乙肝表面抗原在酿酒酵母中的异源表达  

Heterologous Expression of HBsAg in Saccharomyces Cerevisiae

作　　者：邱玲[1,2] 钱俊佳 康振[1,2] 陈坚[2,4] 

机构地区：[1]江南大学工业生物技术教育部重点实验室,江苏无锡214122 [2]江南大学生物工程学院,江苏无锡214122 [3]希森美康生物科技无锡有限公司,江苏无锡214028 [4]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

出　　处：《食品与生物技术学报》2015年第9期906-913,共8页Journal of Food Science and Biotechnology

基　　金：国家863计划重点项目(2011AA100905);教育部长江学者和创新团队发展计划项目(IRT1135);国家博士后基金面上项目(2013 M540414);江苏省优势学科项目

摘　　要：乙肝表面抗原HBs Ag是乙肝疫苗的主要组分。从转录水平、翻译水平以及翻译后水平对其进行优化,实现了HBs Ag在重组酿酒酵母中的高效异源表达。与组成型启动子TEF1、TDH3、HXT7、ADH1相比,诱导型启动子GAL1可大幅度提高HBs Ag的表达;与GCCACC和AAAAAA相比,kozak序列TACACA最有利于HBs Ag的翻译表达;而共表达二硫键异构酶pdi对HBs Ag的活性表达具有显著的促进作用。通过3种水平的优化,HBs Ag活性从最初的8.87 IU/g提高至151.08 IU/g;最终,利用Ni柱亲和层析成功获得纯化的HBs Ag,活性为3.32 IU/m L。Hepatitis B surface antigen（HBsA g） is a major component of the hepatitis B vaccine. In the present work,we achieved its high efficient heterologous expression in the recombinant Saccharomyces cerevisiae by optimization at three levels：transcription,translation and post-translation. Compared with the constitutive promoters TEF1,TDH3,HXT7 and ADH1,the inducible promoter GAL1 substantially increased the expression of HBs Ag. Moreover,compared with GCCACC and AAAAAA,the kozak sequence TACACA is more beneficial to the translation and active expression of HBs Ag. More importantly,co-expression of protein disulfide isomerase（PDI） also significantly increased the expression of HBs Ag. Through three levels of optimizations,the activity of recombinant HBsA g was increased from 8.87 IU/g CDW to 151.08 IU/g CDW.Finally,applying the Ni column affinity chromatography,we successfully obtained the purified HBsA g with an activity of 3.32 IU/m L.

关 键 词：酿酒酵母 乙肝表面抗原 启动子 KOZAK序列 共表达 

分 类 号：R512.62[医药卫生—内科学]