[Modification of hybrid antimicrobial peptide CecA-mil gene and its over-secretion expression in Pichia pastoris].
According to the partiality codon of Pichia pastoris, hybrid antimicrobial peptide CecA-mil gene was reconstructed, synthesized and cloned into pPICZalpha-A to construct the recombinant expression vector pPICZa-A-CM. The pPICZalpha-A-CM was transformed into yeast host strain X-33. Under the control of the promoter AOX1 (alcohol oxidase 1), a approximately 1.9 kD cecA-mil protein was expressed with the high level of 245 microg/mL after optimized the requirements for the flask-shaking culture fermentation of the Pichia pastoris rX-33/pPICZalpha-A-CM. The hybrid antibacterial peptide had a broad spectrum antibacterial activity on both gram-positive and gram-negative bacteria, especially showed potent antibacterial activity against ampicillin-resistant and kanamycin- resistant bacteria, such as Staphylococcus aureus (cowan I) and pathogenic E. coli (O1) from chicken. In addition, the hybrid antibacterial peptide showed an extreme heat-stable and acid-stable characteristic. Based on these characteristics, the recombinant antibacterial peptide CecA-mil display application foreground in the field of antisepsis of food, prevention of disease, additives of animal feedstuff and so on.
Key Laboratory of Animal Disease Diagnosis and Immunology, Ministry of Agriculture, Nanjing Agricultural University, Nanjing 210095, China. zsf780331@tom.com
Su-fang
SF
Zhang
Rui-bing
RB
Cao
Yun
Y
Jia
Bin
B
Zhou
Pu-yan
PY
Chen
D000890:Anti-Infective Agents / Q000737:chemistry / Q000494:pharmacology*; D023181:Antimicrobial Cationic Peptides / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics* / Q000494:pharmacology; D001419:Bacteria / Q000187:drug effects*; D005285:Fermentation; D008826:Microbial Sensitivity Tests; D010843:Pichia / Q000235:genetics*; D014170:Transformation, Genetic
D000890:Anti-Infective Agents; D023181:Antimicrobial Cationic Peptides