Expression in Escherichia coli of a cytochrome P450 enzyme with a cobalt protoporphyrin IX prosthetic group

Methods Mol Biol. 2013:987:107-13. doi: 10.1007/978-1-62703-321-3_9.

Abstract

Unlike many hemoproteins, the prosthetic heme group of most cytochrome P450 enzymes cannot be extracted and replaced by modified heme groups. Here, we describe a procedure for generating a cytochrome P450 enzyme (CYP119) with cobalt protoporphyrin IX as its prosthetic group. This is achieved by expressing the protein in Escherichia coli in iron-limited medium and adding cobalt to the medium at the moment that inducible protein expression is initiated.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Archaeal Proteins / biosynthesis*
  • Archaeal Proteins / chemistry*
  • Archaeal Proteins / genetics
  • Cytochrome P-450 Enzyme System / biosynthesis*
  • Cytochrome P-450 Enzyme System / chemistry*
  • Cytochrome P-450 Enzyme System / genetics
  • Escherichia coli / genetics*
  • Gene Expression
  • Mass Spectrometry
  • Protein Engineering / methods*
  • Protoporphyrins / metabolism*
  • Temperature

Substances

  • Archaeal Proteins
  • Protoporphyrins
  • cobaltiprotoporphyrin
  • Cytochrome P-450 Enzyme System
  • CYP119 protein, Sulfolobus solfataricus