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Title of Journal: Biotechnol Lett

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Abbravation: Biotechnology Letters

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Springer Netherlands

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DOI

10.1002/chin.199628274

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1573-6776

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Isotopic labeling of the heme cofactor in cytochro

Authors: David Bryson PekLang Lim Alin Lawson Sandhya Manjunath Gregory M Raner
Publish Date: 2011/06/10
Volume: 33, Issue: 10, Pages: 2019-2026
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Abstract

A recombinant bacterial expression system that generates 13Clabeled heme or 15Nlabeled heme in functional cytochrome P450 enzymes and other hemecontaining systems is reported here using a mutant strain of Escherichia coli HU227 in which the HemA gene is inactive By synthesizing several isotopomers of aminolevulinic acid with 13C or 15N at different locations isotopes have been incorporated with high abundance into the heme cofactor of five different cytochrome P450 isoforms along with one peroxidase Confirmed both 13C and 15Nincorporation spectral and catalytic assays show the labeled enzymes produced in this system are functional

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