title: Cloning of the gene cluster responsible for the biosynthesis of brasilicardin A, a unique diterpenoid.
abstract: Brasilicardin A (BCA), produced by Nocardia brasiliensis IFM 0406 (currently referred to as N. terpenica), has a unique structure consisting of a diterpene skeleton with L-rhamnose, N-acetylglucosamine, amino acid, and 3-hydroxybenzoate moieties, and exhibits potent biological activities. To understand the biosynthetic machinery of this unique compound, we have cloned the corresponding gene cluster. Firstly, we cloned a gene by PCR that encodes geranylgeranyl diphosphate synthase (GGPPS), which produces a direct precursor of diterpene compounds. We obtained four candidate genes and one of the genes was confirmed to encode a GGPPS. By sequence analysis of regions flanking the GGPPS gene, we identified eleven genes (bra1-11), all oriented in the same direction. We did not, however, detect any genes related to L-rhamnose and N-acetylglucosamine biosyntheses in the flanking regions. A gene disruption experiment did indeed show that this gene cluster was responsible for BCA biosynthesis.
authors.affiliation: Biotechnology Research Center, Toyama Prefectural University, Toyama, Japan.
authors.forename: Yutaka
authors.initials: Y
authors.lastname: Hayashi
authors.forename: Nobuyasu
authors.initials: N
authors.lastname: Matsuura
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Toshima
authors.forename: Nobuya
authors.initials: N
authors.lastname: Itoh
authors.forename: Jun
authors.initials: J
authors.lastname: Ishikawa
authors.forename: Yuzuru
authors.initials: Y
authors.lastname: Mikami
authors.forename: Tohru
authors.initials: T
authors.lastname: Dairi
mesh_terms: D000595:Amino Acid Sequence; D000617:Aminoglycosides / Q000096:biosynthesis* / Q000235:genetics*; D003001:Cloning, Molecular; D004224:Diterpenes / Q000737:chemistry*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D005810:Multigene Family; D009615:Nocardia / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D000617:Aminoglycosides; D004224:Diterpenes; C118259:brasilicardin A