LAB REPORT 4 1

Lab report 4 - Enzyme Activity Lab report

Enzyme Activity Lab report

February 19, 2023

Research Question:

In what way does the amount of catalase as well as the rate of reaction are different in

regards to various concentrations of hydrogen peroxide (0%, 0.75%, 1.5%, 3%, 5%)?
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Personal Engagement:

This lab was an enlightening experience in the sense that I haven't performed a lab in a

long period of time. I felt a wave of excitement and ambition toward performing this experiment.

This lab was performed and conducted in a group setting which brought a need for and sense of

collaboration. Within the collaboration of my classmates and the joint critical thinking, we

wanted to see how the amount of catalase as well as the rate of reaction differ when in different

concentrations of hydrogen peroxide, such as 0%, 0.75%, 1.5%, 3%, 5%. This lab was split into

four trials which made this experiment twice as interesting when comparing the results with my

classmates afterward and seeing how different the reactions were between the trials. Each

member of the group has a specific role within the experiment and engaging with each other to

make sure the work that we were doing was precise, through timing and measuring certain

things, has really strengthened my teamwork and collaboration skills. This lab widened my

knowledge about enzymes through hands-on work that was truly engrossing and strengthened

my ambition towards performing more labs in the future. Overall this was a very interesting

experiment and helped me see how my skills in the lab improved from when it was my very first

time performing an experiment in the lab ever.

Background:

Enzymes are chemicals in living organisms that serve as a catalyst (Enzyme | Definition,

Mechanisms, & Nomenclature | Britannica, n.d.). They control the rate at which chemical

reactions occur while remaining unaffected by the process (Enzyme | Definition, Mechanisms, &

Nomenclature | Britannica, n.d.). Enzymes are proteins that aid in the speeding up of

metabolism, or the chemical events that occur in our bodies (Enzymes: What Are Enzymes,

Pancreas, Digestion & Liver Function, 2021). Some chemicals are created while others are
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destroyed. Enzymes are found in all living organisms and are naturally produced by our bodies

(Enzymes: What Are Enzymes, Pancreas, Digestion & Liver Function, 2021). Within living

organisms many biological processes take place, which is referred to as chemical reactions, those

reactions happen to be regulated by enzymes, which prove to show their significance (Enzyme |

Definition, Mechanisms, & Nomenclature | Britannica, n.d.). Enzymes have the ability to

catalyze every part of cell metabolism, such as food digestion, chemical energy transformation as

well as its conservation, along with the process of constructing cellular macromolecules from

smaller antecedents (Enzyme | Definition, Mechanisms, & Nomenclature | Britannica, n.d.).

Enzymes are proteins that are made up of amino acids that are joined with one another towards

one or numerous polypeptide chains (Biochemistry, Proteins Enzymes - StatPearls, 2022). The

primary structure of a polypeptide chain is the sequence of amino acids (Biochemistry, Proteins

Enzymes - StatPearls, 2022). Which indicates the enzyme's three-dimensional structure, such as

the active site's structure (Biochemistry, Proteins Enzymes - StatPearls, 2022). The function of an

enzyme is inextricably tied to its three-dimensional structure, which determines how it

accomplishes substrate binding, catalysis, and control (What Is Enzyme Structure And

Functions?, n.d.). When an enzyme comes across certain changes in temperature or pH

fluctuations, the protein structure will denature, meaning that it could lose its integrity as well as

its enzymatic capacity (Enzyme | Definition, Mechanisms, & Nomenclature | Britannica, n.d.).

Certain examples of enzymes include when lipase breaks down fats into fatty acids when

protease breaks down protein into amino acids, as well as when carbohydrase breaks down

carbohydrates into sugars (Enzymes: What Are Enzymes, Pancreas, Digestion & Liver Function,

2021). Many factors influence enzyme activity, including substrate concentration as well as the

presence of inhibitory substances (Enzyme | Definition, Mechanisms, & Nomenclature |

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Britannica, n.d.). Temperature, pH, and concentration are all parameters that can influence

enzyme activity (Enzymes Review (Article), n.d.). Enzymes function best in specified

temperature and pH ranges, and poor circumstances can lead the enzyme to decrease its ability to

attach to a substrate (Enzymes Review (Article), n.d.). Enzymes also have the ability to denature.

Enzyme denaturation occurs when a protein's structure and functional group experiences

conformational changes, which cause inactivity (What Is Meant by Denaturation of Enzyme?,

n.d.). Denatured proteins tend to acquire a looser, more irregular structure along with them being

mostly insoluble (What Is Meant by Denaturation of Enzyme?, n.d.). As a harmful byproduct of

regular cellular processes, cells produce hydrogen peroxide (H2O2) (Genovesi et al., 2021).

Catalase is an enzyme that swiftly degrades hydrogen peroxide into water and oxygen (Genovesi

et al., 2021). One catalase enzyme molecule can act on 40 million molecules of hydrogen

peroxide every second (Genovesi et al., 2021). When catalase comes into contact with hydrogen

peroxide, the hydrogen peroxide combines with the catalase enzyme in the injured cells to

generate oxygen foam (Genovesi et al., 2021). The oxygen gas bubbles when the catalase

reaction is carried out in a test tube (Genovesi et al., 2021). The quantity of catalase activity

present is indicated by the height of the foam (Genovesi et al., 2021). When hydrogen peroxide

is added to a solution, its reaction rate increases because more substrate molecules may collide

with the enzyme, resulting in more product (Lohner, 2016). As you raise the amount of H2O2 in

the reaction, the amount of foam created increases (Lohner, 2016).

Hypothesis:

The higher the concentration of hydrogen peroxide, the more foam generated, indicating

that more catalase was required to catalyze hydrogen and oxygen (Lohner, 2016). Catalase
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breaks down hydrogen peroxide into hydrogen and oxygen, with a higher concentration of

hydrogen peroxide yielding more oxygen due to a faster reaction (Lohner, 2016).

Requirements/Materials:

Potato extract (catalase)

Hydrogen peroxide
Forceps
Water
Beakers
Vials
Water bath
Ruler
Stopwatch

Procedure:

Effect of substrate concentration on the rate of the reaction

1. Prepare 1 vial of catalase with a 100 units/mL concentration.

2. Dilute the substrate so you end up with 5 different cups with the following hydrogen peroxide

concentrations: 3.0%, 1.5%, 0.75%, 0.38%, and 0%.

3. Measure and record the depth of the hydrogen peroxide in the vials using a ruler.

4. Add 2 cubes of potato to 20ml of 3.0% hydrogen peroxide solution..

5. Leave the solution to stand until no more foam forms and then record the new height of the

hydrogen peroxide solution and the time(s) taken to reach that height.

7. Repeat the procedure for each of the other hydrogen peroxide concentrations.

8. Do 3 trials for each step and record the results in a table.

Safety/ethical /environmental considerations:

1. Wear lab coat at all times

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2. Do not eat or drink the solanum tuberosum solution

3. Hair that is long should be tied

Variables

Table 1: Variables of the experiment; Independent Variables, Dependant Variables, Constants, as

well as the Controlled Variables

Independent Dependant Constants Controlled Variables

Variable Variables

The different Quantity of catalase Stopwatch and The temperature of

concentrations of activity. testtubes. the room as well as
hydrogen peroxide the amount of
(0%, 0.75%, 1.5%, catalase.
3%, 5%).

Data Processing:

Qualitative Data

Figure 1: Trial 1 - before the catalase Figure 2: Trial 1 - after the catalase

Figure 3: Trial 2 - before the catalase Figure 4: Trial 2 - after the catalase
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0% - 0.75% - 1.5% - 3% - 5% 0% - 0.75% - 1.5% - 3% - 5%

Figure 5: Trial 3 - before the catalase Figure 6: Trial 3 - after the catalase

0% - 0.75% - 1.5% - 3% - 5% 0% - 0.75% - 1.5% - 3% - 5%

Figure 7: Trial 4 - before the catalase Figure 8: Trial 4 - after the catalase
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0% - 0.75% - 1.5% - 3% - 5% 0% - 0.75% - 1.5% - 3% - 5%

Quantitative Data

Table 2: Raw Data - Concentrations, Initial Height, Final Height, and Change in Height for Trial
1,2,3, and 4

Trial 1 Trial 2 Trial 3 Trial 4

conce Initial Final Chan Initial Final Chan Initial Final Chan Initial Final Chan
ntrati height height ge in height height ge in height height ge in height height ge in
on (cm ± (cm ± heig (cm ± (cm ± heigh (cm ± (cm ± height (cm ± (cm ± heigh
0.5) of 0.5) of ht 0.5) of 0.5) of t (cm 0.5) of 0.5) of (cm ± 0.5) of 0.5) of t (cm
hydroge hydroge (cm hydrog hydrog ± 0.5) hydrog hydrog 0.5) hydrog hydrog ±
n n ± en en (final en en (final en en 0.5)
peroxid peroxid 0.5) peroxid peroxi - peroxid peroxid - peroxi peroxi (final
e e (final e de initial e e initial de de -
(substra (substra - (substra (substr heigh (substr (substr height (substr (substr initial
te) te) after initia te) ate) t) ate) ate) ) ate) ate) heigh
before adding l before after before after before after t)
adding solanu heig adding adding adding adding adding adding
solanu m ht) solanu solanu solanu solanu solanu solanu
m tuberos m m m m m m
tuberos um tuberos tubero tuberos tuberos tuberos tubero
um extract um sum um um um sum
extract (enzym extract extract extract extract extract extract
(enzym e) (enzym (enzy (enzym (enzym (enzym (enzy
e) e) me) e) e) e) me)

0% 3.5 4.5 1.0 4.1 4.2 0.1 3.9 5.0 1.1 4.1 5.5 1.4

0.75 3.0 4.0 1.0 4.6 5.2 0.6 3.2 4.7 1.5 4.3 5.3 1.0
%

1.5% 3.5 5.1 1.6 4.3 5.5 1.2 3.6 4.5 0.9 4.2 5.4 1.2

3% 3.6 5.0 1.4 4.2 5.3 1.1 3.8 4.9 1.1 4.2 5.7 1.5
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5% 3.6 4.9 1.3 4.1 5.0 0.9 3.7 4.8 1.1 4.1 5.5 1.4

Processed Data:

Table 3: Processed Data - Concentration, Mean Change in Height, Rate of Change rounded to the
Nearest Hundredth, and Standard Deviation Rounded to the Nearest Hundredth

Concentra Mean change in Rate of change / rate of reaction (m/s) rounded to the nearest
tion height (cm ± 0.5) hundredth

(ΔHt1 + ΔHt2 + ΔHt3 mean change in height

+ ΔHt4 ------------------------------
--------------------- average time in mins it took the foam to stop (time in trial 1 + time in
4) trial 2 + time in tiral 3 + time in trial 4 / 4)

0% 0.9 Average time (mins): 4.25

0.9 / 4.25 = 0.21 cm/min

= 3.5 x 10-5m/s

0.75% 1.025 Average time (mins): 3.75

1.025 / 3.75 = 0.27 cm/min

= 4.5 x 10-5 m/s

1.5% 1.225 Average time (mins): 4

1.225 / 4 = 0.31 cm/min

= 5.17 x 10-5 m/s

3% 1.275 Average time (mins): 4.25

1.275 / 4.25 = 0.3 cm/min

= 5 x 10-5 m/s

5% 1.175 Average time (mins): 4.5

1.175 / 4.5 = 0.26 cm/min

= 4.33 x 10-5 m/s

Standard deviation rounded to the nearest hundredth

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0% 0.48

0.75% 0.32

1.5% 0.25

3% 0.18

5% 0.19

Figure 9: Each trial's rate of reaction (m/s) after adding solanum tuberosum extract to each
concentration of hydrogen peroxide.

Figure 10: Standard deviation rounded to the nearest hundredth

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Analysis of Collected Data:

Through analyzing the data collected, it has been determined that the different

concentrations of hydrogen peroxide result in a range of various heights, rates of reactions, and

changes, as well as its standard deviation. For the 0% percent concentration, the mean change in

height (cm ± 0.5) was 0.9, while the rate of change and reaction (m/s) rounded to the nearest

hundredth was 3.5 x 10-5m/s. The average time (mins) for it was 4.25. The standard deviation of

the 0% concentration rounded to the nearest hundredth was 0.48. The 0.75% hydrogen peroxide

concentration had a mean change in height (cm ± 0.5) of 1.025, rate of change and reaction (m/s)

rounded to the nearest hundredth was 4.5 x 10-5 m/s, with an average time (mins) of 3.75. The

standard deviation rounded to the nearest hundredth for the 0.75% concentration of hydrogen

peroxide is 0.32. For the 1.5% concentration of hydrogen peroxide, it had a mean change in

height (cm ± 0.5) of 1.225 along with its rate of change and reaction (m/s) rounded to the nearest

hundredth being 5.17 x 10-5 m/s with an average time of 4 minutes, as well as its standard

deviation rounded to the nearest hundredth being 0.25. The 3% hydrogen peroxide

concentration’s mean change in height (cm ± 0.5) was 1.275 and its rate of change and reaction

(m/s) rounded to the nearest hundredth was 4.33 x 10-5 m/s with its average time (mins) was 4.33

x 10-5 m/s. The standard deviation rounded to the nearest hundredth of the 3% hydrogen peroxide

concentration is 0.18. The 5% hydrogen peroxide concentration has a mean change in height (cm

± 0.5) of 1.175 as well as 4.33 x 10-5 m/s being the rate of change and reaction (m/s) rounded to
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the nearest hundredth, while its average time (mins) is 4.5. The standard deviation rounded to the

nearest hundredth for the 5% concentration is 0.19.

Conclusion:

To sum up everything that has been stated, the hypothesis conducted was correct. That

the higher the concentration of hydrogen peroxide, the more foam will be generated, indicating

that more catalase was required to catalyze hydrogen and oxygen (Lohner, 2016). The catalase

breaks down hydrogen peroxide into hydrogen and oxygen, with a higher concentration of

hydrogen peroxide yielding more oxygen due to a faster reaction (Lohner, 2016). The results of

this experiment aligned with the hypothesis, as anticipated, and proved to show its accuracy and

in fact that the mean of the various trials the height of the foam had increased, the higher the

concentration of hydrogen peroxide, concentrations being (0%, 0.75% 1.5%, 3%, and 5%). The

highest concentration of 5% had the highest mean change in height (rounded to the nearest

hundredth) (cm ± 0.5) while the lowest concentration of 0% had the lowest mean change in

height (rounded to the nearest hundredth) (cm ± 0.5). While the rest of the concentrations that

mean change in height kept increasing as the concentration got higher played a factor in proving

that the higher the concentration the higher the change in height (Lohner, 2016). Not only does

the change in height get higher the higher the concentration is, but also the rate of change of the

solution. The rate of change slowly decreased at first, but then increased again as the

concentration of hydrogen peroxide got higher. Overall, this proves the total hypothesis to be

accurate due to the data shown. This experiment was similar to another piece of work conducted

by Jessica Lee. Lee’s experiment was similar to the one conducted and showed very similar

evidence that as the concentration increased so did the rate of reaction (Lee, 2013). In
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conclusion, this lab was a very successful one that proved valid results and showed the

hypothesis to be accurate.

Evaluation (strengths/weaknesses/ suggestions for improvement):

Table 4: Evaluation (strengths/weaknesses/ suggestions for improvement)

Strengths Weaknesses Suggestions for

Improvement

Collaborated with classmates Difficulties in getting Trying to be as precise as

well in regard to conducting accurate measurements possible with the timing and
this experiment. (multiple attempts to get measurements.
accurate measurements took
place).

Continued on with this The pipette was not rinsed in Being more precise when
experiment cautiously and between some uses. retrieving the catalase,
carefully. making sure it's the correct
amount.
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References

Biochemistry, Proteins Enzymes - StatPearls. (2022, April 28). NCBI. Retrieved February 19,

2023, from https://www.ncbi.nlm.nih.gov/books/NBK554481/

Enzyme | Definition, Mechanisms, & Nomenclature | Britannica. (n.d.). Encyclopedia Britannica.

Retrieved February 19, 2023, from https://www.britannica.com/science/enzyme

Enzymes review (article). (n.d.). Khan Academy. Retrieved February 19, 2023, from

https://www.khanacademy.org/science/ap-biology/cellular-energetics/environmental-impa

cts-on-enzyme-function/a/hs-enzymes-review

Enzymes: What Are Enzymes, Pancreas, Digestion & Liver Function. (2021, May 12). Cleveland

Clinic. Retrieved February 19, 2023, from

https://my.clevelandclinic.org/health/articles/21532-enzymes

(2013, October 23). Lee 1 Jessica Lee AP Biology Mrs. Kingston 23 October 2013 Abstract: The

purpose of this lab is to investigate the impact of temperature, substrate concentration.

Retrieved February 22, 2023, from

https://www.region18.org/uploaded/student_stand-outs/jessica_lee_enzyme_lab.pdf

Genovesi, E., Blinderman, L., & Natale, P. (2021, August 1). 6.2: Catalase. Biology LibreTexts.

Retrieved February 19, 2023, from

https://bio.libretexts.org/Learning_Objects/Laboratory_Experiments/General_Biology_L

abs/Unfolding_the_Mystery_of_Life_-_Biology_Lab_Manual_for_Non-Science_Majors

_(Genovesi_Blinderman_and_Natale)/06%3A_Enzymes/6.02%3A_Catalase
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Lohner, S. (2016, November 10). Exploring Enzymes. Scientific American. Retrieved February

19, 2023, from https://www.scientificamerican.com/article/exploring-enzymes/

Lohner, S. (2016, November 10). Exploring Enzymes. Scientific American. Retrieved February

22, 2023, from https://www.scientificamerican.com/article/exploring-enzymes/

What Is Enzyme Structure And Functions? (n.d.). Infinita Biotech. Retrieved February 19, 2023,

from https://infinitabiotech.com/blog/enzyme-structure-and-function/

What is meant by denaturation of enzyme? (n.d.). Byju's. Retrieved February 19, 2023, from

https://byjus.com/question-answer/what-is-meant-by-denaturation-of-enzyme/