A simple method is presented for removing protease from commercial sources of Streptornyces hyaluronidase derived from Streptomyces hyalurolyticus using a column of insolubilized chicken ovomucoid. The protease binds to ovomucoid; 91% of the hyaluronidase and no more than 0.7% of the protease passes through the column. A calcium chloride solution is used to remove the bound protease. Purified hyaluronidase cleaves the hyaluronic acid of proteoglycan aggregates without affecting the size of nroteoglycan monomers. The ability of digested monomer to bind hy~uronic acid, however, is somewhat diminished.
Hyaluronate lyase (EC 4.2.2.1) fromStreptomyces hyalurolyticus hydrolyzes hyaluranic acid, producing oligosaccharides that contain nonreducing terminal A4a5unsaturated glucopy~nuronic acid units. Other glycosaminoglycans are neither substrates nor inhibitors of this reaction (1).
Proteoglycans isolated from hyaline cartilage contain protein with covalently attached glycosaminoglycan chains and a region which binds specifically to hyaluronic acid (2). Native proteoglycans are usually aggregated to hyaluronic acid. An enzyme able to cleave the hyaluronic acid portion of this aggregate, without disrupting the protein core or the other giycosaminoglycans, would provide a useful tool in studies separating proteoglycan aggregation from other functions of these molecules.
However, the usefulness of Streptomyces hyaluronidase for this purpose is limited because the commercial preparation of this enzyme also contains significant proteolytic activity (3).