Identification of a new class of cytochrome P450 from a Rhodococcus sp

Gareth A Roberts, Gideon Grogan, Andy Greter, Sabine L Flitsch, Nicholas J Turner

Research output: Contribution to journalArticlepeer-review


A degenerate set of PCR primers were used to clone a gene encoding a cytochrome P450 (the P450RhF gene) from Rhodococcus sp. strain NCIMB 9784 which is of unique primary structural organization. Surprisingly, analysis of the translation product revealed that the P450 is fused to a reductase domain at the C terminus which displays sequence conservation for dioxygenase reductase proteins. The reductase partner comprises flavin mononucleotide- and NADH-binding motifs and a [2Fe2S] ferredoxin-like center. The gene was engineered for heterologous expression in Escherichia coli, and conditions were found in which the enzyme was produced in a soluble form. A recombinant strain of E. coli was able to mediate the O dealkylation of 7-ethoxycoumarin in good yield, despite the absence of any recombinant redox proteins. This unprecedented finding leads us to propose that P450RhF represents the first example of a new class of cytochromes P450 in which the reducing equivalents are supplied by a novel reductase in a fused arrangement.

Original languageEnglish
Pages (from-to)3898-908
Number of pages11
JournalJournal of Bacteriology
Issue number14
Publication statusPublished - Jul 2002


  • Amino Acid Sequence
  • Base Sequence
  • Cloning, Molecular
  • Cytochrome P-450 Enzyme System/chemistry
  • Molecular Sequence Data
  • Polymerase Chain Reaction
  • Rhodococcus/enzymology

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