大肠杆菌表达腺苷蛋氨酸合酶及产酶条件优化  

Heterologous expression of adenosylmethionine synthase in Escherichia coli and optimization of enzyme-producing conditions

作　　者：毛职医 谈新苑 曹蓉 张晓娟[1,2] 付静 徐建国 徐国强 张晓梅[3] 许正宏 MAO Zhiyi;TAN Xinyuan;CAO Rong;ZHANG Xiaojuan;FU Jing;XU Jianguo;XU Guoqiang;ZHANG Xiaomei;XU Zhenghong(The Key Laboratory of Industrial Biotechnology,Ministry of Education,School of Biotechnology,Jiangnan University,Wuxi 214122,China;National Engineering Laboratory for Cereal Fermentation Technology,Jiangnan University,Wuxi 214122,China;School of Pharmaceutical Sciences,Jiangnan University,Wuxi 214122,China;Wuxi Fuqi Pharmaceutical Co.,Ltd.,Wuxi 214100,China)

机构地区：[1]江南大学生物工程学院,工业生物技术教育部重点实验室,江苏无锡214122 [2]江南大学,粮食发酵工艺与技术国家工程实验室,江苏无锡214122 [3]江南大学药学院,江苏无锡214122 [4]无锡福祈制药有限公司,江苏无锡214100

出　　处：《食品与发酵工业》2020年第10期8-13,共6页Food and Fermentation Industries

基　　金：江苏省自然基金面上项目(BK20191333)。

摘　　要：腺苷蛋氨酸(adenosymethionine,SAM)是人体中重要的代谢中间体,在医疗上被广泛应用于治疗肝炎,抑郁症以及关节炎等疾病。酶法进行腺苷蛋氨酸的生产具有良好的工业前景。将来源于酿酒酵母BY4741中的腺苷蛋氨酸合酶基因(sam2)进行密码子优化后,与p ET-28a(+)和p ET-3b(+)进行连接获得重组质粒p ET-28a(+)-sam2和p ET-3b(+)-sam2,分别实现其在E. coli BL21(DE3)和E. coli Rosetta(DE3)中的异源表达,结果表明,重组菌E. coli BL21/p ET-28a(+)-sam2的SAM酶活较高,达到0. 184 U/m L。对该重组菌株进行发酵条件优化,在最优条件下,SAM合酶最高酶活为0. 245 U/m L,比优化前提高34%。Adenosylmethionine(SAM) plays a vital role in the organisms’ metabolic activity as an important intermediate metabolite and has been widely used in pharmaceutical industry involving the therapies for liver,nervous systems,osteoarthritis and so on. Thus,production of SAM by enzyme transformation process exhibits potential industrial prospects. Codon optimization of the adenosylmethionine synthase gene sam2 derived from S. cerevisiae BY4741 was performed,and the gene was ligated to pET-28 a( +) and pET-3 b( +),resulting in recombinant plasmids p ET-28 a( +)-sam2 and pET-3 b( +)-sam2,respectively,and was heterologous expressed in E. coli BL21( DE3) and E.coli Rosseta( DE3) respectively. Results showed that the recombinant strain E. coli BL21/pET28 a( +)-sam2 exhibited higher enzymatic activity up to 0. 184 U/m L. Thus,the recombinant strain E.coli BL21/p ET-28 a( +)-sam2 was chosen for the optimization of fermentation condition to further improve the enzymatic activity. Under optimal conditions,the enzyme activity of the recombinant adenosylmethionine synthase reached 0. 245 U/m L,which was 34% higher than that before optimization.

关 键 词：大肠杆菌 S-腺苷蛋氨酸合酶 异源表达 酶法合成 诱导条件优化 

分 类 号：TQ925[轻工技术与工程—发酵工程]