Activity 6

CATALASE ENZYME

I. Background Information: Liver and other living tissues contain the enzyme catalase. This enzyme breaks down
hydrogen peroxide, which is a harmful by-product of the process of cellular respiration if it builds up in
concentration in the cells. If we use potato or other tissue containing this enzyme, we can use this to measure
the relative influence of varying several different factors on the activity of enzymes in living tissue.
In order to obtain energy and building blocks from food, the digestive system must break down proteins, fats and
carbohydrates. In this process, specific enzymes catalyze hydrolysis reactions in which food polymers are broken
up into monomers.

II. Introduction: What would happen to your cells if they made a poisonous chemical? You might think that they
would die. In fact, your cells are always making poisonous chemicals. They do not die because your cells use
enzymes to break down these poisonous chemicals into harmless substances. Enzymes are proteins that speed up
the rate of reactions that would otherwise happen more slowly. The enzyme is not altered by the reaction. You
have hundreds of different enzymes in each of your cells.
Each of these enzymes is responsible for one particular reaction that occurs in the cell. In this lab, you will study
an enzyme that is found in the cells of many living tissues. The name of the enzyme is catalase (KAT-uh-LAYSS); it
speeds up a reaction which breaks down hydrogen peroxide, a toxic chemical, into 2 harmless substances--water
and oxygen.
The reaction is as follows: 2H2O2 ----> 2H2O + O2
This reaction is important to cells because hydrogen peroxide (H 2O2) is produced as a byproduct of many normal
cellular reactions. If the cells did not break down the hydrogen peroxide, they would be poisoned and die. In this
lab, you will study the catalase found in liver cells. You will be using chicken or beef liver. It might seem strange to
use dead cells to study the function of enzymes. This is possible because when a cell dies, the enzymes remain
intact and active for several weeks, as long as the tissue is kept refrigerated.

III. Objectives: Demonstrate the activity of an enzyme in living tissues and examine the effects of
environmental changes on enzymatic activity.

IV. Pre-lab: Answer the following questions.

1. What type of biological molecule are enzymes?

2. Describe how enzyme works and its importance to living organisms.
3. Define the following terms as they apply to enzymes:
a. Active Site
b. Substrate
c. Denature
V. Materials:

 6 Test tubes and Test tube holder

 10-ml Graduated cylinder
 Scissors and Forceps
 Stirring Rod
 Beaker
 Hot Plate
 Hydrogen peroxide
 Ammonia
 Fresh Liver

VI. Procedure

PART A - Observe Normal Catalase Reaction

1. Using forceps and scissors, cut a small piece of liver and put it into the test tube. Push it inside the test
tube using a stirring rod.
2. Add 2 ml of hydrogen peroxide into the test tube. Observe the bubbles. Throughout this investigation,
estimate the rate of the reaction (how rapidly the solution bubbles) by using the following legend below
and record it on the data table provide below:
5 – Extremely active bubbling; foamy
3 – Moderately active bubbling
1 – Less active bubbling
0 – non-reactive; no bubbling

3. Feel the test tube. Note that the test tube feels warmer after the reaction. Record this
observation on the data table.

Data Table A. Catalase Reaction

Content of Test Tube Rate of Reaction Remarks/Observation

Normal Liver

Photo Documentation
PART B – Is Catalase Reusable?
1. Place a small piece of liver into a clean test tube and add 2 ml of hydrogen peroxide solution. Observe
the reaction. Record the rate of reaction and your observation into the data table provided below.

2. Pour off the liquid into a second test tube. Add a piece of liver in this test tube and observe the
reaction. Record the rate of reaction and your observation into the data table provided below.

3. In the first test tube with remaining liver, add another 2 ml of hydrogen peroxide and observe the
reaction. Record the rate of reaction and your observation into the data table provided below.

Data Table B. Catalase Reaction

Content of Test Tube Rate of Reaction Remarks/Observation

Normal Liver

Liver Added to Used

Peroxide

Used Catalase

Photo Documentation:

PART C - What is the Effect of Temperature on Catalase Activity?

1. Put a piece of liver inside a beaker and soak it in distilled water for several minutes. After soaking,
transfer the piece of liver using forceps into a clean test tube and add 2 ml of hydrogen peroxide
in it. Observe the reaction. Record the rate of reaction and your observation into the data table
provided below.

2. Put a piece of liver inside a test tube, add distilled water and place this test tube in a boiling water
bath for 5 minutes. After which, remove the test tube from the hot water bath, allow it to air
cool, then drain out the water. Add 2 ml of hydrogen peroxide. Observe the reaction. Record the
rate of reaction and your observation into the data table provided below.

Data Table C. Catalase Reaction

Content of Test Tube Rate of Reaction Remarks/Observation

Liver soaked in distilled

water

Boiled liver
Photo Documentation:

PART D - What is the Effect of pH on Catalase Activity?

1. Put a piece of liver inside a beaker and soak it in ammonia for several minutes. After
soaking, transfer the piece of liver using forceps into a clean test tube and add 2 ml of
hydrogen peroxide in it. Observe the reaction. Record the rate of reaction and your
observation into the data table provided below.

Data Table D. Catalase Reaction

Content of Test Tube Rate of Reaction Remarks/Observation

Liver soaked in ammonia

Photo Documentation:

VII. Analysis

PART A

1. What gas is being released after the hydrogen peroxide was added to the piece of liver? How
can we confirm it?

2. Is the catalase reaction endothermic or exothermic? Explain.

PART B

1. What is happening inside the test tube after the hydrogen peroxide was added to the piece
of liver?

2. Assuming that the reaction is complete in Procedure 2, what do you think is the liquid
you poured into the other test tube composed of? How do you know this?

3. After completing procedure 3, do you think catalase is reusable?

PART C

1. Describe the reaction of the hydrogen peroxide and boiled liver. What is the scientific
explanation for this?
 2. What do you think will happen to the reaction if the liver was kept in a freezer for so long?

3. How does temperature affect the activity of catalase?

4. What is the optimum temperature for catalase?

PART D

1. Describe the reaction of the hydrogen peroxide and liver soaked in ammonia. What is the
scientific explanation for this?

2. What do you think will happen to the reaction if the liver was soaked in a strong acid like
hydrochloric acid?

3. How does pH affect the activity of catalase?

4. Does there appear to be an optimum pH for catalase to properly function? What is it?

VIII. Generalization: What have you learned from this activity?