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

Abstract：

: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 approximately 1 mg 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 microM. 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.

journal_name

Protein Expr Purif

journal_title

Protein expression and purification

authors

Cipáková I,Hostinová E,Gasperík J,Velebný V

doi

10.1016/j.pep.2004.04.024

keywords:

subject

Has Abstract

pub_date

2004-09-01 00:00:00

pages

207-12

issue

1

eissn

1046-5928

issn

1096-0279

pii

S1046592804001226

journal_volume

37

pub_type

杂志文章

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