Expression in Escherichia coli of a Cytochrome P450 Enzyme with a Cobalt Protoporphyrin IX Prosthetic Group

  • Wesley E. Straub
  • Clinton R. Nishida
  • Paul R. Ortiz de Montellano
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 987)

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.

Key words

Cobalt protoporphyrin IX Cobalt heme CYP119 Heme modification Bacterial ferrochelatase 

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Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  • Wesley E. Straub
    • 1
  • Clinton R. Nishida
    • 2
  • Paul R. Ortiz de Montellano
    • 2
  1. 1.Life TechnologiesSan FranciscoUSA
  2. 2.Department of Pharmaceutical ChemistryUniversity of CaliforniaSan FranciscoUSA

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