An Investigation into the effect of temperature on the rate of catalase

activity of kale.

Introduction: Enzymes are protein-based compounds which act as catalysts for biological
reactions in the bodies of living organisms. The process in which enzymes fasten the process
of biological reaction involves the substrate fitting and binding to the active site of the
enzyme. Catalase is a type of enzyme which is found in numerous tissues around the body.
As for the case of catalase, the substrate which it breaks down is hydrogen peroxide
( 2H2O2). “During respiration, the body sometimes produces hydrogen peroxide. However,
as hydrogen peroxide is highly toxic, catalase is required to quickly break down the
hydrogen peroxide ( 2H2O2) into hydrogen (2H2O) and oxygen (O2) molecules.”1 Similar to
other enzymes, the rate of catalase activities is highly dependent on four main factors:
temperature, PH levels, concentration of enzymes, and concentration of substrates. This
investigation will look into the effect of temperature on the rate of catalase activity of kale.
The catalase activity of the kale refers to the catalase in the kale breaking down the
hydrogen peroxide inserted into the beaker with the kale. Kale was chosen for the
investigation because it is a “vegetable with a high level of catalase” 2 and hence there would
be an easier, clearer, differences in reading during the experiment.

Research question: What is the effect of temperature on the rate of catalase activity?

Hypothesis: I predict that as the temperature increases, the rate of catalase activity of kale
will also increase up to the optimum point of 37 °C (normal body temperature) and then the
catalase activity will decrease. This is because, as temperature increase, it causes the
hydrogen bonds to loosen which makes it easier for the catalase to break down the
hydrogen peroxide molecules, and in turn fasten the rate of catalase activity. Every enzyme
has optimum temperatures and PH levels, for catalase, the optimum temperature is the
body temperature of 37 °C. I predicted that the catalase activity will decrease after the
temperature reaches 37 degrees Celsius because after the enzyme is in its optimum
temperature of 37 degrees Celsius, the temperature becomes too high for the enzyme to
function and so the enzyme will begin to denature. When the enzyme denatures, the shape
of the enzyme becomes distorted and the active site of the enzyme is no longer able to bind
with the substrate. Hence, I predict that the catalase activity will decrease after the
temperature reach 37 °C.

Predicted graph:

1
Smith, B. 2018. “How does temperature affect catalase enzyme activity?”. Sciencing. 30 th April 2018. Accessed
on 25th May 2022. https://sciencing.com/temperature-affect-catalase-enzyme-activity-7776025.html
2
Amerman, D. 2018. “Vegetarian sources of catalase”. SF Gate. 27 th December 2018. Accessed on 25th May
2022. https://healthyeating.sfgate.com/vegetarian-sources-catalase-3693.html
Table of variables:
type of
variable variable method
 To change and set the temperature of kale throughout the
experiment, the glassware with the kale, buffer solution
and the hydrogen peroxide solution inside will be placed
into an electric water bath (not submerged and the
materials inside the conical flasks and measuring cylinders
independen  Temperature of will not be in direct contact with the water bath) and set at
t kale 5 different temperatures of 23, 30, 37, 44, and 51 °C.
 The catalase activity refers to the catalase breaking down
the hydrogen peroxide( 2H2O2) into hydrogen (2H2O) and
oxygen (O2) molecules. To measure the rate of catalase
activity, the amount of oxygen produced after 5 minutes of
 Rate of catalase inserting the hydrogen peroxide solution into the conical
dependant activity flask with the kale will be measured.
controlled  The mass of kale affects the concentration of catalase
mixed into the buffer solution in the conical flask which in
turn affects the rate of catalase activity and hence, needs to
be controlled. In order to control the mass of kale, 10g of
 Mass of kale kale will be measured and used for each trial.
 Concentration  The increase of hydrogen peroxide increases the number
of hydrogen of hydrogen peroxide substrates which collide with the
peroxide catalase enzymes, and hence increase the rate of the
catalase activity. In order to control the concentration of
hydrogen peroxide, 15ml of 10% hydrogen peroxide
solution will be inserted for each trial.
  When the pH level of catalase distances from its optimum,
the shape of the catalase becomes increasingly more
distorted and its function to bind with and break down the
hydrogen peroxide substrates weaken. To control the pH
 pH level of level in the experiment, the buffer solution will all have the
buffer solution optimal pH for catalase which is 7.5 pH.
 When the glassware such as the conical flasks and
measuring cylinders which have the buffer solution and kale
inside changes, the surface area of the kale changes and
can therefore affect the rate of catalase activity. To control
the glassware used, the same 100cm^3 conical flasks and
 Glassware used 50cm^3 measuring cylinders will be used for each trial.

Reagents:
5 x 10g minced kale
5 x 50ml buffer solution of 7.5 pH
5 x 10ml 10% hydrogen peroxide solution

Apparatus:
1 x timer
1 x measuring cylinder (50cm^3)
1 x measuring cylinder (100cm^3)
2 x conical flask (100cm^3)
1 x 50cm rubber tubing
1 x pneumatic trough
1 x electric water bath
1 x 2-holed rubber bung
1 x 10cm^3 syringe
1x knife
1 x cutting board

Method:

1) Set the water bath at a temperature of 23°C

2) Measure 100ml 7.5 pH buffer solution using measuring cylinder (100cm^3)
3) Mince the kale into very small pieces using the knife on the cutting board
4) Pour the buffer solution and 10g of minced kale into the 100cm^3 conical flask and
cover the conical flask with the 2-holed rubber bung
5) Insert the 50cm rubber tubing into one of the holes in the rubber bung
6) Half-fill the pneumatic trough with water
7) Fill the 50cm^3 measuring cylinder with water and invert it and place it inside the
pneumatic trough with the open-end under the surface of the water in the
pneumatic trough
8) Place the other end of the rubber tubing in the measuring cylinder in order for the
oxygen to be transported to the measuring cylinder
9) Place all the equipment prepared into the water bath
 10) Measure 10ml of the hydrogen peroxide solution into the 10cm^3 syringe and put
the syringe into the other hole of the rubber bung and start the timer of 5 minutes
11) After 5 minutes, record the volume of oxygen in the measuring cylinder
12) cleanse and sterilise all of the apparatus
13) repeat steps 1 to 10 for all the different temperatures

diagram:

Hazard risk prevention

Hydrogen peroxide
Knife
Glassware (conical flask)
Warm water bath
May cause chemical harm
when in contact with skin
May cause cuts and
bleeding
Will break when dropped
When the dangerous hot
water is in contact with skin,
there is risk of skin burn
- Wear eye goggles to
protect eyes
- Wear disposable
gloves to protect
hands
- Wear lab coat to
reduce skin exposure
- Be cautious when
using the knife
- Be cautious when
carrying the flasks
around
- Be cautious when
placing the
equipment in the
water bath
- Do not place finger
in the water bath
Environmental considerations:
- All of the measuring cylinders and conical flasks are washed and reused
- The kale can be disposed as food waste and compostable

Bibliography:
1) Amerman, D. 2018. “Vegetarian sources of catalase”. SF Gate. 27 th December 2018.
Accessed on 25th May 2022. https://healthyeating.sfgate.com/vegetarian-sources-catalase-
3693.html
2) Smith, B. 2018. “How does temperature affect catalase enzyme activity?”. Sciencing. 30 th
April 2018. Accessed on 25th May 2022. https://sciencing.com/temperature-affect-catalase-
enzyme-activity-7776025.html