Josephine Davina Putri

Biology B
The Effect Of Substrate Concentration On The Activity Of
Enzymes
1. Introduction
o Problem
Does an increase in substrate concentration increase the
rate of reaction?
o Hypothesis
When there is an increase in substrate concentration
added to enzymes, the rate of reaction will become
faster until it reaches the optimum concentration where
all the active sites are occupied and the activity is
maximum.
o Background Information
An enzyme is a catalyst that speeds up the rate of
chemical reaction. A substrate is a substance that an
enzyme acts upon. If there is an increase in the amount
of enzyme, there will be a continuous increase in the
rate of chemical reaction. The result for this reaction will
be a linear graph which continuously increase. In this
experiment, potatoes are used as an enzyme, and the
amount of oxygen produced is measured to determine
enzyme activity. The amount Catalase that is contained
in potatoes can catalyze the conversion of hydrogen
peroxide into oxygen and water. The enzyme catalase
catalyzes the reaction:
2H2O2 =2H2O + O2.
Hydrogen peroxide is harmful for cells, so these cells
produce the catalase enzyme to remove it from the
body. In the human body, catalase is produced in the
liver to break down Hydrogen peroxide (IB Biology). In
this experiment, we can observe the decomposition
reaction by catalase to break down hydrogen peroxide
(Rembac). When substrate concentration is increased,

Josephine Davina Putri

Biology B
rate of reaction also increases, but once all the active
sites of the enzyme are fully occupied by substrates, the
rate of reaction is at maximum and would not increase
since the active sites are all occupied. (Aubusson,
Kennedy, and Hickman 288-289). The graph of
substrate concentration can be seen below:

The main objective of this quantitative experiment is to

determine the activity of catalase enzyme or the effect of increasing
the amount of hydrogen peroxide, which is the substrate in this
experiment. During the experiment, factors that must be kept
constant include the size of potato used, concentration and type of
measurement.

Six variations of hydrogen peroxide concentrations

were varied. The independent variable is the amount of hydrogen

peroxide and water in the test tube, and the dependent variable is
the activity of the enzyme. The experiment is repeated once to
make sure it is reliable.
2. Materials
o Hydrogen Peroxide
o Distilled Water
o Test Tubes
o 10ml pipettes with automatic filler
o Cork borer
o Potato
o Ruler
o Volumetric Flask
o Knife
o Stopwatch
3. Procedures

Josephine Davina Putri

Biology B
o 10% of hydrogen peroxide solution is prepared by
mixing 25ml of 40% hydrogen peroxide with 75% of
distilled water using the 10ml pipettes to fill the
volumetric flask to the 100ml line.
o The potato is cut using a cork borer into cylinder shapes
and those potatoes are measured and cut into the same
lengths.
o The 10% hydrogen peroxide solution was mixed with
different volumes of distilled water and put into 6
different test tubes according to the table below. Each
test tube are labeled to differentiate one from each
other.
Test tube

1
2
3
4
5
6

Amount of Water

Amount of

(mL)

Hydrogen Peroxide

0
0.5
1
1.5
2
2.5

(mL)
3
2.5
2
1.5
1
0.5

o The potatoes are put inside those solutions and height

of bubbles are measured using a ruler and time is
recorded using a stopwatch every 8 minutes.
o The experiment was repeated once.
4. Safety Precautions
o Always handle Hydrogen Peroxide with gloves because it
is harmful to the skin
o Be careful not to spill the hydrogen peroxide, because it
is a transparent liquid that can be mistaken as water
o Keep the hydrogen peroxide closed, so that it is not
exposed to light
o Be careful when cutting with a knife, because the hands
could be cut
5. Results

Josephine Davina Putri

Biology B
o How materials and equipments are set

o Record of data with time intervals: 8 minutes

o Table
Test

Volume

Volume

Percentag Heigh

Heigh

Averag

of

t at

hydroge

distille

concentra 8.00

16.00

height

d water tion

peroxide
0.5
1
1.5
2
2.5
3

2.5
2
1.5
1
0.5
0

4.3
4.1
5
4.7
4.5
5

3.89
3.9
4.65
4.6
4.65
5

Tube of

1
2
3
4
5
6

1.67%
3.33%
5%
6.67%
8.33%
10%

t at

3.6
3.7
4.3
4.5
4.8
5

o Duration of experiment: 16 minutes

o Graph

Josephine Davina Putri

Biology B

The Effect Of Substrate Concentration On The Activity Of Enzymes

1.67

3.33

5.0

6.67

8.33

10.0

percentage of substrate concentration (%)

6. Discussion
According to the results of the experiment, it is proven that
our hypothesis was right. When the substrate concentration
increases, the rate of reaction will be faster. However, the
reliability of this experiment is a problem, since this
experiment was only repeated once. It is better if the
experiment is repeated twice to get a more accurate average.
According to the research from second hand sources such as
the Internet and the textbook, there should be a point where
the activity is maximum and the graph shows a straight line.
In our experiment, it seems like the activity has not yet
reached maximum activity. Adding more concentration of
hydrogen peroxide can show the maximum activity and make
the data more accurate. Our data can also be not valid, since
we missed several seconds when measuring the height of
bubbles. A few seconds difference can change the data, and
make the data unreliable.
7. Conclusion
o Our hypothesis is correct, where if there is an increase
in substrate concentration added to enzymes, the rate
of reaction will become faster until it reaches the

Josephine Davina Putri

Biology B
optimum concentration where all the active sites are
occupied and the activity is maximum.
o More repetitions must be used so the data can be more
reliable
o The time must be observed carefully to improve
accuracy of the experiment.

Josephine Davina Putri

Biology B
8. References
"IB Biology on Decomposition of Hydrogen Peroxide by Enzyme
Catalase." IB Biology on Decomposition of Hydrogen
Peroxide by Enzyme Catalase. IB Biology, n.d. Web. 20 Nov.
2014.
Rembac, Justin. "Catalase and Hydrogen Peroxide Experiment."
Education.com. Education, n.d. Web. 20 Nov. 2014.