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Enzyme-catalyzed reaction kinetics. The reaction above (sec question #2) is cata

ID: 959237 • Letter: E

Question

Enzyme-catalyzed reaction kinetics. The reaction above (sec question #2) is catalyzed by an enzyme called triose phosphate isomerase (TPI). This enzyme accelerates both the forward and reverse reactions, using either DHAP or GAP as a substrate. Both reactions follow Michaelis-Menten kinetics, with kinetic parameters K_M = 1.2 times 10^-3 M and k_cat = 6.5 times 10^4 min^-1. Assuming that TPI binds and releases its substrates rapidly (compared to the isomerization chemistry), which substrate binds to the enzyme with higher affinity? Under the following conditions: [TPI] = 1 times 10^-6 M, [DHAP] = 2 x 10^-3 M, and [GAP] = 0 M. What is the initial reaction velocity (V_o) for the forward reaction?

Explanation / Answer

A) As kcat=Turn over frequency=Vmax/[Eo]   Vmax=maximum reaction rate,[Eo]=initial enzyme concentration.For same initial enzyme concentration, both substrate has same Vmax.Also they have same km=michaelis –menten constant ,so rate of rxn for both substrate is the same.

Rate=v=Vmax[S]/Km+[S]

They bind with equal affinity.

(B)initial reaction velocity for forward=vo= Vmax[So]/Km+[So] [So]=initial substrate concentration

Kcat*[Eo]=Vmax

Vmax=6.5*10^-4 min-1 *1*10^-6 M=6.5*10^-10 Mmin-1

Vo=(6.5*10^-10 Mmin-1) (2*10^-3)/(1.2*10^-3M)+( (2*10^-3)

      =13*10^-13/3.2*10^-3 Mmin-1

       =4.0625 *10^-10 Mmin-1(Answer)