High-level expression and purification of a recombinant hBD-1 fused to LMM protein in Escherichia coli

被引：44

作者：

Cipáková, I ^{[1
]}

Hostinová, E

Gasperík, JG

Velebny, V

机构：

[1] CPN Spol SRO, Dolni Dobrouc 56102 401, Czech Republic

[2] Slovak Acad Sci, Inst Mol Biol, Ctr Excellence Mol Med, Bratislava 84551, Slovakia

来源：

PROTEIN EXPRESSION AND PURIFICATION | 2004年 / 37卷 / 01期

关键词：

D O I：

10.1016/j.pep.2004.04.024

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

In this work, we present the production of an active 43 aa recombinant human beta-defensin-1 (rhBD-1(43)) in Escherichia coli AD202 cells using specific pLMM1-rhBD-1 expression system. Unique solubility properties of the C-terminal fragment of light meromyosin (LMM) allowed us to overcome foreseeable problems with isolation procedures and toxicity caused by rhBD-1 to the host organism. As a result, the majority of fusion protein (LMM-rhBD-1(43)) was obtained in the soluble state, isolated by a low salt-high salt treatment of total cell protein. The rhBD-1(43) Was cleaved from the fusion with Protease 4 and purified on CM Sepharose Fast Flow column with the yield of similar to 1mg rhBD-1(43) from 6 g of wet weight cells. Purified rhBD-1(43) showed antimicrobial activity against E coli ML-35p at a concentration of 129 M. The procedure of rhBD-1 expression and purification we present can provide a reliable and simple method for production of different cationic peptides for biological studies. (C) 2004 Elsevier Inc. All rights reserved.

引用

页码：207 / 212

页数：6