title: Expression in Escherichia coli of a cytochrome P450 enzyme with a cobalt protoporphyrin IX prosthetic group.
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.
authors.affiliation: Life Technologies, San Francisco, CA, USA.
authors.forename: Wesley E
authors.initials: WE
authors.lastname: Straub
authors.forename: Clinton R
authors.initials: CR
authors.lastname: Nishida
authors.forename: Paul R Ortiz
authors.initials: PR
authors.lastname: de Montellano
mesh_terms: D019843:Archaeal Proteins / Q000096:biosynthesis* / Q000737:chemistry* / Q000235:genetics; D003577:Cytochrome P-450 Enzyme System / Q000096:biosynthesis* / Q000737:chemistry* / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics*; D015870:Gene Expression; D013058:Mass Spectrometry; D015202:Protein Engineering / Q000379:methods*; D011524:Protoporphyrins / Q000378:metabolism*; D013696:Temperature
chemical_list: D019843:Archaeal Proteins; D011524:Protoporphyrins; C007095:cobaltiprotoporphyrin; D003577:Cytochrome P-450 Enzyme System; C408874:CYP119 protein, Sulfolobus solfataricus