Source: Tice, Shannon: Catalase Lab, Biology 412. http://chem.lapeer.org/Bio2Docs/CatalaseLab.html
Lab to Investigate the action of the enzyme Catalase
Introduction:
Catalase is an enzyme normally found in many plant and animal tissues. Its purpose is to destroy
toxic substances which may be introduced into cells. Also, some cells use catalase to destroy
cellular debris or worn out organelles. In this lab, a catalase solution from potatoes will be used to
determine the effect of temperature and pH on the action of this enzyme. The substrate of the
enzyme will be 3% hydrogen peroxide (H2O2). Catalase works by the following mechanism:
2 H2O2 ------------------>2 H2O + O2
Enzymes are Proteins, they are:
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

biological catalysts,
are denatured by heat
act on a specific substrate



require a specific pH
act best at a specific temperature
are not used up in the reaction
Obtaining the catalase:
Take some potato and grind it with some sand and water in a mortar and pestle. Decant the liquid
into a labeled test tube. Use a pipette to draw up the enzyme solution.
Materials:
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HCl
6 test tubes
hot water bath (250mL beaker)
graduated cylinder
NaOH
test tube rack
ice bath (250mL beaker)
beakers
MSDS sheets for HCl, NaOH, & H2O2
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H2O2
test tube holder
thermometer
mortar and pestle
fine sand
pipette
ruler
Safety:
HCl and NaOH are strong acids and bases. If you spill these chemicals on your skin,
immediately wash in running water. Wear goggles at all times. Be careful of hot water and
hot plates.
Source: Tice, Shannon: Catalase Lab, Biology 412. http://chem.lapeer.org/Bio2Docs/CatalaseLab.html
Procedure:
Part A:
1. Label 3 test tubes hot, cold and room temp. Prepare a water bath with a temperature of at
least 70oC
2. Place 3ml of potato catalase into each test tube.
3. Place the test tube labeled “hot” in the hot water bath for 3 minutes.
4. Place the test tube labeled “cold” in the cold water (This should be at 0oC) bath for 3 minutes
and the test tube labeled room temperature leave in the test tube rack. Record the exact
temperature of each water bath.
5. Create another set of test tubes to act as “controls”. (These tubes will be identical except you
will add H2O (water) to the tubes instead of H2O2 ( hydrogen peroxide))
6. After 3 minutes if the temperatures are at the correct temperature add 3ml of H2O2 to each of
the first set of three tubes. Add 3ml of water to the other “control” set of tubes.
7. Wait one minute and record the height of the bubbles in each tube in cm. Record in Table 1.
Measure the radius of the test tube and record in Table 1.
Part B:
1. Label 3 test tubes acid, base and water.
2. Place 3ml of potato catalase into each test tube.
3. Add 10 drops of HCl to the tube labeled acid, 10 drops of NaOH to the tube labeled base and 10
drops of water to the tube labeled water. Mix contents by gently swirling. (vortex mixing)
4. Wait 2 minutes then add 3ml of H2O2 to each tube.
5. Wait 1 minute for reaction to occur, then measure the height of the bubbles in cm. Record in
Table 2.
6. Measure the radius of the test tube and record in Table 2.
Source: Tice, Shannon: Catalase Lab, Biology 412. http://chem.lapeer.org/Bio2Docs/CatalaseLab.html
Part C:
1. Calculate the volume of each reaction using the equation
 r h.
2
Record in data table.
Data:
Table 1: Effect of Temperature on Catalase Reactions
Catalase
Height of Bubbles
(cm)
Radius of Test
Tube (cm)
Volume of
Reaction (cm3)
Hot
Room Temp
Cold
Control
Height of Bubbles
(cm)
Radius of Test
Tube (cm)
Volume of
Reaction (cm3)
Hot
Room Temp
Cold
Other observations:
________________________________________________________________________________
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Source: Tice, Shannon: Catalase Lab, Biology 412. http://chem.lapeer.org/Bio2Docs/CatalaseLab.html
Table 2: Effect of pH on Catalase Reactions
Height of Bubbles
(cm)
Radius of Test
Tube (cm)
Volume of
Reaction (cm3)
Acid
Neutral
Base
Water
Error analysis:
Human
Equipment
Source: Tice, Shannon: Catalase Lab, Biology 412. http://chem.lapeer.org/Bio2Docs/CatalaseLab.html
Analysis:
1. What is the substrate in this experiment?
2. What is the enzyme in this experiment?
3. What effect did heat have on the activity of the enzyme? What effect did cold have? Room
Temperature?
4. What type of biological molecule are enzymes?
5. Describe how an enzyme works and its importance to all living organisms.
6. What effect did lowering the pH have on the activity of the enzyme? Raising the pH?
7. What were the controls in this experiment?
Source: Tice, Shannon: Catalase Lab, Biology 412. http://chem.lapeer.org/Bio2Docs/CatalaseLab.html
Write up a complete lab report for this lab.
Due:______________________ You must include:
Cover page: correctly set out.
Title
Name:
Subject
Date:
Submitted to:
Title:
Identify the dependent and independent variables
Introduction:
Outline of the concepts covered in this lab
Key terms and their meanings:
Enzyme, denatured, substrate, variable, catalase
globular protein, pH, catalyst.
Hypothesis:
Using an If…. Then…. Statement that should be linked to the
independent and dependent variables
Materials, safety, Procedure:
These can be done using the traditional method or the flow
chart method. You should also include MSDS template for the
chemicals used.
Observations:
Quantitative data tables, Qualitative descriptions.
Error analysis:
human and equipment error
Conclusion:
Should relate back to the hypothesis and show whether it was
correct or not. You should also suggest next steps.
Citation of resources:
Should include text, lab handout and any other resources used
Correct APA/MLA format.
Source: Tice, Shannon: Catalase Lab, Biology 412. http://chem.lapeer.org/Bio2Docs/CatalaseLab.html
Name: _________________________________________ Date: _______________________________
Please include this copy with your lab report stapled to the back of the lab.
Lab Rubric:
0
Criteria
Marks
Cover page
Missing
Title
Does not identify
variables at all
Introduction
1
2
3
4
Considerable
attempt to follow
format
variables identified
correctly. Title
wordy rather than
concise.
Considerable
coverage of
concepts/ minimal
errors
All terms present
& correct
Correct format
Not connected to
title.
Correct format
No personal
pronouns
Past tense passive
voice
Complete
observations, all
correct forms
followed
Extensive attempt
to follow format
Limited attempt to
follow required
format
Limited attempt to
identify variables
1 variable
Some attempt to
follow format
missing
Limited attempt to
cover concepts
Some attempt to
cover concepts/
Some errors
Key terms & meanings
missing
Hypothesis
missing
Missing 5 or less
Major errors
Incorrect format
Missing 3 or less
Some errors
Partially correct
Materials/safety/procedure
Use of personal
pronouns
incomplete
Limited use of
correct format
Incorrect tense
Some use of
correct format
Incorrect tense
missing
Limited attempt to
show observations
Diagrams missing
or drawn in pen
Some attempt to
show observations
Diagrams partially
incorrect
Conclusion
missing
Limited ability to
connect to
hypothesis
Some attempt to
link to hypothesis
Considerable
connection to
hypothesis
References
missing
No correct format
Partially correct
format
Neatness & order
Messy incorrect
order
Headings not
underlined
Some incorrect
order
Some Headings
not underlined
Orderly
presentation
Correct format
3 references
Including textbook
All headings
underlined.
Orderly
presentation
Observations
Total marks possible = _________________40
Both variable
identified but
incorrect
Variables
identified correctly
title concise and to
the point.
Extensive
coverage of
concepts no
errors.
All present extra
terms included.
Correct format
Correctly
connected to title.
Flow chart used to
high degree of
Effectiveness.
Complete &
correct
observations and
detailed
descriptive
observations
included
Extensive
connection to
hypothesis
Makes connections
to next steps
Correct format
more than 3
references.
Typed, all
headings
underlined, no
errors of
presentation