title: Functional analyses of cytochrome P450 genes responsible for the early steps of brassicicene C biosynthesis.
abstract: We previously revealed that Orf8 and Orf6, which were identified in the brassicicene C biosynthetic gene cluster in Alternaria brassicicola strain ATCC96836, were fusicoccadiene (FD) synthase and 16-O-methyltransferase, respectively. In the present Letter, the early biosynthetic steps after the formation of FD were investigated. Plasmids carrying the FD synthase gene, one (or two) of five cytochrome P450 genes (orf1, orf2, orf5, orf7, and orf11) identified in the cluster and a cytochrome P450 reductase gene cloned from strain ATCC96836 were constructed and introduced into Saccharomyces cerevisiae. Based on the structures of the compounds produced by the transformants, Orf1 is suggested to be an 8beta-hydroxylation enzyme that yields FD 8beta-ol (4), followed by 16-hydroxylation by Orf7 to produce FD 8beta16-diol (5).
authors.affiliation: Biotechnology Research Center, Toyama Prefectural University, Imizu, Toyama 939-0398, Japan. mhashimo@iwate-med.ac.jp
authors.forename: Makoto
authors.initials: M
authors.lastname: Hashimoto
authors.forename: Yusuke
authors.initials: Y
authors.lastname: Higuchi
authors.forename: Shunji
authors.initials: S
authors.lastname: Takahashi
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Osada
authors.forename: Toshiyuki
authors.initials: T
authors.lastname: Sakaki
authors.forename: Tomonobu
authors.initials: T
authors.lastname: Toyomasu
authors.forename: Takeshi
authors.initials: T
authors.lastname: Sassa
authors.forename: Nobuo
authors.initials: N
authors.lastname: Kato
authors.forename: Tohru
authors.initials: T
authors.lastname: Dairi
mesh_terms: D000528:Alternaria / Q000235:genetics; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics* / Q000378:metabolism; D004224:Diterpenes / Q000737:chemistry / Q000378:metabolism*; D006900:Hydroxylation; D005810:Multigene Family; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D004224:Diterpenes; D011994:Recombinant Proteins; C539991:brassicicene C; D003577:Cytochrome P-450 Enzyme System