Rapid Reconstitution of Biosynthetic Machinery for Fungal Metabolites in <i>Aspergillus oryzae</i>: Total Biosynthesis of Aflatrem

Koichi Tagami, Atsushi Minami, Ryuya Fujii, Chengwei Liu, Mizuki Tanaka, Katsuya Gomi, Tohru Dairi, Hideaki Oikawa

doi:10.1002/cbic.201402195

<jats:title>Abstract</jats:title><jats:p>Reconstitution of the biosynthetic machinery for fungal secondary metabolites in <jats:italic>Aspergillus oryzae</jats:italic> 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 <jats:italic>A. oryzae</jats:italic> 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.</jats:p>

Rapid Reconstitution of Biosynthetic Machinery for Fungal Metabolites in <i>Aspergillus oryzae</i>: Total Biosynthesis of Aflatrem

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