Functional analyses of cytochrome P450 genes responsible for the early steps of brassicicene C biosynthesis.
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).
Hashimoto
Makoto
M
Biotechnology Research Center, Toyama Prefectural University, Imizu, Toyama 939-0398, Japan. mhashimo@iwate-med.ac.jp
Higuchi
Yusuke
Y
Takahashi
Shunji
S
Osada
Hiroyuki
H
Sakaki
Toshiyuki
T
Toyomasu
Tomonobu
T
Sassa
Takeshi
T
Kato
Nobuo
N
Dairi
Tohru
T
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
D004224:Diterpenes; D011994:Recombinant Proteins; C539991:brassicicene C; D003577:Cytochrome P-450 Enzyme System