Investigation: Enzymes
Enzyme Lab Teacher's Guide
Objectives
Measure the effects of changes in temperature, pH, and enzyme concentration 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 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; it speeds
up a reaction which 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.
The reaction is: 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 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.
MATERIALS:
6 Test tubes / Test tube holders
3% Hydrogen peroxide
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Ice / Hot water
Straight-edged razor blade
Scissors and Forceps
Measuring Pipettes
Stirring rod
Fresh liver, Apple, and Potato, Yeast Learn more
Vinegar / Baking Soda
HCL and NaOH
pH paper (optional)
PART A - Observe Normal Catalase Reaction
1. Place about 2 ml of the 3% hydrogen peroxide solution into a clean test tube.
2. Using forceps and scissors cut a small piece of liver and add it to the test tube. Push it into the hydrogen
peroxide with a stirring rod. Observe the bubbles.
What gas is being released? (consider the equation above) _____
Throughout this investigation you will estimate the rate of the reaction (how rapidly the solution
bubbles) on a scale of 0-5 (0=no271
reaction, 1=slow, ..... 5= very fast). Assume that the reaction in
step 2 proceeded at a rate of "4"
Shares
Recall that a reaction that absorbs heat is endothermic; a reaction that gives off heat is exothermic. Feel the
temperature of the test tube with your hand.
Has it gotten warmer or colder?
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Is the reaction endothermic or exothermic?
3. Pour off the liquid into a second test tube.
Assuming the reaction is complete, what is this liquid composed of?
What do you think would happen if you added more liver to this liquid?
Test this and record the reaction rate. Reaction Rate:
4. Add another 2ml of hydrogen peroxide to the liver remaining in the first test tube. What is the reaction rate?
Synthesis -- Answer the question: Is catalase reusable?
CLAIM:
EVIDENCE
REASONING.
Part B - What Tissues Contain Catalase?
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You will now test for the presence of catalase in tissues other than liver. Place 2 ml of hydrogen peroxide in
each of 3 clean test tubes and then add each of the three test substances to the tubes. As you add each test
substance, record the reaction rate (0-5) for each tube.
Substance Apple Potato Yeast
Rate of
Reaction (0-5)
Synthesis -- Do all living tissues contain catalase?
Claim:
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Evidence:
Reasoning:
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PART C - What is the Effect of Temperature on Catalase Activity?
1. Put a piece of liver into the bottom of a clean test tube and cover it with a small amount of water. Place this
test tube in a boiling water bath for 5 minutes.
2. Remove the test tube from the hot water bath, allow it to air cool, then pour out the water. Add 2 ml of
hydrogen peroxide. CAUTION: Use a test-tube holder for hot test tubes.
What is the reaction rate for the boiled liver and peroxide? __________
3. Put equal quantities of liver into 2 clean test tubes and 1 ml H2O2 into 2 other test tubes. Put one test tube of
liver and one of H2O2 into an ice bath. Place the other set in a warm water bath (not boiling).
After 3 minutes, pour each tube of H2O2 into the corresponding tube of liver and observe the reaction
What is the reaction rate for the cold liver/peroxide? _____
What is the reaction rate for the warm liver/peroxide? ____
Synthesis -- How does temperature affect the catalase enzyme?
Claim:
Evidence:
Reasoning:
PART D - What is the Effect of pH on Catalase Activity
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1. Add 2 ml hydrogen peroxide to 4 clean test tubes, then add:
Tube 1 – add 3 drops of acetic acid (vinegar) pH =_______
Tube 2 – add 3 drops of sodium bicarbonate (base) pH =______
Tube 3 – add 3 drops of water (neutral) pH =_____
Tube 4 -- add 3 drops of 1M NaOH pH = _____
Now add liver to each of the test tubes (try to do it all at about the same time, so you can easily compare)
Rate of Reaction for:
Strong Acid (HCL) ____ Acid _____ Neutral ______ Base_____ Strong base (NAOH) _____
1. How does pH affect the reaction rate of catalase? Propose a way to refine your experiment to find
the exact, or OPTIMAL pH and temperature of catalase.
2. The following graph shows reaction rates of various enzymes in the body. Pepsin is found in the stomach,
amylase in the saliva, and phosphatase in the liver.
Synthesis: How does pH affect the activity of enzymes?
Claim:
Evidence:
Reasoning:
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Part E - Design an Experiment
Lactaid is a product designed to help people who cannot digest milk sugar (lactose) because they are missing
the enzyme lactase. Many people are lactose-intolerant, a condition that is mainly genetic. Lactase breaks
down lactose into two subunits: glucose and galactose.
To test for the presence of monosaccharides and reducing disaccharide sugars in food, the food sample is
dissolved in water, and a small amount of Benedict's reagent is added. The solution should progress in the
colors of blue (with no glucose present), green, yellow, orange, red, and then brick red when there is a large
amount of glucose present. (Google benedict's test to see the way this looks.)
Design an experiment where you would determine how quicly lactaid works to break down milk sugar at
different temperatures.. Be specific in your description, use drawings if necessary.
Other Resources on Enzymes
Analyzing Graphics - Enzymes - shows substrates and enzyme interactions and explores competitive inhibition
Observe Catalase Activity in Yeast - create sodium alginate spheres to observe how catalase breaks down
hydrogen peroxide
Enzyme Activity Using Toothpickase - simulate the activity of an enzyme by breaking toothpicks.
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