Liver Enzyme Lab

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 liver or other
tissue containing this enzyme, we can measure the effect of varying temperature and pH on enzyme activity. The
reaction for the break-down of hydrogen peroxide (by catalase) into water and oxygen gas is given below.
2H
2
O
2
2H
2
O + O
2
Hypothesis: If we increase the temperature the efficiency of the catalase enzyme will go up until
we reach the temperature at which the enzyme denatures.

Materials:
7 test tubes
1 test tube rack
Hydrogen Peroxide (H
2
O
2
)
Distilled water (H
2
O)
Raw Calf Liver
Boiling water bath (Hot plate, water, beaker)
Ice Bath (ice, water, cooler)


Procedure:
1. You will prepare the contents as outlined below for each corresponding test tube:
a. Test tube #1- Water
b. Test tube #2- Water & Liver
c. Test tube #3- Water, Liver, & H
2
O
2

d. Test tube #4- Water, Liver (that has been boiled several minutes or strong acid) & H
2
O
2

e. Test tube #5- Water, Liver (that has been cooled on ice for several minutes or strong base) & H
2
O
2

2. Make a data table.
3. Record a hypothesis for each test tube set-up before beginning the lab.
4. The volume of water will always be 5mL.
5. The size of the piece of liver should be approximately the same and relatively small.
6. Place test tubes 4 & 5 in ice/water bath as soon as you prepare it.
7. Test one test tube at a time.
a. Set-up your test tube.
b. Add 5mL of H
2
O
2

c. Record your observations for 3 minutes.
d. Interpret what you think has happened to the enzyme.
8. Clean up





#5
Water,
Liver(cooled) ,
H
2
O
2

No reaction
will occur
Bubbling;
reaction
occured
Liver rose to top
and rapid
bubbling


Methods Summary Chart:
Independent Variable
Temperature

Levels of the Independent Variable (if applicable)
Room temperature, Hot, Cold

Dependent Variable
Rate of oxygen production

Method for Measuring Changes in the Dependent
Variable

Amount of bubbles

Control Group (if applicable)
Room temperature and test tube #1 with just
water

Constants (factors that stay the same between your
control group and your experimental groups)

Amount of H
2
O
2,
type of liver, amount of water


Number of Trials 1 per treatment


Test Tube Contents Hypothesis Observation Interpretation
#1

Water
No Reaction Liver just sits
there
Its just water
#2

Water & liver

No reaction Liver just sits
in the water
Water is neutral
(pH 7)


#3
Water, Liver,
H
2
O
2

Reaction
occurs
Bubbles and
overflowed
out of tube
Hydrogen
peroxide is being
decomposed to
oxygen and water.
#4
Water,
Liver(boiled),
H
2
O
2

Reaction
occurs
Liver is
cooking; no
reaction
Liver enzymes
were denatured
in boiled water
No reaction occurred until H2O2 was added. Once the liver was boiled, no reaction
occurred. The hypothesis agreed with the results for all the tests except the tests that
included the boiled and cooled liver. For the boiled liver we hypothesized that a reaction
would occur, but none did. For the cooled liver, we hypothesized that a reaction would not
occur, but one did. Some potential errors could be timing procedures and another error could
have been that some solutions didnt have enough time for the reaction to actually occur. A
way to fix this would be by giving each at least thirty more minutes to react because some
reactions occur slower than others. To fix the timing procedures it would be best to start the
reactions all at the same time and have someone focus on each one of the test tubes so no
reaction occurs longer than the other ones.