Diterpene Synthase-Catalyzed Biosynthesis of Distinct Clerodane Stereoisomers

Chembiochem. 2019 Jan 2;20(1):111-117. doi: 10.1002/cbic.201800580. Epub 2018 Nov 28.

Abstract

The diterpene synthase clerodienyl diphosphate synthase 1 (PvCPS1) from the crop plant switchgrass (Panicum virgatum) stereoselectively converts (E,E,E)-geranylgeranyl diphosphate (GGPP) into the clerodane natural product, cis-trans-clerodienyl diphosphate (CLPP, 1). Structure-guided point mutations of PvCPS1 redirected product stereoselectivity toward the formation of a rare cis-clerodane diastereomer, cis-cis-CLPP (2). Additionally, an alternative cis-clerodane diastereomer, (5S,8S,9R,10R)-13Z-CLPP (3), was produced when treating PvCPS1 and select variants thereof with the cis-prenyl substrate (Z,Z,Z)-nerylneryl diphosphate (NNPP). These results support the hypothesis that substrate configuration and minor active-site alterations impact precatalysis substrate folding in the stereoselective biosynthesis of clerodane diterpenoid scaffolds, and can be employed to provide enzymatic access to a broader range of bioactive clerodane natural products.

Keywords: clerodane diterpenes; diterpene synthase; plant natural products; protein mutagenesis; terpenoid metabolism.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Alkyl and Aryl Transferases / chemistry*
  • Alkyl and Aryl Transferases / genetics
  • Biocatalysis
  • Catalytic Domain
  • Diterpenes, Clerodane / chemistry
  • Diterpenes, Clerodane / metabolism*
  • Models, Chemical
  • Panicum / enzymology
  • Plant Proteins / chemistry*
  • Plant Proteins / genetics
  • Point Mutation
  • Quantum Theory
  • Stereoisomerism
  • Thermodynamics

Substances

  • Diterpenes, Clerodane
  • Plant Proteins
  • Alkyl and Aryl Transferases