title: Production of antibacterial peptide from bee venom via a new strategy for heterologous expression.
abstract: Honey bee is important economic insect that not only pollinates fruits and crops but also provides products with various physiological activities. Bee venom is a functional agent that is widely applied in clinical treatment and pharmacy. Secapin is one of these agents that have a significant role in therapy. The functions of secapin from the bee venom have been documented, but little information is known about its heterologous expression under natural condition. Moreover, few scholars verified experimentally the functions of secapin from bee venom in vitro. In this study, we successfully constructed a heterologous expression vector, which is different from conventional expression system. A transgenic approach was established for transformation of secapin gene from the venom of Apis mellifera carnica (Ac-sec) into the edible fungi, Coprinus cinereus. Ac-sec was encoded by a 234 bp nucleotide that contained a signal peptide domain and two potential phosphorylation sites. The sequence exhibited highly homology with various secapins characterized from honey bee and related species. Southern blot data indicated that Ac-sec was present as single or multiple copy loci in the C. cinereus genome. By co-transformation and double-layer active assay, Ac-sec was expressed successfully in C. cinereus and the antibacterial activity of the recombinants was identified, showing notable antibacterial activities on different bacteria. Although Ac-sec is from the venom of Apidae, phylogenetic analysis demonstrated that Ac-sec was more closely related to that of Vespid than to bee species from Apidae. The molecular characteristics of Ac-sec and the potential roles of small peptides in biology were discussed.
authors.affiliation: College of Food Science, South China Agricultural University, Guangzhou, 510642, China.
authors.forename: Chunsheng
authors.initials: C
authors.lastname: Hou
authors.forename: Liqiong
authors.initials: L
authors.lastname: Guo
authors.forename: Junfang
authors.initials: J
authors.lastname: Lin
authors.forename: Linfeng
authors.initials: L
authors.lastname: You
authors.forename: Wuhua
authors.initials: W
authors.lastname: Wu
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000900:Anti-Bacterial Agents; D001514:Bee Venoms / Q000235:genetics*; D001516:Bees; D003001:Cloning, Molecular; D003302:Coprinus / Q000235:genetics*; D008969:Molecular Sequence Data; D030781:Organisms, Genetically Modified; D010802:Phylogeny; D011994:Recombinant Proteins / Q000096:biosynthesis; D016415:Sequence Alignment; D019076:Transgenes
chemical_list: D000900:Anti-Bacterial Agents; D001514:Bee Venoms; D011994:Recombinant Proteins; C029737:secapin