Srivastava P N, Farooqui A A
Biochem J. 1979 Dec 1;183(3):531-7. doi: 10.1042/bj1830531.
Bull seminal-plasma hyaluronidase was purified 180-fold by chromatography on concanvalin A-Sepharose, heparin Sepharose, Sephadex G-200 and Sephacryl S-200. With hyaluronic acid as the substrate, the specific activity and turnover number of purified hyaluronidase were 3.63 mumol/min per mg (104000 National Formulary units/mg of protein) and 214 min-1 (mol of product formed/mol of enzyme per min) respectively. Polyacrylamide-gel electrophoresis indicated that the purified enzyme migrated as a single band on 7.5 and 10% (w/v) gels at pH 4.3 and 5.3. Bull seminal-plasma hyaluronidase was markedly inhibited by hydroxylamine, phenylhydrazine and semicarbazide. Purified hyaluronidase (1.25 munits; 1 unit = 1 mumol of N-acetylglucosamine liberated/min at 37 degrees C) dispersed the cumulus clot of rabbit ova in 1 h at 22 degrees C.
通过在伴刀豆球蛋白A - 琼脂糖、肝素琼脂糖、葡聚糖凝胶G - 200和Sephacryl S - 200上进行层析，公牛精浆透明质酸酶被纯化了180倍。以透明质酸为底物时，纯化后的透明质酸酶的比活性和转换数分别为每毫克3.63微摩尔/分钟（104000美国国家处方集单位/毫克蛋白质）和214分钟-1（每分钟形成的产物摩尔数/酶摩尔数）。聚丙烯酰胺凝胶电泳表明，纯化后的酶在pH 4.3和5.3的7.5%和10%（w/v）凝胶上迁移时呈现为单一条带。公牛精浆透明质酸酶受到羟胺、苯肼和氨基脲的显著抑制。纯化后的透明质酸酶（1.25微单位；1单位 = 在37℃下每分钟释放1微摩尔N - 乙酰葡糖胺）在22℃下1小时内分散了兔卵丘凝块。
