For an enzyme that follows Michealis-Menten kinetics, the initial velocity at different substrate concentration has been determined.
|
[S] (mM)
|
0.1
|
0.3
|
1.0
|
3.0
|
10.0
|
30.0
|
|
v0 (mM/min)
|
0.21
|
0.50
|
0.94
|
1.25
|
1.42
|
1.47
|
a. Determine KM and VMAX
b. The experiments are replicated with addition of a reversible inhibitor (the new results are shown in below table). Determine the effect of the inhibitor on KM and VMAX and the type of inhibition.
|
[S] (mM)
|
0.1
|
0.3
|
1.0
|
3.0
|
10.0
|
30.0
|
|
v0 (mM/min)
|
0.12
|
0.30
|
0.68
|
1.07
|
1.34
|
1.44
|
c. Calculate the enzyme concentration in the reaction mix, when the turnover number of the enzyme, kcat is 250 s-1.