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Enzyme Concentration and Activity Core Practical

The experiment investigates how enzyme concentration affects reaction rate by measuring oxygen produced from the reaction between hydrogen peroxide and the potato enzyme catalase. It was hypothesized that increasing catalase concentration would increase the reaction rate and oxygen volume within 60 seconds. Potato slices were added to hydrogen peroxide solutions in increasing amounts from 0 to 10 slices. Oxygen volumes produced were highest with 10 slices, supporting that higher enzyme concentration leads to greater reaction rates. The experiment could be improved with more precise measurements and controlling external factors.

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Agnes Okanlawon
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9K views

Enzyme Concentration and Activity Core Practical

The experiment investigates how enzyme concentration affects reaction rate by measuring oxygen produced from the reaction between hydrogen peroxide and the potato enzyme catalase. It was hypothesized that increasing catalase concentration would increase the reaction rate and oxygen volume within 60 seconds. Potato slices were added to hydrogen peroxide solutions in increasing amounts from 0 to 10 slices. Oxygen volumes produced were highest with 10 slices, supporting that higher enzyme concentration leads to greater reaction rates. The experiment could be improved with more precise measurements and controlling external factors.

Uploaded by

Agnes Okanlawon
Copyright
© Attribution Non-Commercial (BY-NC)
Available Formats
Download as DOCX, PDF, TXT or read online on Scribd
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Enzyme Concentration and activity Core Practical

Aim:
The aim of the experiment is to investigate how enzyme concentration can affect the initial
reaction rate by measuring the amount of oxygen produced as a by product of the reaction
between hydrogen peroxide and Catalase an enzyme found in potatoes.

Hypothesis
As the amount of the enzyme of Catalase increases so will the rate of reaction and the
volume of oxygen produced in the 60 seconds. This is because as the amount of enzyme
increases it allows the substrate more of a chance of reacting with the active site as there
are more enzyme for them to collide with. The initial rate of reaction is directly proportional
to the enzyme concentration therefore the more catalase enzyme present, the greater the
number of active sites that are available to form enzyme-substrate complexes.

Procedure
1. Set up the apparatus so that the test tube remains in the water and the syringe
barrel collecting tube stays under water in separate beakers. Place the delivery tube
into the test tube and the bottom of the syringe. Also place the screw clip over the
tip of the syringe.
2. Cut up the potato discs 0.2mm thick ready to place into the test tube in twos.
3. Carry out a control by pouring 5cm3 of buffer solution and 5cm3 of hydrogen
peroxide in order to check that it is the result of adding the catalase for the potato
that causes the oxygen to be produced.
4. Afterwards put in 2 potato discs into the solution and record the amount of oxygen
gas produced by measuring the amount of oxygen bubbles produced in the syringe.
5. Immediately place the bung and delivery tube firmly into the boiling tube. Place the
other end of the delivery tube under the collecting tube.
6. Start the stop clock as soon as the first bubble of oxygen enters the collecting tube
from the delivery tube. Collect any gas produced in the first 60 seconds.
7. Measure the volume of oxygen produced in the 60 seconds; raise the collecting tube
so that the water level water in the tube is level with the surrounding water in the
beaker.
8. Wash out the boiling tube thoroughly.
9. Repeats steps 4 to 7 with 4 slices and 6 slices and 8 slices and then finally 10 slices of
potato.

The independent variable in the experiment was the amount of the enzyme catalase in the
form of slices of potato this determined the changes in the initial rates of reactions. The
dependant variable is the amount of oxygen produced as a bi-product of the reaction
2H₂O₂ 2H₂O + O₂.
The controlled variables were the amount of time the samples were left, this is because no
matter how much enzyme you add eventually the substrate would be used up and the
reaction would become saturated and slow down regardless of the amount of enzyme
present. We also controlled the amount of hydrogen peroxide staying at a constant 5cm₃
per sample and the temperature of the water the test tube and syringe was placed in. Also
the Ph was controlled by adding 5cm₃ of buffer solution in each sample in order to keep the
ph of the potatoes used affecting the rate of initial reaction.
In order to make sure that the volume of oxygen produced is all due to the enzyme-
substrate complex we must carry out a control which in this case is to show that the
hydrogen peroxide will not produce oxygen bubbles by itself without the catalase enzyme.
This control solution of hydrogen peroxide and buffer solution was also left for 60 seconds
and no oxygen was produced.
We made sure our results were reliable and precise by measuring using a syringe barrel
collecting tube to measure the volume and also repeated the experiment so that we were
able to have an average measurement which cancelled out any anomalies or errors in
calculation or contaminations of the different solutions. We only measured the initial rate of
reaction because after a certain period of the time the enzyme reaction would slow down
naturally as the substrate would become used up until all of it would be gone.

Risks and safety procedures


Hydrogen peroxide is corrosive; therefore we must avoid contact with eyes, skin and
clothing by wearing goggles and not letting it touch skin by wearing gloves if possible and
use great care to protect from other parts of the body and ingesting and drinking.

Results

Amount of Potato Volume of oxygen Volume of oxygen Volume of oxygen


discs produced (cm₃) 1 produced (cm₃) 2 produced (cm₃)
Average
0 0 0 0
2 0.25 0.2 0.225
4 1 0.8 0.9
6 1.5 1.4 1.45
8 2 1.8 1.9
10 4 3.4 3.7
Conclusion
The amount of enzyme increased directly caused the initial rate of reaction to increase you
can see this in the results as the amount of oxygen produced in 60 seconds increased. Just
as stated in the hypothesis this happens because the amount of enzymes allows more active
site available for the hydrogen peroxide to combine with. So as we added more potato discs
with the catalase enzyme there was more reaction taking place in the 60 seconds.

Alterations
In order to make the results more reliable we would carry out more readings with more
precise measurement by going up in 1 unit of slices instead of two. Also the thickness of the
discs couldn’t really be measure to that accuracy so would have been better if we had used
a potato slicer. Also the set up of the apparatus was very fiddly and the oxygen bubbles
were lost when we emerged it in the water. Also the water in the water bath started to cool
as the experiment ran on. However all the results turned out as we predicted with the same
trend we would expect.

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