Catalase Practical

Alysia Ninham
SACE #465217G
Practical Partners: Courtney Bronca and Jordan Graf
Abstract
The practical experiment conducted on a specific catalase, being
yeast, in reaction time with hydrogen peroxide (H2O2). The purpose
of the scientific experiment was to observe how concentration of a
changing substrate, hydrogen peroxide, affects the reaction rate of
the yeast enzyme catalyst. Techniques involved in this practical
included the precise measurement of hydrogen peroxide in each
concentration, accurate timing of reaction rates, numeracy skills
including the averaging of replicates, weight and measurement
calculations using scales and equipment. The method on the paper
disks is called the floating disk method. The results obtained from
this test conclude that with higher concentrations of a solution, the
reaction time will be quicker. This can be explained using the
collision theory of particles. The results reflected a very noticeable
relationship between concentration and reaction rate. The trend
could indicate that with higher concentrations of solutions, the
reaction rate is quicker.
Introduction
Macromolecules (DOnofrio, 2009) are important in all biological
processes. They are separated into 4 main groups; proteins, nucleic
acids, lipids and carbohydrates.
Catalases (Britannica, 2014) are important enzymes, a type of
protein, used to help reactions in biological processes. Enzymes
(Crierie, A., 1999) are important protein macromolecules. Catalases
(Dean, C., 2015) are found in a lot of living cells. This includes yeast.
Without the use of enzymes it would take your body days to digest a
single meal.
In reference to the reactions in this practical, the design can be
referred to the particle collision theory. Collision theory (Clark, 2002)
features a theory that says when the concentration of a substance is
higher, the reaction rate will be quicker. This is because in equal
volumes, a higher concentration has more particles, therefor more
chances to collide with other particles to react with.
The practical generally involves one main equation for scientific
purposes of the experiment.
H2O2

catalase

H20 + O2

This equation says that when a catalase, like yeast, is added to the
hydrogen peroxide, the products that are formed are water ad
oxygen. In the purposes of this practical, there is no way we can test

Catalase Practical
Alysia Ninham
SACE #465217G
Practical Partners: Courtney Bronca and Jordan Graf
for the appearance of water, so we are testing the formation on
oxygen.
In this practical an enzyme, yeast, acting as a catalase is placed in a
solution of hydrogen peroxide, acting as a substrate. In this practical
the hydrogen peroxide binds to the active sites of the catalase. The
oxygen bubbles produced by the reaction should cause the paper
disks to rise to the surface of the solution, thus representing a
reaction.
For this practical, the suspected hypothesis is that if the
concentration of the hydrogen peroxide increases, then the reaction
time will be quicker. The hypothesis will be supported if the graph
shows a noticeable trend between the increase of concentration
with a faster reaction time. The hypothesis will not be supported if
the results show no relationship.
The independent variable in this practical is the concentration of the
hydrogen peroxide solutions. This is manipulated by using different
dilution rates of the substance.
The dependent variable is the reaction time taken for the paper disk
to float in the solution. This is measured by a stopwatch and
recorded. Averages of the times were calculated to reduce the
effects of random error.
A control group was used, represented by the 0% concentration of
hydrogen peroxide. This was used to demonstrate that the disks
only reacted when there was a substrate present.
Materials and Procedure
1 packet (7g) yeast
teaspoon sugar
Stirring rod
7 250ml beakers
100mL water at 30 Celsius
40mL of each level of concentration of hydrogen peroxide
(H2O2) (0, 0.1, 0.2, 1, 3, 5)
18 paper disks
Paper towel
Forceps
Stopwatch
1. A yeast suspension was created using the 7g of yeast, tsp.
of sugar and 100mL of warm water in a beaker.

2.
3.
4.
5.
6.
7.

Catalase Practical
Alysia Ninham
SACE #465217G
Practical Partners: Courtney Bronca and Jordan Graf
40mL of each concentration of hydrogen peroxide was
measured into 6 of the remaining beakers.
Using the forceps the paper discs were dipped into the yeast
and were placed, one at a time, at the bottom of each
solution.
The time taken for the disk to rise (floating disk method) in
each solution was recorded.
If disc did not rise after 5 minutes, the measurement was
recorded as >300 seconds.
Steps 3-5 were repeated for each solution including 3
replicates.
The reaction rate between the catalase (yeast) and the
hydrogen peroxide was calculated and graphed using the
formula below.

Reaction rate =

1
Average reaction time( s)

Results
Hydrogen
peroxide
concentrati
on (%)
0

Reactio
n
Replicat
e 1 (s)
>300.00

Raw Data
Reactio
Reactio
n
n
Replicat Replicat
e 3 (s)
e 3 (s)
>300.00 >300.00

0.1
0.2
1
3
5

90.56
53.12
16.39
12.46
5.03

30.09
11.23
10.50
3.42
3.12

45.98
10.58
5.47
1.69
1.50

Averag
e (s)

Reactio
n Rate
(s-1)

>300.0
0
55.54
24.98
10.79
5.86
3.22

0.003
0.018
0.040
0.093
0.171
0.311

Catalase Practical
Alysia Ninham
SACE #465217G
Practical Partners: Courtney Bronca and Jordan Graf

Hydrogen Peroxide Concentration Effects On Reaction Rates

The trend of the above shows a direct relationship between the

concentration of hydrogen peroxide and the average times for the
reaction to take place. The graph follows a generally curved trend
with a few outliers. These could be caused by any or multiple errors
discussed below.
The trend also shows that when the concentration increases, the
reaction rate will eventually slow down to a minimal increase. This is
because of the overworking of the one enzyme. Eventually the
enzyme will become tired in a way and fail to increase the reaction
rate.
Discussion
The trend provided by the results is explained by the collision
theory, mentioned in the introduction. The reaction rate was faster
with a higher concentration because there were more particles in
the same volume of solution.
Possible sources of random error may include unknown
contamination, or inhibitors. This could be reflected in some of the
outliers, for example the very first data reading is noticeable higher
than the other readings. The reaction time could be extended if
inhibitors have bound to the activation site, which would stop the
substrate having a reaction with the catalase.
Another random error could be the reuse of the hydrogen peroxide.
Because the same hydrogen peroxide solution was used for each

Catalase Practical
Alysia Ninham
SACE #465217G
Practical Partners: Courtney Bronca and Jordan Graf
replicate there could be a chance of the catalase having an effect on
the following reaction. If any yeast resided in the bottom of the
beaker it may have an influence on acting with the new disk in
creating more oxygen bubbles, thus making the reaction time
increase. This is identified by the increasing trend of reaction times
with every replicate.
The catalase could have also contributed to a source of random
error. If the yeast solution was sat stationary for too long it can
separate creating more of a yeast suspension. If the solution was
not stirred before the paper disk was submerged it may have
affected the yeast on the paper disk. It can be hard to see in the
results if an affect was present.
Another source of random error could be the presence of inhibitors.
These could have come from contamination through direct touching
or contact with amino acids.
No systematic errors can be detected, as there are no true values
for this practical. This reason also means there are no comments
that can be made on the accuracy of the results.
Strengths in the practical design include the variety of
concentrations in the hydrogen peroxide, the replication of tests
reducing the effects of random error and the sterile use of forceps
with the paper disks to reduce contamination. The practical also
includes a high level of accuracy in the time measurement. The
stopwatch was able to measure up to two decimal places. The
calculations of the reaction times then allowed the data to be
increased in decimal places, thus increasing the reliability of the
data.
A weakness includes the size of the paper disk, because they were
so small it made it hard to place them at the bottom of the solution
and keep hold of them when submerged in the yeast.
Some improvements could be to have larger paper disks making it
easy to hold onto. To do this, it may affect the reaction rate time.
This is a high reason why there are no exact values. Another
improvement could be to have to sterilize the container the paper
disks were stored in. Although the paper disks were not directly
touched there can be some chance of contamination from previous
contents of the container. Also, increasing the variety of hydrogen
peroxide concentration. Although there was a wide variety, more
concentration levels may reduce the effects of the random errors by
even more, creating more precise results.

Catalase Practical
Alysia Ninham
SACE #465217G
Practical Partners: Courtney Bronca and Jordan Graf
The results from the practical can help a connection be made as to
why hydrogen peroxide has an effect on the living cells in our body.
The presence of hydrogen peroxide or substrate allows our body to
be alive. When a catalase is added, such as from our food, it allows
a reaction to occur. This reaction allows our cells to do our job. This
is an explanation on how our body digests food, as mentioned in the
introduction.
Conclusion
An evaluation on the results can show that the hypothesis was
supported. This is reflected by the precision in the graph and the
trend that follows the hypothesis statement in the introduction. This
is identified by the relationship between the increasing hydrogen
peroxide levels with the simultaneous increase of the reaction times
of the paper disks.
Reference List
Crierie, A., 1999. Biology SACE 2 Key Ideas. 3rd ed. South Australia:
Greg Eather.
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biochemistry | Encyclopedia Britannica. [ONLINE] Available at:
http://www.britannica.com/EBchecked/topic/99062/catalase.
[Accessed 27 February 2015].
An introduction to the collision theory in rates of reaction. 2015. An
introduction to the collision theory in rates of reaction. [ONLINE]
Available at:
http://www.chemguide.co.uk/physical/basicrates/introduction.html.
[Accessed 27 February 2015].
Building Blocks of Life Study Guide - Biology101.org. 2009. Building
Blocks of Life Study Guide - Biology101.org. [ONLINE] Available at:
http://www.biology101.org/biologystudyguides/buildingblocksoflife.p
hp. [Accessed 27 February 2015].
Collins, P., Penny.Collins@asms.sa.edu.au, 2015. Prac Tomorrow. [Email] Message to BC BiologyClass2015 (ninh0003@asms.sa.edu.au).
Sent 25/02/2015 9:07am. Available at: 25/02/2015 [Accessed 26
February 15].