title: In vitro and in vivo activity of the dimer of PMAP-36 expressed in Pichia pastoris.
abstract: The antimicrobial peptide PMAP-36 exists as a homodimer stabilized by an intermolecular disulfide bridge. The dimer of PMAP-36 exhibits a potent and rapid microbicidal activity against a wide spectrum of microorganisms. The gene encoding the antiparallel dimer (PMAP-36)2 was designed and codon-optimized according to bias of Pichia pastoris. The gene was then expressed in the P. pastoris strain GS115. The concentration of the recombinant product reached 106 mg/l. In vitro activity assays indicated that the recombinant peptide showed antimicrobial activities against Gram-positive and Gram-negative bacteria but did not cause hemolysis of chicken erythrocytes. Subsequently, 120 7-day-old male Arbor Acres broilers were used to evaluate the in vivo activities of the peptide. A prophylactic dose of ciprofloxacin lactate was supplemented as the control. The results showed that recombinant (PMAP-36)2 significantly increased the serum IgM content of the birds (p < 0.05). The recombinant peptide significantly increased the amounts of Bifidobacterium and decreased the amount of Escherichia coli cells in the ceca of the experimental birds (p < 0.05). The results obtained in the present study indicate that the recombinant (PMAP-36)2 has a potent in vitro and in vivo activity and can be used as an alternative to antibiotic treatment.
authors.affiliation: Laboratory of Molecular Nutrition and Immunity, Institute of Animal Nutrition, Northeast Agricultural University, Harbin, PR China.
authors.forename: Liang
authors.initials: L
authors.lastname: Wang
authors.forename: Hao
authors.initials: H
authors.lastname: Zhang
authors.forename: Zhiqiang
authors.initials: Z
authors.lastname: Jia
authors.forename: Qingquan
authors.initials: Q
authors.lastname: Ma
authors.forename: Na
authors.initials: N
authors.lastname: Dong
authors.forename: Anshan
authors.initials: A
authors.lastname: Shan
mesh_terms: D000818:Animals; D023181:Antimicrobial Cationic Peptides / Q000008:administration & dosage* / Q000235:genetics / Q000378:metabolism*; D058491:Bacterial Load; D001644:Bifidobacterium / Q000302:isolation & purification; D002432:Cecum / Q000382:microbiology; D002645:Chickens; D004912:Erythrocytes / Q000187:drug effects; D004926:Escherichia coli / Q000302:isolation & purification; D015870:Gene Expression; D006090:Gram-Negative Bacteria / Q000187:drug effects; D006094:Gram-Positive Bacteria / Q000187:drug effects; D010843:Pichia / Q000235:genetics; D015202:Protein Engineering; D055503:Protein Multimerization; D011506:Proteins / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000008:administration & dosage / Q000235:genetics / Q000378:metabolism
chemical_list: D023181:Antimicrobial Cationic Peptides; C085474:PMAP-36; D011506:Proteins; D011994:Recombinant Proteins