D-Amino Acid Oxidase, Porcine Kidney (D-AAO, D-Amino acid: oxygen oxidoreductase, deaminating, DAO, OXDA)

D-Amino Acid Oxidase is a flavoprotein oxidase, flavin being the only prosthetic group. The enzyme from porcine kidney has been extensively studied. It is isolated as a stable crystalline complex with benzoate, from which the holo and apoenzyme may be prepared. The benzoate is readily exchanged for a substrate. D-Amino acid oxidase catalyzes the oxidation of D-amino acids as shown below: D-AAO RCHNH2COOH + O2 + H2O ------> RCOCOOH + NH3 + H2O2 The D isomers of alanine, methionine, valine, isoleucine, phenylalanine and proline serve as good substrates while the L isomers do not react. D-Amino acid oxidase has several possible applications such as the determination of D-amino acids, the separation of natural L- amino acid isomers from a racemic mixture and in the preparation of keto acids. The usefulness and application of D-amino acid oxidase can be significantly increased if it is available in an immobilized form (Parkin, K. and Hultin, H.O., Biotech. and Bioeng., Vol XXI, 939, 1979). Enzyme Commission (EC) Number: 1.4.3.3 (BRENDA | IUBMB ) MDL number: MFCD00081546 EG/EC Number: 232-563-5 Source: Porcine kidney Form: Supplied as a chromatographically purified, lyophilized powder. Protein: 10 mg/ml (biuret) Activity: ~10-20U/mg Protein Molecular Weight: Monomeric, 38,000-39,000 D Unit Definition: The amount of enzyme that will deaminate by oxidation one micromole of D-alanine to pyruvate per minute at pH 8.3, at 37 degrees C in the presence of catalase. The assay is based on the method described by Bergmeyer (Methods of Enzymatic Analysis, Bergmeyer, H.U. ed. Vol 1, 431, 1974, Academic Press, New York). The decrease in the absorbance at 340nm, due to the oxidation of NADH, is a measure of D-amino acid oxidase activity. Quality Control: SDS-PAGE Unit Definition: One unit will oxidatively deaminate 1umole of D-alanine to pyruvate per minute at pH 8.3 at 25 degrees C, in the presence of a catalase. Storage and Stability: 6 months at -20 degrees C