title: Expression and purification of two different antimicrobial peptides, PR-39 and Protegrin-1 in Escherichia coli.
abstract: To implement coexpression of antimicrobial peptides PR-39 and Protegrin-1 (PG-1) in prokaryotic expression system, a tandem gene fragment encoding PR-39 and PG-1 has been synthesized chemically. The cleavage site (Asn-Gly) of hydroxylamine hydrochloride was introduced between PR-39 and PG-1. The fragment was inserted into vector pGEX-4T-1 and expressed in Escherichia coli. The fusions of single peptides to GST were created at the same time. The fusion protein GST-PR-39-PG-1, purified by affinity chromatography, was cleaved first by hydroxylamine hydrochloride to release recombinant PG-1 and then by enterokinase to release PR-39. Purification of recombinant PR-39 and PG-1 was achieved. About 1.9 mg/l recombinant PR-39 and 1.1 mg/l PG-1 were obtained. The recombinant antimicrobial peptides showed antibacterial activities that were similar to those released from fusions of single peptides to GST.
authors.affiliation: College of Animal Sciences, Zhejiang University, Hangzhou 310029, PR China.
authors.forename: Fu
authors.initials: F
authors.lastname: Fan
authors.forename: Yueming
authors.initials: Y
authors.lastname: Wu
authors.forename: Jianxin
authors.initials: J
authors.lastname: Liu
mesh_terms: D000595:Amino Acid Sequence; D023181:Antimicrobial Cationic Peptides / Q000235:genetics* / Q000302:isolation & purification; D002846:Chromatography, Affinity; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D005822:Genetic Vectors; D005982:Glutathione Transferase / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D023181:Antimicrobial Cationic Peptides; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; C094880:protegrin-1; C072212:PR 39; D005982:Glutathione Transferase