Question 1: The reaction for the cleavage of fructose 1,6-bisphosphate to dihydroxyacetone
phosphate and glyceraldehyde 3-phosphate is catalyzed by the enzyme fructose 1,6-bisphosphate
aldolase and has a standard free energy change of ΔG°′ = +23.8 kJ/mol.
A. What is the Keq of the reaction at 37 oC? (R = 8.314 J/°mol)?
1.
2.
3.
4.
5.
6.
7.
8.
9.
10.
B. If the intracellular concentration of fructose 1,6-bisphosphate at equilibrium (ΔG = 0) was 10
mM, what would be the predicted concentration of the products?
[ ]
1. [ ]
[ ]
2. [ ]
– multiply both sides by 10
3. [ ]
C. In cells, the reaction catalyzed by fructose 1,6-bisphosphate aldolase readily occurs to
produce dihydroxyacetone phosphate and glyceraldehyde 3-phosphate. Why do you think
this is the case especially since the aldolase reaction has a large positive ΔG°′ = 23.8
kJ/mol?
1. Although the ΔG°′ is positive this does not represent physiological conditions. Under
standard physiological conditions, the concentration of fructose 1,6-bisphosphate
aldolase is much higher than the products dihydroxyacetone phosphate and
glyceraldehyde 3-phosphate. This results in a ΔG that is close or below zero, which
allows the reaction to proceed.
Question 2: You have just discovered a new enzyme that catalyzes the degradation of cellulose to
glucose. You hope to commercialize this enzyme ultimately to invent a process for biofuel
manufacture. In order to determine how efficient the enzyme is and how fast the enzyme catalyzes
the reaction, you have to perform some basic enzyme kinetics. Plot the following enzyme data for
the enzyme catalyzed reaction and determine the Km and Vmax. (4 points)
Enzyme Data for the Enzyme Catalyzed Reaction
Substrate (cellulose)
Rate of conversion from cellulose to
Concentration (µ molar 1/S 1/V
glucose (m/min)
concentration)
0 0
6 98 0.1667 0.0102
15 230 0.0667 0.0043
24 390 0.0417 0.0026
50 600 0.0200 0.0017
95 680 0.0105 0.0015
180 700 0.0056 0.0014
Enzyme Data for the Enzyme Catalyzed Reaction
800
Rate of conversion from cellulose to glucose (
700
600
500
um/min)
400
300
200
100
0
0 20 40 60 80 100 120 140 160 180 200
Substrate (cellulose) Concentration ( umolar concentration)
Lineweaver-Burk plot of Enzyme Data for the
Catalyzed Reaction
0.0120
y = 0.0561x + 0.0007
0.0100
0.0080
1/V
0.0060
0.0040
0.0020
0.0000
0.0000 0.0200 0.0400 0.0600 0.0800 0.1000 0.1200 0.1400 0.1600 0.1800
1/S
Determination of Vmax
1. Y-Intercept = 1/Vmax
2. 0.0007 = 1/Vmax
3. Vmax = 1428.57
Determination of Km
1. Slope = Km/Vmax
2. 0.0561 = Km/1428.57
3. Km = 79.316
