ヌクレオシドオキシダーゼを固定したベンゾキノン含有グラファイトペースト電極およびその応用

概要

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Nucleoside oxidase, which catalyzes the oxidation of nucleoside to the corresponding 5'-carboxylic acid with forming hydrogen peroxide, was purified to homogeneity from cells of Flavobacterium meningosepticum. This enzyme was immobilized onto the surface of the graphite paste electrode containing benzoquinone and was covered with a dialysis membrane. This enzyme electrode produced biocatalytic current in buffer solutions of pH 6-10. Responses by adenosine, guanosine and inosine were the same and the magnitudes of responses by uridine, cytidine and thymidine were 4/5 as active as those by inosine. This electrode did not respond to bases, sugars and nucleotides. The activity of 5'-nucleotidase in serum was determined easily by this electrode and the content of nucleotides in food also determined by the use of nucleotidase. These methods were based on the measurement of the increase in anodic current by the nucleoside that was released enzymatically, and so were not interfered with by the oxidizable compounds in a sample.
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