CPB 30103: Biochemical Engineering

By Law Jeng Yih

Tutorial 2

Section A

1. _________ speed up chemical reactions.

a. Enzymes
b. Catalysts
c. Both A and B
d. Neither A nor B

2. The amount of energy needed for a specific chemical reaction to take

place is known as________.
a. reaction energy
b. chemical energy
c. energy of absorption
d. energy of activation

3. Inhibitor molecules that bind at a location other than the active site
are ________.
a. competitive inhibitors
b. noncompetitive inhibitors
c. active inhibitors
d. passive inhibitors

4. For the reversible enzyme-catalyzed reaction, A <===> B of the

following parameters is dependent upon the enzyme concentration?
a. the maximum velocity, Vmax
b. the Michaelis constant, Km
c. the equilibrium constant, Keq
d. Both A and B

5. In the derivation of the Michaelis-Menten equation, the steady-state

assumption refers to the condition that:
a. The concentration of any particular enzyme form (e.g., EX) remains
constant and steady over the time course of the experiment.
b. The rate of increase of product concentration with time is steady.
c. The reaction conditions (pH, ionic strength, temperature, etc.) are
steady over the time course of the experiment.

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 CPB 30103: Biochemical Engineering
By Law Jeng Yih

Section B

1. Define the following terms:

a) enzyme
b) enzyme kinetics

2. What is the turnover number of an enzyme?

3. Identify the class of enzymes that

a) split peptide bonds or glycosidic bonds with water
b) add water, ammonia or carbon dioxide across double bonds, or
remove these elements to produce double bonds

4. In the Lock-and-Key model of enzyme action, the part of the enzyme

that recognizes the substrate is known as the ________________.

5. If a compound is a ____________ inhibitor, the addition of more

substrate reverses the inhibition.

6. Match the type of reaction with the enzymes:

● aminase ●dehydrogenase
● Isomerase ● synthetase

a) _____________ converts a cis-fatty acid to trans.

b) _____________ removes 2 H atoms to form double bond
c) _____________ combine two molecules using ATP
d) _____________ adds NH3

7. The optimum temperature and pH for trypsin are 37°C and 8.2
respectively. Determine the effect of the following on its rate of
reaction.
a) Increasing the concentration of protein
b) Changing the pH to 4
c) Running the reaction at 70°C

8. If the activation energy for an uncatalyzed reaction is + 25 kJ/mol,

then what can you say about the activation energy for the enzyme
catalyzed reaction?

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 CPB 30103: Biochemical Engineering
By Law Jeng Yih

9. Determine Km and Vm from the graph below.

10. Determine the Km and Vm initial rates of an enzyme-catalyzed reaction

for substrate concentration in the table below.

So, mol/L v, mol/(L.min)

20.0 1.14

10.0 0.87

6.7 0.70

5.0 0.59

4.0 0.50

3.3 0.44

2.9 0.39

2.5 0.35

Using Eadie-Hofftee plot to determine Km and Vm.

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 CPB 30103: Biochemical Engineering
By Law Jeng Yih

11. Using the data shown below

a) Draw 1/vo vs 1/[S] plots. Determine the Km and Vmax from the
double reciprocal plot.
b) Decide what type of inhibitors "x" and "z" are
c) Calculate the Ki for "x" and "z"
Competitive Inhibitor : Km' = Km (1 + [I]/Ki)
Noncompetitive Inhibitor: Vmax' = Vmax / (1 + [I]/Ki)

12. What is non competetive inhibition? Draw the chemical scheme

(species, reaction steps and parameters) for the reversible mixed
inhibition of a Michaelis-Menten enzyme.

13. Triose phosphate isomerase catalyzes the conversion of

glyceraldeydes-3-phosphate to dihydroxyacetone phosphate. The Km
of this enzyme for glyceraldeydes-3-phosphate is 1.8x10-5M when
[glyceraldeydes-3-phosphate] is 30µM, the rate of the reaction, v was
82.5 µmole.mL-1sec-1.
a) Calculate Vmax for the enzyme
b) Assuming 3nmoles per mL of enzyme was used in the experiment
and [Etotal] = 3nmoles/mL, what is kcat for the enzyme?
c) What is the catalytic efficiency (kcat/Km) for triose phosphate
isomerase?

14. Identify each statement as describing an inhibitor that is

●Competitive ●Noncompetitive
a) Binds to enzyme, not active site
b) Structure is similar to substrate
c) Inhibition is not reversed with substrate

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 CPB 30103: Biochemical Engineering
By Law Jeng Yih

15. Linearize the Michaelis-Menten Equation for the following plots.

a) Lineweaver-Burk plot