CH 600 Lc

PROBLEM SET
ENZYMES

NAME:
YR. & SEC:

ANSWER THE FOLLOWING QUESTIONS. Use yellow pad for your answers.
1. Define the following terms:
A. Oxygenase
B. Epimerase C. Protease
D. Hydroxylase
E. Oxidase
2. Histidine is frequently used as a general acid or base in enzyme catalysis. Considering the pKa value of the side
chain, suggest a reason why is this so?
3. How is the cooperative behavior of allosteric enzymes reflected in a plot of reaction rate against substrate
concentration ?
4. What is the metabolic role of of aspartate transcarbamoylase?
5. What properties of metal make them useful co-factots?
6. You are a physician with a patient who is suffering from premature pancreatic zymogen activation, which means
that the pancreatic enzymes are being acrivated in the pancreas (rather than in the small intestine) and pancreatic
tissue is being damaged as result. What would be most effective: chymotrypsin inhibitor, a trypsin inhibitor, or an
elastase inhibitor? Why?
7. A bacterial enzyme catalyzes the hydrolysis of maltose as shown in the reaction given below:
Maltose + H2O 2 glucose
If the reaction has a Km of 0.135 mM and a V max of 65 mol/min. What is the reaction velocity when the
concentration of maltose is 1.0 mM?
8.

Use the plot provided to estimate Km and Vmax values of the enzyme catalyzed reactions.

9.

When[S] = 5 KM, how close is vo to Vmax? When [S] = 20 KM, how close is vo to Vmax? What do these results tell
you about the accuracy of estimating Vmax from a plot of vo versus [S]?
Substrate

KM (M)

N-Acetylvaline ethyl ether

N-Acetyltyrosine ethyl ether

8.8 X 10 -2
6.6 X 10-4

10. The KM values for the reaction of chymotrypsin with two different substrates are given:

A. Which substrate has the higher apparent affinity for the enzyme? Explain.
B. Which substrate is likely to give a higher value for Vmax?

11. The enzyme deoxyribokinase catalyzes the conversion of 2-deoxyribose to ADP. A study of this enzyme
produced the following data:
Exp #
1
2
3
4
5
6
7

[2-deoxyribose] rate of ADP formation

mmol/L
(mole/10min)
1.00
0.124
1.60
0.147
2.56
0.191
5.12
0.255
6.88
0.274
15.0
0.294
20.0
0.300

Use V versus [S] and a 1/V versus 1/[S] plots to graphically determine the K M and Vmax for this enzyme.

12. The following data were obtained on isocitrate lyase from an algal species. Deduce the K m and Vmax for the

enzyme, and determine the Ki for OAA and the nature of the inhibition by 0.5 mM oxaloacetate (OAA). See
solutions for info.
[S]m
rate Glyoxalate formation
(mole/10min)
0.0318
0.0420
0.0464
0.0583
0.0593
0.0700
0.1185
0.0955
0.2222 0.1167

Rate with OAA present

0.0040
0.0055
0.0075
0.0131
0.0233

13. The kinetics of an enzyme are measured as a function of substrate in the presence and the in absence of 2mM
inhibitor (I).
Velocity (mol/minute)
No inhibitor
Inhibitor
10.4
4.1
14.5
6.4
22.5
11.3
33.8
22.6
40.5
33.8

[S], (M)
3
5
10
30
90

A. What are the values of Vmax and KM in the absence of inhibitor? In its presence? In its presence?

B. What is the type of inhibition?

14. The following kinetic data were obtained for an enzyme in the absence of an inhibitor (1) and in the presence of
two different inhibitors (2) and (3) at 5mM concentration.
[S], mM
1
2
3
8
12

(mol/mL/sec)
(1)
12
20
29
35
40

(mol/mL/sec)
(2)
4.3
8
14
21
26

(mol/mL/sec)
(3)
5.5
9
13
16
18

A. Determine the Vmax and KM of the enzyme

B. Determine the type of inhibition.

11. Why is it surprising that some RNA molecules may have enzymatic activity?
12. Explain, on the basis of diffusion, why there is a limit to the maximum size of the turnover number of any
enzyme.
13. Prove that Km equals the substrate concentration at Vmax .
14. Why do structural analogs if the transition-state intermediate of an enzyme inhibit the enzyme competitively and

with low Ki values (binds tightly)?

15. Inhibitors of acetylcholinesterase, such as edrophonium, are used to treat Alzheimers disease. The substrate for
acetylcholinesterase is acertlcholine. Structures are given below:
H3C

O
O
Acetylcholine

CH3
+
N CH3
CH3

H3C
+
H3C N
H3C

OH

Edrophonium

A. What kind of inhibitor is edrophonium? Explain.

B. Can inhibition by edrophonium be overcome in vitro by increasing the substrate concentration? Explain.
C. Does this inhibitor bind reversibly or irreversibly to the enzyme? Explain.