Characterization of the gene cluster CYP264B1-geoA from Sorangium cellulosum So ce56: biosynthesis of (+)-eremophilene and its hydroxylation

Chembiochem. 2015 Jan 19;16(2):337-44. doi: 10.1002/cbic.201402443. Epub 2014 Dec 12.

Abstract

Terpenoids can be found in almost all forms of life; however, the biosynthesis of bacterial terpenoids has not been intensively studied. This study reports the identification and functional characterization of the gene cluster CYP264B1-geoA from Sorangium cellulosum So ce56. Expression of the enzymes and synthesis of their products for NMR analysis and X-ray diffraction were carried out by employing an Escherichia coli whole-cell conversion system that provides the geoA substrate farnesyl pyrophosphate through simultaneous overexpression of the mevalonate pathway genes. The geoA product was identified as a novel sesquiterpene, and assigned NMR signals unambiguously proved that geoA is an (+)-eremophilene synthase. The very tight binding of (+)-eremophilene (∼0.40 μM), which is also available in S. cellulosum So ce56, and its oxidation by CYP264B1 suggest that the CYP264B1-geoA gene cluster is required for the biosynthesis of (+)-eremophilene derivatives.

Keywords: Sorangium cellulosum So ce56; cytochromes; eremophilenes; hydroxylation; terpenes.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alkyl and Aryl Transferases / genetics
  • Alkyl and Aryl Transferases / metabolism
  • Cloning, Molecular
  • Cytochrome P-450 Enzyme System / genetics*
  • Cytochrome P-450 Enzyme System / metabolism*
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Fungal Proteins / genetics
  • Fungal Proteins / metabolism
  • Gas Chromatography-Mass Spectrometry
  • Hydroxylation
  • Magnetic Resonance Spectroscopy
  • Mevalonic Acid / analogs & derivatives
  • Mevalonic Acid / metabolism
  • Molecular Structure
  • Multigene Family*
  • Myxococcales / genetics*
  • Myxococcales / metabolism*
  • Retinoic Acid 4-Hydroxylase
  • Sesquiterpenes / chemistry
  • Sesquiterpenes / metabolism*
  • X-Ray Diffraction

Substances

  • Fungal Proteins
  • Sesquiterpenes
  • mevalonolactone
  • Cytochrome P-450 Enzyme System
  • Retinoic Acid 4-Hydroxylase
  • Alkyl and Aryl Transferases
  • terpene synthase
  • Mevalonic Acid