High-Yield Recombinant Expression of the Chicken Antimicrobial Peptide Fowlicidin-2 in Escherichia coli

文献类型： 外文期刊

第一作者： Feng, Xingjun

作者： Feng, Xingjun;Xu, Wenshan;Qu, Pei;Li, Xiaochong;Xing, Liwei;Liu, Di;Jiao, Jian;Wang, Jue;Li, Zhongqiu;Liu, Chunlong;Liu, Chunlong

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关键词： recombinant expression;antimicrobial peptides;fowlicidin-2;cyanogen bromide;Escherichia coli;antibacterial activity

期刊名称：BIOTECHNOLOGY PROGRESS （ 影响因子：2.681； 五年影响因子：2.691 ）

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收录情况： SCI

摘要： The antimicrobial peptide fowlicidin-2 identified in chicken is a member of the cathelicidins family. The mature fowlicidin-2 possesses high antibacterial efficacy and lipopolysaccharide (LPS) neutralizing activity, and also represents an excellent candidate as an antimicrobial agent. In the present study, the recombinant fowlicidin-2 was successfully produced by Escherichia coli (E. coli) recombinant expression system. The gene encoding fowlicidin-2 with the codon preference of E. coli was designed through codon optimization and synthesized in vitro. The gene was then ligated into the plasmid pET-32a(+), which features fusion protein thioredoxin at the N-terminal. The recombinant plasmid was transformed into E. coli BL21(DE3) and cultured in Luria-Bertani (LB) medium. After isopropyl--D-thiogalactopyranoside (IPTG) induction, the fowlicidin-2 fusion protein was successfully expressed as inclusion bodies. The inclusion bodies were dissolved and successfully released the peptide in 70% formic acid solution containing cyanogen bromide (CNBr) in a single step. After purification by reverse-phase high-performance liquid chromatography (RP-HPLC), approximate to 6.0 mg of fowlicidin-2 with purity more than 97% was obtained from 1 litre of bacteria culture. The recombinant peptide exhibited high antibacterial activity against the Gram-positive and Gram-negative bacteria, and even drug-resistant strains. This system could be used to rapidly and efficiently produce milligram quantities of a battery of recombinant antimicrobial peptides as well as for large-scale production. (c) 2015 American Institute of Chemical Engineers Biotechnol. Prog., 31:369-374, 2015

分类号： Q

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