Purification and refolding optimization of recombinant bovine enterokinase light chain overexpressed in Escherichia coli
文献类型：期刊论文
| 作者 | Tan, Haidong; Wang, Jinxia; Zhao, Zongbao (Kent) |
| 刊名 | protein expression and purification
 |
| 出版日期 | 2007-11-01 |
| 卷号 | 56期号:1页码:40-47 |
| 关键词 | bovine enterokinase fusion protein inclusion body overexpression protein purification refolding |
| 产权排序 | 1;1 |
| 通讯作者 | 赵宗保 |
| 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. |
| 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 |
| 关键词[WOS] | enteropeptidase catalytic subunit ; saccharomyces-cerevisiae ; porcine enterokinase ; pichia-pastoris ; expression ; protease ; gene |
| 收录类别 | SCI |
| 原文出处 | true |
| 语种 | 英语 |
| WOS记录号 | WOS:000250639100006 |
| 公开日期 | 2010-11-30 |
| 源URL | [http://159.226.238.44/handle/321008/99367]  |
| 专题 | 大连化学物理研究所_中国科学院大连化学物理研究所 |
| 作者单位 | Chinese Acad Sci, Dalian Inst Chem Phys, Dalian 116023, Peoples R China |
| 推荐引用方式 GB/T 7714 | 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. |
| APA | Tan, Haidong,Wang, Jinxia,&Zhao, Zongbao .(2007).Purification and refolding optimization of recombinant bovine enterokinase light chain overexpressed in Escherichia coli.protein expression and purification,56(1),40-47. |
| MLA | Tan, Haidong,et al."Purification and refolding optimization of recombinant bovine enterokinase light chain overexpressed in Escherichia coli".protein expression and purification 56.1(2007):40-47. |
入库方式： OAI收割
来源：大连化学物理研究所
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