1. Start with the Michaelis-Menten equation and convert it to a double-reciprocal equation. Show how you would make a linear Lineweaver-Burk plot using this equation. 2. Based on th chart make a Lineweaver-Burk plot and later use the plot to complete the information in the table.
1. Start with the Michaelis-Menten equation and convert it to a double-reciprocal equation. Show how you would make a linear Lineweaver-Burk plot using this equation.
2. Based on th chart make a Lineweaver-Burk plot and later use the plot to complete the information in the table.
![[S],
V, mm/sec
(no inhibitor)
mM
.5
12
2.0
20
5.0
29
13.5 35
45.0
40
V, mm/sec
(+inhibitor)
5.5
9
13
16
18
Reaction
No inhibitor
+ inhibitor
Type of
inhibition
Best fit line
equation
Km or
Km (app)
Vmax or
Vmax (app)](/v2/_next/image?url=https%3A%2F%2Fcontent.bartleby.com%2Fqna-images%2Fquestion%2F4bc2db8d-b0f4-497e-9433-81c9ca5ab613%2Fdb4991a9-7e98-443b-863b-b5155d1f8a2b%2Fm38cu3h_processed.png&w=3840&q=75)
For a one-substrate enzyme-catalyzed reaction, the Michaelis-Menton equation shows the quantitative relationship between the initial velocity (Vo), maximum velocity(Vmax), the initial substrate concentration ([S]) and enzyme's affinity(Km).
Michaelis Menton equation --> vo = vmax · [S] / Km + [S]
Values for vo and substrate concentration can be obtained experimentally but values like Vmax and Km cannot be obtained experimentally.
To obtain these values, we plot the vo vs [S] on a graph, which is called a MM plot. From the graph, we can easily determine the value of Km.
Theoretically, when Km = [S], vo= vmax /2
But since the MM plot is a curve, the value of Km obtained from the graph is only an approximation.
To obtain a more accurate value of Km, we plot what is called a double reciprocal plot. This double reciprocal plot is also called the Lineweaver Burk Plot.
Lineweaver Burk Plot
MM equation can be transformed into an equation that is much easier to plot experimentally. We simply take the reciprocal on both sides.
vo = vmax · [S] / (Km + [S]) becomes 1/vo = ( Km / Vmax · 1 / [S] ) +1/vmax which looks very similar to y = mx +c (the straight line equation)
where y is 1/vo,
- x is 1/[S]
- m, the slope is Km/Vmax
- c, the y-intercept is 1/vmax
1/vo = ( Km / Vmax · [S] ) +1/vmax is called the Lineweaver Burk Equation
When we plot 1/Vo on the y-axis vs 1/[S] on the x-axis, the plot is called a Lineweaver Burk Plot.
Lineweaver Burk Plot enables us to accurately determine the Km and Vmax values.
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