title: High-level production of a novel antimicrobial peptide perinerin in Escherichia coli by fusion expression.
abstract: Perinerin is a small antimicrobial peptide (AMP) isolated from an Asian marine clamworm, Perinereis aibuhitensis Grube. It shows marked activity in vitro against both Gram-negative and Gram-positive bacteria. To obtain it in large amounts, the coding sequence of perinerin was cloned into pET32a(+) vector and expression as a Trx fusion protein in Escherichia coli. The soluble fusion protein collected from the supernatant of the cell lyste was separated by Ni(2+)-chelating chromatography. The purified protein was then cleaved by Factor Xa protease to release mature perinerin. Final purification was achieved by ion-exchange chromatography. Recombinant perinerin exhibited a similar antimicrobial activity to the native perinerin. These works might provide a significant foundation for the following research on the action of mechanism of marine AMPs.
authors.affiliation: Department of Marine Biochemistry Engineer, School of Life Science and Technology China Pharmaceutical University, Nanjing, 210009, People's Republic of China.
authors.forename: Qing-Feng
authors.initials: QF
authors.lastname: Zhou
authors.forename: Xue-Gang
authors.initials: XG
authors.lastname: Luo
authors.forename: Liang
authors.initials: L
authors.lastname: Ye
authors.forename: Tao
authors.initials: T
authors.lastname: Xi
mesh_terms: D000818:Animals; D000890:Anti-Infective Agents / Q000378:metabolism* / Q000494:pharmacology; D015153:Blotting, Western; D004262:DNA Restriction Enzymes / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism; D006090:Gram-Negative Bacteria / Q000187:drug effects; D006094:Gram-Positive Bacteria / Q000187:drug effects; D008826:Microbial Sensitivity Tests; D008957:Models, Genetic; D010455:Peptides / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification / Q000378:metabolism* / Q000494:pharmacology
chemical_list: D000890:Anti-Infective Agents; D010455:Peptides; C494852:Perinerin; D011993:Recombinant Fusion Proteins; D004262:DNA Restriction Enzymes