The present invention provides an industrially advantageous process for producing glyoxylic acid by a biochemical procedure using oxidase. In particular, the present invention provides novel (D)-2-hydroxy-acid oxidase acting on a 2-hycoxy acid to produce a corresponding 2-keto acid, and a process for producing glyoxylic acid in which the 2-hydroxy-acid oxidase, a culture solution of microorganisms capable of producing the enzyme, microorganism cells isolated from the culture solution, or a product of treatment of microorganism cells acts on glycolic acid to convert the glycolic acid to glyoxylic acid.
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