Purification and characterization of hyaluronidase from Streptococcus dysgalactiae.

Streptococcus dysgalactiae IID 678, belonging to group C of the streptococci, secreted a large amount of hyaluronidase (hyaluronate lyase, EC 4.2.2.1) into a culture medium containing hyaluronic acid. The purification procedures of hyaluronidase were 70% ammonium sulfate precipitation, ECT EC) LA-eel hi lose chromatography, phospho-cellulose chromatography, and gel filtration on Sephacryl S-300. The hyaluronidase was purified approximately 27, 000-fold from the culture filtrate. The purified enzyme was homogeneous by SDS-polyacrylamide gel electrophoresis. The enzyme degradated only hyaluronic acid and chondroitin to Δ4, 5-unsaturated disaccharides and did not act on other glycosaminoglycans containing sulfate groups, while the degradation rate of chondroitin was about 1/60 of that of hyaluronic acid. The optimum pH was wide, from pH 5.8 to pH 6.6, and the optimum temperature was 37°C. Fe2+, Cu2+, Pb2+, and Hg2+ ions inhibited the activity strongly and Zn2+ inhibited it by half. The molecular weight of the enzyme was estimated to be 125, 000 by gel filtration and 117, 000 by SDS-polyacrylamide gel electrophoresis. The enzyme was different immunochemically from the hyaluronidase from Streptococcus pyogenes belonging to group A.