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

Hui Yeng Yeannie Yap, Mariano Jordi Muria-Gonzalez, Boon Hong Kong, Keith A. Stubbs, Chon Seng Tan, Szu Ting Ng, Nget Hong Tan, Peter S. Solomon, Shin Yee Fung, Yit Heng Chooi

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Background: Genome 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. Results: Metabolite 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). Conclusions: This demonstrates that yeast-based genome mining, guided by transcriptomics, is a promising approach for uncovering bioactive compounds from medicinal mushrooms.

Original languageEnglish
Article number103
JournalMicrobial Cell Factories
Volume16
Issue number1
DOIs
Publication statusPublished - 12 Jun 2017

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Sesquiterpenes
Agaricales
Yeast
Genes
Yeasts
Genome
Inhibitory Concentration 50
Terpenes
Basidiomycota
Stereoisomerism
MCF-7 Cells
Cytotoxicity
Metabolites
Adenocarcinoma
Breast
Alcohols
Cells
Nuclear magnetic resonance
Cell Line
cadinol

Cite this

Yap, Hui Yeng Yeannie ; Muria-Gonzalez, Mariano Jordi ; Kong, Boon Hong ; Stubbs, Keith A. ; Tan, Chon Seng ; Ng, Szu Ting ; Tan, Nget Hong ; Solomon, Peter S. ; Fung, Shin Yee ; Chooi, Yit Heng. / Heterologous expression of cytotoxic sesquiterpenoids from the medicinal mushroom Lignosus rhinocerotis in yeast. In: Microbial Cell Factories. 2017 ; Vol. 16, No. 1.
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abstract = "Background: Genome 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. Results: Metabolite 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). Conclusions: This demonstrates that yeast-based genome mining, guided by transcriptomics, is a promising approach for uncovering bioactive compounds from medicinal mushrooms.",
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author = "Yap, {Hui Yeng Yeannie} and Muria-Gonzalez, {Mariano Jordi} and Kong, {Boon Hong} and Stubbs, {Keith A.} and Tan, {Chon Seng} and Ng, {Szu Ting} and Tan, {Nget Hong} and Solomon, {Peter S.} and Fung, {Shin Yee} and Chooi, {Yit Heng}",
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Heterologous expression of cytotoxic sesquiterpenoids from the medicinal mushroom Lignosus rhinocerotis in yeast. / Yap, Hui Yeng Yeannie; Muria-Gonzalez, Mariano Jordi; Kong, Boon Hong; Stubbs, Keith A.; Tan, Chon Seng; Ng, Szu Ting; Tan, Nget Hong; Solomon, Peter S.; Fung, Shin Yee; Chooi, Yit Heng.

In: Microbial Cell Factories, Vol. 16, No. 1, 103, 12.06.2017.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Heterologous expression of cytotoxic sesquiterpenoids from the medicinal mushroom Lignosus rhinocerotis in yeast

AU - Yap, Hui Yeng Yeannie

AU - Muria-Gonzalez, Mariano Jordi

AU - Kong, Boon Hong

AU - Stubbs, Keith A.

AU - Tan, Chon Seng

AU - Ng, Szu Ting

AU - Tan, Nget Hong

AU - Solomon, Peter S.

AU - Fung, Shin Yee

AU - Chooi, Yit Heng

PY - 2017/6/12

Y1 - 2017/6/12

N2 - Background: Genome 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. Results: Metabolite 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). Conclusions: This demonstrates that yeast-based genome mining, guided by transcriptomics, is a promising approach for uncovering bioactive compounds from medicinal mushrooms.

AB - Background: Genome 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. Results: Metabolite 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). Conclusions: This demonstrates that yeast-based genome mining, guided by transcriptomics, is a promising approach for uncovering bioactive compounds from medicinal mushrooms.

KW - (+)-Torreyol

KW - Heterologous expression

KW - Lignosus rhinocerotis

KW - Saccharomyces cerevisiae

KW - Sesquiterpene synthase

KW - Sesquiterpenoid

KW - Tiger milk mushroom

KW - α-Cadinol

UR - http://www.scopus.com/inward/record.url?scp=85020387800&partnerID=8YFLogxK

U2 - 10.1186/s12934-017-0713-x

DO - 10.1186/s12934-017-0713-x

M3 - Article

VL - 16

JO - Microbial Cell Factories

JF - Microbial Cell Factories

SN - 1475-2859

IS - 1

M1 - 103

ER -