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Novel fructosyl amino acid oxidase

An amino acid and fructosyl technology, applied in the field of preparing this oxidase, can solve the problems of low heat resistance, storage stability of fructosyl amino acid oxidase and the like

Inactive Publication Date: 2004-03-31
KIKKOMAN CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to its low heat resistance, fructosyl amino acid oxidase has problems in its storage stability when it is used as an enzyme for clinical diagnosis in kit reagents

Method used

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  • Novel fructosyl amino acid oxidase
  • Novel fructosyl amino acid oxidase
  • Novel fructosyl amino acid oxidase

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Experimental program
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Embodiment

[0085] The present invention is more specifically described using the following examples. These examples are not intended to limit the technical field of the present invention.

[0086] (A) Preparation of recombinant plasmids

[0087] Recombinant plasmid DNAs of oxidase genes (genes having the nucleotide sequence of SEQ ID NO: 3, genes encoding the amino acid sequence of SEQ ID NO: 1) and recombinant plasmid DNAs having oxidase genes were prepared respectively. , wherein the nucleotide sequence region encoding the amino terminal of the gene has been modified to be suitable for Escherichia coli using codons, thereby increasing the expression level of the gene in Escherichia coli. For the preparation of the oxidase in the present invention described below, any plasmid can be used. The use of recombinant plasmid DNA having a modified oxidase gene is effective in mass production of the oxidase in the present invention.

[0088] (1) Preparation of recombinant plasmid pFA5 DNA

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Abstract

A novel fructosyl amino acid oxidase which is excellent in heat stability. Moreover, a fructosyl amino acid oxidase gene encoding this novel fructosyl amino acid oxidase; a recombinant DNA in which this gene is inserted into a vector; a transformant or a transductant containing this gene; and a process for producing the novel fructosyl amino acid oxidase which comprises culturing the above transformant or transductant in a medium and collecting the novel fructosyl amino acid oxidase from the culture.

Description

field of invention [0001] The present invention relates to a novel fructosyl amino acid oxidase with stability at high temperature, a gene encoding the novel fructosyl amino acid oxidase, and a method for preparing the oxidase. Background of the invention [0002] In the presence of oxygen, fructosyl amino acid oxidase catalyzes the production of glyoxylic acid or α-ketoaldehydes, α-amino acids and hydrogen peroxide by oxidizing iminodiacetic acid or its derivatives (also known as "Amadori compounds") reaction. Fructosyl amino acid oxidase is found in fungi and bacteria. The present invention proposes analytical methods for measuring Amadori compounds contained in foods and produced in vivo using these enzymes (Japanese Patent Laid-Open (Kokoku) No. 5-33997, Japanese Patent Laid-Open (Kokoku) No. 6-65300, Japanese Patent Laid-Open (Kokoku) No. Kokai) No.2-195900, Japanese Patent Publication (Kokai) No.3-155780, Japanese Patent Publication (Kokai) No.4-4874, Japanese Patent...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/06
CPCC12N9/0022
Inventor 阪上了一梶山直树
Owner KIKKOMAN CORP