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ENZYME LAB: (Formal Lab Report Required)

ABSTRACT (Write this last, it should include all parts, except background info & hypotheses, and be less
than 150 words.)
BACKGROUND INFORMATION: (Include some of the following information about catalase plus a
description of what an enzyme and catalyst is, how they act and how enzymes are affected by different
factors. Include in your background the role of an enzyme in the living cell. Also define denaturation as it
applies to enzymes and describe factors that may cause it. Briefly give an overview of what will be done in
the lab. Suggest what should happen based on research you have provided in your background information.)
Living cells must be able to decompose hydrogen peroxide (H 2O2), a dangerous chemical which is a
by-product of reactions in normal cells. The breakdown of hydrogen peroxide is aided by the
enzyme catalase, always present in plant and animal tissue.
PURPOSE/QUESTION: In this lab you will examine some properties of enzymes, examine a non-protein
catalyst and observe these under various conditions. (Briefly state what factors are going to be examined and
the purpose of different parts of the lab. Indicate what material will be used in each part.)
HYPOTHESES: (These should be written in the form of If then statements. You need separate
hypotheses for each of the parts of the lab.)
MATERIALS:This section should include labeled diagrams of the different set-ups for each part and all
materials used. Do NOT give me a list of materials. This list tells you what you need for the lab:
Manganese dioxide
Sand
Rubber stoppers
3% hydrogen peroxide
Large test tubes
Hot plate
pieces of raw liver and potato
Test-tube rack
Thermometer
0.1
M NaOH solution
Glass stirring rod
0.1
M HCl solution
Wood splints
Distilled water
pH paper
CAUTION: Hydrogen peroxide at this concentration can cause serious burns to the skin or will damage
clothing. Wash off any spills with large volumes of tap water. Goggles should be worn.
PROCEDURE (Rewrite the procedure steps in past tense passive voice. Describe what was done in a
paragraph for each part, not numbered points.)
Use the following code for estimating the rate of the reaction: 0 no reaction; 1 slow; 2 moderate; 3
rapid; and 4 very rapid. This rating scale needs to be explained at the beginning of observations.)

Part 1: The Effect of a Catalyst


Pour hydrogen peroxide into each of two test tubes to a depth of about 2 cm. Into one test tube, sprinkle a
small amount of sand, and in the other about the same amount of manganese dioxide. Quickly
cover test tubes lightly with rubber stoppers. Remove stopper and test both tube contents with a glowing
splint at the mouth of the test tube. Observe, record the rate of reaction taking place and describe the
reactions in both tubes.

Part 2:The Effect of an Enzyme


Pour hydrogen peroxide into each of two clean test tubes to a depth of about 2 cm (about 2 mL). Place a
piece of raw liver about the size of a rice grain into one test tube and a piece of potato of the same size into
the other test tube. Cover test tubes lightly with rubber stoppers. Observe the rate of reaction for each tube
and test each tube with a glowing splint and observe what happens.
Part 3: Reusing the Enzyme
Divide the liquid contents of the liver + H2O2 (from Part 2 above) into two clean test tubes. Cut the piece of
liver from the liver + H2O2 (from Part 2 above) into two equal parts and place one half in each of the test
tubes. Into one of the test-tubes, add a fresh piece of raw liver. Into the other test-tube add 1 mL of fresh
hydrogen peroxide. Observe the rate of reaction for each test tube and record.
Part 4: The Effect of Particle Size
Place a small piece of liver about the size of a rice grain into one test tube and a piece of potato of the same
size into another test tube. Sprinkle some sand (the same amount you used in Part 1) into each tube and
CAREFULLY crush the material with a stirring rod. (DO NOT MAKE A HOLE IN THE BOTTOM OF
THE TEST TUBE.) Pour about 2 mL of hydrogen peroxide into each of these test tubes. (Try to match the
final depth of potato + H2O2 from Part 2.) Rate the speed, comparing the results obtained using the
uncrushed pieces of liver and potato in Part 2.

Part 5: The Effect of Temperature (You need 3 test tubes for this part.)
Test-tube 1: Place a small piece of boiled liver (prepared by the teacher) in the bottom of a test tube. Then
add 2 mL of hydrogen peroxide. Observe and rate the speed and record your results.
Test-tube 2: Pour 2 mL of hydrogen peroxide into a test-tube and place in a warm water bath (37 C) for 2
minutes. Then add a small piece of liver (size of rice grain) into the peroxide. Record the temperature of
your hydrogen peroxide and rate the speed of the reaction.
Test-tube 3: Pour 2 mL of hydrogen peroxide into a test-tube and place in the ice bath for 1 minutes. Then
add a small piece of liver (size of rice grain) into the peroxide. Record the temperature of your hydrogen
peroxide and rate the speed of the reaction.
Part 6: The Effect of pH (You need 3 test tubes.)
Into each of three clean test tubes place a small piece of liver. Sprinkle some sand into each tube and use a
stirring rod to CAREFULLY crush the liver. Into Test-tube 1, add 2 mL of distilled water. Into Test-tube 2,
add 2 mL of sodium hydroxide solution. Into Test-tube 3, add 2 mL of hydrochloric acid. Test the pH of
each of the tubes. Then pour 2 mL of hydrogen peroxide into each of the test-tubes. Observe, rate the speeds
of the reactions and record the results.
OBSERVATIONS (You need observation tables for each of the parts of the lab.)
ANALYSIS: (Answer the following questions in complete sentences include the questions in the answers.
The numbers of the questions refer to the corresponding parts of the procedure. When questions ask you to
account for differences to theoretical results, comment on errors or any difficulties you had in the lab. )

1. a) What was the reason for adding the sand to the hydrogen peroxide?
b) What was the reason for adding the manganese dioxide to the hydrogen peroxide? What is the
role of the MnO2 (manganese dioxide)?
c) What was the reason for testing both test-tubes with a glowing splint?
d) What is the gas that was produced?
e) Write a balanced chemical equation for the breakdown of hydrogen peroxide with MnO 2 .
2. a) What was the reason for adding the piece of fresh liver to the test-tube of hydrogen peroxide?
b) What was the reason for adding the potato to the test tube of hydrogen peroxide?
c) What enzyme is found in potato and liver?
d) Why do you think there should or was a difference in the rate of reaction between H 2O2 and liver
vs. H2O2 and potato?
e) What gas was produced?
f) Write a balanced chemical equation for the breakdown of hydrogen peroxide with catalase.
3. a) Theoretically what should happen when a fresh piece of liver is added to the test tube with the
used liver and used hydrogen peroxide?
b) Did your actual observations agree or disagree with expected results in 3a)? How do you account
for any differences?
c) Theoretically what should happen when fresh hydrogen peroxide is added to the test tube with the
used liver and used hydrogen peroxide?
d) Did your actual observations agree or disagree with expected results in 3c) ? How do you
account for any differences?
e) Overall, can enzymes be reused or are they used up according to your results?
4. a) What is the reason for crushing the piece of liver and potato in this part of the experiment?
b) Theoretically, in comparison to your results from part 2 of the experiment, what did you expect to
observe when you added hydrogen peroxide to the crushed potato and liver?
c) Did your actual results agree or disagree with your expected results? How do you account for
this?
d) Overall, what is the effect of particle size on rate of enzyme action shown by this lab?
5. a) Why was the piece of liver used in Test-Tube 1 boiled?
b) What did you expect to observe when you added hydrogen peroxide to the boiled liver?
c) What process should have occurred to the liver?
d) Did your results agree or disagree with your expectations? How do you account for differences?
e) Theoretically, how would you expect the rate of the reaction of test-tube 2 from the 37 C water
bath compare to the hydrogen peroxide in test-tube 3 kept in the ice bath compare? Why?
g) Did your expectations agree or disagree with your actual results? How do you account for this?
h) Overall, what was the effect of temperature on the rate of enzyme action shown by this lab?
i) What should be the ideal operating temperature for enzymes and why?
6. a) Theoretically how did you expect the rates of reaction to compare among the distilled water,
sodium hydroxide and hydrochloric acid?
b) Did your actual results agree or disagree with your expectations? How do you account for this?

c) What process has occurred with the enzyme in your test-tube if it no longer appears to work?
d) Overall, what was the effect of pH on the rate of enzyme action shown by this lab?
CONCLUSION (Summarize what happened in each of the parts of the lab, indicating the effects of the
different factors on the enzyme. Restate your hypotheses, commenting on whether they were rejected or
supported.)
GRAPHICAL ANALYSIS
1. a) Plot the following 2 sets of data on a single graph using pH on the x-axis and Enzyme Activity on
the y-axis. Graphs must be done in pencil by hand on graph paper. (not by computer) The values in the rows
beside A represent Enzyme Activity (Quantity of product formed) for Pancreatic Lipase at the different pH
values. The numbers in the row beside B represent the Enzyme Activity for Pepsin (a gastric protease).
pH
A
B

0
12

1
16

2
19

3
15

4
0
9

5
3
4

6
7
1

7
10
0

8
15
-

9
18
-

10
14
-

11
7
-

12
0
-

b) What is the ideal pH for Gastrin and Pancreatic Lipase? How do you know?
2. Study the graph below showing the enzymatic action at different temperatures and answer the questions:
a) Which temperature provides the greatest initial rate of reaction?
b) At which temperature can the greatest quantity of product be formed indefinitely?
c) What do the horizontal lines labeled A and B on the graph indicate about the enzyme rate of reaction
at these points?
d) Discuss what is happening at 25 C?
e) What is the ideal rate for this enzyme on the graph and why?