Investigation

13
Enzyme Activity

How do abiotic and biotic factors influence the rates of enzymatic 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 substanceswater and
oxygen.
This reaction is important to cells because hydrogen peroxide (H2O2) is produced as
a byproduct of many normal cellular reactions. If the cells did not break down
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:
Reaction chamber and Scissors
stopper Hydrogen peroxide
Plastic pan Hole punch
50ml graduated cylinder Disc paper
Thermometer Syringe
3 Plastic Cups Tweezers
Plastic baggie Timer
Chicken or beef liver Water

Procedure:
1. At the lab bench you will find a round vial with a rubber stopper top. This is
called the reaction chamber. You will also find a 50ml graduated cylinder
and a plastic pan which will be used as a water bath.
2. Fill the pan full of tap water. Allow the water to come to room
temperature.
3. Submerge the 50ml graduated cylinder to fill it with water. Turn the
graduated cylinder upside down , keeping the open end under water, so as to
keep it filled with water.
 4. The open end of the graduated cylinder is about 3cm above the bottom of the
pan.
5. Place the thermometer in the pan and record the temperature of the water.
6. Prepare three cups. Label them: water, catalase, and H2O2.
7. Prepare your catalase by placing a piece of liver into a plastic baggie. Zip the
baggie and pulverize for about 2 minutes. Using scissors, cut open one end of
the baggie and pour contents into its cup labeled catalase. Discard any
lumps. *Be sure to use the appropriate trash identified by your teacher.
8. Pour approximately 50 ml of the assigned solutions into the other cups.

Part A. The time course of enzyme activity (control)
1. Obtain your reaction chamber. Using a syringe, pour 5ml of Hydrogen
peroxide into the reaction chamber. Using a hole punch, prepare 8 discs
and dip each in water and place them on the edge of your reaction
chamber. Stopper the reaction chamber and invert.
2. IMMEDIATELY, submerge it in the water bath and place the plastic tubing
into the mouth of the graduated cylinder, so all the bubbles formed in the
reaction chamber are captured by the inverted graduated cylinder.
3. Measure the gas levels in the graduated cylinder at 10-second intervals
for 5 minutes. Record the levels in a data table of your own design.
4. If some bubbles were lost in the water bath, repeat the steps again.
5. Collect the data in your lab book.

Part B. The effect of enzyme concentration on enzyme activity.
1. Repeat the experiment from Part A, using 3 different levels of enzyme
concentration. You may easily do this by using the following procedures:
a. Follow the procedures from Part A, but use 2 paper discs
submerged in catalase in the reaction chamber.
b. Follow the procedures from Part A, but use 6 paper discs
submerged in catalase in the reaction chamber.
c. Follow the procedures from Part A, but use 8 paper discs
submerged in catalase in the reaction chamber.
2. You should conduct 2 trails for each concentration and find the average.
3. Record all data in a data table of your own design.
4. Plot the data on the same graph as Part A. Dont forget to clearly label the
enzyme concentrations on your plotted lines.

Part C. Inquiry. Pick a variable to test.