title: Production of small antibacterial peptides using silkworm-baculovirus protein expression system.
abstract: The recombinant proteins with strong antimicrobial activity are known to be very difficult to express using bacterial expression system. Here, human β-defensin (DEFB) 1, DEFB2, and DEFB3 were successfully produced using a silkworm-baculovirus protein expression system. We have generated four baculoviruses for each DEFB protein to compare the effect of different peptide tags in secretion into silkworm larval hemolymph. Interestingly, the best performing peptide tags for the secretion were different among DEFBs: C-terminal GST-H8 tag for DEFB1, N-terminal H8 tag for DEFB2, and C-terminal H8 tag for DEFB3, respectively. In addition, the colony count assay demonstrated that the recombinant DEFB2 s showed antimicrobial activities against Escherichia coli, Pseudomonas aeruginosa, and Paenibacillus thiaminolyticus.
authors.affiliation: Laboratory of Silkworm Science, Kyushu University Graduate School of Bioresource and Bioenvironmental Sciences, Fukuoka, Japan.
authors.forename: Mai
authors.initials: M
authors.lastname: Fukushima
authors.forename: Kazuhiro
authors.initials: K
authors.lastname: Iiyama
authors.forename: Jun
authors.initials: J
authors.lastname: Yamashita
authors.forename: Masutaka
authors.initials: M
authors.lastname: Furue
authors.forename: Gaku
authors.initials: G
authors.lastname: Tsuji
authors.forename: Shigeo
authors.initials: S
authors.lastname: Imanishi
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Mon
authors.forename: Jae Man
authors.initials: JM
authors.lastname: Lee
authors.forename: Takahiro
authors.initials: T
authors.lastname: Kusakabe
mesh_terms: D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D001709:Biotechnology / Q000379:methods*; D012831:Bombyx / Q000235:genetics*; D015169:Colony Count, Microbial; D004926:Escherichia coli / Q000187:drug effects; D006458:Hemolymph / Q000378:metabolism; D007814:Larva; D008826:Microbial Sensitivity Tests; D015202:Protein Engineering / Q000379:methods*; D011550:Pseudomonas aeruginosa / Q000187:drug effects; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D023083:beta-Defensins / Q000096:biosynthesis / Q000235:genetics* / Q000494:pharmacology*
chemical_list: C406407:DEFB1 protein, human; C422950:DEFB103A protein, human; C106641:DEFB4A protein, human; D011993:Recombinant Fusion Proteins; D023083:beta-Defensins