LAB REPORT- ENZYME ACTIVITY 13.03.

2022
Drnda Medina IBDP III2

Personal engagement:
All biological activities in living creatures involve chemical reactions, and enzymes manage
the majority of them. Having this in mind, enzymes are globular proteins that work as a
catalyst in living organisms. When it acts as a catalyst, the enzyme will speed a chemical
reaction- it will control chemical pace without changing itself. Catalase is one of these
enzymes. Catalase1 is an enzyme that initiates and serves as a catalyzer in the decomposition
of hydrogen peroxide into water and oxygen. It is only abundant in organisms that exist in the
presence of oxygen, and it inhibits the buildup of peroxide, which is continually created by
multiple metabolic processes, and protects cellular organelles and tissues from harm. Catalase
functions as a major regulator in the metabolism of hydrogen peroxide. Catalase is also
engaged in managing the concentration of hydrogen peroxide, which is important in the
signaling process, according to certain research.2 Catalase protects live cells against oxidative
damage, which may happen when cells or other molecules in the body come into contact with
oxidative chemicals. This harm is a normal byproduct of chemical processes within your
cells. By-products of the reactions can include dangerous substances like hydrogen peroxide,
just as a by-product of a pleasant campfire might be undesired smoke that makes you cough
or hurts your eyes. The catalase enzyme assists in the removal of toxic chemicals by breaking
down hydrogen peroxide (H2O2) into harmless water and oxygen. 3 In the following
breakdown reaction, the catalase enzyme converts the substrate, peroxide, to water and
oxygen.
Catalase
2H 2 O 2 
 
 2H 2 O  O 2 (gas)
(substrate) (enzyme) (products) 4

The structure of an enzyme's active site is also affected by changes in pH. Each enzyme
performs optimally at a certain pH level. The optimal pH for an enzyme is determined by the
environment in which it operates. The pH of the reaction solution has a significant impact on
enzyme activity and stability. Amino acids containing ionic groups such as carboxyl group,
amino group, thiol group, imidazole group, phenolic hydroxyl group, and so on make up the
active sites of enzymes. The structure of the active site, the affinity for substrate, and the
catalytic activity are all affected by the dissociation state of amino acid residues.5 The rate of
enzyme activity increases as the pH rises. At the enzyme's optimal pH, maximum activity is
achieved. As the pH rises, the structure of the enzyme's active site changes, resulting in a
1
https://www.britannica.com/science/catalase (11th March 2021)
2
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6885225/#:~:text=Catalase%20is%20one%20of%20the
%20most%20important%20antioxidant%20enzymes.,diseases%20as%20a%20therapeutic%20agent. (11th
March 2021)
3
https://www.scientificamerican.com/article/exploring-enzymes/ (11th March 2021)
4
https://2019.igem.org/wiki/images/0/04/T--Technion-Israel--design-catalase-scheme.png (11th March 2021)
5
https://www.sciencedirect.com/topics/materials-science/enzyme-activity#:~:text=The%20specific%20activity
%20of%20an,concentration%20and%20the%20substrate%20concentration. (11th March 2021)
significant drop in activity. That is called denaturation, which might be permanent and
irrevocable. Plants have catalase as well. Catalase is found primarily in potatoes, which is
unusual because plants do not filter poisons from food. In potatoes as well, catalase breaks
down hydrogen peroxide into water and oxygen gas. Catalase, on the other hand, is engaged
in photorespiration, which explains its presence but not its abundance. Photorespiration is a
mechanism that helps plants avoid creating hydrogen peroxide, which can oxidize plant
materials and kill the plant if the ratio of absorbed light to water intake is too high.6

Research question:
How will different hydrogen peroxide (H2O2 – 110ml) concentrations (with Sodium
hydroxide (NaOH); Sulfuric acid (H₂SO₄) and water (H2O) affect the pH rate (pH meter) of
catalase?

Hypothesis 0 (null hypothesis):

The catalase in potato will not have an effect on any concentration with H2O2
Hypothesis 1:
Enzymes will operate as catalysts by speeding up chemical processes within living
organisms. We can see how enzymes like catalase execute decomposition, or the breaking
down, of other compounds using a potato and hydrogen peroxide.

VARIABLES:

Range Units
Independent variable NaOH+H202- 50+110= 160 ml
sodium peroxide
H2S04+H202- 50+110=160 ml
piranha solution
H20+H202- 50+110=160 ml
hydrogen peroxide
water
Dependent variable pH Colors on pH meter pH meter
Table 1: Presenting independent and dependent variables

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https://education.seattlepi.com/catalase-enzymes-potatoes-4108.html (11th March 2021)
CONTROLLED HOW IS IT POSSIBLE EFFECTS ON
VARIABLES CONTROLLED? RESULTS
Time used to measure All samples will have same Potatoes in each beaker must
reaction measuring time- 5 minutes be kept for the same time
The outcomes of the
intervals and trials will be
comparable, allowing a
conclusion to be drawn.
Type of potato Same type and length of There are different types of
potato potatoes and ways of them
having an effect on some
substances, so it is important
to use the same one during
the entire experiment. In this
experiment, one normal
potato was used and from it,
samples were taken so that
each beaker has same type
of potato. pH values would
not be same as different
types of potato might have
different reactions to
different solutions.
Amount of substances 110 ml of H2O2 in every To set concentrations of
beaker with 50ml of solutions more simply and
substances in each beaker accurately, as well as to
with H202 (water, H2SO4 make the volume of enzyme
and NaOH) and substrate for the reaction
similar, the amount of
solution used in all test tubes
must be equal. If varied
quantities of solutions are
used, there is room for
mistake since the volume of
enzyme may not be adequate
to react with all of the
solutions, even if the
substrate-enzyme ratio is
made.
Type of enzyme Catalase Catalase is an enzyme found
in potassium that transforms
hydrogen peroxide to water
and oxygen. The catalase in
potato juice accelerates the
breakdown of hydrogen
peroxide. Due to this fact,
potatoes + catalase of
course, will be constant in
each trial of samples.
Exposure to environment- Same temperature and pH Temperature has an effect
temperature and pH on enzyme activity, as it is
already known. As a result,
temperature of the
surroundings and samples
must be maintained, which
was accomplished by taking
temperature readings with a
thermometer during the
experiment and shielding
everything from heating and
cooling. Extremely high
temperatures can denature
an enzyme, causing it to lose
its function.
pH: Each enzyme has a
specific pH range that it
prefers. Enzyme activity will
be slowed if the pH is
changed outside of this
range. Having this in mind,
to make sure results are
occurring naturally, both
temperature and pH are
constant in this experiment.
Type of hydrogen peroxide H2O2 5mol in all trials Hydrogen peroxide had to
be the same as different
solutions react differently
with hydrogen solution.
Taking this into
consideration, hydrogen
peroxide had to be kept
same in all trials as it could
lead to inaccurate results in
our experiment and true
reactions would become
unknown.
Type of beaker 3x3 beakers made from It's critical to use identical
same material containers for all
concentrations since some
materials have distinct
properties, such as thermal
insulation, chemical
resistance, and so on, which
might alter the pH
indirectly. To acquire
reliable findings in the end,
we must supply the same
environment for all chemical
solutions. The glass beakers
were used in this
 experiment.
Table 2: Presenting controlled variables

MATERIALS EQUIPMENT
- Potato - Beaker
- Water| H2O 50 ml - Ruler
- Hydrogen peroxide 110ml 3x - Paper
- Sodium hydroxide | NaOH 50ml - Pen
- Sulfuric acid | H2SO4 50ml - pH meter
- Knife
- Stopwatch
- Tweezers
- Pipette
Table3: Materials and equipment needed

Procedure and methodology:

1. Prepare your potato by having 12 samples. Three samples per three beaker. Each
potato strip must be 3cm long.
2. Have 12 beakers, 3 for each trial. In each beaker add 110ml of hydrogen peroxide.
First sample: 3 beakers with 110ml of hydrogen peroxide with 50ml of Sulfuric acid;
Second sample: 3 beakers with 110ml of hydrogen peroxide with 50ml of sodium
hydroxide; Third sample: 3 beakers with 110ml of hydrogen peroxide with 50ml of
water. Use pipette for accurate measurement.
3. Label beakers with corresponding name for the solution: with sulfuric acid, with
sodium hydroxide and with water
4. Add one potato strip in each beaker- first do sample one, then two than three
5. Start the stopwatch- for 5 minutes and observe bubbling
6. Measure the height difference in solution- how much did it bubble
7. Measure pH value of solution with the pH meter
8. Write down measures
9. Clean up afterwards using gloves to protect yourself from acids and use dish soap

Safety, ethical, and environmental concerns:

Sulfuric acid can cause permanent blindness if it comes into close contact with the eyes. This
chemical can induce internal burns, irreparable organ damage, and even death if consumed.
Sulfuric acid aerosols cause significant eye and respiratory tract irritation, as well as tissue
damage, when inhaled at high concentrations.7 In the event of skin contact, ocular contact,
ingestion, and/or inhalation, strong bases are extremely dangerous. Eyes and skin can be
corroded by strong bases. To prevent any harmful outcomes, interaction with the face and

7
https://www.ehs.com/2014/07/sulfuric-acid-safety-tips-sulfuric-acid-msds-information/ (11th March 2021)
eyes was avoided at all costs, and hands were cleaned with soap after each time the skin came
into contact with any, since this may have triggered an allergic reaction or irritated the skin.
Every student also wore protective coats. The experiment was carried out with extreme
caution to ensure that no ingredients spilled on the garments or on the floor, and that no
beakers or other glass equipment were broken. Our teacher helped us pour the acid solutions
as she is more professional and careful which led to having no problems. In terms of ethics,
none of the features of this experiment were unethical. There were no animals involved, and
the environment was not harmed in any manner.

Analysis
Raw Data:

Beaker Time it took to Bubbling- how pH color

bubble (seconds) many ml did it reach
#1- water with 11s 60ml Brown (0.4)
hydrogen peroxide
#2-sulfuric acid with 24s 12ml Light Purple (0.11)
hydrogen peroxide
#3-sodium 7s 15ml Pink (7.02)
hydroxide with
hydrogen peroxide
Table4: First sample and results of bubbling and pH

Beaker Time it took to Bubbling- how pH color

bubble (seconds) many ml did it reach
#1- water with 10s 55ml Brown (0.3)
hydrogen peroxide
#2-sulfuric acid with 26s 12ml Light Purple (0.12)
hydrogen peroxide
#3-sodium 7s 16ml Orange (0.2)
hydroxide with
hydrogen peroxide
Table5: Second sample and results of bubbling and pH

Beaker Time it took to Bubbling- how pH color

bubble (seconds) many ml did it reach
#1- water with 8s 52ml Brown (0.5)
hydrogen peroxide
#2-sulfuric acid with 25s 10ml Light Purple(0.12)
hydrogen peroxide
#3-sodium 9s 16ml Yellow/light orange
hydroxide with (0.7)
hydrogen peroxide
 Table6: Third sample and results of bubbling and pH
Processing raw data:

Evaluation:
It took 7seconds for bubbles to appear in sodium hydroxide solution with hydrogen peroxide
and bubbles raised for 15ml, in sulfuric acid with hydrogen peroxide bubbles reached 12ml in
span of 24 seconds. Lastly, water solution with hydrogen peroxide had bubbles appear in
11seconds and they reached 60ml

Strengths & Limitations and their improvements:

This experiment's benefits include the fact that it does not require many complicated
instruments, making it very simple to carry out. The controlled environment conditions are
simple to maintain, for example, concentrations of different solutions and the use of the same
types of potato and hydrogen peroxide. Most of these were measured quickly and simply with
simple instruments. Another advantage is that the data is easily quantifiable, as the study
topic concerns how pH would change in various settings, which was measured using
bubbling quantities, implying that the raw data will include time calculations. While
composing the lab, I saw that I had improved significantly in terms of calculating various
portions and constructing the final graph with all required segments. I had to perform more
study for working in Excel in this section, which helped me better for all subsequent lab
reports. The graph was the area where I made the most progress. I utilized the pH meter for
the first time, and it was quite helpful in terms of assessing my data and evaluating my
hypothesis – both H1 and H0. This showed that there was a substantial relationship between
bubbling and pH readings. In addition, I was considerably more prepared for this lab than I
had been for the last one.The experiment has little ethical, safety, or environmental problems,
therefore it is rather safe with proper precautions.
Evaluation & Conclusion:
Changes in ph levels and hydrogen peroxide concentrations can impact catalase activity in
potatoes, according to the research. Looking at the graphs of both variables, it is clear that
increasing the ranges used in the experiment would have resulted in a decrease in the volume
of oxygen gas produced, as well as free active sites with no substrate to occupy them, limiting
the reaction and radically slowing the rate of reaction. Here we see the pH of the water,
NaOH and HCL. The pH of the water was brownish color aprox. 0.4, NaOH was ranging
from pink which was around 7.4 to yellow and orange which was 0.7 and HCL was light
purple, about 0.12. The water was somewhat acidic, but closer to neutral, and the NaOH is
extremely basic and the HCL is quite acidic. The water-filled test tube was the one that
created the most bubbles. It created bubbles in the amount of 50ml. The NaOH test tube got
roughly 10 ml, whereas the HCL test tube yielded 15 ml. Because the catalase operated the
best, this test tube with water created the most. In addition, the catalase worked effectively in
water.
When talking about limitation, there is a slight possibility that equipment was not cleaned
thoroughly, meaning someone else might have use pipette and beakers I used where they
used different concentrations and chemicals which might have stayed inside the beaker and
pipette. This might have messed up my reaction and whole experiment as there was a mixture
of different chemicals that were not needed. A good solution for this would be to use one-use
pipettes and immediately throw them away once an experiment is finished. As for the
beakers, the best solution would be to clean them before starting with the experiment with
dish soap and drying them good. Another limitation might be the sample for potato strips,
meaning they were not cut equally which could have damaged my results and reaction with
catalase. Solution would be to use either a measuring scale or perfectly measure potato strips
in grams so that they are all equal, or to have more than three samples, and then calculate the
mean of all results.
Bibliography:

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https://www.britannica.com/science/catalase. (11th March 2022)

Buddies, Science. “Exploring Enzymes.” Scientific American, Scientific American, 10 Nov.

2016, https://www.scientificamerican.com/article/exploring-enzymes (11th March 2022)

Cascio, Christopher. “Catalase Enzymes in Potatoes.” Education, 29 Sept. 2016,

https://education.seattlepi.com/catalase-enzymes-potatoes-4108.html (11th March 2022)

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