乳房链球菌透明质酸酶的分离与鉴定

Isolation and characterization of hyaluronidase from Streptococcus uberis.

作者信息

Schaufuss P, Sting R, Schaeg W, Blobel H

机构信息

Institut für Bakteriologie und Immunologie, Fachbereich Veterinärmedizin, Justus-Liebig-Universität Giessen.

出版信息

Zentralbl Bakteriol. 1989 May;271(1):46-53. doi: 10.1016/s0934-8840(89)80052-0.

DOI:10.1016/s0934-8840(89)80052-0

PMID:2765091

Abstract

All tested cultures of Streptococcus uberis produced free hyaluronidase. Hyaluronidase could be isolated by ammonium sulfate precipitation and was further purified by chromatography on DEAE-cellulose, gelfiltration on ultragel ACA44 and isoelectric focusing. The purification factor was estimated to be 1689. The purified hyaluronidase had an isoelectric point at pH 4.9 and a molecular weight of approximately 54000 D. It showed maximal enzyme activity at pH 6.0 and 45 degrees C. The Michaelis constant was estimated to be 7.0 X 10(-2) mg/ml. Hyaluronidase activity was stimulated by Ca++, Mg++, Mn++, Co++, Li+, and K+ and inhibited by Zn++ and Cd++ at final concentrations of 10 mmol/l, respectively.

摘要

所有检测的乳房链球菌培养物均产生游离透明质酸酶。透明质酸酶可通过硫酸铵沉淀分离，并通过DEAE - 纤维素柱层析、在超凝胶ACA44上进行凝胶过滤和等电聚焦进一步纯化。纯化因子估计为1689。纯化的透明质酸酶的等电点为pH 4.9，分子量约为54000 D。它在pH 6.0和45℃时表现出最大酶活性。米氏常数估计为7.0×10(-2)mg/ml。透明质酸酶活性分别受到终浓度为10 mmol/l的Ca++、Mg++、Mn++、Co++、Li+和K+的刺激，而受到Zn++和Cd++的抑制。