How does temperature

affects catalase’s rate of

reaction
Science CBA 1

Katie Zheng [8 April 2024 - - April 2024]

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 ! Table of Contents !
! Table of Contents !
2

" Research Question"

3

# Introduction #
3

$ Considerations$
3

ℹ Background informationℹ
4

& My Hypothesis&
7

' Equipment and Materials '

8

⚠ Safety hazards ⚠
9

) Variables)
9

* Method*
10

+ Table of results+
11

, Analysing the results,

11

- Anomalous Results / Sources of Error-

13

. Conclusion.
16

2
✏ Recommendations/ Changes I would make✏
16

0 References and Sources 0

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" Research Question"

I researched the question, “Does temperature affect the rate of

reaction between catalase and hydrogen peroxide?”

# Introduction #
For my CBA, I chose to do an experiment on catalase. I decided to do it
because I found enzymes and the way they work really interesting when I studied
them for the digestive system. I wanted to further research another enzyme that
is part of our body. I decided to choose catalyse because it was easily available to
buy (from pig liver, celery, potatoes etc….)

$ Considerations$
I had a limited amount of time (3-4 weeks) so I had to choose a simple
research question, that wouldn’t take a long period of time to complete. I had to
make sure I had enough time to complete the experiment as well as conduct
research and conclude my findings
I was only able to acquire basic lab equipment and materials, so I chose an
experiment with materials I had in the lab and at home.
I also had a tight budget so anything I did need to buy had to be relatively
cheap, this restricted the experiments I could do.
I also had to keep safety in mind as I didn’t want get hurt performing the
experiment and I definitely didn’t want to damage the surroundings or injure
people in my vicinity. This meant no experiments with combustion etc….

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 ℹ Background informationℹ
I am doing my CBA 1 on the effect of temperature on catalase, the enzyme is
found in the tissues of many organisms both plant and human, it is one of the
most common enzymes, and is found in almost all living things. It acts as a
catalyst for the conversion of hydrogen peroxide into water and oxygen.

Hydrogen peroxide is a harmful byproduct of many normal metabolic

processes eg. when the immune system is activated in response to bacteria, large
amounts of hydrogen peroxide is produced by certain cells to fight the infection.
To prevent damage to cells and tissues, it must be quickly converted into other,
less dangerous substances. Catalase is frequently used by cells to rapidly
catalyse the decomposition of hydrogen peroxide into less-reactive oxygen and
water molecules, to protect the body from further harm. A high concentration of
catalase can be found in the liver of mammals which is why I will be using pig
liver as my source of catalase for this experiment.

We know catalase is an enzyme but what is an enzyme? An enzyme is a large

protein molecule, that acts as a catalyst in biochemical reactions that occur in
living things, eg. Lipase which is made in the pancreas to speed up the
breakdown of fats. A catalyst is a specific type of protein that increases the rate of
a chemical reaction, making it occur faster. Enzymes work by lowering the
amount of activation energy needed to get the reaction to start, which helps the
reaction occur faster.

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 Diagram on how activation energy works:
In a reaction where an enzyme acts as a catalyst, the substance that the
enzyme acts upon is known as the substrate. Each enzyme is specifically
designed for a particular substrate, because the shape of an enzyme active site.
The active site is the location where the enzyme binds to the substrate. The
shape of the active site for catalase (the enzyme I am investigating) is matched up
to the shape of hydrogen peroxide (which is my substrate) so it can bind, like
pieces of a puzzle. Each enzyme is only able to bind with a substrate that has a
complementary shape.

When an enzyme and substrate bind it creates an enzyme-substrate complex,

where the enzyme changes the substrate from in original form. In my case when
catalase and hydrogen bind, catalase converts hydrogen peroxide into water
(H2O) and oxygen (O2). After creating the product, enzymes are recycled and used
again.

3D rendering of a catalase enzyme ⬇

Fun fact ' : Catalase has one of the

highest turnover rates of all enzymes,
one catalase molecule can convert
millions of hydrogen peroxide
molecules into water and oxygen
each second!

The active site is what determines an enzymes ability to function properly, as

it is a highly specific 3D structure for a particular substrate. There are
environmental conditions that can change the shape of an enzyme, if the shape of
an enzyme is changed its activity decreases or can even completely stop , we call
this being denatured. The 2 main factors that can affect the shape of an enzyme
(denature it) are pH and temperature. I chose to investigate temperature and not
pH as too many variables may confuse my results.

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 For most human enzymes optimal pH is 7 (neutral) and optimal temperature is
37° (average body temperature). Temperatures of 40-50°C can denature most
human enzymes. Some can still function under this temperature but almost all
enzymes denature under 70-80°C. The experiment I conducted tests how
effective catalase is in different temperatures, which helps us see how much
catalase denatures under different temperatures.

Since I only have a limited time to run the experiment I have to make sure the
reaction happens within 3 minutes, so I decided to use small chunks of pig liver
instead of adding a whole chunk because it would increase the surface area. The
larger the surface area the more space there is for the hydrogen peroxide
particles and the catalase particles to collide and react. This increases the chance
of a successful collision.

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 & My Hypothesis&

My hypothesis is that if the temperature is below or above 37°C, the rate of the
catalytic action of catalase will decrease. I think the rate of catalytic will be higher
in average body temperature (37°C) because catalase occurs naturally in the
body, and has evolved to work under body temperature. I think the rate of
catalytic action will decrease in temperatures above or below average body
temperature because enzymes denature in overly cold / hot temperatures.

Expected results ⬆

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 ' Equipment and Materials '
Equipment :
- iPad to measure the time, and to take pictures of my experiment
- Measuring spoon to add the liver solution into the test tubes, and to

measure the amount of liver

- Test tubes to hold the hydrogen peroxide solution
- Rubber bung with tubing to seal the test tubes
- Tubing to transport the gas from the test tube into the beaker
- Graduated cylinder to accurately measure the solutions
- Beakers to hold the water
- Hot plate to heat the water
- Freezer to freeze the water
- Blender to make the liver solution
- Thermometer to measure the temperature?

Materials :
- 20ml of Hydrogen Peroxide (5% concentration)
- 1L of Water
- 4.5g of Liver solution (Blended pig liver)
- 200g of Ice

Diagram of experiment

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 ⚠ Safety hazards ⚠
- I made sure all glass equipment was stored safely, to stop them from
shattering and causing and accident.
- I kept the hot plate away from the edge of the table, and avoided touching
the hot area
- I stored hydrogen peroxide safety, and disposed of it responsibly as it is a
corrosive. I was careful to not touch it with my bare skin
- I made sure to cut the liver with care, to not cut my hands. I wore gloves
when cutting the liver
- I wore safety goggles to prevent any solution getting into my eyes
- I wore a lab coat to prevent materials from getting on my clothes
- I performed the experiment standing up to to avoid getting hot water on my
legs, and so I could move away from any dangerous components
- I used a low concentration hydrogen peroxide to avoid corrosion on lab
surfaces and my skin

) Variables)
Independent Variables :
The temperature of the water is my only independent variable as too many
would make the experiment messy. I will be conducting this experiment at 3
different temperatures; 5°C, 37°C and 70°C

Dependent Variables :
The rate of reaction between catalase and hydrogen peroxide. (The amount
of bubbles produced as a result of this reaction 3 )

Controlled Variables :
- The amount of liver added
- The size of each piece of equipment
- The concentration of the hydrogen peroxide (5%)
- The amount of hydrogen peroxide
- The time each test gets (1 minutes)
- The pH of the solution

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 * Method*
This experiment will have 3 iterations :
- Heat 20mL of water in a beaker on the hot plate till 80°C
- Heat 20mL of water in a beaker on the hot plate until 37°C
- Chill the water by adding ice in a beaker until 5°C

1. Measure 10mL of water using a graduated cylinder from the beaker

2. Place 10 mL of water into a test tube A.

3. Place 0.5g of liver into test tube A.

4. Attach the rubber tubing to the glass bend on the rubber test tube stopper, place the
end of the tubing in a 250 mL beaker containing 100mL of water.

5. Measure out 2 mL of hydrogen peroxide in a graduated cylinder.

6. Place 2 mL of hydrogen peroxide (H2O2) into the test tube A then immediately seal
the test tube with the stopper that has the glass bend and rubber tubing attached. The
end of the tube should be fully submerged in water in the other beaker.

7. Allow the reaction to proceed for 1 minute. Count the bubbles that begin to form in the
water over this period of time.

8. At the end of the minute, record the total number of bubbles you counted in the
correct data table.

9. Rinse, clean and dry all the equipment then repeat the experiment until the result
table is full.

Image of equipment used ⬇

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 Image of what my setup looked like ⬇

+ Table of results+
Experiment Amount of bubbles per 1 minute

Temperature C° Trial 1 Trial 2 Trial 3 Average of 3

trials

Ice Water 4 5°C 31 19 20 23.33

Body 37°C 72 53 46 57
temperature

Hot Water 5 70°C 16 10 17 14.33

, Analysing the results,

From the results I could tell that the 36°C experiment reacted faster, and with
more force than the other 2 temperatures, it had more bubbles on average This
proves my hypothesis as the results align with my theory. The results make sense
as enzymes denature (change shape) in overly hot or cold temperatures. There
were some variance in the amount of bubbles (the biggest difference between
trials being a 26 bubble difference between trial 1 of the 37°C water and trial 3 of the
37°C water), but on average over the 3 trials the results were to be expected and
fulfilled my expectations.

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 The results also showed that the enzymes performed better in 5°C water
than 70°C, even though it denatures at both of those temperatures. I think this is
because 5°C is significantly closer to 37°C (32°C difference) in comparison to 70°C
which is 33°C hotter. This means the higher the difference the lower the amount
of bubbles produce within the 1 minute timeframe.

From the bar chart you can tell that overall in each trial the 37°C experiment
consistently produced more bubbles than the 5°C or 70°C ones. On average the

5°C 36°C 70°C

80

60

40

20

0
Trial 1 Trial 2 Trial 3 Average

body temperature had 150.74% more bubbles than the ice water and 297% more
bubbles than the hot water.

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 - Anomalous Results / Sources of Error-

The first time I ran the experiment, I accidentally used a rubber bung with
another extra tube. This was human error as I chose the wrong piece of
equipment. I assumed it would work the same as a rubber bung with 1 glass tube,
but the extra tube let out a majority of the gas. This meant the amount of bubbles
was incorrect as most of the gas had escaped. The hole also allowed for slighted
faster cooling which meant the temperatures adjusted to room temperature too
quickly. This lead to only 1 or 2 bubbles forming the beaker so I decided to do it
again but when I repeated the experiment I had issues with temperature.

The issue was the time between heating up / cooling down the water and
then using it in the experiment. If I heated/cooled the water too early and took too
long to complete the rest of the steps the water would revert to room
temperature. This was environmental error which was caused by the
temperature of the lab which influenced the temperature of the water
overtime.
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 During the very first round of the experiment I heated / cooled the water first
and let it sit as I completed the other preparation, this created anomalous results
in comparison to the other results as I think the water adjusted to room
temperature creating more bubbles than there should be for both cold and hot
water.

To minimise this error I decided to continuously assess the temperature of

the water by keeping the thermometer inside the beaker, and after finishing other
prep I checked to see it the water was still the correct temperature and adjusted
the temperature accordingly so the results were more accurate.

However I wasn’t able to fix it perfectly as the temperature still changed while
the experiment was happening in the test tube. Another issue related to
temperature during the experiment was the temperature of the test tubes, I
realised the hot water after entering the test tube left it quite hot, even after I
washed it. This meant if I put my cold water into the test tube after the hot water
experiment was performed in that test tube the temperature of the water would
rise because the glass would heat it. To stop this from occurring and skewing my
results I used a separate test tubes for each temperature.

From this I realised that temperature is a very easily changed variable that is
very sensitive. The temperature needed a lot of monitoring which meant it was
impossible to do 2 variables as I had previously intended. When I first decided to
do an experiment on catalase I wanted to use 2 variables, pH and temperature
but due to a lack of time and I had to simplify it, reflecting on it I’m glad I did
simplify but I regret not choosing pH as my variable, as using buffer solution to

Temperature C° Anomalous Trial 1 Trial 2 Trial 3 Average of 3 Average of 3

Trial (trial 0) trials trials (not
(Before any ( including including
adjustments trial 0) trial 0)
were made)

Ice Water 4 5°C 44 31 19 20 28.5 23.33

Body 37°C 50 72 53 46 55.25 57

temperature

Hot Water 70°C 56 16 10 17 24.75 14.33

5

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 5°C 36°C 70°C

80

60

40

20

0
Trial 1 Trial 2 Trial 3 Avg. without trial 0 Avg. including trial 0 Trial 0

keep the pH the same would have probably been simpler and easier but I still
enjoyed experimenting with the temperature.
From the bar chart you can see that no matter the starting temperature
the amount of bubbles produced were very similar. I think this is because
the water had adjusted to room temperature before I performed the
experiment causing all 3 temperatures to actually be almost room
temperature. Comparing the anomalous results from trial 0 you can see that
the amount of bubbles produced by the 5°C water and the 70°C were far
higher than in trial 1,2,3 or the average of the 3 trials

After realising including trial 0 would project a false average where both
cold and hot water (5°C and 70°C ) performed

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 . Conclusion.
In conclusion, I was overall very happy with the results of the CBA and my
experiment as a whole. I learnt a lot about using the scientific method as well as
practicing writing a report on my findings. Although I struggled with carrying out
the experiment I enjoyed the process. Learning about catalase was interesting
and now I know much more about enzymes and the way the work. The results
aligned with my hypothesis (

✏ Recommendations/ Changes I would make✏

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 0 References and Sources 0
https://www.theconicalflask.ie/teacher-corner/cbas
https://www.reading.ac.uk/ - 3D rendering of catalase
https://www.hopkinsmedicine.org - Uses of enzymes
https://www.sciencedaily.com/releases/2008/ - Role of hydrogen peroxide in
cell health
https://pubmed.ncbi.nlm.nih.gov/16618987/#:~:text=Catalase - What is
catalase?
https://www.rcboe.org/cms/lib010/GA01903614/Centricity/Domain/1472/IA
How Temperature Affects the Rate of Enzyme Activity.docx#:~:

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