title: Production and purification of a novel antibiotic peptide, adenoregulin, from a recombinant Escherichia coli.
abstract: Adenoregulin is a member of dermaseptin family which are vertebrate antibiotic peptides having lethal effects against a broad spectrum of bacteria, fungi and protozoa. The 99 bp adenoregulin gene was cloned in the expression vector pET32a and transformed into Escherichia coli BL21(DE3). In fed-batch cultivation of BL21(DE3)/pET32a-adr, an exponential feeding strategy was applied to gain 60 g dry cells l-1. The recombinant fusion protein Trx-ADR was expressed in a soluble form. The fusion protein was isolated by Ni2+-chelating chromatography, cleaved with CNBr and purified to homogeneity through reverse phase-HPLC and size exclusion-HPLC. The purified recombinant adenoregulin had antibacterial activity against Escherichia coli K12D31 with apparent Mr of 3.4 kDa, identical to the anticipated value.
authors.affiliation: State Key Laboratory of Bioreactor Engineering New World Institute of Biotechnology, East China University of Science and Technology, Shanghai, 200237, P.R. China.
authors.forename: Yu-Xun
authors.initials: YX
authors.lastname: Zhou
authors.forename: Wei
authors.initials: W
authors.lastname: Cao
authors.forename: Qing-Ping
authors.initials: QP
authors.lastname: Luo
authors.forename: Yu-Shu
authors.initials: YS
authors.lastname: Ma
authors.forename: Jin-Zhi
authors.initials: JZ
authors.lastname: Wang
authors.forename: Dong-Zhi
authors.initials: DZ
authors.lastname: Wei
mesh_terms: D029845:Amphibian Proteins / Q000096:biosynthesis / Q000235:genetics; D000818:Animals; D023181:Antimicrobial Cationic Peptides / Q000096:biosynthesis* / Q000235:genetics; D001001:Anura; D019149:Bioreactors / Q000382:microbiology; D002455:Cell Division / Q000187:drug effects; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000166:cytology / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000302:isolation & purification / Q000494:pharmacology; D013879:Thioredoxins / Q000235:genetics
chemical_list: D029845:Amphibian Proteins; D023181:Antimicrobial Cationic Peptides; D011993:Recombinant Fusion Proteins; C485283:dermaseptin B2, Phyllomedusa bicolor; D013879:Thioredoxins