title: Recombinant expression, purification, and antimicrobial activity of a novel hybrid antimicrobial peptide LFT33.
abstract: With great therapeutic potential against antibiotic-resistant bacteria, viruses, and even parasites, antimicrobial peptides (AMPs) have received increased interest as pharmaceutical agents in recent years. It is a worthy yet challenging work to carry out the implement and improvement of AMPs production using bioengineering techniques. In the present study, a novel hybrid peptide LFT33 was designed derived from LfcinB and thanatin. The cDNA fragment encoding LFT33 with preferred codons of Escherichia coli was chemically synthesized and ligated into the vector pET32a(+) to express the LFT33 fusion protein. The fusion protein was successfully expressed in soluble form in E. coli induced under optimized conditions. After purification by affinity chromatography, the fusion protein was cleaved successfully by enterokinase and released the peptide LFT33. About 0.5 mg of the recombinant LFT33 was obtained by reversed-phase high performance liquid chromatography from 1 l of culture medium. Mass spectrometry analysis of the purified recombinant LFT33 demonstrated that the molecular weight perfectly matched the calculated mass (4,195 Da). The recombinant peptide LFT33 caused an increase in antimicrobial activity (IC(50) = 16-64 μg/ml) against given strains and did not show hemolytic activity for human erythrocytes. The results indicated that the hybrid peptide LFT33 could serve as a promising candidate for pharmaceutical agents.
authors.affiliation: College of Animal Science and Technology, Northeast Agricultural University, 59 Mucai Street, Xiangfang District, Harbin, 150030, China. fengxingjun2008@163.com
authors.forename: Xingjun
authors.initials: X
authors.lastname: Feng
authors.forename: Chunlong
authors.initials: C
authors.lastname: Liu
authors.forename: Jiayin
authors.initials: J
authors.lastname: Guo
authors.forename: Xueying
authors.initials: X
authors.lastname: Song
authors.forename: Jing
authors.initials: J
authors.lastname: Li
authors.forename: Wenshan
authors.initials: W
authors.lastname: Xu
authors.forename: Zhongqiu
authors.initials: Z
authors.lastname: Li
mesh_terms: D023181:Antimicrobial Cationic Peptides / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000494:pharmacology*; D001419:Bacteria / Q000187:drug effects; D002176:Candida albicans / Q000187:drug effects; D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid; D004912:Erythrocytes / Q000187:drug effects; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D005822:Genetic Vectors; D020128:Inhibitory Concentration 50; D013058:Mass Spectrometry; D008970:Molecular Weight; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000494:pharmacology
chemical_list: D023181:Antimicrobial Cationic Peptides; D011993:Recombinant Fusion Proteins