A MILD PURIFICATION METHOD FOR POLYSACCHARIDE BINDING MEMBRANE-PROTEINS - PHASE-SEPARATION OF DIGITONIN EXTRACTS TO ISOLATE THE HYALURONATESYNTHASE FROM STREPTOCOCCUS SP IN ACTIVE FORM

A new method was developed to purify the streptococcal hyaluronate syn thase in active form to electrophoretic homogeneity. The method is bas ed on the extraction of protoplast membranes with digitonin and a phas e separation into an aqueous and a detergent phase induced by addition of polyethylene glycol 6000 at 0 degrees C. Proteins bound to hyaluro nate were enriched in the aqueous phase, whereas other membrane protei ns resided in the detergent phase. Final purification of the hyalurona te synthase was achieved by ion exchange chromatography. (C) 1996 Acad emic Press, Inc.