Name : ______________________________________ Class: 1 2 3 4 5 6 7 Date: _____________
Enzyme Catalysis Lab
Learning Objectives
● Measure the effects of change in temperature and pH on reaction rates of an enzyme,
● Explain how environmental factors affect the rate of enzyme-catalyzed reactions.
INTRODUCTION
What would happen to your cells if they made a toxic 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 catalyze (speed up) the rate of biochemical reactions that would
otherwise happen more slowly. They do this by decreasing the activation energy (amount of
energy required to start) for a metabolic reaction. The enzyme is not changed or used 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; it speeds up a reaction that breaks down hydrogen peroxide, a toxic
chemical, into 2 harmless substances--water and oxygen. Light can also break down H2O2
which is why the chemical is sold in dark containers.
2H2O2 → 2H2O + O2
This reaction is important to cells because hydrogen peroxide 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 yeast or potato cells.
EXPERIMENTAL DESIGN
The experimental variables for this experiment are explained below.
Test Tube A Test Tube B Test Tube C Test Tube D Test Tube E
Independent variables Control Exp: Heated Exp: Cold Exp: Acid Exp: Base
Dependent variable Measure the height of a foam column produced to compare the rates of
reaction of catalase enzyme hydrolysis of hydrogen peroxide (substrate).
HYPOTHESES
Predict what will happen to the rate of the reaction if you change the temperature and pH of the solution
before adding the substrate. Write your hypothesis statements as “If...., then…, because…”
Hypothesis 1
(effect of
temperature)
Hypothesis 2
(effect of pH)
MATERIALS
● 8 Test tubes ● Potato puree (filtered)
● Test tube holder ● Chicken liver
● 3% H2O2 (hydrogen peroxide) ● Ice bath
● 0.1 M HCL (hydrochloric acid) ● Hot water bath
● 0.1 M NaOH (sodium hydroxide) ● Measuring pipettes with bulb
● pH paper ● Digital thermometer
● Matches & wooden splints
METHODS
PART A - THE CONTROLS: Observe a Normal Catalase Reaction
1. Use a pipette, add a small piece of chicken liver (or potato) to a clean test tube labeled A & C.
2. Use a pipette to transfer 3 ml of the 3% hydrogen peroxide solution into test tube B & C.
3. REACTION: Observe the reaction and record the reaction in each tube and record your observations below.
a. Tube A (liver only) ______________________________________________________________________________
b. Tube B (H2O2 only) ______________________________________________________________________________
c. Tube C (liver and H2O2) __________________________________________________________________________
4. PRODUCTS: Use a wooden splint and light it with a match. Carefully insert the burning splint into the top of test
tube A. Be careful not to touch the sides because it may be wet. Relight the splint and repeat this step for tubes
B and C. Record your observations below.
a. Tube A (liver only) ______________________________________________________________________________
b. Tube B (H2O2 only) ______________________________________________________________________________
c. Tube C (liver and H2O2) __________________________________________________________________________
A. What gas is being produced by the reaction? Support your answer with evidence.
PART B EXPERIMENTAL TEST: What is the Effect of Temperature on Catalase Enzyme Activity?
1. Transfer 3 ml of potato puree filtrate into clean test tubes labeled A, B and C.
2. Place test tube B into a hot water bath and test tube C into an ice water bath. Leave test tube A at room temp.
3. Wait for 5 minutes.
4. Remove both test tubes using a test tube holder and carefully place them into the test tube rack.
5. Add about 3 ml of the 3% hydrogen peroxide solution to each test tube A, B and C.
6. Observe the reaction and record the reaction rate by measuring the height of the foam column in millimeters.
A. Briefly describe the enzyme reaction rate after NO treatment of the potato.
B. Briefly describe the enzyme reaction rate after HOT treatment of the potato.
C. Briefly describe the enzyme reaction rate after COLD treatment of the potato.
EXPLAIN
What effect does TEMPERATURE have on the rate of reaction for the enzyme catalase?
CLAIM:
EVIDENCE:
PART C QUESTION: What is the Effect of pH on Catalase Enzyme Activity?
1. Transfer 3 ml of potato into clean test tubes labeled A, D and E.
2. Do not add anything to the test tube labeled A.
3. Your instructor will add about 1 ml of 0.1 M HCl (hydrochloric acid) to the test tube labeled D.
4. Your instructor will add about 1 ml of 0.1 M NaOH (sodium hydroxide base) to the test tube labeled E.
5. Use a pH paper test strip and record the pH of each test tube in the data table.
6. Add about 3 ml of the 3% hydrogen peroxide solution to each test tube A, D and E.
7. Observe the reaction and record the reaction rate by measuring the height of the foam column in millimeters.
A. Briefly describe the enzyme reaction rate after NO treatment of the potato.
B. Briefly describe the enzyme reaction when the ACID was added to the potato solution.
C. Briefly describe the enzyme reaction when the BASE was added to the potato solution.
EXPLAIN
What effect does pH have on the rate of reaction for the enzyme catalase?
CLAIM:
EVIDENCE:
DATA TABLE FOR PART B & PART C ONLY
Record your observations in the table below.
Tube Contains Additive or Treatment Reaction Observed
Test Tube A
Test Tube B
Test Tube C
Test Tube D
Test Tube E
ANALYSIS & CONCLUSIONS
Explain the purpose of the chicken liver and potatoes in this experiment.
What is the enzyme for this reaction called? What is the substrate for this reaction?
What are the TWO products for this reaction?
Summarize what happened in each vial containing different mixtures and treatments (acid, base, heated,
cooled, control). Use proper terminology in your descriptions.
● Control ___________________________________________________________________________________________________
___________________________________________________________________________________________________________
● Heated ___________________________________________________________________________________________________
___________________________________________________________________________________________________________
● Cooled ___________________________________________________________________________________________________
___________________________________________________________________________________________________________
● Acid ______________________________________________________________________________________________________
___________________________________________________________________________________________________________
● Base _____________________________________________________________________________________________________
___________________________________________________________________________________________________________
Explain the results of the reaction when hydrogen peroxide was added to each vial. Discuss why the
amount of foam produced was different for each vial.
● Control ___________________________________________________________________________________________________
___________________________________________________________________________________________________________
● Heated ___________________________________________________________________________________________________
___________________________________________________________________________________________________________
● Cooled ___________________________________________________________________________________________________
___________________________________________________________________________________________________________
● Acid ______________________________________________________________________________________________________
___________________________________________________________________________________________________________
● Base _____________________________________________________________________________________________________
___________________________________________________________________________________________________________
Based on the results of this experiment, describe the optimal pH and temperature for catalase enzyme
function. Use evidence from the experiment to support your claims.
ANSWER:
PROVE:
EXPLAIN:
SUMMARIZE:
Describe the term DENATURATION. Explain how this affects the activity (function) of an enzyme. Can this
be reversed for the enzyme catalase? Why or why not?
ANSWER:
PROVE:
EXPLAIN:
SUMMARIZE:
