
BIOL111L .

Lab Manual; Morris, J., Hartl, D., Knoll, A., Lue, R., Michael, M. 2019. “Biology How Life
Works.” 3rd ed. New York (NY): Macmilla.

Audesirk, T., Audesirk, G., Byers, B.E., 2008. Life on Earth 5th Edition. Pearson Benjamin
Cummings. San Francisco, CA.
.
Suffolk County Community College. 2005. Principles of Biology Laboratory Manual.
MacGraw Hill Publishing Boston, MA.