title: Expression and purification of moricin CM4 and human β-defensins 4 in Escherichia coli using a new technology.
abstract: Different strategies have been developed to produce small antimicrobial peptides using recombinant techniques. Here we report a new technology of biosynthesis of moricin CM4 and human β-defensins 4 (HβD4) in the Escherichia coli. The CM4 and HβD4 gene were cloned into a vector containing the tags elastin-like peptide (ELP) and intein to construct the expression vector pET-EI-CM4 and pET-EI-HβD4. All the peptides, expressed as soluble fusions, were isolated from the protein debris by the method called inverse transition cycling (ITC) rather than traditional immobilized metal affinity chromatography (IMAC) and separated from the fusion leader by self-cleavage. Fully reduced peptides that were purified exhibited expected antimicrobial activity. The approach described here is a low-cost, convenient and potential way for generating small antimicrobial peptide.
authors.affiliation: Jiangsu Province Key Laboratory for Molecular and Medical Biotechnology, Life Sciences College, Nanjing Normal University, Nanjing 210046, China.
authors.forename: Yang
authors.initials: Y
authors.lastname: Shen
authors.forename: Hong-Xin
authors.initials: HX
authors.lastname: Ai
authors.forename: Ren
authors.initials: R
authors.lastname: Song
authors.forename: Zhen-Ning
authors.initials: ZN
authors.lastname: Liang
authors.forename: Jian-Feng
authors.initials: JF
authors.lastname: Li
authors.forename: Shuang-Quan
authors.initials: SQ
authors.lastname: Zhang
mesh_terms: D023181:Antimicrobial Cationic Peptides / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification* / Q000494:pharmacology; D001709:Biotechnology / Q000379:methods*; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D005822:Genetic Vectors; D006801:Humans; D019476:Insect Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification* / Q000494:pharmacology; D023083:beta-Defensins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification* / Q000494:pharmacology
chemical_list: D023181:Antimicrobial Cationic Peptides; C106641:DEFB4A protein, human; D019476:Insect Proteins; D023083:beta-Defensins; C097209:moricin protein, Bombyx mori