Calculate the degree of purification

A crude cell-free extract of a skeletal muscle contained 32 mg protein/ml. Ten ul (microlitres) of the extract catalyzed a reaction at rate of 0.14 umole/min under standard optimum assay conditions. Fifty ml of the extract were fractionated by ammonium sulphate precipitation. The supernatant obtained after centrifugation was redissolved in 10ml. This solution was found to contain 50 mg protein/ml. Ten ul (microlitres) of this purified fraction catalyzed the reaction at a rate of 0.65 umole/min.

(a) Calculate the percent recovery of the enzyme in the purified fraction. (Yield = units of enzyme in fraction / units of enzyme in original fraction).

(b) Calculate the degree of purification obtained by the ammonium sulphate fractionation. (Specific activity = total units of enzyme in fraction / total amount of protein in a fraction)(Fold Purification = specific activity of a fraction / specific activity of original fraction).

 

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