IBDP Biology Required Practical

Required
Practical 3
Factor Affecting Enzyme Activity

Skill: Experimental investigation of a

factor affecting enzyme activity.

Ⓒ Power of Science, 2022

 Factor Affecting Enzyme Activity
IBDP Biology Required Practical

Required Practical 3
Skill: Experimental investigation of a factor affecting enzyme activity.

Key Terms
• Enzyme • Product • Inhibitors
• Catalyst • pH • Catalase Enzyme
• Active Site • Temperature • Denaturation
• Surface Area • Saturation of
• Substrate Active Site

There are a range of investigations that can be carried out to investigate a factor that affects
enzyme activity.

Possible investigations:

• Potato catalase (enzyme) from potato tissue on hydrogen peroxide (substrate) –

This is the focus of this document. Catalase can be sourced from a range of biological
tissues including avocado and liver.
Alternatively:

• Bromelin (enzyme), from pineapples, on breakdown of gelatine (substrate) in Jell-O.

Measure the change in mass of gelatine.
• Pectinase (enzyme), from a science supply company, on pectin (substrate) in apples.
Measure the volume of juice produced from pureed apple.
• Pepsin (enzyme), from a science supply company, on albumin (substrate) in egg
white. Measure the change in mass of egg white.
Further research will allow students to design appropriate enzyme experiments. Students
could use one of these experiments for their final Internal Assessment investigation
For their Internal Assessment (IA), ensure that students include:

• At least 5 variations for their selected independent variable.

• At least 5 repeats for each version of the independent variable.

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 Factor Affecting Enzyme Activity
IBDP Biology Required Practical

Potato Catalase Experiment

Possible variables (factors) to be investigated include:

• Concentration of hydrogen peroxide

• Temperature of hydrogen peroxide, using water baths
• pH of hydrogen peroxide (if possible, use pH buffers)
• Volume of potato extract/juice
• Time of reaction
Note: The variable that you choose to investigate is your independent variable, and all of
the other variables will be controlled variables.

Possible methods of measuring catalase activity

Hydrogen peroxide decomposes into oxygen gas and water. The oxygen gas can be
measured by a variety of methods which include:

• Water displacement by oxygen gas

• Measuring volume of oxygen gas produced using a gas syringe
• Measuring mass lost (as O2 is released) - Use relatively large volumes of hydrogen
peroxide (100ml)
• Height of foam produced in a measuring cylinder (add some liquid soap to trap the
oxygen gas)
Consider the following if you have access to data logging equipment:

• Change in concentration of oxygen gas, measured by O2 Probe

• Change in pressure produced by oxygen gas, measured by gas pressure sensor

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 Factor Affecting Enzyme Activity
IBDP Biology Required Practical

Experiment Outline – Paper Disc Method

1. Use a hole punch to cut discs (small circles) of filter paper
2. Pour 30ml of hydrogen peroxide into a test-tube
3. Cut a slice of potato (any size), and crush in a mortar and pestle
4. Place one disc of filter paper on the crushed potato tissue until it is moist
5. Place the moist filter paper disc onto the tip of a stirring rod.
6. Use the stirring rod to push the moist filter paper disc to the bottom of the test tube
of hydrogen peroxide, and start the timer
7. Remove the stirring rod
8. Stop the timer when the filter paper disc floats to the top of the hydrogen peroxide
(when it reaches the top)
9. Repeat steps 4 to 7 at least four times
10. Repeat steps 1 to 8 for each variation of the independent variable

Suggestions for Experiment Set-up

• This method works for investigating concentration of hydrogen peroxide,
temperature of hydrogen peroxide, and pH of hydrogen peroxide
• Use concentrations of 1% or less of hydrogen peroxide if investigating concentration
of hydrogen peroxide – first run a trial, with the minimum time being around 30s
• If investigating any variable other than concentration of hydrogen peroxide, then use
a 0.5% concentration of hydrogen peroxide
[For teachers: It is worth trialing concentrations before students do the experiment, as
catalase concentration can vary in different potatoes.]

• Within the method, include at least 5 variations of the independent variable (factor
affecting catalase activity), with at least five repetitions of each.

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 Factor Affecting Enzyme Activity
IBDP Biology Required Practical

Experiment Report Write-up

(Optional)

This is a good opportunity for you to practice experiment report writing skills in preparation
for completing your Biology Internal Assessment. The following provides guidance for what
should be included in a complete experiment report:

Exploration
(View IB Exploration Criterion Rubric)

Research Question
Write a clear research question linking the independent variable (factor affecting catalase
activity) and the dependent variable (time for filter disc to float to the top of hydrogen
peroxide).

Background information
Include information on the following:

• Explain the structure and function of enzymes.

• Explain how the key variables (Substrate concentration, pH, Temperature) affect
enzymes. [HL students should refer to inhibitors].
• Include specific information concerning the enzyme catalase.
• Explain why the filter discs rise – refer to the production of oxygen as hydrogen
peroxide is decomposed by catalase.
• Explain why hydrogen peroxide is produced in living tissues
• Explain why hydrogen peroxide must be broken down in living tissues
Ensure that scientific literature is used and cited appropriately within this section.

Hypothesis
Answer the research question by predicting the effect of changing the independent variable
(factor affecting catalase activity) on the dependent variable (time for filter discs to float),
supported by cited scientific literature.

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 Factor Affecting Enzyme Activity
IBDP Biology Required Practical

Variables
Independent Variable: (factor affecting catalase activity): Identify the range for the
independent variable, and explain your reason for choosing this range [Trial experiments are
useful here, and this impresses IB Examiners]. Consider doing some research to get an
estimate of the expected NaCl concentration.
Dependent Variable: (Time for filter disc to float): Explain why the filter discs float, and why
the time it takes to float to the top is a measure of catalase activity.
Control Variables: Identify the control variables, and explain how and why each control
variable is controlled. Where possible, include measured values of controlled variables
[Example: Volume of hydrogen peroxide solution = 30ml].
Consider controlling the following:

• All variables listed previously which were not selected as your independent variable
• Volume/Height of hydrogen peroxide in the test tube
• Source of catalase
• Age and health of potato tissue
• Diameter of boiling tube
• Concentration of inhibitors – easily controlled by not adding inhibitors

Material List
Make a clear list of all materials required to carry out the investigation. Be specific regarding
measurements, use metric units, and include precision error of all measuring instruments.
Examples:

• 150 ml of 0.5% hydrogen peroxide solution

• Timer ± 0.01s

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 Factor Affecting Enzyme Activity
IBDP Biology Required Practical

Procedure
Produce a clear numbered step-by-step procedure.
Ensure that you:

• Identify at least five variations of the independent variable (NaCl concentration)

• Clearly state that each variation is repeated at least 5 times.
• State which calculations will be carried out:
o Mean time for filter discs to float for each variation (of your independent
variable)
o Standard deviation of time for filter discs to float for each hydrogen peroxide
concentration.

Safety, Environmental, and Ethical Considerations

• Identify all possible safety concerns, and explain what is done to keep people safe
when doing the experiment.
• Explain all precautions that are taken for all chemicals used in the experiment.
Where possible, include citations.
• Explain how all materials are safely disposed of after the experiment.
• Pay particular attention to the use and disposal of hydrogen peroxide.

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 Factor Affecting Enzyme Activity
IBDP Biology Required Practical

Analysis
(View IB Analysis Criterion Rubric)

Qualitative Data
Include a list of relevant qualitative data which can help in answering the research question.
Consider:

• Movement of filter discs as they rise

• Any bubbles forming within the hydrogen peroxide solution
• General trend regarding time

Raw Data and Processed data

Raw and Processed data table can be combined into one data table.
Sample Table:
Table 1: Statement linking your independent variable to time for filter discs to rise to the top

Independent Time taken for filter discs to rise to the top / ±0.01s
variable / Standard
I II III IV V Mean
unit deviation

Ensure that you include the following for the raw data section of the table:

• Titles, including the independent variable (factor affecting catalase activity) and the
dependent variable (time for filter discs to reach the top)
• Column and row headings
• Units and precision error shown for all measured values
• Consistent precision (# of decimals) in data, which matches precision error
Ensure that you include the following for the processed data (Mean and Standard Deviation)
section of the table:

• Calculated mean time for filter discs to rise to the top when submerged in each
version of the dependent variable (factor affecting enzyme activity)
• Calculated standard deviation for your version of your dependent variable
• Units and precision error shown for all measured and calculated values
• Consistent precision in processed data, which matches precision error
• Mean and standard deviation, correctly calculated
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 Factor Affecting Enzyme Activity
IBDP Biology Required Practical

Graph

Use the processed data for your factor affecting catalase activity on the X-axis, and mean
time for filter discs to rise to the surface on the Y-axis, to produce a scatter graph.

The graph should include the following:

• Axes titles with units and precision error

• Correctly plotted data points
• Appropriate line of best fit (trend line)
• Correctly constructed error bars using the calculated standard deviations
• Correctly calculated R2 values

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 Factor Affecting Enzyme Activity
IBDP Biology Required Practical

Evaluation
(View IB Evaluation Criterion Rubric)

Conclusion
Describe the trend on the graph, identifying the relationship between the independent
variable (factor affecting catalase activity) and dependent variable (time for filter discs to
rise to the surface). Explain the trend using scientific theory, supported by citations from
scientific literature.
The conclusion should include the following relevant to the research question:

• A correct description of the trend (if any) in the graph

• An explanation of the trend using an understanding of biology
• An attempt to answer the research question
• A comment on the implications of the data in regards to the hypothesis
• A description of the implications of the error bars
• A description of the significance of the R2 values
The conclusion should include the following in relation to scientific literature:

• An explanation of the trend using relevant (cited) biological theory

• A comparison of results to scientific theory with citations [if possible, refer to
scientific journals]
• Suggested explanations for differences between your results and biological theory

Evaluation of the Experimental Procedure

Include strengths, problems and improvements.
Strengths
Possible strengths include:

• Sufficient data to identify a trend on a graph – 5 data points are required to identify
a trend
• Sufficient data to carry out statistical analysis - Standard Deviation requires 5 repeats
• Sufficient data to answer the research question – 5 variations of the independent
variable and 5 repeats
• Data recorded by measuring instruments allows the research question to be
answered – in this case, the stopwatch.
• Variables were effectively controlled, and it is possible to explain how these
variables were able to be kept constant.

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 Factor Affecting Enzyme Activity
IBDP Biology Required Practical

Comment on the range of the data. If the standard deviations are small, the data is reliable
and this is a strength. If the standard deviations are large, the data is not reliable and this is
a weakness.

Problems and Improvements

Identify any problems (Qualitative observations can help identify problems) that may affect
the quality of the data collected (the time for the disc to rise to the top). Explain the impact
of each problem on the data.
Suggest improvements to attempt to solve each identified problem. Explain how each
suggested improvement would improve the quality of the data collected (the change in the
time for the disc to rise to the top).
Possible problems for this investigation include:

• Temperature may change during the reaction

• The concentration of each hydrogen peroxide solution decreases as additional filter
discs are inserted
• There are challenges in ensuring that the filter disc is at the bottom of the test tube
when the timer is started

Extensions
Suggest and explain extensions to the experiment.
Extensions to this experiment could include:

• Repeating the investigation with different sources of catalase.

• Using a different method (outline the method) to investigate the effect of the factor
affecting catalase activity

Works Cited
• Use one recognized system for documenting sources – consider using an online tool
such as MyBib.
• Include in-text citations within the report, including all graphics used.
• Include a complete, alphabetical works cited list at the end of the report, for all of
the in-text citations
• All internet sources should include date accessed in the works cited list.
…………………………………………………………………………………………………………………………………………………………….

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 Factor Affecting Enzyme Activity
IBDP Biology Required Practical

Communication
(View IB Communication Criterion Rubric)

Communication is assessed holistically, with the IB Examiner making a judgement based on

the whole essay.
The Examiner will be looking for the following:

• Appropriate titles for each section

• The report is 6-12 A4/Letter pages long
• The variables, procedure, data and conclusion relate to the research question
• In-text citations are effectively used throughout the report
• Correct alphabetical works cited (including date accessed for online sources) is
included for all in-text citations
• All graphics include captions and are referred to in the text
• SI units are used throughout the report
• Biological terminology is correctly used throughout the report

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 Thank You!
Factor Affecting Enzyme Activity
IBDP Biology Required Practical

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