Biosynthesis of Terpenoid–Pyrrolobenzoxazine Hybrid Natural Product CJ‐12662

Abstract

The fungal natural product CJ-12662 is a structurally complex terpene-amino acid hybrid, and is a potent anthelmintic compound. The biosynthetic pathway of CJ-12662 is elucidated based on metabolite analysis from heterologous expression. We demonstrate the terpene portion is derived from successive P450-catalyzed oxidations of amorpha-4,11-diene, while three flavin-dependent enzymes are involved in morphing the esterified tryptophan into a chlorinated pyrrolobenzoxazine, utilizing a cascaded [1,2]-Meisenheimer rearrangement.

Deciphering chemical logic of fungal natural product biosynthesis through heterologous expression and genome mining

“…10C). 134 Amorpha-4,11-diene, a famous plant terpene that is the precursor to artemisinin, is synthesized by the TC ThmB and triply hydroxylated by three P450 enzymes in the BGC. The resulting triol is esterified regioselectively at the C 2′ alcohol with l -tryptophan by the NRPS ThmA.…”

Biosynthesis of pyrroloindoline-containing natural products

“…terpene cyclase (ThmB) cyclizes the farnesyl pyrophosphate (FPP) to form the amorphadiene (116), which is subsequently converted by ThmI, ThmG, ThmF, ThmK, and ThmJ into the sesquiterpene (120). Notably, three hydroxylation reactions on the sesquiterpene, mediated by three different P450s, follow the same catalytic mechanism (one-electron and radical-based oxidation process).…”

“…Through heterologous expression and in vitro assays, they firstly illustrated the biosynthetic pathways of 49 and 48 , which include a number of enzymes, such as NRPS (ThmA), PKS (ThmK), terpene cyclase (TC, ThmB), P450s (ThmF, ThmG and ThmI), FMOs (ThmC, ThmH and ThmM) and MT (ThmD). 116 Although many enzymes are involved in their biosynthetic pathways, their biosynthesis can be mainly divided into three stages (Fig. 14), as follows.…”

Depside Bond Formation by the Starter-Unit Acyltransferase Domain of a Fungal Polyketide Synthase

“…This possible pathway is supported by the fact that an in vitro enzymatic reaction of purified DrcB with 1 , l -threonine, adenosine triphosphate (ATP), and magnesium chloride afforded 2 (Figure S5). Although single-module NRPSs with an A–T–C T domain configuration have been identified in fungal natural product pathways, to the best of our knowledge, the biosynthesis of 2 is the first example of a pathway in which a single-module NRPS is coupled with an NR-PKS.…”