3.An enzyme catalyzed reaction is studied and the following kinetic analysis is obtained: (S), mM 0.050 0.075 0.125 0.175 | 0.935 v, (µM min') 0.93 1.264 1.77 2.14 3.7 a. Using Excel, make a fully labeled Lineweaver-Burk plot and determine the Km and Vmax for the enzyme b. The reactions were set up dissolving 1 mg of the enzyme (MW- 10000 Da) in 100 ml of final reaction buffer. Determine the turnover number for the enzyme assuming 1 active site exists per enzyme molecule? c. Determine the catalytic efficiency for the enzyme

Biochemistry
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ISBN:9781319114671
Author:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.
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Chapter1: Biochemistry: An Evolving Science
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3.An enzyme catalyzed reaction is studied and the following kinetic analysis is obtained:
|[S), mM
0.050
0.075
v, (µM min")
0.93
1.264
1.77
2.14
3.7
0.125
0.175
0.935
a. Using Excel, make a fully labeled Lineweaver-Burk plot and determine the Km and
Vmax for the enzyme
b. The reactions were set up dissolving 1 mg of the enzyme (MW= 100000 Da) in 100 ml of
final reaction buffer. Determine the turnover number for the enzyme assuming 1 active site
exists per enzyme molecule?
c. Determine the catalytic efficiency for the enzyme
d. The same reactions are performed in presence of an inhibitor A and the resulting
velocities determined:
v plus inhibitor, (µM
min)
0.272
0.37
0.518
0.626
|1.08
|[S), mM
0.050
0.075
0.125
0.175
0.935
Plot these data on the same graph as above and determine the new Km and Vmax and the type
of inhibitor (competitive, non-competitive).
e. Can the effects of the inhibitor be over-ridden by adding more substrate? Why?
Transcribed Image Text:3.An enzyme catalyzed reaction is studied and the following kinetic analysis is obtained: |[S), mM 0.050 0.075 v, (µM min") 0.93 1.264 1.77 2.14 3.7 0.125 0.175 0.935 a. Using Excel, make a fully labeled Lineweaver-Burk plot and determine the Km and Vmax for the enzyme b. The reactions were set up dissolving 1 mg of the enzyme (MW= 100000 Da) in 100 ml of final reaction buffer. Determine the turnover number for the enzyme assuming 1 active site exists per enzyme molecule? c. Determine the catalytic efficiency for the enzyme d. The same reactions are performed in presence of an inhibitor A and the resulting velocities determined: v plus inhibitor, (µM min) 0.272 0.37 0.518 0.626 |1.08 |[S), mM 0.050 0.075 0.125 0.175 0.935 Plot these data on the same graph as above and determine the new Km and Vmax and the type of inhibitor (competitive, non-competitive). e. Can the effects of the inhibitor be over-ridden by adding more substrate? Why?
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