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A mild purification method for polysaccharide binding membrane proteins: phase separation of digitonin extracts to
1Institut für Physiologische Chemie und Pathobiochemie, Münster, Germany.
Protein Expression and Purification
|June 1, 1996
Abstract:
A new method was developed to purify the streptococcal hyaluronate synthase in active form to electrophoretic homogeneity. The method is based on the extraction of protoplast membranes with digitonin and a phase separation into an aqueous and a detergent phase induced by addition of polyethylene glycol 6000 at 0 degree C. Proteins bound to hyaluronate were enriched in the aqueous phase, whereas other membrane proteins resided in the detergent phase. Final purification of the hyaluronate synthase was achieved by ion exchange chromatography.