Enzyme Kinetics - PPT, IBT, Biotechnology, Engineering, Semester Notes | EduRev

: Enzyme Kinetics - PPT, IBT, Biotechnology, Engineering, Semester Notes | EduRev

 Page 1


Chapter 5 (part 3)
Enzyme Kinetics (cont.)
Page 2


Chapter 5 (part 3)
Enzyme Kinetics (cont.)
Michaelis-Menton
E + S ES E + P
•V
max
•K
m
•K
cat
•K
cat
/K
m
k
1
k
-1
k
cat
•Vo = Vmax [S]
Km + [S]
•V
max
•K
m
•k
cat
•k
cat
/K
m
Page 3


Chapter 5 (part 3)
Enzyme Kinetics (cont.)
Michaelis-Menton
E + S ES E + P
•V
max
•K
m
•K
cat
•K
cat
/K
m
k
1
k
-1
k
cat
•Vo = Vmax [S]
Km + [S]
•V
max
•K
m
•k
cat
•k
cat
/K
m
How do you get values for V
max
, K
m
and k
cat
?
• Can determine K
m
and V
max
experimentally
• Km can be determined without an absolutely 
pure enzyme
• K
cat
values can be determined if V
max
is known 
and the absolute concentration of enzyme is 
known (V
max
= k
cat
[E
total
]
Page 4


Chapter 5 (part 3)
Enzyme Kinetics (cont.)
Michaelis-Menton
E + S ES E + P
•V
max
•K
m
•K
cat
•K
cat
/K
m
k
1
k
-1
k
cat
•Vo = Vmax [S]
Km + [S]
•V
max
•K
m
•k
cat
•k
cat
/K
m
How do you get values for V
max
, K
m
and k
cat
?
• Can determine K
m
and V
max
experimentally
• Km can be determined without an absolutely 
pure enzyme
• K
cat
values can be determined if V
max
is known 
and the absolute concentration of enzyme is 
known (V
max
= k
cat
[E
total
]
Lineweaver-Burke Plots
(double reciprocal plots)
•Plot 1/[S] vs 1/V
o
•L-B equation for straight 
line
•X-intercept = -1/Km
•Y-intercept = 1/Vmax
•Easier to extrapolate 
values w/ straight line vs
hyperbolic curve
Page 5


Chapter 5 (part 3)
Enzyme Kinetics (cont.)
Michaelis-Menton
E + S ES E + P
•V
max
•K
m
•K
cat
•K
cat
/K
m
k
1
k
-1
k
cat
•Vo = Vmax [S]
Km + [S]
•V
max
•K
m
•k
cat
•k
cat
/K
m
How do you get values for V
max
, K
m
and k
cat
?
• Can determine K
m
and V
max
experimentally
• Km can be determined without an absolutely 
pure enzyme
• K
cat
values can be determined if V
max
is known 
and the absolute concentration of enzyme is 
known (V
max
= k
cat
[E
total
]
Lineweaver-Burke Plots
(double reciprocal plots)
•Plot 1/[S] vs 1/V
o
•L-B equation for straight 
line
•X-intercept = -1/Km
•Y-intercept = 1/Vmax
•Easier to extrapolate 
values w/ straight line vs
hyperbolic curve
B
B
B
B
B
B
B
0
0.05
0.1
0.15
0.2
0.25
012345 678910
Vo
[S]
[S] Vo
0.5 0.075
0.75 0.09
20.152
40.196
60.21
80.214
10 0.23

V max
Km
Km ~ 1.3 mM
Vmax ~ 0.25 
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