粘质沙雷氏菌作为一种产软骨素酶细菌的分离及一种新型软骨素酶AC的纯化。

Isolation of Serratia marcescens as a chondroitinase-producing bacterium and purification of a novel chondroitinase AC.

作者信息

Ke Tao, Zhangfu Long, Qing Gao, Yong Tao, Hong Jin, Hongyan Ran, Kun Liu, Shigui Liu

机构信息

National Laboratory of Grassland Biocontrol Engineering, Sichuan University, Chengdu 610064, China.

出版信息

Biotechnol Lett. 2005 Apr;27(7):489-93. doi: 10.1007/s10529-005-2538-7.

DOI:10.1007/s10529-005-2538-7

PMID:15928855

Abstract

A strain of Serratia marcescens that produced chondroitinase was isolated from soil. It produced a novel chondroitinase AC, which was purified to homogeneity. The enzyme was composed of two identical subunits of 35 kDa as revealed by SDS-PAGE and gel filtration. The isoelectric point for the chondroitinase AC was 7.19. Its optimal activity was at pH 7.5 and 40 degrees C. The purified enzyme was active on chondroitin sulfates A and C and hyaluronic acid, but was not with chondroitin sulfate B (dermatan sulfate), heparin or heparan sulfate. The apparent K(m) and V(max) of the chondroitinase AC for chondroitin sulfate A were 0.4 mg ml(-1) and 85 mmol min(-1) mg(-1), respectively, and for chondroitin sulfate C, 0.5 mg ml(-1) and 103 mmol min(-1) mg(-1), respectively.

摘要

从土壤中分离出一株产软骨素酶的粘质沙雷氏菌。它产生了一种新型软骨素酶AC，并纯化至同质。通过SDS-PAGE和凝胶过滤显示，该酶由两个相同的35 kDa亚基组成。软骨素酶AC的等电点为7.19。其最佳活性在pH 7.5和40℃。纯化后的酶对硫酸软骨素A和C以及透明质酸有活性，但对硫酸软骨素B（硫酸皮肤素）、肝素或硫酸乙酰肝素无活性。软骨素酶AC对硫酸软骨素A的表观K(m)和V(max)分别为0.4 mg ml(-1)和85 mmol min(-1) mg(-1)，对硫酸软骨素C分别为0.5 mg ml(-1)和103 mmol min(-1) mg(-1)。