硫酸软骨素酶的分离纯化及部分性质

journal6 ›› 2004, Vol. 25 ›› Issue (4): 44-48.

硫酸软骨素酶的分离纯化及部分性质

（1.四川大学生命科学学院，四川成都 610064 ；2.成都川大创新生物技术研究所，四川成都 610064）

出版日期:2004-12-15 发布日期:2012-09-22
作者简介:陶科（1978-），女，四川大学生命科学学院博士研究生，主要从事遗传学、微生物学方面的研究与资源开发.

Isolation,Purification and Properties of Chondroitinase From Serratia Marcescens

(1.College of Life Science Sichuan University,Chengdu 610064，China;2.Chengdu Chuangxin Bio-technology Institute,Sichuan University,Chengdu 610064,China)

Online:2004-12-15 Published:2012-09-22

摘要/Abstract

摘要：通过细胞破碎、硫酸铵分级沉淀、DEAE-Sepharose FF和Sephacryl S-200柱层析，首次从粘质沙雷氏菌中提取出硫酸软骨素酶.纯化倍数11.46,比活为81.23 U/mg蛋白.提取液经PAGE和SDS-PAGE检测，呈现一条带.该酶相对分子质量为71.2 KD,亚基分子质量为35 KD,该酶为2个相同亚基组成.等电点为7.19，最适pH值7.5，最适温度为40 ℃，以4-硫酸软骨素为底物测得Km值为3.98×10^-4mol/L.

关键词： 沙雷氏菌, 硫酸软骨素酶, 分离纯化

Abstract: Chondroitinase was isolated and purified from Serratia marcescens and its kinetic property was examined.The Chondroitinase was purified by salting-out,DEAE-Sepharose FF ion-exchange chromatogram,and gel filtrition with Sephacryl S-200 chromatogram.The purified enzyme moved as a single electrophoretic band in PAGE.The specific activity was 81.23 U/mg.It’s subunit weight was 35 KD with SDS-PAGE,moleculer weight was 70.20 KD with gel filtrition on Sephacryl S-300(AKTA FPLC).The optimum pH value for the enzyme was 7.5.The optimum temperature for the enzyme was about 40 ℃.The Michealis-Menton constant(Km) was 3.98×10^-4mol/L with 4-Chondroitin Sulfate as its substrate.

Key words: Serratia marcescens, chondroitinase, isolation and purification

陶科, 龙章富, 刘昆, 陶勇, 金虹, 冉红艳, 高庆, 刘世贵. 硫酸软骨素酶的分离纯化及部分性质[J]. journal6, 2004, 25(4): 44-48.

TAO Ke, LONG Zhang-Fu, LIU Kun, TAO Yong, JIN Hong, RAN Hong-Yan, GAO Qing, LIU Shi-Gui. Isolation,Purification and Properties of Chondroitinase From Serratia Marcescens[J]. journal6, 2004, 25(4): 44-48.

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