AFFINITY-CHROMATOGRAPHY OF PROTEINASES USING BACITRACIN IMMOBILIZED TO POROUS-GLASS BEADS

This study describes an affinity chromatography procedure for proteina se purification using bioselective binding to immobilized bacitracin. By coupling bacitracin to controlled-pore glass (CPG) beads, an affini ty matrix was obtained that permitted rapid purification of proteinase s under conditions that minimize autolysis. Bacitracin-CPG was used to bioselectively adsorb the extracellular proteinase secreted by Entero coccus faecalis var. liquefaciens IFPL 383, The overall purification o btained with this procedure was 5149-fold. The ability of bacitracin-C PG to bind other proteinases was examined using various commercial pro teinases, The specific activities of subtilin BPN' and proteinase K we re increased by bioselective adsorption and excellent recoveries of al l proteinases applied were obtained.