Genome mining in Trichoderma viride J1-030: discovery and identification of novel sesquiterpene synthase and its products

Abstract

Sesquiterpene synthases in Trichoderma viride have been seldom studied, despite the efficiency of filamentous fungi for terpenoid production. Using the farnesyl diphosphate-overexpressing Saccharomyces cerevisiae platform to produce diverse terpenoids, we herein identified an unknown sesquiterpene synthase from T. viride by genome mining and determined the structure of its corresponding products. One new 5/6 bicyclic sesquiterpene and its esterified derivative were characterised by GC–MS and 1D and 2D NMR spectroscopy. To the best of our knowledge, this is the first well-identified sesquiterpene synthase from T. viride to date.

Molecular methods unravel the biosynthetic potential of Trichoderma species

“…47,52 Terpenes are considered the most abundant and chemically diverse class of natural products, with an estimated number of more than 80 000 different compounds. [53][54][55] These highly diverse natural products are derived from the cyclization of polyprenyl diphosphates such as geranyl diphosphate (GPP), farnesyl diphosphate (FPP), geranylgeranyl diphosphate (GGPP) and geranylfarnesyl diphosphate (GFPP) through the action of terpene cyclases (TC). [53][54][55][56] Several terpenoids have been characterized and reported from different Trichoderma spp., however only few of their responsible biosynthetic gene clusters have been experimentally characterized.…”

“…[53][54][55] These highly diverse natural products are derived from the cyclization of polyprenyl diphosphates such as geranyl diphosphate (GPP), farnesyl diphosphate (FPP), geranylgeranyl diphosphate (GGPP) and geranylfarnesyl diphosphate (GFPP) through the action of terpene cyclases (TC). [53][54][55][56] Several terpenoids have been characterized and reported from different Trichoderma spp., however only few of their responsible biosynthetic gene clusters have been experimentally characterized. 53,57 Nevertheless the interest in the terpene synthases inventory of different Trichoderma spp.…”

“…This produced a new 5/6 bicyclic brasilane-type sesquiterpene 72 (Scheme 5). 55 In a parallel experiment by a different group, Murai et al 54 identied a terpene cyclase gene in T. atroviride FKI-3849 (TaTS) that is not clustered with other biosynthetic genes. Heterologous expression of this gene in A. oryzae also resulted in the production of the brasilane-type sesquiterpene trichobrasilenol 72.…”

Biosynthesis of oxygenated brasilane terpene glycosides involves a promiscuous N-acetylglucosamine transferase

“…[3][4][5][6][7][8] Recently, two groups have independently reported the characterisation of the terpene cyclase responsible for brasilane biosynthesis in Trichoderma spp. 9,10 These reports facilitated the search for the underlying biosynthetic gene clusters in the Hypoxylaceae which led us to investigate the participating tailoring enzymes.…”

“…Recently, two independent studies found and characterized the terpene cyclases (TaTC6, Tvi09626) responsible for brasilane backbone assembly in Trichoderma spp. 9,10 Using BLASTp homology searches with Tvi09626 as a template, we found a candidate biosynthetic gene cluster (BGC, bra) in the A. truncatum genome containing five genes termed braA-E (Scheme 1). BraA was identified as a terpene cyclase showing 70.9% similarity to Tvi09626, while BraB was annotated as a putative O-glycosyltransferase (O-GT) with homology to the O-GT PaGT involved in fusicoccin biosynthesis in Diaporthe (Phomopsis) amygdali.…”