Cloning of the gene cluster responsible for the biosynthesis of brasilicardin A, a unique diterpenoid.
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.
Hayashi
Yutaka
Y
Biotechnology Research Center, Toyama Prefectural University, Toyama, Japan.
Matsuura
Nobuyasu
N
Toshima
Hiroaki
H
Itoh
Nobuya
N
Ishikawa
Jun
J
Mikami
Yuzuru
Y
Dairi
Tohru
T
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
D000617:Aminoglycosides; D004224:Diterpenes; C118259:brasilicardin A