title: [Secreted expression of the combinant antimicrobial peptide PL in Pichia pastoris and its antibacterial activity in vitro].
abstract: In order to obtain a high activity antibacterial peptide, An expression vector pPICZalphaA-pl is constructed with a tandem of four antimicrobial peptides in the same direction,which includes Protegrin-1 (PG-1), Scorpion Defensin (SD), Metalnikowin-2A and Sheep Myeloid Antibacterial Peptide (SMAP-29) (serial number in GenBank are AAB27599, AAAB27538, P80409 and P49928 respectively). At the same time the expression vector pPICZalphaA-sd which express Scorpion Defensin was contructed. The expression vectors of pPICZalphaA-pl and pPICZalphaA-sd were linearized and transformed into the yeast host strain X-33 respectively. Under the control of the promoter AOX1 (alcohol oxidase1), the peptides PL and SD were secreted expressed. Their heat-stable property, acid-stable property and MIC were detected in vitro. The results suggest the peptides PL and SD have good heat-stable and acid-stable properties, and the combinant PL peptide showes higher antibacterial activity against several Gram-positive bacteria (G+) and Gram-negative bacteria (G-) than the peptide SD, especially against Escherichia coli. The antibacterial activity of combinant antimicrobial peptide PL shows its far exploiting perspective.
authors.affiliation: Key Laboratory of Animal Disease Diagnosis and Immunology, Ministry of Agriculture, Nanjing Agricultural University, Nanjing 210095, China.
authors.forename: Ming-Fu
authors.initials: MF
authors.lastname: Niu
authors.forename: Xiang
authors.initials: X
authors.lastname: Li
authors.forename: Rui-Bing
authors.initials: RB
authors.lastname: Cao
authors.forename: Bin
authors.initials: B
authors.lastname: Zhou
authors.forename: De-Sheng
authors.initials: DS
authors.lastname: Chen
authors.forename: Pu-Yan
authors.initials: PY
authors.lastname: Chen
mesh_terms: D000818:Animals; D000890:Anti-Infective Agents / Q000378:metabolism* / Q000494:pharmacology*; D023181:Antimicrobial Cationic Peptides / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D001412:Bacillus subtilis / Q000187:drug effects / Q000254:growth & development; D001798:Blood Proteins / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D054804:Cathelicidins; D023082:Defensins / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D004925:Escherichia / Q000187:drug effects / Q000254:growth & development; D006863:Hydrogen-Ion Concentration; D010843:Pichia / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology*; D012475:Salmonella / Q000187:drug effects / Q000254:growth & development; D012605:Scorpions / Q000378:metabolism; D012756:Sheep / Q000378:metabolism; D013211:Staphylococcus aureus / Q000187:drug effects / Q000254:growth & development; D013997:Time Factors
chemical_list: D000890:Anti-Infective Agents; D023181:Antimicrobial Cationic Peptides; D001798:Blood Proteins; D054804:Cathelicidins; D023082:Defensins; D011993:Recombinant Fusion Proteins; C403305:SMAP29 protein, Ovis aries; C094880:protegrin-1