calculate Kcat from Vmax
pp_fon
Dear all
i wonder how to calculate a kcat from Vmax cause i get the Vmax value by
use a Michalis-Menten plot and the Unit of Vmax value is
umol/min/mg(enzyme). and i use 0.0004 mg of enzyme per reaction and the MW
of enzyme is 64 kDa. How can i calculate the Vmax value
thank=^D
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Dr Engelbert Buxbaum
> i wonder how to calculate a kcat from Vmax cause i get the Vmax value by
> use a Michalis-Menten plot and the Unit of Vmax value is
> umol/min/mg(enzyme). and i use 0.0004 mg of enzyme per reaction and the
> MW of enzyme is 64 kDa. How can i calculate the Vmax value
Vmax you get from non-linear curve fitting of your v vs [S] data to the
Henri-Michaelis-Menten equation v = Vmax * [S] / (Km + [S]). Note that
using linearisation (Lineweaver-Burk or the like) causes a lot of
statistical problems, which in the days of supercomputer power on every
desk we no longer need to accept. The unit of the velocity is mol/s = kat.
Since Vmax / [E] = kcat you need to calculate the molar concentration of
enzyme (from molecular weight and the weight/volume concentration) and
divide your Vmax by it. The result will have the unit of a 1st order rate
constant, 1/s.
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