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Enzyme Lab:

Procedure for Part 5:


1. Place a piece of liver about the size of a pea in a test tube.
2. Place 1 mL of hydrogen peroxide in the test tube with the liver.
3. Wait for the reaction to cease.
4. Pour the water that was produced from the original reaction in a different
test tube.
5. Add another 1 mL of fresh hydrogen peroxide into the first test tube that
contains the already reacted liver.
6. Add a fresh piece of pea-sized liver into the second test tube with the old
water.
7. Record which test tube reacts and how quickly each reaction occurs.
The Results and Observations For Each Trial Testing the Productivity of Enzymes
Reactants with H2O2
Glass Beads

Rate of Reaction
No Reaction

Manganese Dioxide MnO2

Fast reaction
Small bubbles
Fizzing

Potato

Slight fizziness
Slower reaction
Less bubbling
Foams very quickly
Large bubbles
Large air pockets

Liver

Create Your Own

Used Liver and Fresh


Hydrogen Peroxide:
Fast reaction, large
bubbles
H2O and O2 created

Ground Liver

Reacted Hydrogen
Peroxide and Fresh Liver:
No reaction
Slower reaction

Heated Liver

No reaction

Cooled Liver

Very fast reaction

Other Observations
The three glass beads do
not contain an enzyme
If more hydrogen
peroxide is added, more
bubbles produced and the
fizzing continues.
Tiny air pockets formed
Filled half of the test tube
The liquid formed was
water and oxygen
Fastest and largest
reaction for the entire
experiment

Smaller and fewer bubbles


created
The Liver looks green,
grey and is hard
Not as fast as regular liver

Liver in HCl

Small bubbles
No reaction

The liquid in the test tube


looked murky as if the
liver was degrading

Trial 1:
The three glass beads did not cause a reaction with hydrogen peroxide because
there was no enzyme. The substrate, hydrogen peroxide, did not have anything to
catalyze it because the glass beads do not contain an enzyme or a catalyst. Even
though hydrogen peroxide was in the test tube with the beads, a reaction would be
too slow to occur without an enzyme present.
Trial 2:
Manganese dioxide is a catalyst because it speeds up the rate of a reaction, lowers
activation energy, and does not get used up in a reaction. However, manganese
dioxide is not an enzyme because enzymes are only produced in living organisms.
Enzymes are proteins that are created in the body of living things. When more
hydrogen peroxide is added to the reaction after it occurred, more bubbles were
created because another reaction occurred. This happened because the substrate
was fully used up by the catalyst causing the first reaction to stop. After the first set
of reactants was catalyzed, the manganese dioxide stopped reacting because all of
the substrates had been changed into water and oxygen. This allowed the
manganese dioxide to be able to catalyze more substrates after the original reaction
was completed. Due to the fact that catalysts are never used up in a reaction, adding
more substrate causes the reaction to continue and the catalyst to keep functioning
and making more products.
Trial 3:
A potato does contain an enzyme because a small reaction did occur when hydrogen
peroxide was added with it in a test tube. This reaction occurred slowly because the
potatos enzyme is not as efficient as the enzyme in manganese dioxide or liver. This
could be due to competitive inhibitors that cause fewer substrates to be catalyzed at
a time. There also may have be fewer enzymes in a potato than in liver that would
cause it to take longer to catalyze the same amount of hydrogen peroxide. The
potato catalyzed the hydrogen peroxide and created oxygen and water.
Trial 4:
Liver does contain an enzyme because the products oxygen and water were
produced. The hydrogen peroxide was catalyzed by the enzyme in the liver, so at the
end of the reaction, the only substances left in the test tube were liver, water, and
oxygen. The hydrogen peroxide was completely converted to the products because
the enzyme in liver was able to speed up the reaction and decrease the activation
energy to start the reaction. The chemical equation for this reaction is:

2H2O2+ enzyme O2 +2H2O + enzyme


Based upon the above equation, the liver must contain the enzyme because all other
reactants do not appear at the end of the reaction. Due to the fact that liver appears
on both sides of the equation, it is the enzyme.
Trial 5:
The reaction between the liver and the hydrogen peroxide stopped because the
hydrogen peroxide changed. This is because the liver completely catalyzed all of the
substrate so the enzyme no longer needed to function. This is proven because
adding more hydrogen peroxide to the old liver from the trial 4 also caused a
reaction. Meaning that the liver did not change and that the enzyme is in the liver.
This is known because the enzyme was never used up even though the liver was
already used in another reaction. The liquid produced at the end of trial 4 was
combined with fresh liver but no reaction occurred. This is due to the fact that the
hydrogen peroxide was completely catalyzed and only water and oxygen remained.
Thus, there was no substrate for the enzyme to react with. This concludes that the
enzyme is in the liver and the hydrogen peroxide was the reactant that changed.
Trial 6:
This reaction occurred slower and produced less than the intact liver. The reaction
should have been the same speed as trial 5 because a physical change does not
denature an enzyme or stop it from functioning. Additionally, there were no
noncompetitive or competitive inhibitors in this reaction that would have caused
the productivity of the enzyme to change. The reason this may have occurred was
because there was less liver than in the previous trial, therefore, there would have
been fewer enzymes to catalyze the same amount of substrates. Fewer enzymes
would cause the reaction to occur more slowly than in Trial 5, even though there
was no real change to the enzyme when the liver was ground up.
Trial 7:
Heat is one factor that denatures proteins. The enzyme in liver did not function
because it was denatured. Denaturation occurs when high heat causes the protein to
unfold and bonds between polypeptide chains to break. Hydrogen bonds can be
broken in denaturation so the enzyme uncoils. However, heat cannot denature
peptide bonds so the amino acid sequence, the primary structure, remains in tact.
Additionally, a denatured enzyme can no longer function.
Trail 8:
While high heat denatures a protein, the cold did not completely stop the reaction.
The liver was not cold enough to fully denature the enzyme. While the cooler
temperature slowed the reaction, it did not completely stop the function of the
enzyme. Not all of the bonds were broken so the protein still functioned, however,

the speed of the reaction was decreased and there were fewer products. Cold
temperature does denature proteins if it is low enough to truly affect the bonds.
Trial 9:
There was no reaction because the acid caused the protein to denature. Liver had a
pH of seven and the hydrochloric acid had a pH of two. The highly acidic liquid
denatured the enzyme causing it not to function. None of the substrates were
catalyzed thus causing no new products to be formed. No bubbles appeared in the
test tube so no oxygen was created. Again, the tertiary, quaternary, and secondary
structures were all affected and unfolded. This was because the hydrogen bonds
were disturbed by the addition of H+ ions from the acid. These ions caused the
bonds to break and thus the protein unfolded. The primary structure was still intact
because the peptide bonds were not affected by increased acidity.