The following is lineweaver-Burke plot of an experimental result for determinati

Question

The following is lineweaver-Burke plot of an experimental result for determination of k_, k_m and V_max of an enzyme that catalyze the transformation of its substrate (S). The reaction rates were measured in unit of mu M/min and concentration of substrate is in mM, The equation shows the mathematical between1/v(Y) vs 1/(S) (x). Make sure the unit for each of the following parameter from your calculation is clearly shown. a. Calculate V_max b. Calculate K_m c. If the concentration of the enzyme in the enzymatic reaction mixture is 30 nM, Calculate k_ d. What would be the initial reaction rate if the concentration of substrate is 100 mu M

Explanation / Answer

a. the intercept (when x=0) gives 1/Vmax , So, 0.0001=1/Vmax gives Vmax=10000

b. The x-axis intercept (when y=0) gives -1/Km, so -0.20 = -1/Km, gives Km= 5

c. the picture is not clear, I assume the last word is kcat)Vmax/2 = Km, Vmax=10 , Vmax=kcat*[E], ==> Kcat=Vmax/[E] = 10/30

d. V0 = Vmax [S]/KM , 10000*100/5 = 200000

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