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题名: Purification and refolding optimization of recombinant bovine enterokinase light chain overexpressed in Escherichia coli
作者: Tan, Haidong;  Wang, Jinxia;  Zhao, Zongbao (Kent)
通讯作者: 赵宗保
关键词: bovine enterokinase ;  fusion protein ;  inclusion body ;  overexpression ;  protein purification ;  refolding
刊名: PROTEIN EXPRESSION AND PURIFICATION
发表日期: 2007-11-01
DOI: 10.1016/j.pep.2007.07.006
卷: 56, 期:1, 页:40-47
收录类别: SCI
文章类型: Article
部门归属: 18
项目归属: 1816
产权排名: 1;1
WOS标题词: Science & Technology ;  Life Sciences & Biomedicine
类目[WOS]: Biochemical Research Methods ;  Biochemistry & Molecular Biology ;  Biotechnology & Applied Microbiology
研究领域[WOS]: Biochemistry & Molecular Biology ;  Biotechnology & Applied Microbiology
英文摘要: The nucleotide sequence encoding bovine enterokinase light chain (EK) from Chinese northern yellow bovine was isolated. Two single-nucleotide mutations, namely, C245G and A528T were identified. The gene encoding the Pro82Arg/Glu176Asp variant of known bovine EK was fused with glutathione S-transferase and overexpressed mainly as an inclusion body in Escherichia coli BL21 (DE3), upon induction with IPTG and glucose. Effective fusion protein purification, refolding, auto-catalytic cleavage and mature EK recovery were described. The specific activity of the purified EK was determined as 1 10 +/- 10 U/mg, which was comparable to a specific activity of >= 20 U/mg of the E. coli expressed EK sample provided by Sigma (Cat. No. E4906). This procedure produced approximately 53 mg of EK per 500 mL of cell culture, which was much higher than previous reports, thus providing a basis for large-scale production of EK and for further applications in biotechnology. (c) 2007 Elsevier Inc. All rights reserved.
关键词[WOS]: ENTEROPEPTIDASE CATALYTIC SUBUNIT ;  SACCHAROMYCES-CEREVISIAE ;  PORCINE ENTEROKINASE ;  PICHIA-PASTORIS ;  EXPRESSION ;  PROTEASE ;  GENE
语种: 英语
原文出处: 查看原文
WOS记录号: WOS:000250639100006
Citation statistics: 
内容类型: 期刊论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/99367
Appears in Collections:中国科学院大连化学物理研究所_期刊论文

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作者单位: 1.Chinese Acad Sci, Dalian Inst Chem Phys, Dalian 116023, Peoples R China

Recommended Citation:
Tan, Haidong,Wang, Jinxia,Zhao, Zongbao . Purification and refolding optimization of recombinant bovine enterokinase light chain overexpressed in Escherichia coli[J]. PROTEIN EXPRESSION AND PURIFICATION,2007,56(1):40-47.
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