title: Reconstitution of biosynthetic machinery for indole-diterpene paxilline in Aspergillus oryzae.
abstract: Indole-diterpenes represented by paxilline share a common pentacyclic core skeleton derived from indole and geranylgeranyl diphosphate. To shed light on the detailed biosynthetic mechanism of the paspaline-type hexacyclic skeleton, we examined the reconstitution of paxilline biosynthetic machinery in Aspergillus oryzae NSAR1. Stepwise introduction of the six pax genes enabled us to isolate all biosynthetic intermediates and to synthesize paxilline. In vitro and in vivo studies on the key enzymes, prenyltransferase PaxC and cyclase PaxB, allowed us to elucidate actual substrates of these enzymes. Using the isolated and the synthesized epoxide substrates, the highly intriguing stepwide epoxidation/cyclization mechanism for the construction of core structure has been confirmed. In addition, we also demonstrated "tandem transformation" to simultaneously introduce two genes using a single vector (paxG/paxB, pAdeA; paxP/paxQ, pUNA). This may provide further option for the reconstitution strategy to synthesize more complex fungal metabolites.
authors: Division of Chemistry, Graduate School of Science, Hokkaido University, Sapporo 060-0810, Japan.
authors: Koichi
authors: K
authors: Tagami
authors: Chengwei
authors: C
authors: Liu
authors: Atsushi
authors: A
authors: Minami
authors: Motoyoshi
authors: M
authors: Noike
authors: Tetsuya
authors: T
authors: Isaka
authors: Shuhei
authors: S
authors: Fueki
authors: Yoshihiro
authors: Y
authors: Shichijo
authors: Hiroaki
authors: H
authors: Toshima
authors: Katsuya
authors: K
authors: Gomi
authors: Tohru
authors: T
authors: Dairi
authors: Hideaki
authors: H
authors: Oikawa
mesh_terms: D001236:Aspergillus oryzae / Q000737:chemistry / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D004224:Diterpenes / Q000737:chemistry / Q000378:metabolism*; D007211:Indoles / Q000737:chemistry / Q000378:metabolism*; D015394:Molecular Structure
chemical_list: D004224:Diterpenes; D007211:Indoles; C048220:paxilline