The use of organic solvents at low temperature for the separation of enzymes. Application to aqueous rabbit muscle extract

Abstract

A fractionation of the water-soluble proteins of rabbit muscle was carried out using organic solvents at low temps. and low ionic strengths. Enzyme estimations were used to follow the separation of the proteins. The following conditions were best for sharp separation with the least denaturation: precipitation with acetone at pH 6.5, at very low ionic strengths (around 0.03) and low temps. (0[degree] and below). The results obtained were easily reproducible and a 4- to 5-fold purification in one stage was achieved on the avg. The procedure compared favorably with other methods of enzyme purification. Most of the glycolysis enzymes separated out free from adenylic deaminase and adenosinetriphosphatase. Creatinephosphokinase was obtained free from myokinase.