谷氨酸棒杆菌表达大肠杆菌来源海藻糖酶  

Expression of Trehalase from Escherichia coli by Corynebacterium glutamicum

作　　者：满在伟 崔慧慧 李锦 张迎阳[1] 张建波 张建涛 MAN Zai-wei;CUI Hui-hui;LI Jin;ZHANG Ying-yang;ZHANG Jian-bo;ZHANG Jian-tao(College of Biological and Food Engineering,Changzhou University,Changzhou 213164,Jiangsu,China;Shandong Hengren Gongmao Co.,Ltd.,Zaozhuang 277533,Shandong,China)

机构地区：[1]常州大学生物与食品工程学院,江苏常州213164 [2]山东恒仁工贸有限公司,山东枣庄277533

出　　处：《食品研究与开发》2022年第24期181-187,共7页Food Research and Development

基　　金：国家自然科学基金青年科学基金项目(31700075);山东省重大科技创新工程项目(2019JZZY020605)。

摘　　要：对谷氨酸棒杆菌(Corynebacterium glutamicum,C.glutamicum)表达大肠杆菌(Escherichia coli,E.coli)来源海藻糖酶进行研究。构建重组C.glutamicum菌株Cgtrl表达E.coli BL21来源海藻糖酶。菌株Cgtrl海藻糖酶最适表达条件:诱导剂异丙基-β-D-硫代半乳糖苷(isopropyl-β-D-thiogalactopyranoside,IPTG)添加量为0.5 mmol/L,诱导温度为22℃,诱导时间为12 h。在最适表达条件下重组海藻糖酶酶活为9.1 U/mL。重组海藻糖酶酶学性质研究表明,未经纯化的重组海藻糖酶具有较好的底物特异性能,专一性水解海藻糖,海藻糖水解率在96%以上,低温条件下稳定性较好,能够长时间低温保存,最适作用温度为45℃,最适作用pH值为6.6。The expression of trehalase from Escherichia coli BL21 by the constructed recombinant Corynebacterium glutamicum was studied.The optimal conditions for trehalase expression by Cgtrl were 0.5 mmol/L of isopropyl-β-D-thiogalactopyranoside as the inducer,induction temperature of 22℃,and induction time of 12 h.The recombinant trehalase activity reached 9.1 U/mL under the optimal expression conditions.Unpurified recombinant trehalase specifically hydrolyzed trehalose.The hydrolysis ratio of trehalose exceeded 96%.The recombinant trehalase was stable and could maintain enzyme activity for a long period at low temperature.The enzyme activity was optimal at 45℃and pH 6.6.

关 键 词：谷氨酸棒杆菌 大肠杆菌 海藻糖酶 表达条件优化 酶学性质 

分 类 号：TQ925[轻工技术与工程—发酵工程]