Name: Date: Period:

Catalase Lab Activity

Introduction
Do you ever wonder why hydrogen peroxide bubbles when you pour it on your open
wound? Catalase, an enzyme made in many cells like those of your blood, as well as in
turnips, liver, and potatoes, causes the decomposition of hydrogen peroxide. In our
cells, catalase breaks down hydrogen peroxide to form oxygen gas and water.

2H2O2  O2 (g) + 2H2O

Students, you will investigate the presence of catalase in potatoes and examine the
effect of temperature, as in cooking, on the ability of enzyme to bind to and cause a
change in the substrate.

Caution should be taken when handling extremely hot water and chemicals like
hydrogen peroxide.

Materials
 Potato puree* (see note below)
 Fresh 3% hydrogen peroxide**
 5 large test tubes, 25 x 200 cm
 Disposable plastic transfer pipettes
 Sharpie or other marker
 Bunsen burner or coffee maker for hot water
 Ruler
 Stirrers

* Potato puree can be prepared by blending 1 cup of diced potato with 1 cup of
water. Blend until potato pieces are smaller than ¼ inch.

** Dilute hydrogen peroxide solution to 3% if a higher-concentration solution is

available.

Procedure
Measuring potato catalase activity:
1. Place a mark on a test tube at 2 cm and 5 cm from the tube bottom.

2. Transfer potato puree using a scoopula to the 2 cm mark on the test tube. (You
can also use a popsicle stick to transfer the potato puree.)

3. Add approximately 3 cm of hydrogen peroxide using a disposable plastic transfer

pipet (DPTP) or some other pipet. Stir the contents with a stirring rod.

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 4. Observe the formation of the oxygen gas bubbles that develop from the action of
the potato enzyme acting on the hydrogen peroxide.

5. Wait one minute and then measure the height of the foam by measuring from the
top of the liquid to the top of the foam.

6. Now measure the height of the foam each minute for a total of 5 minutes.

7. Record the height of the foam onto Table 1.

8. Construct a graph of foam height (y-axis) vs. time (x-axis). Label the graph as
Graph #1.

Measuring the effect of temperature on catalase activity:

1. Label four more test tubes: 1, 2, 3, and 4.

2. Place potato puree 2 cm deep in each test tube as before.

3. Place each test tube at a constant temperature for 3 minutes:

a. Test tube #1 - room-temperature (20-25C)

b. Test tube #2 - ice water bath (0-4 C) (Put ice in water for 5 minutes and
measure temperature until stable)

c. Test tube #3 - body-temperature water bath (37-40 C) (Use tap water if a
water bath is not available; measure temperature with a thermometer so
that the temperature is consistent throughout the class).

d. Test tube #4 – boil water with a Bunsen burner (or bring a coffee maker to
produce hot water); record temperature with a thermometer that can
withstand temperatures greater than 100 C.

4. Add 3 cm of 3% hydrogen peroxide per test tube. Continue incubating potato

puree in the presence of hydrogen peroxide at their respective temperatures.

5. After 5 minutes, compare the rate of oxygen production by measuring the foam
height in each tube. Enter data in Table 2.

6. Construct a graph of foam height (y-axis) vs. temperature (x-axis). Label the
graph as Graph #2.

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Name: Date: Period:

Data Analysis
Measuring Potato Catalase

Table #1
1 minute 2 minutes 3 minutes 4 minutes 5 minutes
Foam Height
(cm)

Graph #1

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3
 Data Analysis

The Effect of Temperature on Enzyme Action

Table #2
Test Tube #1 Test Tube #2 Test Tube #3 Test Tube #4
Foam Height (cm)

Graph #2

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 Data Analysis

1. Describe what happens in the first five minutes after potato pulp at room
temperature is added to hydrogen peroxide.

2. Examine the results from other groups and compare the rates of oxygen production
at different temperatures. Explain any differences between groups.

3. Why do you think that the height of the foam varied with both increasing and
decreasing temperature?

4. Why do you think it is important to decompose hydrogen peroxide produced in

cells?

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5
 Answer Key

1. Describe what happens in the first five minutes after potato pulp at room
temperature is added to hydrogen peroxide. Potatoes contain the enzyme
catalase, which reacts with hydrogen peroxide to form oxygen and water.
Potato cells release their contents as they are broken up by blending and
catalase binds to its substrate, hydrogen peroxide, producing the gas. The foam
is a result of the gas and this is what the students are measuring. The height of
the foam is an indirect measure of the gas produced during the chemical
reaction.

2. Examine the results from other groups and compare the rates of oxygen production
at different temperatures. Explain any differences between groups. Small
inaccuracies in measuring out the potato puree and the hydrogen peroxide will
likely occur and these may cause a slight variability in the foam height.
Another source of error to be aware of is that the potato puree needs to be
homogeneous so that all students will receive a representative sample. Make
sure the students stir the potato puree before removing their sample to avoid
having sample differences.

3. Why do you think that the height of the foam varied with both increasing and
decreasing temperature? The speed of an enzymatic reaction is dependent on
movement of molecules to interact. Molecular diffusion slows down with
decreasing temperature. Therefore, in lower temperatures, the enzyme and
substrate (catalase and hydrogen peroxide) will not interact as frequently.
With high temperatures, the hydrogen bonds that stabilize the structure of the
catalase will be disrupted causing a change the enzyme shape. This is
especially relevant when the conformation of the active site is altered. The
enzyme loses its ability to bind to the substrate, thus losing its ability to
produce product by the reaction.

4. Why do you think it is important to decompose hydrogen peroxide produced in

cells? The students may not realize that hydrogen peroxide can participate in
oxidative damage to cells during oxidative stress caused by pollutants, tobacco,
and radiation. Catalase is an enzyme that acts to reduce damage by converting
hydrogen peroxide to oxygen and water.

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