Diterpene synthases of the biosynthetic system of medicinally active diterpenoids in Marrubium vulgare

Plant J. 2014 Sep;79(6):914-27. doi: 10.1111/tpj.12589. Epub 2014 Jul 23.


Marrubium vulgare (Lamiaceae) is a medicinal plant whose major bioactive compounds, marrubiin and other labdane-related furanoid diterpenoids, have potential applications as anti-diabetics, analgesics or vasorelaxants. Metabolite and transcriptome profiling of M. vulgare leaves identified five different candidate diterpene synthases (diTPSs) of the TPS-c and TPS-e/f clades. We describe the in vitro and in vivo functional characterization of the M. vulgare diTPS family. In addition to MvEKS ent-kaurene synthase of general metabolism, we identified three diTPSs of specialized metabolism: MvCPS3 (+)-copalyl diphosphate synthase, and the functional diTPS pair MvCPS1 and MvELS. In a sequential reaction, MvCPS1 and MvELS produce a unique oxygenated diterpene scaffold 9,13-epoxy-labd-14-ene en route to marrubiin and an array of related compounds. In contrast with previously known diTPSs that introduce a hydroxyl group at carbon C-8 of the labdane backbone, the MvCPS1-catalyzed reaction proceeds via oxygenation of an intermediate carbocation at C-9, yielding the bicyclic peregrinol diphosphate. MvELS belongs to a subgroup of the diTPS TPS-e/f clade with unusual βα-domain architecture. MvELS is active in vitro and in vivo with three different prenyl diphosphate substrates forming the marrubiin precursor 9,13-epoxy-labd-14-ene, as identified by nuclear magnetic resonance (NMR) analysis, manoyl oxide and miltiradiene. MvELS fills a central position in the biosynthetic system that forms the foundation for the diverse repertoire of Marrubium diterpenoids. Co-expression of MvCPS1 and MvELS in engineered E. coli and Nicotiana benthamiana offers opportunities for producing precursors for an array of biologically active diterpenoids.

Keywords: Marrubium vulgare; diterpene synthase; diterpenoid biosynthesis; marrubiin; medicinal plants; natural product biosynthesis; plant specialized metabolism.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alkyl and Aryl Transferases
  • Amino Acid Sequence
  • Diterpenes / metabolism*
  • Escherichia coli / genetics
  • Marrubium / chemistry
  • Marrubium / enzymology*
  • Marrubium / genetics
  • Molecular Sequence Data
  • Organophosphates
  • Phylogeny
  • Plant Proteins
  • Plants, Medicinal


  • Diterpenes
  • Organophosphates
  • Plant Proteins
  • copalyl diphosphate
  • labdane
  • manoyl oxide
  • miltiradiene
  • marrubiin
  • Alkyl and Aryl Transferases
  • ent-kaurene synthetase A
  • ent-kaurene synthetase B

Associated data

  • GENBANK/KJ584450
  • GENBANK/KJ584451
  • GENBANK/KJ584452
  • GENBANK/KJ584453
  • GENBANK/KJ584454