|Table of Contents|

Secretory expression of bile salt hydrolase gene from Lactobacillus plantarum BBE7 in Pichia pastoris(PDF)

Chinese Journal of Applied & Environmental Biology[ISSN:1006-687X/CN:51-1482/Q]

Issue:
2014 01
Page:
107-111
Research Field:
Articles
Publishing date:

Info

Title:
Secretory expression of bile salt hydrolase gene from Lactobacillus plantarum BBE7 in Pichia pastoris
Author(s):
DONG Zixing ZHANG Juan LEE Byonghoon DU Guocheng CHEN Jian LI Huazhong
1 School of Biotechnology, Jiangnan University, Wuxi 214122, China 2 Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, China 3 National Engineering Laboratory for Cereal Fermentation Technology, Jiangnan University, Wuxi 214122, China 4 Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, China
Keywords:
Lactobacillus plantarum BBE7 bile salt hydrolase Pichia pastoris secretory expression orthogonal experimental design
CLC:
Q936 : Q786
PACS:
DOI:
10.3724/SP.J.1145.2014.00107
DocumentCode:

Abstract:
When bile salt hydrolase (bsh) genes from Lactobacillus plantarum are expressed intracellularly and extracellularly by Escherichia coli, they usually form inclusion bodies. Therefore, Pichia pastoris, an efficient expression system, was used to achieve the secretory expression of bsh. Firstly, signal peptide α-factor was ligated with bsh by fusion PCR, and the fragment obtained was then cloned into vector pPIC9K, resulting in a recombinant plasmid pPIC9K-α-factor-BSH. Finally, this plasmid was linearized by Sal I and transformed into Pichia pastoris GS115. As analyzed by SDS-PAGE and BSH assay, the molecular weight and extracellular activity of recombinant BSH was determined to be 37.0 × 103 and 1.08 U mL-1, respectively. After optimization of fermentation conditions using orthogonal experimental design (L9 (34)),the extracellular BSH activity was 2.69 U mL-1, 1.49 times higher than before. This lays a solid foundation for purification, characterization and large-scale production of BSH.

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Last Update: 2014-03-06