食品科学

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假肠膜明串珠菌甘露醇脱氢酶基因的克隆及表达

程雅韵,王 新,郑 琳1,李官浩,崔虎山,金 清   

  1. 1.延边大学农学院,吉林 延吉 133002;2.延边大学附属医院西区,吉林 延吉 133000
  • 出版日期:2016-07-15 发布日期:2016-07-26
  • 通讯作者: 金 清
  • 基金资助:

    国家自然科学基金地区科学基金项目(31260362);吉林省教育厅“十二五”科学技术研究项目(吉教科合字[2015第40号])

Cloning and Expression of Mannitol Dehydrogenase Gene from Leuconostoc pseudomesenteroides

CHENG Yayun, WANG Xin, ZHENG Lin, LI Guanhao, CUI Hushan, JIN Qing   

  1. 1. Agricultural College of Yanbian University, Yanji 133002, China;
    2. West District of Affiliated Hospital of Yanbian University, Yanji 133000, China
  • Online:2016-07-15 Published:2016-07-26
  • Contact: JIN Qing

摘要:

利用聚合酶链式反应从假肠膜明串珠菌中扩增出甘露醇脱氢酶(mannitol dehydrogenase,MDA)基因的结构基因,克隆入表达载体pETDuet-1,构建了甘露醇脱氢酶表达质粒pETDuet-1-mdh,将其转化进入大肠杆菌BL21(DE3),经异丙基-β-D-硫代半乳糖苷诱导表达。假肠膜明串珠菌甘露醇脱氢酶结构基因长度为1 017 bp,重组甘露醇脱氢酶基因在大肠杆菌内成功表达,其蛋白质分子质量为36.0 kD;重组甘露醇脱氢酶活力为0.15 U/mg pro,高于假肠膜明串珠菌中甘露醇脱氢酶活力0.03 U/mg pro。

关键词: 甘露醇, 甘露醇脱氢酶, 假肠膜明串珠菌, 基因克隆, 表达

Abstract:

The structural gene encoding mannitol dehydrogenase (MDH) from Leuconostoc pseudomesenteroides was
amplified by PCR and cloned into the vector pETDuet-1. As a result, the plasmid pETDuet-1-mdh was constructed and
transformed into E. coli BL21(DE3) for MDH expression induced by isopropyl-β-D-thiogalactoside (IPTG). The length
of the mdh structural gene was l 017 bp. The recombinant mdh gene was successfully expressed in Escherichia coli and
the molecular weight of the expressed protein was 36.0 kD. The activity of recombinant mannitol dehydrogenase was
0.15 U/mg pro, which was higher than the MDH activity of Leuconostoc pseudomesenteroides (0.03 U/mg pro).

Key words: mannitol, mannitol dehydrogenase, Leuconostoc pseudomesenteroides, gene cloning, expression

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