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Bchm2000 PX
Bchm2000 PX
(1)
Sample Questions x
Kinetic graphs
[Substrate], mM
0.02
0.04
0.07
0.1
0.15
0.2
0.3
0.5
0.7
You are trying to reproduce experimental data from the previous student in
the lab. He/she had reported that the enzyme under investigation has a
kcat of 1875 s-1 and a catalytic efficiency of 7.5 107 M-1 s-1.
(a) What is the KM of this enzyme?
(b) In repeating the measurement, you want to have a vmax of 10 mM s-1
(as this is easy to measure). How much enzyme should you use?
Provide your result in M.
(c) Calculate at which substrate concentrations in M you should measure
the initial rate v0 to obtain the following values:
v0, mM s-1
[Substrate], M
2.50
4.50
6.00
9.00
(d) Create a predicted Michaelis-Menten plot for your enzyme.
(e) Draw a predicted Lineweaver-Burke plot for your enzyme.
Biochemistry 2000
Sample Questions x
Kinetic graphs
Answers:
(1)
(a)
70
vmax
v0, mM/s
60
50
40
30
20
10
0
0.0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9
KM
[Substrate], mM
(b)
0.10
-1
1/v0, mM s
0.08
0.06
0.04
1/vmax
0.02
-1/KM
-20
-10
10
20
30
40
50
60
1/[Substrate], mM-1
(c)
KM and vmax can be most precisely determined from the intercepts in the
Lineweaver-Burke plot:
1/vmax = 0.015385 mM-1 s vmax = 65 mM s-1
-1/KM = -10 mM-1
KM = 0.1 mM
(d)
Biochemistry 2000
Sample Questions x
Kinetic graphs
vmax
25
KM
50
[Substrate], M
(e)
1/[Substrate],
M-1
0.1200
0.0489
0.0267
0.0044
1/v0, mM-1 s
0.4000
0.2222
0.1667
0.1111
0.45
0.40
0.35
0.30
-1
11
10
9
8
7
6
5
4
3
2
1
0
0
1/v0, mM s
v0, mM/s
0.25
0.20
0.15
1/vmax
0.10
0.05
-1/KM
-0.04 -0.02
0.00
0.02
0.04
0.06
0.08
-1
1/[Substrate], M
0.10
0.12
0.14