Development of a high-throughput screening method for racemase activity and its application to the identification of alanine racemase variants with activity towards l-arginine

Simon C. Willies; Jemma L. White; Nicholas J. Turner

doi:10.1016/j.tet.2012.06.062

Development of a high-throughput screening method for racemase activity and its application to the identification of alanine racemase variants with activity towards l-arginine

Simon C. Willies, Jemma L. White, Nicholas J. Turner

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

Abstract

A high-throughput solid phase screening method has been developed for alanine racemase. Conversion of an l-amino acid to the d-amino acid is detected by a d-amino acid oxidase, which is utilised as a reporter enzyme along with horseradish peroxidase and a colourimetric dye. This system has successfully been used to identify a mutant with increased activity towards arginine. Kinetic analysis and modelling studies have been performed to understand the effect of the mutations found. © 2012 Elsevier Ltd. All rights reserved.

Original language	English
Pages (from-to)	7564-7567
Number of pages	3
Journal	Tetrahedron
Volume	68
Issue number	37
DOIs	https://doi.org/10.1016/j.tet.2012.06.062
Publication status	Published - 16 Sept 2012

Keywords

Amino acid
Biocatalysis
High-throughput screening
Protein engineering
Racemase

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10.1016/j.tet.2012.06.062

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title = "Development of a high-throughput screening method for racemase activity and its application to the identification of alanine racemase variants with activity towards l-arginine",

abstract = "A high-throughput solid phase screening method has been developed for alanine racemase. Conversion of an l-amino acid to the d-amino acid is detected by a d-amino acid oxidase, which is utilised as a reporter enzyme along with horseradish peroxidase and a colourimetric dye. This system has successfully been used to identify a mutant with increased activity towards arginine. Kinetic analysis and modelling studies have been performed to understand the effect of the mutations found. {\textcopyright} 2012 Elsevier Ltd. All rights reserved.",

keywords = "Amino acid, Biocatalysis, High-throughput screening, Protein engineering, Racemase",

author = "Willies, {Simon C.} and White, {Jemma L.} and Turner, {Nicholas J.}",

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AU - Willies, Simon C.

AU - White, Jemma L.

AU - Turner, Nicholas J.

PY - 2012/9/16

Y1 - 2012/9/16

N2 - A high-throughput solid phase screening method has been developed for alanine racemase. Conversion of an l-amino acid to the d-amino acid is detected by a d-amino acid oxidase, which is utilised as a reporter enzyme along with horseradish peroxidase and a colourimetric dye. This system has successfully been used to identify a mutant with increased activity towards arginine. Kinetic analysis and modelling studies have been performed to understand the effect of the mutations found. © 2012 Elsevier Ltd. All rights reserved.

AB - A high-throughput solid phase screening method has been developed for alanine racemase. Conversion of an l-amino acid to the d-amino acid is detected by a d-amino acid oxidase, which is utilised as a reporter enzyme along with horseradish peroxidase and a colourimetric dye. This system has successfully been used to identify a mutant with increased activity towards arginine. Kinetic analysis and modelling studies have been performed to understand the effect of the mutations found. © 2012 Elsevier Ltd. All rights reserved.

KW - Amino acid

KW - Biocatalysis

KW - High-throughput screening

KW - Protein engineering

KW - Racemase

U2 - 10.1016/j.tet.2012.06.062

DO - 10.1016/j.tet.2012.06.062

M3 - Article

SN - 0040-4020

VL - 68

SP - 7564

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JO - Tetrahedron

JF - Tetrahedron

IS - 37

ER -

Development of a high-throughput screening method for racemase activity and its application to the identification of alanine racemase variants with activity towards l-arginine

Abstract

Keywords

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