2. experiments measuring the initial You purify an enzyme and perform a series o

Question

2. experiments measuring the initial You purify an enzyme and perform a series of Michaelis-Menten kinetics enzyme velocity in the presence of various concentrations of substrate. a. You plot Vo vs. [S] and perform the non-linear obtaining the l Vass for your enzyme preparation? regression, obtaining the following curve. What are Ka and r09827 4.0774x 1.5091+x Ke and b. You also prepare a double reciprocal (Lineweaver-Burk) plot of the same data. Using it, w r-0.9963 y 0.4623x+0.1867 c. Which values are more reliable? Why?

Explanation / Answer

a. Michaelis–Menten kinetics is given by

V0 = Vmax [S]/([S] + KM)

Comparing with the equation given in the graph,

Vmax = 4.0774 and Km = 1.5091

b. In reciprocal form, Michaelis–Menten kinetics is given by

1/v0 = km /Vmax 1/[S] + 1/Vmax

Comparing the fitting equation with the above equation

1/Vmax = 0.1867 and Km/Vmax = 0.4623

=> Vmax = 5.3562 and Km = 0.4623 x Vmax = 0.4623 x 5.3562 = 2.4762

c. Certianly, the values obtaining using the Lineweaver Burk plot will provide the more reliable value. The reason is that the fitting of linear equation provides a better fitting equation than fitting the non-linear equation.

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