The In Vitro Production of prFMN for Reconstitution of UbiD Enzymes

Stephen A Marshall, Karl Fisher, David Leys

Research output: Contribution to journalArticlepeer-review


Prenylated flavin (prFMN) is a modified FMN cofactor, the isoalloxazine is extended by an additional six membered nonaromatic ring. The modification confers azomethine ylide characteristics on the oxidised prFMN, allowing it to support the reversible nonoxidative decarboxylation of unsaturated acids by the UbiD family of decarboxylases. In absence of a chemical synthesis route for prFMN, enzymatic production by the flavin prenyltransferase, UbiX, is required for in vitro reconstitution of prFMN-dependent enzymes. Here we provide an overview of the methods for producing prFMN in vitro using the flavin prenyltransferase UbiX, and the subsequent reconstitution and activation of UbiD enzymes.

Original languageEnglish
Pages (from-to)219-227
Number of pages9
JournalMethods in molecular biology (Clifton, N.J.)
Publication statusPublished - 23 Mar 2021


  • Bacterial Proteins/metabolism
  • Carboxy-Lyases/metabolism
  • Enzyme Activation
  • Flavins/metabolism
  • Oxidation-Reduction
  • Prenylation
  • Pseudomonas aeruginosa/enzymology


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