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Name: Mesk ELsherbiny DP / Grade : 11

Subject: BIOLOGY HL

Research question: What is the effect of increasing the mass of zucchini [ 1,3,6,9,12 grams]
on the activity of catalase in catalyzing the decomposition of 10mL of 10% Hydrogen
peroxide measured by the volume of oxygen gas released using the water displacement
method ?

AIM : The aim of this investigation is to discover how changing the mass of zucchini ( 1,3,6,9,12 grams ) affect
the rate at which Hydrogen peroxide is decomposed into water and oxygen gas. This will be done by placing
the different mass of zucchini with 10 ml of 10% H2O2. The volume of oxygen released from the
decomposition of H2O2 will be measured using the water displacement method .

RATIONAL : Catalase, an enzyme found in many different tissues ( both animal and plant), catalyzes the
breakdown of hydrogen peroxide into water and oxygen:
2H 2O2 ------ > 2H2O + O2

Hydrogen peroxide is a toxic substance that can be formed during aerobic respiration and the enzyme catalase
removes this product. Catalase activity will change with the change of many factors, including temperature, pH
of solution, substrate and enzyme concentrations . This activity can be measured by finding the rate of oxygen
release from hydrogen peroxide using the water displacement method [using a 250 ml graduated cylinder (±2
ml)]

The source of catalase in this investigation will be taken from zucchini. This is known to have a large
concentration of catalase compared to other plant tissues.
The hydrogen peroxide will be set at 10 ml with a 10% concentration. All trials will be performed at room
temperature of 25 degrees and a pH of 7.

HYPOTHESIS : [ state a valid hypothesis for this investigation based on your knowledge of enzymes] [3]

It is predicted that if the mass of the zucchini increases, the catalase enzyme concentration increases. Thus,
the volume of oxygen in the water displacement set up would increase as well when set a specific time interval
of 3 minutes using a stopwatch. This will be observed by recording the water uptake every 30 seconds while
continuously stirring the substrate (hydrogen peroxide) and the zuchinni containing catalase enzyme.

HL BIOLOGY/ SAHAR JABAGI pg. 1

 A. VARIABLES :

Independent variable [ state it and discuss how it is manipulated]:

The independent variable in this investigation is increasing the mass of zucchini in order to increase the
concentration of catalase enzyme, using the following increments ( 1,3,6,9,12 grams ) measured using a digital
balance ±0.01. [2]

Dependent variable : [ state it and discuss how it is measured]: [2]

The dependent variable of this experiment is measuring the volume of oxygen every 30 seconds for 3 minutes
using a stopwatch. The volume of oxygen will measured using water displacement method. Water
displacement’s setup is
1-inverted cylinder
Tube
Ceramic stabilizer with a space fro the tube
The conical flask with the substrate and enzyme
A cork
Water
And a big bowl containing the water.

Controlled variable : [ discuss how and why you have to control these variables] [5]

Controlled variable How is it manipulated? Why is it manipulated?

The temperature of the enzyme- Through using a thermostatic If the temperature is not
substrate solution at 25 degree water bath, monitored by a controlled, it affects the rate at
Celsius. thermometer in degree Celsius. which oxygen is produced.

In which at this optimum

temperature, the average kinetic
energy of both the catalase
enzyme and hydrogen peroxide
particles, resulting in more
frequent successful collisions,
hence enzyme-substrate
complexes are made more
frequently, resulting in an
increased rate at which oxygen is
produced.
Meanwhile, if the temperature is
above this optimum, the enzyme
will denature and lose it is 3-
dimensional active site, which
means that the substrate wouldn’t
be able to fit into it anymore, due
to the breakage of the bonds
holding the enzyme structure.
Thus, no substrate-enzyme

HL BIOLOGY/ SAHAR JABAGI pg. 2


The pH
The volume of hydrogen peroxide
10ml
Same initial volume of water in the
inverted cylinder
Time left 3 minutes for 30 seconds
using a stopwatch
The concentration of hydrogen
peroxide 10%
HL BIOLOGY/ SAHAR JABAGI
Using 20ml of pH7 buffer,
measured using a 50ml graduated
cylinder.
Through measuring 10ml of
hydrogen peroxide using a
measuring cylinder ±0.1
Measuring the oxygen produced
every 30 seconds for 180 seconds
using a stopwatch ±0.05s
complexes would be produced,
and the oxygen production would
reach a minimal or no production
at all.
However, if the temperature is too
low, the kinetic energy of the
substrate and enzyme wouldn’t be
enough to work as fast as the
optimum temperature, resulting in
a decline of the volume of oxygen.
Hence, temperature should be
controlled.
The pH has a significant effect in
the rate at which oxygen is
produced in cm3.
The volume of hydrogen peroxide
is essential to control since it
determines the substrate
concentration in this enzyme
catalysed reaction. Hence, if more
hydrogen peroxide is added to
different trials, the frequency of
successful collisions increases,
thus more enzyme-substrate
complexes would form and that
would affect the rate at which
oxygen is produced in the water
displacement setup.
It would be a reference point for all
trials to ensure it is a fair test.
If the reaction is left for more time,
more hydrogen peroxide
substrates would bind to the active
site of the catalase enzyme
forming more enzyme-substrate
complexes. Inversely, if the
reaction is left for less time, fewer
enzyme-substrate complexes
would be formed. Thus, the results
of the investigation would be
inaccurate in each trial.
pg. 3
 B. METHOD

Materials:
100 ml of 10% hydrogen peroxide
1 25 ml plastic syringe
Plant tissues (zucchini)
Grater
Spatula and weighing bottle
Digital balance
250 ml graduated cylinder [±2 ml]
Delivery tube and trough
5X 100 ml Conical flasks
Marker
Stop watch
10 ml graduated cylinder
20 ml buffer 7

Method :
1. Assemble the water displacement method
2. Using the marker, mark the initial level of the water in the inverted cylinder
3. Measure 1 grams of one of the zucchini tissue using the digital balance.
4. Using the spatula, transfer the plant tissue in the conical flask
5. Using a 10 ml graduated cylinder, measure and Add 3 ml of buffer 7 on the zucchini.
6. Measure 10 ml of the hydrogen peroxide using the syringe.
7. Pour the hydrogen peroxide over the plant tissue and immediately close the mouth of the flask with the
stopper.
8. Start measuring the volume of the oxygen released every 30 seconds for a total of 3 minutes.
9. Repeat steps 2 to 7 for the rest of the masses of zucchini [ 3,6,9 and 12 grams]

HL BIOLOGY/ SAHAR JABAGI pg. 4

C. ANALYSIS

1. The data from all the class will be pooled. You will use 3 trials only.
2. Calculate the rate (slope) of oxygen production for each trial using excel.

3. Explain by giving a sample calculation how the rate was calculated.

The rate at which oxygen is produced from the enzyme catalyzed reaction can be calculated using the slope
equation. Y2-y1/x2-x1 or directly on excel using (slope) function. The slope helps formulate a relationship
between the two variables, hence this is an example of how much oxygen was produced using 9 grams of
zucchini calculated on excel. The time being the x-axis and the oxygen production being the y-axis, are
essential key points to establish before using the excel formula. After identifying the axis, as shown in the
screenshot, I selected the two columns, reaching a final answer of 0.80 g for the first trial. Accordingly, I
calculated the rate for each trial and mass of zucchini.

4. Calculate The mean rate of oxygen production from all the increments

Average rate of oxygen production= 0.80+0.75+0.75/3= 0.77 grams. The mean is necessary to the analysis of
the experiment since it sums up the data in a coherent way.

HL BIOLOGY/ SAHAR JABAGI pg. 5

5. Explain and Calculate the standard deviation of all the increments
The standard deviation shows the accuracy of each trial in this experiment out of 1. In other words it
could be a measure of how dispersed the data is in relation to the mean. Low standard deviation
means data are clustered around the mean, and high standard deviation indicates data are more
spread out. According to the mean calculated for each trial for every mass, the standard deviation was
extremely low, indicating that the trials were accurate since the increments are not far fro each other
and are all less than 1. Refer to the table for the standard deviation for every masses of zucchini.

The standard deviation shows the accuracy of the trials conducted in this experiment.
6. Construct a table to show your processing. Present the mean rate of oxygen production [ + STDEV ]
[4]
The effect of increasing the mass of zucchini/g on the average rate of oxygen production/cm3

Mass Average
of rate of
zucch oxygen
ini in production/ Stdev
grams cm3 / cm3
1 0.30 0.01
3 0.42 0.07
6 0.72 0.03
9 0.77 0.03
12 0.89 0.05

7. Present the rate of oxygen production for the effect of changing the mass of zucchini on the activity of
catalase in a suitable graph and give a brief interpretation of the graph.

The effect of increasing the concentration of enzyme

throughing increasing the mass of zucchini on the av-
erage production of oxygen/ cm3
1
0.89
average rate of production of oxygen

0.9 R² = 0.933744873670022
0.77
0.8 0.72
0.7
0.6
0.5 0.42
0.4 0.3
0.3
0.2
0.1
0
0 2 4 6 8 10 12 14
mass of zucchini in grams

As shown in the graph above, there is a clear direct relationship between increasing the mass of
zucchini in grams and the average rate of oxygen production. For instance, at mass 3 grams, the
average rate of oxygen production was 0.42 cm3. However, as the mass increased to 12 g, the average
HL BIOLOGY/ SAHAR JABAGI pg. 6
oxygen production noticeably increased up to 0.89 cm3. Thus, validating the interpretation made. As
for the R2 value being 0.9337, it acts as an indication that the independent variable (mass of zucchini)
has a direct relationship with the dependent variable (volume of oxygen), since it is extremely close to
1. Thus, there is a positive and strong correlation between these two variables. Moreover, in response
to the error bars in this experiment, they are short, hinting that the results gathered from each trial is to
an extent accurate. However, it is different in every gram of zucchini. In which, in gram 12 and 3, the
standard deviation error bars were the longest, concluding that they both have the highest
inaccuracies in the readings of the volume of oxygen produced which means more trials can be
conducted. [6]
8. Give an evaluation to your method by describing some errors that may have affected your data and
suggest a way to improve these errors. [4]
1- water displacement method is subjected to human error, in which it takes time to record the oxygen
released every 30 seconds for 180 minutes whilst reading the meniscus of the water correctly, thus, an
oxygen sensor would provide more precise data.
2- how much enzyme is in each zucchini cannot be determined, thus it is merely based on assumption
and that affects how many enzyme-substrate complexes could be produced in the reaction and thus
limiting the oxygen as a product of this experiment.
3- the connecting wire could have holes, so the oxygen could have escapes proving imprecise data.
Hence, using petroleium gel would prevent the gase from escaping from the wire whenput 3alih khalas.

HL BIOLOGY/ SAHAR JABAGI pg. 7