一种嗜热细菌来源角质酶的分离纯化及酶学性质  

Purif ication and Characterization of Cutinase from a Thermophilic Bacterium

作　　者：陈晟[1,2] 陈坚[1,2] 吴敬[1,2] 

机构地区：[1]江南大学生物工程学院,江南大学工业生物技术教育部重点实验室,无锡214122 [2]江南大学食品科学与技术国家重点实验室,无锡214122

出　　处：《应用与环境生物学报》2009年第6期846-850,共5页Chinese Journal of Applied and Environmental Biology

基　　金：教育部新世纪优秀人才支持计划资助项目(No.NCET-06-0486);国家高技术研究发展计划资助项目(863计划;No.2009AA02Z204);江苏省六大人才高峰支持计划(No.08-B-吴敬);食品科学与技术国家重点实验室科研基金(No.SKLF-MB-200802);江南大学创新团队项目(No.2008CXTD01)资助~~

摘　　要：通过跟踪发酵液中pNPB水解酶活性,对角质诱导的Thermobifida fusca发酵液进行分离纯化.采用活性炭脱色、硫铵沉淀、Phenyl HP疏水色谱、DEAE Sepharose阴离子交换色谱等方法,分离纯化得到电泳纯pNPB水解酶.该酶水解角质可得到角质单体,是一种角质酶.SDS-PAGE电泳结果显示,角质酶表观分子量约为29×103.该酶的最适温度为60℃,在40℃和60℃下均具有良好的热稳定性.最适pH为8.0,pH稳定范围为6.0～9.0.该角质酶的生化性质适合在纺织工业中应用.A pNPB hydrolase was purified from Thermobifida fusca culture supernatant induced with cutin by active carbon,ammonium sulfate precipitation,phenyl HP sepharose chromatography and DEAE sepharose chromatography through monitoring the pNPB hydrolyzing activity. The pNPB hydrolase could hydrolyze cutin and release cutin monomers,so it is a kind of cutinase. The enzyme was homogeneous by SDS-PAGE analysis and showed a molecular weight of 29 ×103. The cutinase exhibited its maximum activity at 60 ℃ and pH 8.0, and showed its thermostability at 40-60 ℃ and pH 6.0-9.0.These properties indicate that the enzyme has potential application in textile industry. Fig 8, Tab 2, Ref 17

关 键 词：嗜热细菌 角质酶 分离纯化 酶学性质 热稳定性 

分 类 号：Q814[生物学—生物工程]