A novel, simple, and sensitive colorimetric method to determine aromatic amino acid aminotransferase activity using the Salkowski reagent

This study describes the development of a new colorimetric assay to determine aromatic amino acid aminotransferase (ArAT) activity. The assay is based on the transamination of l-tryptophan in the presence of 2-oxoglutarate, which yields indole-3-pyruvate (IPyA). The amount of IPyA formed was quantif...

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Detalles Bibliográficos
Autores principales: Szkop, Michał, Sikora, Piotr, Orzechowski, Sławomir
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer Netherlands 2011
Materias:
Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3297745/
https://www.ncbi.nlm.nih.gov/pubmed/22130693
http://dx.doi.org/10.1007/s12223-011-0089-y
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author Szkop, Michał
Sikora, Piotr
Orzechowski, Sławomir
author_facet Szkop, Michał
Sikora, Piotr
Orzechowski, Sławomir
author_sort Szkop, Michał
collection PubMed
description This study describes the development of a new colorimetric assay to determine aromatic amino acid aminotransferase (ArAT) activity. The assay is based on the transamination of l-tryptophan in the presence of 2-oxoglutarate, which yields indole-3-pyruvate (IPyA). The amount of IPyA formed was quantified by reaction with the Salkowski reagent. Optimized assay conditions are presented for ArAT isozymes isolated from Pseudomonas putida. For comparative purposes, ArAT activity was also determined by high-performance liquid chromatography. ArAT activity staining in polyacrylamide gels with the Salkowski reagent is also presented.
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spelling pubmed-32977452012-03-21 A novel, simple, and sensitive colorimetric method to determine aromatic amino acid aminotransferase activity using the Salkowski reagent Szkop, Michał Sikora, Piotr Orzechowski, Sławomir Folia Microbiol (Praha) Article This study describes the development of a new colorimetric assay to determine aromatic amino acid aminotransferase (ArAT) activity. The assay is based on the transamination of l-tryptophan in the presence of 2-oxoglutarate, which yields indole-3-pyruvate (IPyA). The amount of IPyA formed was quantified by reaction with the Salkowski reagent. Optimized assay conditions are presented for ArAT isozymes isolated from Pseudomonas putida. For comparative purposes, ArAT activity was also determined by high-performance liquid chromatography. ArAT activity staining in polyacrylamide gels with the Salkowski reagent is also presented. Springer Netherlands 2011-12-01 2012 /pmc/articles/PMC3297745/ /pubmed/22130693 http://dx.doi.org/10.1007/s12223-011-0089-y Text en © The Author(s) 2011 https://creativecommons.org/licenses/by-nc/4.0/ This article is distributed under the terms of the Creative Commons Attribution Noncommercial License which permits any noncommercial use, distribution, and reproduction in any medium, provided the original author(s) and source are credited.
spellingShingle Article
Szkop, Michał
Sikora, Piotr
Orzechowski, Sławomir
A novel, simple, and sensitive colorimetric method to determine aromatic amino acid aminotransferase activity using the Salkowski reagent
title A novel, simple, and sensitive colorimetric method to determine aromatic amino acid aminotransferase activity using the Salkowski reagent
title_full A novel, simple, and sensitive colorimetric method to determine aromatic amino acid aminotransferase activity using the Salkowski reagent
title_fullStr A novel, simple, and sensitive colorimetric method to determine aromatic amino acid aminotransferase activity using the Salkowski reagent
title_full_unstemmed A novel, simple, and sensitive colorimetric method to determine aromatic amino acid aminotransferase activity using the Salkowski reagent
title_short A novel, simple, and sensitive colorimetric method to determine aromatic amino acid aminotransferase activity using the Salkowski reagent
title_sort novel, simple, and sensitive colorimetric method to determine aromatic amino acid aminotransferase activity using the salkowski reagent
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3297745/
https://www.ncbi.nlm.nih.gov/pubmed/22130693
http://dx.doi.org/10.1007/s12223-011-0089-y
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