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Dear Hari, You might look at Lucy Waskell's experiences with full length mammalian Cytochrome P450 2B4. While she got >50 mg of purified protein per liter of culture, many other heme proteins are much harder to express in E. coli with heme assembly proteins, chaperones, etc. You just have to try it. However, for cytochrome P450s, you might want to resort to some molecular engineering to increase expression (a la Eric Johnson and co.; look of cytochrome P450 2C9 structure for example). Saribas, Gruenke, and Waskell (2001) Overexpression and Purification of the Membrane-Bound Cytochrome P450 2B4. Protein Expression and Purification 21, 303–309. Good luck, Michael **************************************************************** R. Michael Garavito, Ph.D. Professor of Biochemistry & Molecular Biology 513 Biochemistry Bldg. Michigan State University East Lansing, MI 48824-1319 Office: (517) 355-9724 Lab: (517) 353-9125 FAX: (517) 353-9334 Email: [log in to unmask] **************************************************************** On Aug 26, 2010, at 12:56 PM, Hari Namboodiri wrote: > Hi CCPers > > Can anyone provide insights about expressing heme containing > proteins in E.col? Does E.coli need any porphyrin precursor during > expression or you need special E.coli strains. I have references > mentioning delta-aminolevulinic acid for one ortholog but none for > another. The enzyme is CYP51. > > > Thanks > Hari