FHSC1214 FUNDAMENTALS OF CELL BIOLOGY

FOUNDATION IN SCIENCE
TRIMESTER 1

Name Dk Hanis Batrisyia Binti Pengiran Sabdin

Partner’s name Yong Hok Nian
Practical No. 3&4
Experiment The Investigation of the effects of catalase
concentration on Hydrogen Peroxide and
the investigation of enzymatic effects of
materials on hydrogen peroxide
Group 1
Date 12/02/2015 – 26/02/2015
Lecturer Ms Nicole Ngai Suet Loo
Marking Scheme for Full Report
Content Description of content Marks
Title Able to make a title representing 2 experiments that have been /1
conducted
What you did?
Objectives Able to identify objectives of the 2 experiments /1
What is supposed to be accomplished through an
experiment?
Apparatus Compile 2 experiments /1
Simple list of everything needed to complete the experiment
Materials Compile 2 experiments /1
Simple list of everything needed to complete the experiment
Procedures 1. Describes the process in chronological order /2
2. Includes all the information necessary to carry out the
experiment
3. May include diagrams / images to illustrate the apparatus
used & how it was set up
4. Should be written as a description of “what you did”, not as a
“set of instructions”
Results Table 1 with title (page 36) /2
Graph with appropriate title (page 36) /3
Table 2 with title (page 42) /7
Discussion 1. The first paragraph with One to three sentences to introduce /26
what had been done
2. A brief explanation of concept used in the experiment
3. Explain how the result demonstrated these principles. Use
the important data and results (Table) to demonstrate these
principles.
4. Explain how the independent variables affected the
dependent variables, you may use equations provided and
show the dependent/independent variables.
5. Analysis of graph
6. Sources of error are offered that are consistent with the
experimental results.
7. You may offer a suggestion for improving the experiment, but
it must focus on the most prominent error and be consistent
with the sources of errors.
8. Precautions

Citation /1

Conclusions Able to identify conclusion each for 2 experiments /2

References Must have reference from book, journal or scientific article /2
Must be written in the proper format
Format /1
Total /50
Plagiarism Similarity of the discussion part of your report must be less than
50%
-10
For not following this instruction,
Title : The Study of Enzyme

Objective : To investigate the effects of catalase concentration on hydrogen peroxide and the
enzymatic effects of materials on hydrogen peroxide

Apparatus : 5 boiling tubes, 1 beaker (500cm3), 1 beaker (250cm3), 1 rubber bung with delivery
tube, 4 test tubes, pen knife, white tile, pestle and mortar, weighing boat, filter funnel and filter
paper, hand-held pipette, beaker, test tubes, water bath

Materials : potato cubes, hydrogen peroxide, fresh liver, manganese dioxide, wood splints

First, the experiment to investigate the effects of catalase concentration on hydrogen peroxide
was conducted.

Procedures :

1. Firstly, an electric water bath of 37 ̊ C was set up.

2. A boiling tube is selected with a rubber bung and was labelled as tube A.
3. The potato sample given was cut with a pen knife and was weighed 5g using a weighing
balance on a weighing boat.
4. The potato samples were cut into smaller pieces to enable an easier mashing using the
pestle and mortar. 6cm3 of distilled water was added to the potato samples after
mashing.
5. The solid mashed potato was separated by filtering the mashed potato sample with filter
paper and funnel and the filtrate was collected in a test tube.
6. An empty test tube was filled up with tap water.
7. 5cm3 of hydrogen peroxide was added into Tube A using the hand-held pipette provided.
8. 1cm3 of the filtrate from the mashed potato samples was drawn and added to Tube A.
The test tube was immediately closed with the rubber bung with the furthest end of the
delivery tube from the bung sealed with parafilm.
9. The parafilm was then removed and the tube was immediately immersed in the water
bath. The number of gas bubbles were recorded for 2 minutes.
10. Step 7 to 9 was repeated to get a 2nd measurement.
11. Steps 2 to 10 was repeated with 10g of potato, then 15g of potato and finally 20g of
potato.
12. The data was then recorded.

The 2nd experiment to investigate the enzymatic effects on hydrogen peroxide was conducted.

Procedure :

1. 6 boiling tubes were labeled as 1, 2, 3, 4, 5, 6, and was stood in a test tube rack.
2. The liver was cut into several pieces of roughly 0.8 cm x 0.8 cm x 0.5 cm with a razor
blade.
3. One piece of liver was placed into Tube 1.
4. The second piece of liver was placed into the bottom of tube 2.The liver was gently
spread using a wooden splint without mashing it, over as wide an area as possible of the
 bottom of the boiling tube. Then, tube 2 was placed in water bath of 95 ̊ C for about 5
minutes.
5. Two portions of 0.5g of manganese dioxide powder was measured out onto a weighing
boat using a weighing balance. Each portion was poured into tube 5 and tube 6
respectively.
6. Test tube 6 was put into a water bath of 95 ̊ C for five minutes.
7. Tube 2 and 6 was let cool after 5 minutes.
8. The 3rd piece of liver was then put into boiling tube 3. It was mashed gently into a pulp
with the wooden stick.
9. Potato cubes were cut roughly 0.8 cm x 0.8 cm x 0.5 cm. 1 cube was placed into tube 4.
10. 6 fresh empty test tubes were prepared and was stood on a test tube rack, and was
added 5 cm3 of hydrogen peroxide into each of them.
11. Next, hydrogen peroxide was added into boiling tubes 1, 2, 3, 4, 5, and 6.
12. Parafilm was used to seal the mouth of the boiling tubes by stretching the film over it to
prevent the parafilm from being displaced as a lot of gas is produced.
13. The tubes are left for 20 minutes as a lot of gas was produced in the boiling tubes as
evidenced by the bulging of parafilm from the boiling tube mouths.
14. Once enough gas has accumulated in the boiling tubes, a glowing splint was inserted
into each tube one at a time by penetrating the parafilm with it.
15. The observations were recorded in a table.
Results :

Table 1 – The number of gas bubbles produced

The effects of catalase concentration on H2O2

5g 10g 15g
st nd rd st
Number of 1 2 3 1 2nd 3rd
1 st
2nd 3rd
Attempt
Number of gas
bubbles 51 49 54 79 83 84 109 117 113
produced

Graph 1 – The number of gas bubbles produced against the amount of potato samples used.
Table 2 –

Test Contents with Observations before Observations after using wood

tube 5cm3 hydrogen using wood splint. splint
peroxide
1 Fresh liver Gas bubbles were A light ‘pop’ sound was heard.
produced. Effervescence The glowing wooden splinter
occurred. reignites. White fumes were
produced.
2 Boiled liver A small amount of The glowing wooden splinter
(cooled) bubbles were produced. remains glowing.
Less effervescence
occurred.
3 Pulped liver A lot of gas bubbles were A loud ‘pop’ sound was heard.
produced. The liver The glowing wooden splinter
residue was floating on reignites. White fumes were
the bas bubbles. produced.
Effervescence occurred.
4 Potato cubes Gas bubbles were The glowing wooden splinter
produced. Effervescence remains its glow.
occurred.
5 Manganese Effervescence occurred. A loud ‘pop’ sound was heard.
dioxide The glowing wooden splinter
(untreated) reignites. White fumes were
produced.
6 Boiled Effervescence occurred. A loud ‘pop’ sound was heard.
manganese The glowing wooden splinter
dioxide (cooled reignites. White fumes were
after heating) produced.
Discussion :

Catalase is a common enzyme which is found in almost all living organisms that is
exposed to oxygen. Catalase catalyzes the decomposition of hydrogen peroxide to water and
oxygen which can be shown in the equation : 2H2O2 → 2H2O + O2
Catalase is an enzyme that acts as a cellular defense for oxidative damage that is
caused by reactive oxygen species (ROS). ROS are high reactive ions and free radicals that
involves oxygen. ROS can impair the DNA, RNA, and proteins, in plants. ROS are produced as
a regular product of cellular metabolism. ROS is produced from aerobic organisms. Oxygen ions
and peroxides contributes to ROS. Catalase enzyme converts the hydrogen peroxide to water
and oxygen. (Chidrawi, Downs, & Robson, 2008)
Catalase is a tetramer of four polypeptide chains, with each more than 500 amino acids
long. It comprises of four porphyrin heme(iron) groups that enables the enzyme to react with the
hydrogen peroxide. (Boon & Aaron Downs, 1997-2008)
Two experiments were conducted to investigate the effects of catalase concentration on
hydrogen peroxide and the enzymatic effects of materials on hydrogen peroxide respectively.
The effects of catalase concentration was studied by using potato samples of different
amounts, which was 5g, 10g and 15g that was tested with hydrogen peroxide. Potato was
chosen due to its high presence of catalase. Potato is involved in photorespiration which is a
process that prevents an unwanted ratio of absorbed light to water consumption from producing
hydrogen peroxide, which can oxidize plant matter and kill the plant. Catalase converts the
hydrogen peroxide into water and oxygen which can be seen through the formation of gas
bubbles.
In this experiment, different amount of potato samples, 5g, 10g and 15g samples were
tested 3 times each to get the average number of gas produced.
The enzymatic effects of materials on hydrogen peroxide was conducted with liver,
manganese dioxide, and potato samples once again. Liver was chosen because it is to
represent the presence of catalase in the body and the potato in the plants. Manganese
peroxide was chosen due to its availability to react with hydrogen peroxide to produce oxygen.
When involving manganese dioxide, the equation formed is 2H2O2+ MNO2 2H2O + O2, hence
the product formed is water and oxygen because manganese does not take part in the reaction.
It only acts as a catalyst that speeds up the reaction.
Both of the experiment demonstrated the principle of catalase catalyzing the
decomposition of hydrogen peroxide to water and oxygen.
In the 1st experiment, the constant variable was the concentration of hydrogen peroxide.
The manipulated variable is the amount of catalase which was shown by using different
amounts of potato samples. The responding variable is the number of gas bubbles produced
when reacted. Based on table 1, three attempts were made for each of the amount of the potato
samples. As the amount of potato samples increased, so did the number of gas produced. This
is mainly because of the catalase in the potato catalyzed the decomposition of hydrogen
peroxide to water and oxygen. The product of the decomposition is oxygen. Hence, the number
of gas bubbles were measured.
 The data from graph 1 was obtained from the data table 1. The average amount of the
number of gas bubbles was calculated as the responding variable on the y-axis while the
amount of potato samples used was as the manipulated variable on the x-axis. The line graph of
graph 1 shows that the number of gas bubbles increased due to the amount of oxygen
collected.
In the 2nd experiment, different enzymatic effects of materials on hydrogen peroxide was
studied. The constant variable is the concentration of the hydrogen peroxide, the manipulated
variable is the materials used and the responding variable is the effects on hydrogen peroxide.
The materials that were involved were fresh liver, boiled liver, pulped liver, potato cubes, treated
manganese dioxide and boiled manganese dioxide.
When the liver had been pulped, the reaction was more vigorous due to the increased
surface area for the enzyme to act on. The greater the surface area, the more exposed it is to
the reactant. Meanwhile, the boiled liver produced less result because enzyme had denatured
due to high temperature. This can be explained as enzyme is a protein that is held by weak
bonds and the molecular motion changes. This different molecular motion changed its 3-D
shape of the enzyme and thus it lost its ability to bind with other substrate due to the enzyme is
not complement to its active site. The fresh liver produced more gas bubbles compared to
potato which shows that the liver has more catalase activity than in potato. This is explainable
due to the different functions of liver and potato in nature. Catalase in liver detoxifies chemicals
while the catalase in potato prevents the oxidation of starch. The reaction in the manganese
dioxide was greater than the boiled liver. When boiling a liver, the enzyme structure is denatured
due to high temperature as enzyme is protein and has the characteristics of protein. The high
temperature made it lose its 3-D shape and its ability to bind to the active site of the product as
it is no longer complementary. However, enzymes are specific. Boiling the manganese oxide
activates it because it is an inorganic catalyst, thus it does not denature when heated. The
presence of white fumes, the pop sound and wooden splinter igniting shows that there is
presence of oxygen gas.
Several errors could have occurred throughout the experiment. Miscalculations involving
numbers and amounts of solutions would have a severe effect upon the results. Mathematical
errors may also have of occurred. This can be avoided by calculating accurately and checking
the calculations frequently. Parallax error when measuring solutions can also be avoided by
measuring accurately and measure by meniscus reading. However, no error was done when
doing both of the experiments. This may be due to handling the materials and equipment safely
and correctly. The steps of the procedures were also followed perfectly.
Handling the water bath and the wooden splint must be handled carefully as the heat
can damage our skin. A neat and clean working area is needed to when doing experiments.
Hands, eyes and skin from acids, other corrosive substances, and flying shrapnel from broken
canisters and beakers should be protected with gloves, goggles and full-sleeved shirts.
Conclusion :
Catalase, or enzymes, significantly increases the rate of hydrogen peroxide
decomposition. The experiments conducted shows how catalase added to hydrogen peroxide
leads to the release of oxygen, boiled catalase is denatured, and the presence of catalase in
living things can lead to the breaking down of hydrogen peroxide in the body. It was shown that
the natural decomposition hydrogen peroxide is slower than decomposition taking place with the
addition of enzymes. If hydrogen peroxide was required to decompose naturally, life could not
survive. The addition of catalase increases this decomposition rate allowing life to continue.
In the first experiment, we can conclude that different catalase concentration affects the
rate of reaction. Catalase was found in potato samples. The different amount of potato samples
used showed different readings of gas bubbles produced. The more catalase used, the more
the production of oxygen.
In the second experiment, we can conclude that different enzymatic materials have
different effects when reacted with hydrogen peroxide. At high temperature, enzymatic activity is
affected as enzyme is a protein. Protein denatures at high temperature. We can also conclude
that enzymes are highly specific, this was shown when manganese dioxide was reacted with
hydrogen peroxide. Manganese dioxide is an inorganic catalyst.
Both of the experiments were conducted successfully.

References
1. Boon, E. M., & Aaron Downs, D. M. (1997-2008). biology.kenyon.edu. Retrieved from Catalase
H2o2 : H2O2 Oxireductase: http://biology.kenyon.edu/BMB/Chime/catalase/frames/cattx.htm

2. Chidrawi, G., Downs, A., & Robson, M. (2008). Biology in Focus - HSC Course. McGraw-Hill
Australia.