The effect of substrate concentration

Aim:
To test the effect of substrate concentration on the rate of reaction catabolic reaction of Catalase
and Hydrogen Peroxide

Equipment:

5 test tubes
Test tube rack
Marker pen
Potato
Distilled Water
Hydrogen Peroxide
Cork borer
Ruler
Razor blade
Chopping board
Personal Protective Equipment
Pipette
10ml measuring cylinder

Method:
1. Using the cork borer and the razor blade cut 5 pieces of potato at a diameter of roughly
8mm and a length of 10mm.
2. Using the marker number the test tubes 1-5
3. Fill the test tubes with the appropriate volumes of distilled water and Hydrogen Peroxide
(refer to table)
Test Tube
1
2
3
4
5
Volume H2O2 0
2.5
5.0
7.5
10.0
ml
Volume of
10.0
7.5
5.0
2.5
0
Distilled
water ml
4. Place a piece of potato into each of the five test tubes and mark the height of the liquid in
each test tube.
5. After 5 minutes, measure the height of the bubbles of oxygen that have formed. Measure
from the original mark to the top of the bubbles.
6. Record data

Hypothesis:
I hypothesize that as the concentration of the substrate is increased the rate of reaction will
increase retospectivley.

Results:
Height of bubbles
Group
Group
1
2
(mm)
(mm)

Group
3
(mm)

TestTube Vol. H202

Vol. H20
#
(mL)
(mL)
1
0.0
10.0
0
0
0
2
2.5
7.5
17
40*
15
3
5.0
5.0
4*
45
40
4
7.5
2.5
21
65
55
5
10.0
0.0
23
80
70
* Outliers. Were not taken into account when calculating averages

Group
4
(mm)
0
16
22
27
27

Group
5
(mm)

Average
(mm)

0
11
25
30
35

Results
50.0
45.0

Height of bubbles (mm)

40.0
35.0
30.0
25.0
Series1

20.0
15.0
10.0
5.0
0.0
1

Test tube No.

0.0
14.8
33.0
39.6
47.0

Once the potato was added the substances began to react in test tubes 2-5. This resulted in the
production of white bubbles that formed at the top of the liquid. As the volume of hydrogen was
increased the amount of bubbles produced was increased. There were no bubbles produced in test
tube 1 as there was no presence of hydrogen peroxide to react with the substance. Test 5 produced
the largest amount of bubbles as it only contained hydrogen peroxide.

Diagram:

Discussion:
From the results it is clear that there was a correlation between the height of the bubbles produced
and the level of hydrogen peroxide present. This can be seen in test tube 2 which had 2.5 ml of
hydrogen peroxide and test tube 4 which had 7.5ml of hydrogen peroxide. Test tube 4 which had
the greater amount of hydrogen peroxide on average had its bubbles reach a point 24.8mm higher
than test tube 2. This is because the hydrogen peroxide is the substrate on which the enzyme
catalase is acting. This also explains why there was no change in test tube 1 as there was no
substrate for the enzyme to act upon. Overall the experiment was successful in testing the effect of
substrate on the reaction of enzymes.
The experiment did have errors this can be seen in the results which featured two major outliers in
group 1s test tube 3 and group 2s test tube 2. The accuracy of the experiment could be improved
by repeating the experiment to increase the number of results. Also another reason that could have
led to error was the fact that the potato being an organism could have had growth or genetic defects
this could potentially result in certain parts of the potato have a lower concentration of catalase
than others.

Hazard Report:
Hazard
Potato
Hydrogen Peroxide
Razor blade

Risk
Potato is a biological substance
and can be a pathogen. This can
be toxic if it is ingested
Hydrogen peroxide is toxic and
could be dangerous if it
contacts skin or is ingested
The razor blade is a sharp
implement for cutting this can
be dangerous if not handled
correctly

Control
Personal protective equipment
and not ingesting the
substance.
Personal protective equipment
and correct procedure
Correct safety procedure and
use of equipment

Conclusion:
From the results we can conclude that as the concentration of the substrate is increased the rate of
reaction is increased. This can be seen by reviewing the results for test tube 2 which had 2.5ml of
hydrogen peroxide and test tube 5 which had 10ml. test tube 5 had a much greater amount of
bubbles produced than test tube 2. This proves that the greater the concentration of substrate the
greater of reaction (note this is only up until the substance becomes saturated and the enzymes
reach their maximum rate of reaction).