Design of a novel P450: a functional bacterial-human cytochrome P450 chimera

Biochemistry. 1998 Jun 23;37(25):8848-52. doi: 10.1021/bi972775z.


We report the construction of a functional chimera from approximately 50% bacterial (cytosolic) cytochrome P450cam and 50% mammalian (membrane-bound) cytochrome P450 2C9. The chimeric protein shows a reduced CO-difference spectrum absorption at 446 nm, and circular dichroism spectra indicate that the protein is globular. The protein is soluble and catalyzes the oxidation of 4-chlorotoluene using molecular oxygen and reducing equivalents from bacterial putidaredoxin and putidaredoxin reductase. This chimera provides a novel method for addressing structure-function issues and may prove useful in the design of oxidants for benign and stereospecific synthesis, as well as catalysts for bioremediation of polluted areas. Furthermore, these results provide the first evidence that bacterial P450 enzymes and mammalian P450 enzymes are likely to share a common tertiary structure.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Bacterial Proteins / biosynthesis
  • Bacterial Proteins / chemistry*
  • Bacterial Proteins / genetics*
  • Binding Sites
  • Catalysis
  • Circular Dichroism
  • Cytochrome P-450 Enzyme System / biosynthesis
  • Cytochrome P-450 Enzyme System / chemical synthesis*
  • Cytochrome P-450 Enzyme System / genetics*
  • Electron Transport
  • Electrophoresis, Polyacrylamide Gel
  • Gene Expression
  • Humans
  • Models, Molecular
  • Oxygen / metabolism
  • Protein Engineering* / methods
  • Recombinant Fusion Proteins / biosynthesis
  • Recombinant Fusion Proteins / chemical synthesis*
  • Recombinant Fusion Proteins / metabolism*
  • Solubility


  • Bacterial Proteins
  • Recombinant Fusion Proteins
  • Cytochrome P-450 Enzyme System
  • Oxygen