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J.Health Sci., 56(3), 341-346, 2010
-Research Letter-
Utilization of Kynurenic Acid Produced from D-kynurenine in an in Vitro Assay of D-Amino Acid Oxidase Activity
Ziyu Song, Tadahiro Ogaya,
Kana Ishii, Hideaki Ichiba,
Hideaki Iizuka, and Takeshi Fukushima*
Department of Analytical Chemistry, Faculty of Pharmaceutical Sciences, Toho University, 2-2-1 Miyama, Funabashi 274-8510, Japan
D-2-Amino-4-(2-aminophenyl)-4-oxobutanoic acid (D-kynurenine) was used to assay D-amino acid oxidase (DAAO, EC 1.4.3.3) activity. D-Kynurenine is enzymatically converted by DAAO to 4-hydroxyquinoline-2-carboxylic acid (kynurenic acid), which emits fluorescence at 398 nm (excitation wavelength 251 nm) in the presence of zinc ions. Using D-kynurenine as a substrate, fluorescence intensity originating from kynurenic acid can be used to assess DAAO activity. Under enzymatic reaction at pH 8.5 (37°C), the Km and Vmax of D-kynurenine were 148 μM and 5.53 μmol/min per mg, respectively. Inhibition of DAAO by a commercial DAAO inhibitor was clearly observed in this assay.
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