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一种使用 Salkowski 试剂测定芳香族氨基酸氨基转移酶活性的新颖、简单、灵敏的比色法。

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

机构信息

Department of Biochemistry, Faculty of Agriculture and Biology, Warsaw University of Life Sciences, SGGW, Poland.

出版信息

Folia Microbiol (Praha). 2012 Jan;57(1):1-4. doi: 10.1007/s12223-011-0089-y. Epub 2011 Dec 1.

DOI:10.1007/s12223-011-0089-y
PMID:22130693
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC3297745/
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.

摘要

本研究描述了一种新的比色法测定芳香族氨基酸氨基转移酶（ArAT）活性的方法。该测定法基于在 2-氧代戊二酸存在下 L-色氨酸的转氨基作用，生成吲哚-3-丙酮酸（IPyA）。形成的 IPyA 量通过与 Salkowski 试剂反应进行定量。为从假单胞菌属中分离的 ArAT 同工酶呈现了优化的测定条件。为了比较目的，还通过高效液相色谱法测定了 ArAT 活性。还呈现了用 Salkowski 试剂在聚丙烯酰胺凝胶中进行的 ArAT 活性染色。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0035/3297745/d06b1040423e/12223_2011_89_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0035/3297745/f395d4c26423/12223_2011_89_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0035/3297745/c035d4a8eada/12223_2011_89_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0035/3297745/6468ce35ad7b/12223_2011_89_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0035/3297745/d06b1040423e/12223_2011_89_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0035/3297745/f395d4c26423/12223_2011_89_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0035/3297745/c035d4a8eada/12223_2011_89_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0035/3297745/6468ce35ad7b/12223_2011_89_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0035/3297745/d06b1040423e/12223_2011_89_Fig4_HTML.jpg

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