The self‐sufficient P450 RhF expressed in a whole cell system selectively catalyses the 5‐hydroxylation of diclofenac

Research output: Research - peer-reviewArticle

  • External authors:
  • Jan M. Klenk
  • Bernd Nebel
  • Joanne Porter
  • Justyna K. Kulig
  • Shaneela Hussain
  • Sven M. Richter
  • Michele Tavanti
  • Martin A. Hayes
  • Bernhard Hauer


P450 monooxygenases are able to catalyse the highly regio-­‐ and stereoselective oxidations of many organic molecules. However, the scale-­‐up of such bio-­‐oxidations remains challenging due to the often-­‐low activity, level of expression and stability of P450 biocatalysts. Despite these challenges they are increasingly desirable as recombinant biocatalysts, particularly for the production of drug metabolites. Diclofenac is a widely used anti-­‐inflammatory drug that is persistent in the environment along with the 4’-­‐ and 5-­‐hydroxy metabolites. Here we have used the self-­‐sufficient P450 RhF (CYP116B2) from Rhodococcus sp. in a whole cell system to reproducibly catalyse the highly regioselective oxidation of diclofenac to 5-­‐hydroxydiclofenac. The product is a human metabolite and as such is an important standard for environmental and toxicological analysis. Furthermore, access to significant quantities of 5-­‐hydroxydiclofenac has allowed us to demonstrate further oxidative degradation to the toxic quinoneimine product. Our studies demonstrate the potential for gram-­‐scale production of human drug metabolites through recombinant whole cell biocatalysis.

Bibliographical metadata

Original languageEnglish
JournalBiotechnology Journal (Print)
Early online date18 Jan 2017
StatePublished - 2017