Genome mining in Sorangium cellulosum So ce56: identification and characterization of the homologous electron transfer proteins of a myxobacterial cytochrome P450

J Biol Chem. 2009 Oct 16;284(42):28590-8. doi: 10.1074/jbc.M109.021717. Epub 2009 Aug 20.

Abstract

Myxobacteria, especially members of the genus Sorangium, are known for their biotechnological potential as producers of pharmaceutically valuable secondary metabolites. The biosynthesis of several of those myxobacterial compounds includes cytochrome P450 activity. Although class I cytochrome P450 enzymes occur wide-spread in bacteria and rely on ferredoxins and ferredoxin reductases as essential electron mediators, the study of these proteins is often neglected. Therefore, we decided to search in the Sorangium cellulosum So ce56 genome for putative interaction partners of cytochromes P450. In this work we report the investigation of eight myxobacterial ferredoxins and two ferredoxin reductases with respect to their activity in cytochrome P450 systems. Intriguingly, we found not only one, but two ferredoxins whose ability to sustain an endogenous So ce56 cytochrome P450 was demonstrated by CYP260A1-dependent conversion of nootkatone. Moreover, we could demonstrate that the two ferredoxins were able to receive electrons from both ferredoxin reductases. These findings indicate that S. cellulosum can alternate between different electron transport pathways to sustain cytochrome P450 activity.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biotechnology / methods
  • Computational Biology / methods
  • Cytochrome P-450 Enzyme System / genetics*
  • Electron Spin Resonance Spectroscopy
  • Electrons
  • Ferredoxins / chemistry
  • Ferricyanides / chemistry
  • Flavins / chemistry
  • Gene Expression Regulation, Bacterial
  • Genetic Techniques
  • Genome, Bacterial*
  • Glutathione Transferase / metabolism
  • Kinetics
  • Myxococcales / genetics*
  • Protein Structure, Tertiary

Substances

  • Ferredoxins
  • Ferricyanides
  • Flavins
  • hexacyanoferrate III
  • Cytochrome P-450 Enzyme System
  • Glutathione Transferase