Nimisha Sharma

Biology
IB1
Effect of the substrate concentration on enzyme
catalysed reaction

Aim: To investigate how does changing the substrate concentration of

hydrogen peroxide affect the rate of reaction whereby hydrogen peroxide
is broken down to oxygen gas and water by the enzyme catalase,
measured by the volume of oxygen gas released in 30 seconds.

Raw Data
Table 1- Showing volume of Oxygen released at different
concentrations of hydrogen peroxide over 30 seconds.
Concentra
tion of
H2O2 %
(1.0)
Trial
s
1
2
35
3
30
25
20
15
10
5

1
2
3
1
2
3
1
2
3
1
2
3
1
2
3
1
2
3

Volume of O2 released/cm3 0.5

5.0/sec
0.5
5.0
13.0
19.0

10.0/se
c 0.5
25.0
30.0
40.0

15.0/se
c 0.5
42.0
34.0
48.0

20.0/se
c 0.5
61.0
37.0
53.0

25.0/se
c 0.5
79.0
39.0
54.0

30.0/se
c 0.5
88.0
41.0
55.0

2.0
3.0
9.0
3.0
4.0
4.0
2.0
3.0
7.0
3.0
3.0
3.0
0.0
3.0
4.0
0.0
2.0
3.0

18.0
27.0
20.0
6.0
5.0
6.0
4.0
6.0
9.0
5.0
6.0
4.0
0.0
4.0
4.0
0.0
3.0
3.0

42.0
34.0
48.0
15.0
9.0
10.0
5.0
10.0
12.0
7.0
10.0
5.0
0.0
4.0
5.0
0.0
4.0
5.0

65.0
72.0
75.0
23.0
13.0
12.0
10.0
16.0
17.0
10.0
14.0
6.0
0.0
6.0
6.0
0.0
6.0
6.0

78.0
82.0
88.0
36.0
23.0
34.0
14.0
24.0
25.0
12.0
18.0
8.0
0.0
7.0
6.0
0.0
7.0
6.0

85.0
85.0
94.0
52.0
42.0
54.0
19.0
25.0
31.0
15.0
23.0
11.0
0.0
9.0
7.0
0.0
9.0
7.0

Data Processing
Table 2- Showing average volume of oxygen released at each
concentration of hydrogen peroxide
5.0/s
ec
0.5

Volume of
10.0/s 15.0/s 20.0/s
ec
ec
ec
0.5
0.5
0.5

O 2/cm3
25.0/s
ec
0.5

12.3

31.7

41.3

50.3

57.3

61.3

30

1
2
3

4.7

21.7

40.7

70.7

82.7

88.0

25

1
2
3

3.7

5.7

11.3

16.0

31.0

49.3

20

1
2
3

4.0

6.3

9.0

14.3

21.0

25.0

1
2
3

3.0

5.0

7.3

10.0

12.7

16.3

10

1
2
3

2.3

2.7

3.7

4.7

1
2
3

1.7

2.0

3.0

4.0

H2O2
Concentrat
ion % (
1.0)

35

Tria
ls
1
2
3

5.7

4.3

0.5
30.0/s
ec
0.5

6.7

5.3

Table 3- Showing the Error bars (Max-Min/2)

H2O2

5.0/s

Volume of O 2/cm3 0.5

10.0/s 15.0/s 20.0/s 25.0/s 30.0/s

Concentrat
ion % (
1.0)
35
Trial 1
Trial 2
Trial 3
30
Trial 1
Trial 2
Trial 3
25
Trial 1
Trial 2
Trial 3
20
Trial 1
Trial 2
Trial 3
15
Trial 1
Trial 2
Trial 3
10
Trial 1
Trial 2
Trial 3
5
Trial 1
Trial 2
Trial 3

ec
0.5

ec
0.5

ec
0.5

ec
0.5

ec
0.5

ec
0.5

7.0

7.5

7.0

12.0

20.0

23.5

3.5

4.5

10.0

5.0

5.0

4.5

0.5

0.5

3.0

5.5

6.5

6.0

2.5

2.5

3.5

3.5

5.5

6.0

1.0

2.5

4.0

5.0

6.0

2.0

2.0

3.0

4.0

5.0

6.0

1.5

1.5

2.5

3.0

3.5

4.5

0.0

The data collected above had to be processed in order to draw a

meaningful conclusion from it. First, I found the mean volume of Oxygen
released for 3 trials of each different substrate concentration. Then I
calculated the error bars (range) around the mean for all the trials of each
different substrate concentration.
Sample calculations
1. Calculation of average Volume of the Oxygen produced = Sum of all
trials/No. of trials
2. Calculation of error bars (range) of the volume of oxygen produced= MaxMin/2 (cm3)
3. The gradient in each graphs is found by using the Microsoft Excel on the
computer after generating the graphs.

Graph 1

Volume of oxygen produced in thirty seconds when the concentration of hydrogen peroxide is 5%
6
5 f(x) = 0.18x
R = 0.99
4

Volume of oxygen/cm 3
2
1
0
0

10

15

20

25

30

35

Time/seconds1

Graph 2

Volume of oxygen produced in thirty seconds when the concentration of hydrogen peroxide is 10%
8
7

f(x) = 0.23x
R = 0.99

6
5

Volume of Oxygen/cm 4
3
2
1
0

10 15 20 25 30 35

Time/seconds 1

Graph 3

Volume of oxygen produced in thirty seconds when the concentration of hydrogen peroxide is 15 %
20
15

f(x) = 0.52x
R = 1

Volume of Oxygen/cm 10
5
0

10 15 20 25 30 35

Time/seconds 1

Graph 4

Volume of oxygen produced in thirty seconds when the concentration of hydrogen peroxide is 20%
30
25
f(x) = 0.78x
R = 0.99

20

Volume of Oxygen/cm 15
10
5
0

10 15 20 25 30 35

Time/seconds 1

Graph 5

Volume of oxygen produced in thirty seconds when the concentration of hydrogen peroxide is 25%
60
50
40
f(x) = 1.24x

Voume of Oxygen/cm 30 R = 0.91

20
10
0

10 15 20 25 30 35

Time/seconds 1

Graph 6

Volume of oxygen produced in thirty seconds when the concentration of hydrogen peroxide is 30%
100
90
f(x) = 3.06x
80 R = 0.98
70
60
Volume of Oxygen/cm 50
40
30
20
10
0
0 5 10

15

20

Time/seconds 1

25

30

35

Graph 7

Voume of Oxygen produced in thirty seconds when the concentration of hydrogen peroxide is 35%
70

f(x) = 2.32x
60 R = 0.98
50
40

Volume of Oxygen/cm 30
20
10
0

10

15

20

25

30

35

Time/seconds 1

Processed data
Table 4- Showing the gradients found in the graphs 1-7
Concentration of H2O2 (%)
0
5
10
15
20
25
30
35
Graph 8

Rate of Reaction cm3 sec-1

0.0x
0.185x
0.234x
0.519x
0.782x
1.24x
3.06x
2.32x

Effect of the substrate concentration(H2O2) on the rate of reaction

3.5
3
2.5
Rate of reaction cm sec

2
1.5
1
0.5
0

10 15 20 25 30 35 40

Concentration of H2O2

Conclusion
The data obtained from this experiment suggests that as the concentration of
hydrogen peroxide increases, the rate of reaction increases and that my
hypothesis is valid. The data shows that as the substrate concentration of
hydrogen peroxide increases the amount of oxygen production also increases; it
can be seen on the processed data where the average oxygen produced in 5%
concentration of hydrogen peroxide is 5.3cm3 whereas in 30% concentration it is
88.0 cm3. On the other hand, Average volume of Oxygen produced in 35%
concentration of the hydrogen peroxide is lower than 30% concentration of
hydrogen peroxide, which suggests that it might have reached its optimum level
and supports the hypothesis that the rate of the reaction only increases until a
certain point.
The R2 value obtained from each graph shows that there is a positive correlation
between the rate of reaction and the hydrogen peroxide concentration as all of
the graphs have a high R2 value and the trend line crosses most of the points of
each graph. While Graph 5 have a quite low R 2 value of 0.8241, and only few
points crosses the trend line which suggests that there might be some errors.
Final Graph 8 shows that even though due to quite low R value in Graph 5 the
rate of reaction increases each time when the concentration of the substrate
Hydrogen Peroxide increased. Thus, the results lead to the conclusion that the
rate of reaction is directly proportional to the substrate concentration.

Evaluation

Overall the experiment was very good and very well supported the
hypothesis. There were several reliable aspects to this experiment which
contributed to the success of the experiment. First of all, the average
volume of Oxygen helped to deduce that as the concentration of the

substrate increases the rate of reaction also increases. Second of all, the
uncertainty of 0.5 is very low and cannot effect the reaction very much.
Third of all, the R2 value was very reliable as when R2 equals 1.0, all points
lie exactly on a straight line or trend line . The values of R2 in graph 1-7 are
also very close to 1.0 which suggests that the experiment was carried
pretty well and the results are also good. But, even though it was
performed carefully to minimize errors, in graph 5 there are only few
points on the trend line and the R 2 value is lower than the other graphs.
The highest R2 value is in Graph 3 of 0.9938 which is very close to 1 and
the lowest value is in Graph 5 of 0.8241 which is quite far from 1 and
suggests that there were few errors.

Weaknesses
1. In some of the trials, when the
Hydrogen Peroxide was added
into the catalase test tube, the
cork was not sealed quickly.
Therefore, the collected data
may have few errors and some
of the rates of enzyme activity
may be lower than the actual
rate.
2. In some of the trials, the
stopwatch was not started
frequently just after adding the
hydrogen peroxide into the
catalase test tube.
3. Yeast respires anaerobically, so
we did not know that if the
reaction is actually producing
O2 and not CO2.

Improvements
In order to improve this human error,
we can organize things first and start
the experiment after making sure we
have all the equipment ready and we
can also use more improved
apparatus to improve this.

In order to get accurate results, we

can use more advanced apparatus
instead of a stopwatch as it would not
make any error and will help to get a
good result.
By using an Oxygen sensor, we can
differentiate between O2 gas and CO2
gas which would enhance the results
and will make it more accurate.