Mechanistic Characterisation of Collinodiene Synthase, a Diterpene Synthase from Streptomyces collinus

Chemistry. 2023 Nov 16;29(64):e202302469. doi: 10.1002/chem.202302469. Epub 2023 Oct 2.

Abstract

Two homologs of the diterpene synthase CotB2 from Streptomyces collinus (ScCotB2) and Streptomyces iakyrus (SiCotB2) were investigated for their products by in vitro incubations of the recombinant enzymes with geranylgeranyl pyrophosphate, followed by compound isolation and structure elucidation by NMR. ScCotB2 produced the new compound collinodiene, besides the canonical CotB2 product cyclooctat-9-en-7-ol, dolabella-3,7,18-triene and dolabella-3,7,12-triene, while SiCotB2 gave mainly cyclooctat-9-en-7-ol and only traces of dolabella-3,7,18-triene. The cyclisation mechanism towards the ScCotB2 products and their absolute configurations were investigated through isotopic labelling experiments.

Keywords: NMR spectroscopy; biosynthesis; enzyme mechanisms; isotopes; terpenes.

MeSH terms

  • Bacterial Proteins / chemistry
  • Diterpenes* / chemistry
  • Ligases* / chemistry
  • Streptomyces* / enzymology

Substances

  • Diterpenes
  • Ligases
  • Bacterial Proteins

Supplementary concepts

  • Streptomyces collinus