Phylogenomics and metabolic engineering reveal a conserved gene cluster in Solanaceae plants for withanolide biosynthesis

Abstract Withanolides are steroidal lactones from nightshade (Solanaceae) plants with untapped drug potential due to limited availability of minor representatives caused by lack of biosynthetic pathway knowledge. Here, we combine phylogenomics with metabolic engineering to overcome this limitation....

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Main Authors: Samuel Edward Hakim, Nancy Choudhary, Karan Malhotra, Jian Peng, Arne Bültemeier, Ahmed Arafa, Ronja Friedhoff, Maximilian Bauer, Jessica Eikenberg, Claus-Peter Witte, Marco Herde, Philipp Heretsch, Boas Pucker, Jakob Franke
Format: Article
Language:English
Published: Nature Portfolio 2025-07-01
Series:Nature Communications
Online Access:https://doi.org/10.1038/s41467-025-61686-1
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author Samuel Edward Hakim
Nancy Choudhary
Karan Malhotra
Jian Peng
Arne Bültemeier
Ahmed Arafa
Ronja Friedhoff
Maximilian Bauer
Jessica Eikenberg
Claus-Peter Witte
Marco Herde
Philipp Heretsch
Boas Pucker
Jakob Franke
author_facet Samuel Edward Hakim
Nancy Choudhary
Karan Malhotra
Jian Peng
Arne Bültemeier
Ahmed Arafa
Ronja Friedhoff
Maximilian Bauer
Jessica Eikenberg
Claus-Peter Witte
Marco Herde
Philipp Heretsch
Boas Pucker
Jakob Franke
author_sort Samuel Edward Hakim
collection DOAJ
description Abstract Withanolides are steroidal lactones from nightshade (Solanaceae) plants with untapped drug potential due to limited availability of minor representatives caused by lack of biosynthetic pathway knowledge. Here, we combine phylogenomics with metabolic engineering to overcome this limitation. By sequencing the genome of the medicinal plant ashwagandha (Withania somnifera) and comparing it with nine Solanaceae species, we discover a conserved withanolide biosynthesis gene cluster, consisting of two sub gene clusters with differing expression patterns. We establish metabolic engineering platforms in yeast (Saccharomyces cerevisiae) and the model plant Nicotiana benthamiana to reconstitute the first five oxidations of withanolide biosynthesis, catalysed by the cytochrome P450 monooxygenases CYP87G1, CYP88C7, and CYP749B2 and a short-chain dehydrogenase/reductase, producing the aglycone of withanoside V. Enzyme functions are conserved within both sub gene clusters in W. somnifera and between W. somnifera and Physalis pruinosa. Our work sets the basis for biotechnological withanolide production to unlock their pharmaceutical potential.
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spelling doaj-art-4276c67ff34e408db130c9a2a0f1946b2025-08-20T04:03:02ZengNature PortfolioNature Communications2041-17232025-07-0116112010.1038/s41467-025-61686-1Phylogenomics and metabolic engineering reveal a conserved gene cluster in Solanaceae plants for withanolide biosynthesisSamuel Edward Hakim0Nancy Choudhary1Karan Malhotra2Jian Peng3Arne Bültemeier4Ahmed Arafa5Ronja Friedhoff6Maximilian Bauer7Jessica Eikenberg8Claus-Peter Witte9Marco Herde10Philipp Heretsch11Boas Pucker12Jakob Franke13Institute of Botany, Leibniz University HannoverInstitute of Plant Biology & BRICS, TU BraunschweigInstitute of Botany, Leibniz University HannoverInstitute of Botany, Leibniz University HannoverInstitute of Botany, Leibniz University HannoverInstitute of Botany, Leibniz University HannoverInstitute of Plant Biology & BRICS, TU BraunschweigInstitute of Organic Chemistry, Leibniz University HannoverInstitute of Botany, Leibniz University HannoverDepartment of Molecular Nutrition and Biochemistry of Plants, Leibniz University HannoverDepartment of Molecular Nutrition and Biochemistry of Plants, Leibniz University HannoverInstitute of Organic Chemistry, Leibniz University HannoverInstitute of Plant Biology & BRICS, TU BraunschweigInstitute of Botany, Leibniz University HannoverAbstract Withanolides are steroidal lactones from nightshade (Solanaceae) plants with untapped drug potential due to limited availability of minor representatives caused by lack of biosynthetic pathway knowledge. Here, we combine phylogenomics with metabolic engineering to overcome this limitation. By sequencing the genome of the medicinal plant ashwagandha (Withania somnifera) and comparing it with nine Solanaceae species, we discover a conserved withanolide biosynthesis gene cluster, consisting of two sub gene clusters with differing expression patterns. We establish metabolic engineering platforms in yeast (Saccharomyces cerevisiae) and the model plant Nicotiana benthamiana to reconstitute the first five oxidations of withanolide biosynthesis, catalysed by the cytochrome P450 monooxygenases CYP87G1, CYP88C7, and CYP749B2 and a short-chain dehydrogenase/reductase, producing the aglycone of withanoside V. Enzyme functions are conserved within both sub gene clusters in W. somnifera and between W. somnifera and Physalis pruinosa. Our work sets the basis for biotechnological withanolide production to unlock their pharmaceutical potential.https://doi.org/10.1038/s41467-025-61686-1
spellingShingle Samuel Edward Hakim
Nancy Choudhary
Karan Malhotra
Jian Peng
Arne Bültemeier
Ahmed Arafa
Ronja Friedhoff
Maximilian Bauer
Jessica Eikenberg
Claus-Peter Witte
Marco Herde
Philipp Heretsch
Boas Pucker
Jakob Franke
Phylogenomics and metabolic engineering reveal a conserved gene cluster in Solanaceae plants for withanolide biosynthesis
Nature Communications
title Phylogenomics and metabolic engineering reveal a conserved gene cluster in Solanaceae plants for withanolide biosynthesis
title_full Phylogenomics and metabolic engineering reveal a conserved gene cluster in Solanaceae plants for withanolide biosynthesis
title_fullStr Phylogenomics and metabolic engineering reveal a conserved gene cluster in Solanaceae plants for withanolide biosynthesis
title_full_unstemmed Phylogenomics and metabolic engineering reveal a conserved gene cluster in Solanaceae plants for withanolide biosynthesis
title_short Phylogenomics and metabolic engineering reveal a conserved gene cluster in Solanaceae plants for withanolide biosynthesis
title_sort phylogenomics and metabolic engineering reveal a conserved gene cluster in solanaceae plants for withanolide biosynthesis
url https://doi.org/10.1038/s41467-025-61686-1
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