Heterologous expression of cytotoxic sesquiterpenoids from the medicinal mushroom Lignosus rhinocerotis in yeast

Abstract

BackgroundGenome mining facilitated by heterologous systems is an emerging approach to access the chemical diversity encoded in basidiomycete genomes. In this study, three sesquiterpene synthase genes, GME3634, GME3638, and GME9210, which were highly expressed in the sclerotium of the medicinal mushroom Lignosus rhinocerotis, were cloned and heterologously expressed in a yeast system.ResultsMetabolite profile analysis of the yeast culture extracts by GC–MS showed the production of several sesquiterpene alcohols (C15H26O), including cadinols and germacrene D-4-ol as major products. Other detected sesquiterpenes include selina-6-en-4-ol, β-elemene, β-cubebene, and cedrene. Two purified major compounds namely (+)-torreyol and α-cadinol synthesised by GME3638 and GME3634 respectively, are stereoisomers and their chemical structures were confirmed by 1H and 13C NMR. Phylogenetic analysis revealed that GME3638 and GME3634 are a pair of orthologues, and are grouped together with terpene synthases that synthesise cadinenes and related sesquiterpenes. (+)-Torreyol and α-cadinol were tested against a panel of human cancer cell lines and the latter was found to exhibit selective potent cytotoxicity in breast adenocarcinoma cells (MCF7) with IC50 value of 3.5 ± 0.58 μg/ml while α-cadinol is less active (IC50 = 18.0 ± 3.27 μg/ml).ConclusionsThis demonstrates that yeast-based genome mining, guided by transcriptomics, is a promising approach for uncovering bioactive compounds from medicinal mushrooms.Electronic supplementary materialThe online version of this article (doi:10.1186/s12934-017-0713-x) contains supplementary material, which is available to authorized users.

Inhibition of Protein Glycation by Tiger Milk Mushroom [Lignosus rhinocerus (Cooke) Ryvarden] and Search for Potential Anti-diabetic Activity-Related Metabolic Pathways by Genomic and Transcriptomic Data Mining

“…A large number of transcripts responsible for zeaxanthin, antheraxanthin, and violaxanthin interconversion in carotenoid (also known as tetraterpenoid) biosynthesis were observed and this was followed by the biosynthesis of other types of terpenes and terpenoids including hemiterpenes, sesqui-, mono-, di-, and triterpenoids. This was not unexpected, as previous study on L. rhinocerus transcriptome analysis revealed a few highly expressed gene clusters that encode for sesquiterpene synthases which may be responsible for the production of several sesquiterpene alcohols (C 15 H 26 O), including cadinols and germacrene D -4-ol ( Yap et al, 2015a , 2017 ).…”

“…Elucidation of L. rhinocerus genome provided a better insight into its biology and brought new prospects for exploration ( Yap et al, 2014 ). Yeast-based genome mining guided by transcriptomics approach which has been reported recently has enabled the discovery of several bioactive sesquiterpenoids, namely (+)-torreyol and α-cadinol from this mushroom ( Yap et al, 2017 ).…”

Insight into metabolic diversity of the brown‐rot basidiomycete Postia placenta responsible for sesquiterpene biosynthesis: semi‐comprehensive screening of cytochrome P450 monooxygenase involved in protoilludene metabolism

“…S5) (Schmidt et al, 1999;Wawrzyn et al, 2012;Quin et al, 2014). In addition, the PpSTSs found in class-I and -II were unique because these STSs generated multiple products, and this may represent a catalytic feature for members of these clades (Agger et al, 2009;Wawrzyn et al, 2012;Yap et al, 2017). In contrast, PpSTSs found in the class-III clade (PpSTS-08, -09 and -14) produced single major products.…”

“…These cyclization events give rise to several hundred distinct types of sesquiterpene scaffolds (Miller and Allemann, 2012). Recently, fungal genome projects have revealed the presence of a series of STSs in basidiomycetes and some of these STSs have been functionally characterized; thus, providing new insights into the biochemistry and potential biotechnological uses of basidiomycetous STSs (Agger et al, 2009;Engels et al, 2011;Wawrzyn et al, 2012;Yap et al, 2017). In particular, STSs producing protoilludene have been investigated intensively because sesquiterpenoids derived from this compound are potential anticancer, antifungal and antibiotic agents (Abraham, 2001).…”

Identification of sesquiterpene synthases from the Basidiomycota Coniophora puteana for the efficient and highly selective β-copaene and cubebol production in E. coli

“…To provide insights into the structural and functional diversity of STPSs from Basidiomycota, we carried out a comprehensive phylogenetic analysis. This analysis encompassed the STPS sequences of Antrodia cinnamomea [ 39 ], Armillaria gallica [ 40 ], Boreostereum vibrans [ 41 ], Coprinopsis cinereus [ 35 ], Fomitopsis pinicola [ 42 ], Lignosus rhinocerus [ 43 ] Stereum hirsutum [ 37 ] and Omphalotus olearius [ 42 ], which are all functionally characterized. The phylogenetic analysis suggests that Copu1-3 belong to the same STPS clade, sharing a 1,10-cyclization mechanism with other related enzymes in this group (Fig.…”