Mutagenesis of Cytochromes P450cam and b5

Stephen G. Sligar, Djordje Filipovic, Patrick S. Stayton

Research output: Contribution to journalArticlepeer-review


This chapter discusses the mutagenesis of cytochromes P450 cam and b5. The cytochrome P450cam monooxygenase system consists of putidaredoxin reductase, a nicotinamide adenine dinucleotide (NADH)-specific flavoprotein dehydrogenase, putidaredoxin, an intermediary Fe2S2 iron–sulfur electron-transfer protein, and cytochrome P450cam. It catalyzes the regio- and stereospecific hydroxylation of camphor and initiates the complete degradation of this monoterpene to isobutyrate and acetate, thus allowing the soil bacterium Pseudomonas putida to utilize camphor as a sole carbon source. The solubility and availability of cytochrome P450cam have facilitated its biochemical, bioorganic, and biophysical characterization, making it the prototypal model of the cytochrome P450 family. The sequential cytochrome P450cam reaction cycle is rate limited by electron-transfer steps, and the chemical intermediates have been characterized to the point where the input of the second reducing equivalent initiates dioxygen bond scission.

Original languageEnglish (US)
Pages (from-to)31-49
Number of pages19
JournalMethods in enzymology
Issue numberC
StatePublished - Jan 1991

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology


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