The Biosynthesis of Norsesquiterpene Aculenes Requires Three Cytochrome P450 Enzymes to Catalyze a Stepwise Demethylation Process

作者：Chi‐Fang Lee, Lixun Chen, Chen‐Yu Chiang, Chen‐Yu Lai, Hsiao‐Ching Lin · 发表于：Angewandte Chemie International Edition · 年份：2019 · DOI：10.1002/anie.201910200 · 被引用次数：59 · 研究领域：Plant biochemistry and biosynthesis、Natural product bioactivities and synthesis、Microbial Natural Products and Biosynthesis

) that are produced by Aspergillus aculeatus. The nordaucane skeleton in aculenes A-D may be derived from an ent-daucane precursor through demethylation, however, the enzymes involved remain unexplored. We identified the biosynthetic gene cluster and characterized the biosynthetic pathway based on gene inactivation, feeding experiments, and heterologous reconstitution in Saccharomyces cerevisiae and Aspergillus oryzae. We discovered that three cytochrome P450 monoxygenases are required to catalyze the stepwise demethylation process. AneF converts the 12-methyl group into a carboxylic acid and AneD installs the 10-hydroxy group for later tautomerization and stabilization. Finally, AneG installs an electron-withdrawing carbonyl group at the C-2 position, which triggers C-12 decarboxylation to yield the nordaucane skeleton. Additionally, a terpene cyclase (AneC) was found that forms a new product (dauca-4,7-diene).