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Article
Biosynthesis of Diterpenoids in Tripterygium Adventitious Root Cultures
Plant Physiology
  • Fainmarinat S. Inabuy, Washington State University
  • Justin T. Fischedick, Washington State University
  • Iris Lange, Washington State University
  • Michael Hartmann, Washington State University
  • Narayanan Srividya, Washington State University
  • Amber N. Parrish, Washington State University
  • Meimei Xu, Iowa State University
  • Reuben J. Peters, Iowa State University
  • B. Markus Lange, Washington State University
Document Type
Article
Publication Version
Published Version
Publication Date
9-1-2017
DOI
10.1104/pp.17.00659
Abstract

Adventitious root cultures were developed from Tripterygium regelii Sprague & Takeda and growth conditions optimized for the abundant production of diterpenoids, which can be collected directly from the medium. An analysis of publicly available transcriptome data sets collected with T. regelii roots and root cultures indicated the presence of a large gene family (with 20 members) for terpene synthases (TPSs). Nine candidate diterpene synthase genes were selected for follow-up functional evaluation, of which two belonged to the TPS-c, three to the TPS-e/f and four to the TPS-b subfamily. These genes were characterized by heterologous expression in a modular metabolic engineering system in E. coli. Members of the TPS-c subfamily were characterized as copalyl diphosphate (diterpene) synthases and those belonging to the TPS-e/f family catalyzed the formation of precursors of kaurane diterpenoids. The TPS-b subfamily encompassed genes coding for enzymes involved in abietane diterpenoid biosynthesis and others with activities as monoterpene synthases. The structural characterization of diterpenoids accumulating in the medium of T. regelii adventitious root cultures, facilitated by searching the Spektraris online spectral database, enabled us to formulate a biosynthetic pathway for the biosynthesis of triptolide, a diterpenoid with pharmaceutical potential. Considering the significant enrichment of diterpenoids in the culture medium, fast-growing adventitious root cultures may hold promise as a sustainable resource for the large-scale production of triptolide.

Comments

This article is published as Inabuy, F.X., Fischedick, J.T., Lange, I., Hartmann, M., Srividya, N., Parrish, A.N., Xu, M., Peters, R.J., Lange, B.M. (2017) “Biosynthesis of diterpenoids in Tripterygium adventitious root cultures”, Plant Physiol., 175(1):92-103. doi: 10.1104/pp.17.00659. Posted with permission.

Copyright Owner
American Society of Plant Biologists
Language
en
File Format
application/pdf
Citation Information
Fainmarinat S. Inabuy, Justin T. Fischedick, Iris Lange, Michael Hartmann, et al.. "Biosynthesis of Diterpenoids in Tripterygium Adventitious Root Cultures" Plant Physiology Vol. 175 Iss. 1 (2017) p. 92 - 103
Available at: http://works.bepress.com/reuben_peters/74/