@article {Cao:2012:0929-8665:1064,
title = "Expression in Pichia pastoris and Biological Activity of Avian -defensin 6 and its Mutant Peptide Without Cysteines",
journal = "Protein and Peptide Letters",
parent_itemid = "infobike://ben/ppl",
publishercode ="ben",
year = "2012",
volume = "19",
number = "10",
publication date ="2012-10-01T00:00:00",
pages = "1064-1070",
itemtype = "ARTICLE",
issn = "0929-8665",
url = "https://www.ingentaconnect.com/content/ben/ppl/2012/00000019/00000010/art00008",
doi = "doi:10.2174/092986612802762660",
keyword = "Avian β-Defensin 6, recombinant plasmids, pichia pastoris, expression, antimicrobial peptide, disulfide bonds, Cys residues, hemolytic activity",
author = "Cao, Yanping and Ma, Qingquan and Shan, Anshan and Dong, Na",
abstract = "Avian -defensin 6 (AvBD-6) is an antimicrobial peptide that plays significant roles in the innate immunity of chickens. To explore the effects of disulfide bonds on antimicrobial activity of AvBD-6, two peptides with or without Cys residues were designed and expressed in Pichia
Pastoris. The peptide AvBD-6-B was obtained by removing six Cys residues of AvBD-6. According to the codon bias of Pichia Pastoris, the genes of AvBD-6 and AvBD-6-B were synthesized. The Bgl II-linearized recombinant plasmids pGAPHM-AvBD-6 and pGAPHM-AvBD-6-B were transformed into
Pichia Pastoris GS115 by electroporation. The recombinant AvBD-6 and AvBD-6-B were expressed in YPD for 48 h, 72 h and 96 h at 30 \textdegreeC Tricine-SDS-PAGE analysis demonstrated that both AvBD-6 and AvBD-6-B were expressed in Pichia Pastoris. The concentration of recombinant AvBD-6 and AvBD-6-B
reached 114.9 mg/l and 93.8 mg/l, respectively. The expression products exhibited antimicrobial activity against Gram-positive and Gram-negative bacteria. Antimicrobial activity of AvBD-6-B suggests that the removal of six Cys residues had no significant effect on the antimicrobial activity
of avian -defensins. Neither peptide showed hemolytic activity. This study could serve as an impetus for the production of this antimicrobial peptide as a replacement for antibiotics in animal feed.",
}