Nucleoside oxidase and process for making same, and process and kit for using same
First Claim
1. Nucleoside oxidase having the following substrate specificity and enzymatic action:
- substrate specificity on a nucleoside of the formula [I] ##STR8## wherein Base means nucleic acid base residue, enzymatic action to catalyze at least one enzymatic reaction of the formulae [II] and [III] ##STR9##
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Abstract
A microorganism strain B-0781 belonging to the genus Pseudomonas isolated from a soil sample from an onion field in Japan, produces a novel enzyme nucleoside oxidase having substrate specificity on various nucleosides and enzyme action to catalyze enzymatic reactions involving various nucleosides. The novel nucleoside oxidase is produced by culturing a nucleoside-oxidase-producing microorganism belonging to the genus Pseudomonas in a nutrient medium containing assimilable carbon and nitrogen and inorganic salt, and isolating the thus-formed nucleoside oxidase from the cultured cells. Various nucleoside-5'"'"'-carboxylic acids can be produced, by incubating the novel nucleoside oxidase with a nucleoside having a 5'"'"'-hydroxymethyl group, in an aqueous medium under aerobic conditions, and isolating the thus-formed nucleoside-5'"'"'-carboxylic acid from the incubation medium. Assay methods for nucleosides in liquid samples are provided, which comprise incubating the sample with nucleoside oxidase, thereby consuming oxygen and generating hydrogen peroxide and nucleoside-5'"'"'-carboxylic acid by acting on nucleoside, and quantitatively determining consumed oxygen or liberated hydrogen peroxide. A kit for nucleoside assay is provided, which contains the recited ingredients.
8 Citations
15 Claims
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1. Nucleoside oxidase having the following substrate specificity and enzymatic action:
substrate specificity on a nucleoside of the formula [I] ##STR8## wherein Base means nucleic acid base residue, enzymatic action to catalyze at least one enzymatic reaction of the formulae [II] and [III] ##STR9## - View Dependent Claims (2)
- 3. A process for the production of nucleoside oxidase which comprises culturing a nucleoside-oxidase-producing microorganism belonging to the genus Pseudomonas in a nutrient medium containing an assimilable carbon source, assimilable nitrogen source and inorganic salt, and isolating the thus-formed nucleoside oxidase from the cultured cells.
- 5. A process for the production of a nucleoside-5'"'"'-carboxylic acid, which comprises incubating nucleoside oxidase with a nucleoside having a 5'"'"'-hydroxymethyl group, in an aqueous medium under aerobic conditions, and isolating the thus-formed nucleoside-5'"'"'-carboxylic acid from the incubation medium.
- 7. An assay method for a nucleoside in a liquid sample, which comprises incubating the said sample with nucleoside oxidase, which consumes oxygen and generates hydrogen peroxide and nucleoside-5'"'"'-carboxylic acid by acting on nucleoside, and quantitatively determining consumed oxygen or generated hydrogen peroxide.
- 12. A kit for nucleoside assay, which comprises at least nucleoside oxidase which consumes oxygen and generates hydrogen peroxide and nucleoside-5'"'"'-carboxylic acid by acting on a nucleoside.
Specification