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Withanolide biosynthesis recruits both mevalonate and DOXP pathways of isoprenogenesis in Ashwagandha Withania somnifera L. (Dunal)

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Abstract

Withanolides are pharmaceutically important C28-phytochemicals produced in most prodigal amounts and diversified forms by Withania somnifera. Metabolic origin of withanolides from triterpenoid pathway intermediates implies that isoprenogenesis could significantly govern withanolide production. In plants, isoprenogenesis occurs via two routes: mevalonate (MVA) pathway in cytosol and non-mevalonate or DOXP/MEP pathway in plastids. We have investigated relative carbon contribution of MVA and DOXP pathways to withanolide biosynthesis in W. somnifera. The quantitative NMR-based biosynthetic study involved tracing of 13C label from 13C1-d-glucose to withaferin A in withanolide producing in vitro microshoot cultures of the plant. Enrichment of 13C abundance at each carbon of withaferin A from 13C1-glucose-fed cultures was monitored by normalization and integration of NMR signal intensities. The pattern of carbon position-specific 13C enrichment of withaferin A was analyzed by a retro-biosynthetic approach using a squalene-intermediated metabolic model of withanolide (withaferin A) biosynthesis. The pattern suggested that both DOXP and MVA pathways of isoprenogenesis were significantly involved in withanolide biosynthesis with their relative contribution on the ratio of 25:75, respectively. The results have been discussed in a new conceptual line of biosynthetic load-driven model of relative recruitment of DOXP and MVA pathways for biosynthesis of isoprenoids.

Key message The study elucidates significant contribution of DOXP pathway to withanolide biosynthesis. A new connotation of biosynthetic load-based role of DOXP/MVA recruitment in isoprenoid biosynthesis has been proposed.

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Acknowledgments

The authors are thankful to CSIR, New Delhi, for the financial support in the form of a NMITLI Project on Ashwagandha and to Director, CIMAP for providing the requisite facilities. FS also thanks CSIR, India, for the Senior Research Fellowship.

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Author notes

  1. Narayan D. Chaurasiya

    Present address: National Centre for Natural Products Research (NCNPR), Thad Cochran Research Center, School of Pharmacy, University of Mississippi, Mississippi, Oxford, 38677, USA

Authors and Affiliations

  1. Central Institute of Medicinal and Aromatic Plants (Council of Scientific and Industrial Research), P.O. CIMAP, Lucknow, 226015, India

    Narayan D. Chaurasiya, Neelam S. Sangwan, Farzana Sabir, Laxminarain Misra & Rajender S. Sangwan

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  1. Narayan D. Chaurasiya
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  2. Neelam S. Sangwan
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Correspondence to Rajender S. Sangwan.

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Communicated by P. Kumar.

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Chaurasiya, N.D., Sangwan, N.S., Sabir, F. et al. Withanolide biosynthesis recruits both mevalonate and DOXP pathways of isoprenogenesis in Ashwagandha Withania somnifera L. (Dunal). Plant Cell Rep 31, 1889–1897 (2012). https://doi.org/10.1007/s00299-012-1302-4

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  • DOI: https://doi.org/10.1007/s00299-012-1302-4

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