[Secreted expression of the combinant antimicrobial peptide PL in Pichia pastoris and its antibacterial activity in vitro].
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.
Niu
Ming-Fu
MF
Key Laboratory of Animal Disease Diagnosis and Immunology, Ministry of Agriculture, Nanjing Agricultural University, Nanjing 210095, China.
Li
Xiang
X
Cao
Rui-Bing
RB
Zhou
Bin
B
Chen
De-Sheng
DS
Chen
Pu-Yan
PY
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
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