7a. An enzyme simple contains 24 mg protein/mL, Of this simple, 20uL in a standa

Question

7a. An enzyme simple contains 24 mg protein/mL, Of this simple, 20uL in a standard incubation volume of 0.1mL catalyzed the incorporation of glucose into glycogen at a rate of

1.6 nmol/min. Calculate the velocity of the reaction in terms of the following:

a. Micromoles/minute

I got 1.6x10^-3 umol/min

b. Micromoles/liter/minute

16umol/L/min

c. Micromoles/milligrams of protein/minute

0.067umol/mg of protein/min (Correct?)

d. Units/milliliter

9.64x10^15 (Correct?)

e. Units/ milligrams of protein

4.07x10^17 (Correct?)

7b. Of the sample in Problem 7, 50mL were fractionated by ammonium sulfate precipitation. The fraction precipitating between 30 and 50% saturation was re-dissolved in a total volume of 10mL and dialyzed. The solution after dialysis had l2mL and contained 30mg protein/mL; Of the puri?ed fraction, 20ul catalyzed the reaction rate of 5.9 nmol/ min under the standard assay conditions. Calculate the following:

a. The recovery of enzyme after the ammonium sulfate step

b. ?le fold purification after the ammonium sulfate step

Explanation / Answer

a. 1.6 n mol /min = 1.6 *10^-3 micromol/ min

b. 1.6*10^-3 /0.1*10^-3/min = 16micromoles /litre/min

c. 1.6*10^-3/24*10^-3 / min = 0.06667 micromoles/miligrams of protein/min

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