title: Rapid reconstitution of biosynthetic machinery for fungal metabolites in Aspergillus oryzae: total biosynthesis of aflatrem.
abstract: Reconstitution of the biosynthetic machinery for fungal secondary metabolites in Aspergillus oryzae provides an opportunity both for stepwise determination of the biosynthetic pathways and the total biosynthesis of fungal natural products. However, to maximize the utility of the reconstitution system, a simple and rapid strategy for the introduction of heterologous genes into A. oryzae is required. In this study, we demonstrated an effective method for introducing multiple genes involved in the biosynthesis of fungal metabolites by using the expression vectors pUARA2 and pUSA2, each of which contains two cloning sites. The successful introduction of all the aflatrem biosynthetic genes (seven genes in total) after two rounds of transformation enabled the total biosynthesis of aflatrem. This rapid reconstitution strategy will facilitate the functional analysis of the biosynthetic machinery of fungal metabolites.
authors.affiliation: Division of Chemistry, Graduate School of Science, Hokkaido University, Sapporo 060-0810 (Japan).
authors.forename: Koichi
authors.initials: K
authors.lastname: Tagami
authors.forename: Atsushi
authors.initials: A
authors.lastname: Minami
authors.forename: Ryuya
authors.initials: R
authors.lastname: Fujii
authors.forename: Chengwei
authors.initials: C
authors.lastname: Liu
authors.forename: Mizuki
authors.initials: M
authors.lastname: Tanaka
authors.forename: Katsuya
authors.initials: K
authors.lastname: Gomi
authors.forename: Tohru
authors.initials: T
authors.lastname: Dairi
authors.forename: Hideaki
authors.initials: H
authors.lastname: Oikawa
mesh_terms: D001236:Aspergillus oryzae / Q000235:genetics / Q000378:metabolism*; D053898:Biosynthetic Pathways; D005800:Genes, Fungal; D005822:Genetic Vectors / Q000235:genetics*; D007211:Indoles / Q000378:metabolism*; D005810:Multigene Family; D010407:Penicillium / Q000235:genetics / Q000378:metabolism; D014170:Transformation, Genetic
chemical_list: D007211:Indoles; C019239:aflatrem
keywords: Aspergillus oryzae; aflatrem; gene expression; indole diterpene; total biosynthesis