Title: Enzyme Catalysis

Purpose: The purpose of this activity is to determine the rate for the enzyme catalyzed
decomposition of hydrogen peroxide.

Procedures: Described in the lab manual

Materials: Listed in the lab manual

Data:
Titration Syringe

Initial Reading 10.0 mL

Final Reading 2.5 mL

Baseline 7.5 mL

Time (seconds)
10 30 60 120 180
Baseline Volume 7.5 mL 7.5 mL 7.5 mL 7.5 mL 7.5 mL
Initial Volume 10 mL 10 mL 10 mL 10 mL 10 mL
Final Volume 3.1 mL 3.3 mL 3.5 mL 4.7 mL 5.4 mL
Amount of KmnO4
Used (Initial – Final) 6.9 mL 6.7 mL 6.5 mL 5.3 mL 4.6 mL

Amount of H2O2 used

(baseline- KmnO4 0.6 mL 0.8 mL 1.0 mL 2.2 mL 2.9 mL
used)

Time Interval (seconds)

Initial 0 to 10 to 30 30 to 60 60 to 120 120 to 180

10
Rates 3/50 1/100 1/150 1/50 7/600

4. Graph of hydrogen peroxide used

5. Graph –rates
Conclusion:

1. What is the purpose of this activity?

The purpose of this activity is to determine the rate for the enzyme catalyzed
decomposition of hydrogen peroxide.
2. What is the i.v. in this activity? d.v.? some (four) constants?
The independent variable in this activity was water, oxygen, and energy.
3. What is an enzyme? Substrate? Induced fit?
An enzyme is a protein that catalyzes a chemical reaction. A substrate is a
molecule upon which an enzyme acts. Induced fit is the binding of a substrate or
some other molecule to an enzyme which causes a change in the shape of the
enzyme.
4. In this activity, what is the enzyme? The substrate? The product?
The enzyme was the KMnO4. The substrate was H2O2. The product was
5. How could you show that oxygen gas evolved (formed) in the reaction?
When you dropped some KMnO4 in the product there were bubbles.
6. What are some factors that affect the rate of enzyme catalyzed reactions?
( p. 19-20). Describe each one in detail.
Salt concentration, if it is close to 0 the charged amino acid side chains of the
enzyme molecules will attract each other. Enzymes will denature and form an
inactive precipitate. pH logarithmic scale that measure the acidity or H+
concentration in a solution. Temperature: generally, chemical reactions speed up
 as the temperature increases, more of the reacting molecules have enough kinetic
energy to undergo the reaction. Activation and inhibitors: many molecules other
than the substrate may interact with an enzyme. If such a molecule increases the
rate of a reaction it is an activator. If such a molecule decreases the rate of a
reaction it is a deactivator.
7. What is the function of potassium permanganate in this activity? What is titration?
The function of potassium permanganate in this activity was to titrate the solution.
Titration is a common laboratory method of quantitative chemical analysis that is
used to determine the unknown concentration of a known reactant.
8. What reaction would you expect if you added boiled catalase to hydrogen
peroxide? Explain your answer.
I would not expect anything to happen because the enzymes in the boiled catalase
are already denatured. So nothing can really occur.
9. If you boiled a piece of potato or liver, and added hydrogen peroxide, what
reaction would you expect? Explain why.
I would expect no reaction because
boiling denatures the enzymes and when they are denatured no reaction can
occur.
10. In this activity, when is the rate the highest? Explain why.
At the beginning because the concentration of the substrate is at it’s highest
because the catalase is fresh and accepts many substrates.
11. When is the rate slowest? For what reason is the rate slow?
Near the end because less substrates are available and the enzyme becomes
saturated with substrates.
12. Explain the inhibiting effect of sulfuric acid on the function of catalase. Relate
this to enzyme structure and chemistry.
The inhibiting effect of sulfuric acid (H₂SO₄) is mainly because it lowers
the pH of the enzyme. Sulfuric acid is a very strong acid, so it
denatures the protein. If the pH is lowered, the side chains will begin
picking up hydrogen ions from its environment, which might disrupt its
conformation, ultimately inhibiting its function. The secondary and
tertiary structures of the enzyme become altered, thus changing the
behavior of the enzyme. In other words, the substrate may not fit or
the side chains will not interact correctly because of the hydrogen
added by the sulfuric acid.

13. Predict the effect lowering the temperature would have on the rate of enzyme
activity. Explain your prediction.
If you lower the temperature it will hurt the rate of the enzyme activity because
the colder it gets the more likely it is for the enzyme to denature and not function
well.
14. Different enzymes work better under different conditions. Where in a human body
might it be beneficial to have enzymes that work well in acidic environments?
In the stomach.
15. There is a large amount of catalase in the human liver. Does the liver break down
more hydrogen peroxide in the summer or winter? Explain your answer.
 It doesn’t break down more in summer nor winter because the human body is
always in homeostasis so the temperature outside will not affect the temperature
inside and it will not affect the enzymes.
16. Of the thousands of enzymes known, there is a family of enzymes called proteases
that catalyze a reaction breaking down proteins. What do you think would happen
if you added a protease to your sample of catalase before proceeding with your
experiment?
A reaction would occur between protease. The protease acts upon the protein and
catalase which is a protein. The reaction most likely caused the catalase to
denature. And that probably is what caused it to not break down the H2O2.
17. Summary- at least 15 lines typed, 20 handwritten.
Enzymes are biological catalysts that speed up reactions necessary to live. Each enzymes
specificity is determined by the shape of its active sight and clearly represents the
induced fit how a substrate perfectly binds to the active sight only with a slight
change of the enzyme. Substrates or molecules that require a speedier reaction are
introduced to the active sight of the enzyme where after a certain amount of time
the products are released and the enzyme may be used over and over again.
This lab was helpful in showing us how enzyme catalysis really happens in
biology. I believe we went wrong in establishing our baseline; we did not mess up at all,
Although we almost did once, but we managed to make sure we didn’t drop more than
one drop of KMnO4. My hypothesis was correct as the reaction did begin quickly and
slow as the time moved on. Either way, the lab represented success in the fact that it
taught me how enzymes work.