アポ酵素-補酵素反応を利用したフラビンアデニンジヌクレオチドの電気化学的測定法についての検討 (<特集> バイオサイエンスと分析化学)

概要

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We employed the commercially available D-amino acid oxidase (MW=38000g/mol) which requires one flavin-adenine dinucleotide (FAD) as a cofactor of activation. It can enzymatically catalyze the oxidation of D-amino acid to 2-oxoacid in the presence of H_2O and O_2. In addition, this enzyme produces NH_3 and H_2O_2 during enzymatic reactions. This cofactor is essential for the activation of the apoenzyme. The enzyme is reactivated and the active enzyme produces electroactive H_2O_2 and NH_3 when the cofactor is attached to the specific site of the apoenzyme. Therefore we prepared appropriate amounts of apoenzyme and DL-α-alanine in buffer solution (pH 8.30). H_2O_2 was produced by the enzymatic process in response to the addition of FAD. Then the concentration of FAD was estimated from the electrochemical signal of H_2O_2 at +600mV vs. Ag/AgCl. This configuration discriminated between different concentrations of analyte in the range 0.84〜4.2μM.
社団法人日本分析化学会の論文
1995-10-05