Heterologous Biosynthesis of Fungal Indole Sesquiterpene Sespendole

Abstract

Indole sesquiterpene sespendole, which has been isolated from the filamentous fungus Pseudobotrytis terrestris FKA-25, is a specific inhibitor of lipid droplet synthesis in mouse macrophages. The biosynthetic pathway that involves genes encoding six enzymes (spdEMBQHJ) was elucidated through heterologous expression of spd genes in Aspergillus oryzae, biotransformation experiments, and in vitro enzymatic reactions with a recombinant protein, thereby revealing the mechanism underlying the characteristic modification on the indole ring, catalyzed by a set of prenyltransferase (SpdE)/cytochrome P450 (SpdJ) enzymes. Functional analysis of the homologous genes encoding these enzymes involved in the biosynthesis of lolitrem allowed a biosynthetic pathway for the bicyclic ring skeleton fused to the indole ring to be proposed.

Norditerpenoids biosynthesized by variediene synthase-associated P450 machinery along with modifications by the host cell Aspergillus oryzae

“…Considering that these three genes were adjacent to the DvVS gene, we co-expressed them with DvVS and vndE to further verify their functionalities. As many fungal biosynthetic pathways are revealed through heterologous expression of A. oryzae [ 17 , [39] , [40] , [41] , [42] ], these genes were cloned into the expression vectors pUSA2 and pUARA2 for heterologous expression in A. oryzae . Two expression plasmids, pUSA2- orfABC and pUARA2- vndDE , were prepared and transformed into A. oryzae to generate AO- orfABC/vndDE .…”

Biosynthesis of Indole Diterpene Lolitrems: Radical‐Induced Cyclization of an Epoxyalcohol Affording a Characteristic Lolitremane Skeleton

“…Comparing the chemical shifts of 1 H and 13 C NMR spectra with those of synthetic analogues, we determined the relative stereochemistry to be syn (Supporting Information, Tables S2–S4) . Although the NMR spectra of 18 , which was previously identified by the bioconversion of 2 by AO‐ spdE / ltmJ , were nearly identical to those of sespelline ( S4 ), an unnatural IDT produced by the bioconversion of 2 by AO‐ spdE / spdJ , the difference in their retention times suggested a diastereomeric relationship (Supporting Information, Figure S6).…”

Biosynthesis of indole diterpenes: a reconstitution approach in a heterologous host

“…AtmQ 41 and JanQ catalyzed oxidation of 21 at two allylic positions C13 and/or C7 to yield paspalinine ( 18 ) probably via paspalicine (Scheme 5). 42 In IST sespendole/lecanindole biosynthesis, 37 none of PaxP homolog exists on its BGC while PaxQ homolog SpdQ is found. AO- spdE1BM QH afforded lecanindole B 43 ( 31 ), suggesting that SpdQ catalyzes hydroxylation at the C9 position.…”

“…Transformant AO- spdE1MBQH E2J possessing two additional genes generated sespendole ( 30 ) via 32 (Scheme 6). 37…”