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

M. Szkop, P. Sikora, S. Orzechowski

. 2012 ; 57 (1) : 1-4.

Language English Country Czech Republic

Document type Evaluation Study, Journal Article, Research Support, Non-U.S. Gov't

Persistent link   https://www.medvik.cz/link/bmc12035165

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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$2 doi $a 10.1007/s12223-011-0089-y
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$a Szkop, Michał $u Department of Biochemistry, Faculty of Agriculture and Biology, Warsaw University of Life Sciences, SGGW, Poland. michal_szkop@sggw.pl
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$a A novel, simple, and sensitive colorimetric method to determine aromatic amino acid aminotransferase activity using the Salkowski reagent / $c M. Szkop, P. Sikora, S. Orzechowski
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$a 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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$a bakteriální proteiny $x chemie $x metabolismus $7 D001426
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$a kolorimetrie $x metody $7 D003124
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$a enzymatické testy $x metody $7 D057075
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$a indikátory a reagencie $7 D007202
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$a kinetika $7 D007700
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$a Pseudomonas putida $x chemie $x enzymologie $7 D016958
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$a transaminasy $x chemie $x metabolismus $7 D000637
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$a tryptofan $x metabolismus $7 D014364
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$a hodnotící studie $7 D023362
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$a časopisecké články $7 D016428
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$a práce podpořená grantem $7 D013485
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$a Sikora, Piotr $u Department of Biochemistry, Faculty of Agriculture and BiologyWarsaw University of Life Sciences, SGGW
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$a Orzechowski, Sławomir $u Department of Biochemistry, Faculty of Agriculture and BiologyWarsaw University of Life Sciences, SGGW
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$w MED00011005 $t Folia microbiologica $x 1874-9356 $g Roč. 57, č. 1 (2012), s. 1-4
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