›› 2011, Vol. 23 ›› Issue (6): 0-1226.

• 环境科学 •    

胆盐水解酶基因bsh工程菌表达条件的优化

姚晶1,2,任婧1,吴正钧1,郭本恒1,2,*

  

  1. 1 乳业生物技术国家重点实验室 光明乳业股份有限公司,上海 200436;2上海海洋大学 食品学院,上海 201306
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2011-11-25 发布日期:2011-11-25

Optimization of overexpression conditions of Lactobacillus plantarum bsh genes

YAO Jing;REN Jing;WU Zhengjun;GUO Benheng;*   

  1. 1State Key Laboratory of Dairy Biotechnology,Technology Center of Bright Dairy Co., Ltd., Shanghai 200436, China;2College of Food Science and Technology, Shanghai Ocean University, Shanghai 201306, China
  • Received:1900-01-01 Revised:1900-01-01 Online:2011-11-25 Published:2011-11-25

摘要:

对来源于Lactobacillus plantarum STⅢ菌株的胆盐水解酶基因bsh1在工程菌E.coli BL21/pET28bbsh 1 中的表达条件进行了优化。首先通过一系列单因素实验对影响工程菌发酵条件的因素水平进行筛选。然后将诱导培养时间、诱导剂异丙基硫代βD半乳糖苷(isopropylthioDgalactoside, IPTG) 浓度、胆盐浓度、装液量4个影响因素进行正交试验。在最佳工艺条件下得到的胆盐水解酶BSH比酶活达817.73 U。

关键词: 胆盐水解酶, 发酵条件, 单因素实验, 正交试验

Abstract:

The bsh1 gene was subcloned into expression vector pET28b and its product bile sale hydrolase(BSH) was overexpressed in Escherichia coli BL21. The appropriate value of each significant effect was noted with single factor experimental design. Effects of inducing time, isopropylthioDgalactoside. IPTG concentration, bile salt concentration and amount of medium on BSH activity were investigated by orthogonal test. After orthogonal experimental design, under the optimal condition, the specific activity of BSH was 817.73 U.

Key words: bile salt hydrolase, BSH, fermentation condition, single factor experimental design, orthogonal experimental design