Assays of D-amino acid oxidases.

Gabriella Tedeschi, Loredano Pollegioni, Armando Negri

Dipartimento di Patologia Animale, Igiene e Sanità Pubblica Veterinaria - sez. Biochimica, Università degli Studi di Milano, Milano, Italy. grabriella.tedeschi@unimi.it

Methods in molecular biology (Clifton, N.J.) 2012

filter terms:

D-Amino acid oxidase and D-aspartate oxidase are two well-known FAD-containing flavooxidases that catalyze the same reaction (the oxidative deamination) on different D-amino acids. D-aspartate oxidase is specific for acidic D-amino acids (i.e., D-aspartate and D-glutamate) and D-amino acid oxidase is active on neutral and polar D-amino acids (a low activity is also detected on basic D-amino acids). The assay of these flavoenzymes is of utmost importance in different fields because D-amino acids are common constituents of bacterial cell walls, are present in foods and because free D-serine and D-aspartic acid were identified in brain and peripheral tissues of mammals. In this chapter, we report on the most used methods employed to assay the activity of D-amino acid oxidase and D-aspartate oxidase. Interestingly, their activity can be followed using different assays, namely D-amino acid or oxygen consumption, α-keto acid or ammonia production, or using artificial dyes as final indicator of the flavin redox reaction.

Citation

Gabriella Tedeschi, Loredano Pollegioni, Armando Negri. Assays of D-amino acid oxidases. Methods in molecular biology (Clifton, N.J.). 2012;794:381-95