Medium/High Throughput D-Amino Acid Oxidase Colorimetric Method for Determination of D-Amino Acids. Application for Amino Acid Racemases
Armand Berneman, Marcelo Alves-Ferreira, Nicolas Coatnoan, Nathalie Chamond and Paola Minoprio*
Institut Pasteur, Laboratoire des Processus Infectieux à Trypanosoma, Département d’Infection & Epidémiologie, 25, rue du Dr Roux, 75724, Paris, France
- *Corresponding Author:
- Dr. Paola Minoprio
Laboratoire des Processus Infectieux à Trypanosoma
Département d’Infection & Epidémiologie
25, rue du Dr Roux, 75724, Paris, France
E-mail: [email protected]
Received September 30, 2010; Accepted date: October 30, 2010; Published date: November 02, 2010
Citation: Berneman A, Alves-Ferreira M, Coatnoan N, Chamond N, Minoprio P (2010) Medium/High Throughput D-Amino Acid Oxidase Colorimetric Method for Determination of D-Amino Acids. Application for Amino Acid Racemases. J Microbial Biochem Technol 2:139-146. doi:10.4172/1948-5948.1000039
Copyright: © 2010 Berneman A, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution and reproduction in any medium, provided the original author and source are credited.
Free amino acids detection and quantification in samples still calls for elaborated, powerful and usually high cost and sometimes time-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 based on the enzyme D-amino acid oxidase and the generation of stoechiometric amounts of hydrogen peroxide. Inclusion of horseradish peroxidase and OPD allows the determination of D- amino acid concentration through the absorbance of the reaction products as compared to serial dilutions of equimolar amounts of L/D amino acid standards. The methodology is easily adapted for a medium/high throughput analysis of amino acid racemase activities.