The purification of yeast enolase by zone electrophoresis and ion-exchange chromatography, and the existence of several active forms of the enzyme

Abstract

1. 1. A new method for the purification of yeast enolase has been described. The procedure involves zone electrophoresis in cellulose columns, and specific adsorption of the enzyme on the Mg form of a cation exchanger, sulfomethylcellulose. In a typical preparation, 1.7 g. of pure enolase was obtained from 3 kg. of dried yeast. 2. 2. The purified enzyme shows a high degree of homogeneity in zone electrophoresis and in the ultracentrifuge. Its specific activity is the same as that of the enzyme prepared by the method of Warburg and Christian (1). 3. 3. It has been shown that the crystallized enzyme can contain several electrophoretic components, all having the same specific activity and sedimentation constant. 4. 4. Some advantages of the new purification procedure and the possible origin of the different forms of the enzyme have been discussed.