|
KMID : 1025520010430040465
|
|
Journal of Animal Science and Technology
2001 Volume.43 No. 4 p.465 ~ p.476
|
|
|
Expression of Antimicrobial Cationic Peptides by Pichia pastoris(Methylotrophic Yeast)
|
|
|
|
|
Abstract
|
|
|
|
Antimicrobial cationic peptides have been received increasing attention as natural antibiotics for their broad spectrum of antimicrobial activities and remarkably low cytotoxicity against normal mammalian cells. Pichia pastoris, a methylotrophic yeast, is an outstanding host for high degree of heterologous gene expression. Four candidates of antimicrobial cation peptides (CPs; Lactofemicin, Magainin, Protegrin-1 and Indolicidin) were expressed using pPIC9K-MPM vector containing AOXI promoter and MPM (modified Promagainin) as an acidic fusion partner. The MPM-CPs were expressed and induced by methanol induction method. The cell extracts, solubilized, were subjected to cyanogen bromide cleavage. SDS-PAGE (16.5% tricine) analysis showed the size of Protegrin-1 at 2.1kDa and of Indolicidin at 1.4kDa. The antimicrobial activity of Protegrin-1 or Indolicidin gene integrated transformant was observed by measuring clearing zones on 1% bacto-peptone agar plate against Gram-negative bacteria Escherichia coli XL-1 blue (10^5 CFU/㎖) or Gram-positive bacteria Staphylococcus aureus (10^5 CFU/㎖).
|
|
|
KEYWORD
|
|
|
|
|
|
FullTexts / Linksout information
|
|
|
|
|
|
Listed journal information
|
|
|
|