Enzyme Lab Report by Jan Eric De Castro

Purpose: To investigate how changing the external conditions (cold or hot) affect enzymes in
chemical reactions with substrates.
Materials:
- Potato (cubed cut)
- 3% Hydrogen Peroxide
- Thermometer
- 6 test tubes
- Stirring Rod
- Water (H20)
- Hot Plate
- 1 beaker filled with ice cold water
- 1 beaker filled with warm water
Hypothesis: By changing the external external environment of the enzymes, it will change by
either slowing down or speeding up the rate of reaction with a substrate.
Procedures:
Part A: Observe Normal Catalase Reaction
1. Place 2 ml of the 3% hydrogen peroxide solution into a clean test tube.
2. Place cubed-cut potatoes into the test tube. Push it into the hydrogen peroxide with a stirring
rod. Observe the bubbles;
Throughout this investigation we will estimate the rate of the reaction (how rapidly the solution bubbles)
on a scale of 0-5 (0 meaning no reaction, 1 meaning slow, and to 5 being very fast). Assume that the
reaction in step 2 proceeded at a rate of "4"

Answer these following question:

Is Catalase Reusable?
1. Pour off the liquid from your test tube into the sink.
2. Add another 2 ml of hydrogen peroxide to the liver remaining in the first test tube. Make predictions
what will happen. Is there a reaction?

Part B: What is the Effect of Temperature and pH on Catalase Activity?

1. Put one piece of a potato into the bottom of a clean test tube and fill it with a small amount of
distilled water. Place this test tube in a boiling water bath using a hot plate for 5 minutes.
2. Remove after the 5 minutes are up). Add 2 ml of hydrogen peroxide. CAUTION: Use a testtube holder when handling the hot test tubes. Record the reaction rate from slow to fast (0-5) in
the data.
3. Put equal size and quantity of potatoes into 2 clean test tubes and 1 ml H2O2 into 2 other test
tubes. Put one test tube of potato and one of H2O2 into each of the following water baths: Ice bath
and Warm water bath (not boiling, luke).
4. After 3 minutes, pour each tube of H2O2 into the corresponding tube of potato (cold to cold,
warm to warm) and observe the reaction. Record the reaction rates (0-5) in your data.

Data:
Data Table
Rate of
Reaction
(1-5)

Observations & Conclusions

Part A

Potatoes to Peroxide

When we mixed the potato cube with

the peroxide, it immediately started the
reaction by bubbling and releasing
oxygen.
From touching the test tube while the
reaction started, it became colder, making
it an endothermic reaction.

Part A

Potato added to Used

Peroxide

Since not all of the substrates from the

peroxide has been used up, the fresh
catalase from the potato instantly started
reaction, but slowly diminished because of
the little amount of substrates left.

Part A

Reused Catalase

This reaction was even slower because

of the overflow of substrates with the little
amount of enzymes in the potato. The
enzymes cannot react instantly because
there is a back up, so it became slower, but
still happened over time.

From the little test, it shows that

enzymes can be fast and reusable, making
it a very good catalyst.
Part B

Boiled Potato

When we heat the potato with an

optimum temperature of 81 degrees inside
the test tube for 5 minutes, it denatures the
enzymes.
When we poured the hydrogen peroxide
inside the test tube, it had no reaction, or
was so slow that we didnt have a chance
to see it. So the reaction rate is 0.
From this test, we have learned that
enzymes get denatured, or not working in
unfavorable temperatures/environment.

Part B

Ice Bath Potato

We had to place 1 ml of H2O2 and a

potato in an ice bath for about 3-5
minutes. When we mixed the substances
together, it reacted slowly.
As it proceeds slowly at 0 degrees, it
slows down the reaction of enzymes
because it either takes longer for the
reaction to complete or the amount of
energy has been lowered because of the
substances getting slower.
It was almost to a point to becoming
denatured if it stayed within the ice bath
longer. Enzymes cannot work in bad
homeostatic environment.

Part B

Warm Potato

Like the ice bath, this reaction was

very slow. The reaction started very
slowly and had hardly any activity during
the time when we were testing this part.
This occurred because it was in the
brink of getting denatured. It still had a
reaction because it was not hot enough to
fully denature the enzymes, like what
happened in Part A.

Enzymes do not work well in bad

homeostatic environments.
Pictures:
Part A (1): Results (Potato with Peroxide)
As you can see in this picture, when we mixed the potato and the peroxide, it started
immediately and started to produce huge amounts of gas (oxygen, the white bubbles). This was
able to react because of the favored homeostatic environment of the test tube (room temperature,
did not alter the temp to hot or cold).

Part A (2): Results (Reused Catalase)

From this picture, you can tell that the reaction was not as significant as the previous. We
poured out the previous peroxide, and kept the potato with a new batch of H2O2. It started out in
a medium-speed, then quickly slowed down. This was due to the low amounts of substrates left
in the reaction, but this showed that enzymes are fast and reusable.

Part B (1): Results (Boiling Water)

The boiling water and the unfavorable homeostatic environment caused the enzymes
from the potato to be denatured. So the reaction did not happen at all, or was so slow that we
could not observe it from the time we tested it. This shows that enzymes cannot work on high
temperatures.

Part B (2): Results (Ice Bath)

The temperature was approaching 0 degrees, but not fully. It was cold enough to slow
down the reaction rate to 2, making the substances not as fast. That is why the reaction produced
little bubbles (gas). Enzymes cannot work well in unfavorable homeostatic environments.

Part B (3): Results (Warm Bath)

The results were similar to the cold ice bath. The warm temperature of the test tube
caused the enzyme to slow down (almost denatured). Enzymes cannot work in bad homeostatic
environment.

Conclusion:
In our data, we found out that by not affecting the temperature of the enzymes
environment, it was able to react efficiently with a favorable homeostatic environment. While the
ones that was cooled or heated was very slow or had no reaction at all because it was denatured.
Denatured is an enzyme that is not working correctly. Anything that does not have a homeostatic
environment will denature the enzyme. In other words, pH, salt concentration, temperature, or
other aspect of its environment is altered to the point where the proteins shape would unravel
and lose its native shape. By losing its shape, it can no longer function correctly since it cannot
fit substrates to start the process.
In this experiment, temperature was an extremely huge factor in determining if the
reaction was going to be fast or slow. In normal temperature, it had a very favorable
environment, without any factors affecting the rate of reaction. Though in the ice bath, in much
lower temperature, it slowed down the reaction. This was due to slowing down the enzymes and
the amount of collisions, resulting to a decrease of reaction. Heat was also a source of energy for
reactions to start. The warm potato was also a similar case to the ice-bath. It was almost in the
point of being denatured, but was not left in the hot water long enough for that process to
happen. So the factor of time was crucial to determine if there is no reaction or little amounts of
it.
Each enzyme has a specific substrate that they will break up in a reaction. This is called
the one enzyme/ one substrate. Usually a word that ends with -ase is an enzyme. They break up
substrates in a place called active site. Substrates will lock and key to the active site, then will
start the breaking up process (hydrolysis). This is necessary for the body to go under metabolism,
which is building up what the body wants/needs from the product of the chemical reaction in
enzymes and substrates. Sometimes, substrates will not fit in the active site, so coenzymes and
cofactors will change to a shape so substrates will be able to use the lock and key method.
Furthermore, there is also a place in an enzyme called the allosteric site. This happens when your
body has too much of something, and it needs to either stop or slow down. Inhibitors will stick
into the allosteric site and slow down the chemical reaction in the active site. There are two types
of inhibitors, competitive and noncompetitive inhibitors. Competitive inhibitors compete with

the substrates by locking in the active site to prevent reaction. While the noncompetitive inhibitor
will lock in the allosteric site to slow down reactions.
There are two types of metabolic pathways, catabolic and anabolic. Catabolic is to break
up, and is when ATP is produced for the body, while anabolic process is when it builds up and
also when ATP is used up to combine the molecules together. These two things is necessary to
obtain homeostasis.

Above shows two graphs that correlates to chemical reactions with enzymes. The first
one in the left is called the Michaelis-Menten Kinetics Graph. It shows the rate of reaction when
enzymes and substrates combine. The rate of reaction will first spike up as substrates connect to
all of the enzymes, but will slow down over time because of the back up from the number of
substrates waiting to fit in the active site. This graph proves that as you increase concentration,
the enzymes cannot process the huge amount of substrates, and it will get less steeper and more
constant. The graph in the right shows how catalyst (enzymes) affect a chemical reaction. The
blue line rises because of the amount of activation energy needed to start the reaction, but with a
catalyst, the activation energy is cut in half. Having a catalyst will make it quicker to obtain the
products of the reaction.
Finally, is the case study with the jellos with the fruit juice and the fresh fruit. The reason
why the one with fruit juice hardened while the one with fresh fruits did not, is because of the
enzymes. Fresh fruits contain a huge amount of enzymes that will break up proteins, which was
the main ingredient of the jello. Jello is made out of animal fats and other parts, making it filled
with proteins. So as you waited for the jello to harden, the enzymes were breaking up the
substrates within the jello, making it soggy and watery. The fruit juice was pasteurized, meaning
it was heated to remove the bacteria inside the fruit. But the excessive heat also denatured the
enzymes, making it useless in the mix with the jello. Putting food in lower temperatures will help
prevent food spoilage, or slower the process. The freezing temperatures slows down or
completely halts the reaction happening with enzymes so you can have food for years without it
breaking down or ever spoiling.