PURIFICATION AND CHARACTERIZATION OF HYALURONIDASE FROM STREPTOCOCCUS-DYSGALACTIAE
PURIFICATION AND CHARACTERIZATION OF HYALURONIDASE FROM STREPTOCOCCUS-DYSGALACTIAE
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DOI：
10.1080/00021369.1989.10869625
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发表时间：
1989-08-01
期刊：
影响因子：
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通讯作者：
TOKUYASU, K
中科院分区：
文献类型：
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作者：
HAMAI, A;MORIKAWA, K;TOKUYASU, K
停乳链球菌（Streptococcus dysgalactiae）IID 678，属于链球菌的C组，向含有透明质酸的培养基中分泌大量的透明质酸酶（透明质酸裂解酶，EC 4.2.2.1）。透明质酸酶的纯化步骤为70%硫酸铵沉淀、ECTEOLA-纤维素层析、磷酸纤维素层析和Sephacryl S-300凝胶过滤。透明质酸酶从培养物滤液中纯化约27，000倍。纯化的酶经SDS-聚丙烯酰胺凝胶电泳呈均一性。该酶仅将透明质酸和软骨素降解为Δ 4，5-不饱和二糖，不作用于其他含有硫酸根基团的糖胺聚糖，而软骨素的降解率约为透明质酸的1/60。最适pH范围较宽，从pH5.8到pH6.6，最适温度为37 ℃。C. Fe ~（2+）、Cu ~（2+）、Pb ~（2+）、Hg ~（2+）等离子对该酶活性有强烈的抑制作用，Zn ~（2+）对该酶活性的抑制作用为1/2。凝胶过滤法测得酶的分子量为125，000，SDS-聚丙烯酰胺凝胶电泳法测得酶的分子量为117，000。该酶在免疫化学上与A组化脓性链球菌的透明质酸酶不同。
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, ECTEOLA-cellulose chromatography, phosphocellulose 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 .DELTA.4,5-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.degree. 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.