Total biosynthesis of antiangiogenic agent (-)-terpestacin by artificial reconstitution of the biosynthetic machinery in Aspergillus oryzae.

The total biosynthesis of (-)-terpestacin was achieved by heterologous expression of four biosynthetic enzyme genes (tpcA-D) in Aspergillus oryzae. After construction of preterpestacin I by the action of bifunctional terpene synthase (TpcA), two cytochrome P450s (TpcBC) activate inert C-H bond to install three hydroxyl groups on the A-ring in stereo- and regioselective manners. Subsequently, a flavin-dependent oxidase (TpcD) catalyzes oxidation of the vicinal diol moiety to give a alpha-diketone, which undergoes an enolization to furnish terpestacin. The successful synthesis of structurally elaborated terpestacin showed that a reconstitution approach that harnesses several biosynthetic enzyme genes in A. oryzae could be a promising alternative to the current chemical synthesis of natural terpenoids.