Question

Assuming that the experiments performed in the absence of inhibitors were conducted by adding 5 μl of a 2 mg/ml enzyme stock solution to an assay mixture with a 1 ml total volume, and taking into account that xyzase is a monomeric enzyme with a molecular mass of 45,000 daltons, what is the kcat of xyzase given in s-1 (give the answer in two significant figures)? to see a sample kcat calculation, click on hint in the lower left corner to open the lower panel.

Answer 1

Hey there!:

From the given data ;

Reaction volume = 1 mL , enzyme content = 10 ug ( 5 ug in 2 mg/mL )

Enzyme mol Wt = 45,000 , therefore [E]t is 10 ug/mL , this need to be express as "M" So:

[E]t in molar = g/L * mol/g

[E]t = 0.01 g/L * 1 / 45,000

[E]t = 2.22*10⁻⁷

Vmax = 0.758 umole/min/ per mL

= 758 mmole/L/min

=758000 mole/L/min => 758000 M

Therefore :

Kcat = Vmax/ [E]t

Kcat = 758000 / 2.2*10⁻⁷ M

Kcat = 3.41441 *10¹² / min

Kcat = 3.41441*10¹² / 60 per sec

Kcat = 5.7*10¹⁰ s⁻¹

Hence kcat of xyzase is 5.7*10¹⁰ s⁻¹

Hope that helps!