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Medium/High Throughput D-Amino Acid Oxidase Colorimetric Method for Determination of D-Amino Acids. Application for Amino Acid Racemases

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Author(s): Armand Berneman | Marcelo Alves-Ferreira | Nicolas Coatnoan | Paola Minoprio

Journal: Journal of Microbial & Biochemical Technology
ISSN 1948-5948

Volume: 02;
Issue: 06;
Start page: 139;
Date: 2010;
Original page

Keywords: D-amino acids | Proline racemase | D- amino acid oxidase | amino acid racemases | Alzheimer | aging

ABSTRACT
Free amino acids detection and quantification in samples still calls for elaborated, powerful and usually high cost and sometimestime-consuming technologies such as gas chromatography, HPLC or highly selective and sensitive column-switching techniques. Here,we undertook to develop a simple, one-step, microtiter plate method to detect and/or quantify most D-amino acids in solutions basedon the enzyme D-amino acid oxidase and the generation of stoechiometric amounts of hydrogen peroxide. Inclusion of horseradishperoxidase and OPD allows the determination of D- amino acid concentration through the absorbance of the reaction products ascompared to serial dilutions of equimolar amounts of L/D amino acid standards. The methodology is easily adapted for a medium/highthroughput analysis of amino acid racemase activities.
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