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LICRED: a versatile drop-in vector for rapid generation of redox-self-sufficient cytochromes P450

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JournalMethods in Molecular Biology
DatePublished - 5 Feb 2013
Volume987
Number of pages11
Pages (from-to)239-249
Original languageEnglish

Abstract

Cytochromes P450 (P450s) are a family of heme-containing oxidases with considerable potential as tools for industrial biocatalysis. Organismal genomes are revealing thousands of gene sequences that encode P450s of as yet unknown function, the exploitation of which will require high-throughput tools for their isolation and characterization. Here, we describe a new ligation-independent cloning vector (LICRED) that enables the high-throughput generation of libraries of redox-self-sufficient P450s, by fusing a range of P450 heme domains to the reductase of P450RhF (RhF-Red) in a robust and generically applicable way.

    Research areas

  • Biotransformation, Cloning, Molecular, Cytochrome P-450 Enzyme System, Gene Library, Genetic Vectors, Nocardia, Oxidation-Reduction, Time Factors

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