Question

You do an enzyme kinetic experiment and calculate a Vmax of 100 µmol of product per minute. If 0.1 mL of an enzyme solution was used, that had a concentration of 0.2 mg/mL, what is the best approximation of the turnover number if the enzyme had a molecular weight of 128,000 g/mol?

Answer 1

Turnover number, kcat = 1.666 x 10 -6 moles/sec / 1.5625 x 10 -10 moles. = 10679 sec - 1 , which is approximately 10624 sec -1

Step-by-step explanation:

Vmax = 100 \upsilonmol/min.

(We need to convert Vmax into moles/min.)

Vmax = 100 \upsilonmol/min x 1 mol/ 1000000 \upsilonmol.

Vmax = 1 x 10 ⁻⁴ moles/min.

(We need to convert Vmax to moles/sec).Vmax = 1 x 10 ⁻⁴ moles/min x 1 min/60 sec.

Vmax = 1.666 x 10 ⁻⁶ moles/sec.

​[E₀] = 0.1 ml x 0.2 mg/ml x 1 g/1000mg x mol/128000g {unit conversion}

= 1.5625 x 10⁻¹⁰ moles.

Vmax = kcat x [E₀]

kcat = Vmax / [E₀].

kcat = 1.666 x 10 ⁻⁶ moles/sec / 1.5625 x 10 ⁻¹⁰ moles. = 10679 sec - 1 ~ 10624 sec -1we know, Vmax = 100 \upsilonmol/min.

(converting into moles/min.) Vmax = 100 \upsilonmol/min x 1 mol/ 1000000 \upsilonmol.

Vmax = 1 x 10 ⁻⁴ moles/min.

(now, converting to moles/sec).Vmax = 1 x 10 ⁻⁴ moles/min x 1 min/60 sec.

Vmax = 1.666 x 10 ⁻⁶ moles/sec.

​[E0] = 0.1 ml x 0.2 mg/ml x 1 g/1000mg x mol/128000g {unit conversion}

= 1.5625 x 10⁻¹⁰ moles.

Vmax = kcat x [E₀]

kcat = Vmax / [E₀].

Turnover number, kcat = 1.666 x 10 ⁻⁶moles/sec / 1.5625 x 10 ⁻¹⁰moles. = 10679 sec - 1 , which is approximately 10624 sec -1