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Although graphical methods are employed for accurate determination of the V max

ID: 843314 • Letter: A

Question

Although graphical methods are employed for accurate determination of the Vmax and Km of an enzyme-catalyzed reaction, sometimes these quantities can be quickly estimated by inspecting values of the initial rates, v, at increasing [S]. Estimate the Vmax and Km values of the enzyme-catalyzed reaction for the following data shown in the columns below:

                                            

[S] (M)

v0, (moles/min/)

2.5 x 10-6

28

4.0 x 10-6

40

1 x10-5

70

2 x 10-5

95

4 x 10-5

112

1 x 10 -4

128

2 x 10-3

139

1 x 10-2

140

  

[S] (M)

     

v0, (moles/min/)

     

2.5 x 10-6

     

28

     

4.0 x 10-6

     

40

     

1 x10-5

     

70

     

2 x 10-5

     

95

     

4 x 10-5

     

112

     

1 x 10 -4

     

128

     

2 x 10-3

     

139

     

1 x 10-2

     

140

  

Explanation / Answer

Notice how little the velocity changes as the substrate concentration increases by fivefold from 2 to 10 ?M. Thus, we can estimate a Vmax of 140 ?M/min. Km is defined as the substrate concentration that produces a velocity of (1/2 Vmax), or 70 ?M/min. Inspection of the table indicates that this V0 occurs at [S] 1*10^-5 M, thus Km ? 1 *10^-5 M

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