title: A novel, simple, and sensitive colorimetric method to determine aromatic amino acid aminotransferase activity using the Salkowski reagent.
abstract: 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.
authors: Department of Biochemistry, Faculty of Agriculture and Biology, Warsaw University of Life Sciences, SGGW, Poland. michal_szkop@sggw.pl
authors: Michał
authors: M
authors: Szkop
authors: Piotr
authors: P
authors: Sikora
authors: Sławomir
authors: S
authors: Orzechowski
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry* / Q000378:metabolism; D003124:Colorimetry / Q000379:methods*; D057075:Enzyme Assays / Q000379:methods*; D007202:Indicators and Reagents; D007700:Kinetics; D016958:Pseudomonas putida / Q000737:chemistry / Q000201:enzymology*; D000637:Transaminases / Q000737:chemistry* / Q000378:metabolism; D014364:Tryptophan / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D007202:Indicators and Reagents; D014364:Tryptophan; D000637:Transaminases; C019729:aromatic amino acid aminotransferase