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

Federico Sabbadin; Gideon Grogan; Neil C Bruce

doi:10.1007/978-1-62703-321-3_20

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

Federico Sabbadin, Gideon Grogan, Neil C Bruce

Research output: Contribution to journal › Article › peer-review

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.

Original language	English
Pages (from-to)	239-249
Number of pages	11
Journal	Methods in Molecular Biology
Volume	987
DOIs	https://doi.org/10.1007/978-1-62703-321-3_20
Publication status	Published - 5 Feb 2013

Keywords

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

Access to Document

10.1007/978-1-62703-321-3_20

Cite this

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

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.",

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

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AU - Grogan, Gideon

AU - Bruce, Neil C

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AB - 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.

KW - Biotransformation

KW - Cloning, Molecular

KW - Cytochrome P-450 Enzyme System

KW - Gene Library

KW - Genetic Vectors

KW - Nocardia

KW - Oxidation-Reduction

KW - Time Factors

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DO - 10.1007/978-1-62703-321-3_20

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JO - Methods in Molecular Biology

JF - Methods in Molecular Biology

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