title: Recombinant expression and biological characterization of the antimicrobial peptide fowlicidin-2 inPichia pastoris.
abstract: Fowlicidins are a group of cathelicidin antimicrobial peptides that were initially identified in chickens. Fowlicidin-2, which is composed of 31 amino acids, is widely expressed in the majority of tissues in chickens and has an important role in innate immunity. In the present study, a recombinant expression system for fowlicidin-2 was successfully constructed usingPichia pastorisX-33 and the expression vector pPICZα-A. Under the optimized fermentation conditions, 85.6 mg fowlicidin-2 with >95% purity was obtained from 1 liter culture medium following purification by ion exchange chromatography and reversed phase high performance liquid chromatography. The recombinant fowlicidin-2 exhibited broad spectrum antimicrobial activity and had a minimum inhibitory concentration ranging from 1 to 4 µM. Furthermore, recombinant fowlicidin-2 exhibited hemolytic activity, promoting 50% human erythrocyte hemolysis in the concentration range of 128-256 µM, and anticancer activity, resulting in the death of 50% of A375 human malignant melanoma cells in the concentration range of 2-4 µM. The results of the present study suggest that recombinant fowlicidin-2 may be a promising candidate for therapeutic applications.
authors.affiliation: Department of Animal Nutrition and Feed Science, College of Animal Science and Technology, Northeast Agricultural University, Harbin, Heilongjiang 150030, P.R. China.
authors.forename: Li-Wei
authors.initials: LW
authors.lastname: Xing
authors.affiliation: Department of Animal Nutrition and Feed Science, College of Animal Science and Technology, Northeast Agricultural University, Harbin, Heilongjiang 150030, P.R. China.
authors.forename: Shi-Xun
authors.initials: SX
authors.lastname: Tian
authors.affiliation: Department of Animal Nutrition and Feed Science, College of Animal Science and Technology, Northeast Agricultural University, Harbin, Heilongjiang 150030, P.R. China.
authors.forename: Wei
authors.initials: W
authors.lastname: Gao
authors.affiliation: Department of Animal Nutrition and Feed Science, College of Animal Science and Technology, Northeast Agricultural University, Harbin, Heilongjiang 150030, P.R. China.
authors.forename: Na
authors.initials: N
authors.lastname: Yang
authors.affiliation: Department of Animal Nutrition and Feed Science, College of Animal Science and Technology, Northeast Agricultural University, Harbin, Heilongjiang 150030, P.R. China.
authors.forename: Pei
authors.initials: P
authors.lastname: Qu
authors.affiliation: Department of Animal Nutrition and Feed Science, College of Animal Science and Technology, Northeast Agricultural University, Harbin, Heilongjiang 150030, P.R. China.
authors.forename: Di
authors.initials: D
authors.lastname: Liu
authors.affiliation: Department of Animal Nutrition and Feed Science, College of Animal Science and Technology, Northeast Agricultural University, Harbin, Heilongjiang 150030, P.R. China.
authors.forename: Jian
authors.initials: J
authors.lastname: Jiao
authors.affiliation: Department of Animal Nutrition and Feed Science, College of Animal Science and Technology, Northeast Agricultural University, Harbin, Heilongjiang 150030, P.R. China.
authors.forename: Jue
authors.initials: J
authors.lastname: Wang
authors.affiliation: Department of Animal Nutrition and Feed Science, College of Animal Science and Technology, Northeast Agricultural University, Harbin, Heilongjiang 150030, P.R. China.
authors.forename: Xing-Jun
authors.initials: XJ
authors.lastname: Feng
keywords: Pichia pastoris; antibacterial activity; anticancer activity; antimicrobial peptides; fowlicidin-2; recombinant expression