Alpha-glycated amino acid releasing enzyme

• US 20040247587A1
• Filed: 06/04/2004
• Published: 12/09/2004
• Est. Priority Date: 04/12/1999
• Status: Abandoned Application

• Current Assignee: ARKRAY, Inc
• Original Assignee: ARKRAY, Inc
• Inventors: Yonehara, Satoshi, Ishimaru, Kaori, Yagi, Masayuki
• Application Number:
  US10/861,280
• Publication Number:
  US 20040247587A1
• Time in Patent Office: Days
• US Class Current: 424/94.63
• CPC Class Codes: C12N 1/205 Bacterial isolates
• CPC Class Codes: C12N 9/48 acting on peptide bonds (3.4)
• CPC Class Codes: C12R 2001/01 Bacteria or Actinomycetales ; using bacteria or Actinomycetales

Abstract

The present invention relates to a novel enzyme (α-GARE) which releases an amino acid residue having a glycated α-amino group (α-GA) from a glycated protein etc. and to bacterial strains producing the same. Examples of the bacterial strains include Sphingomonas parapaucimobilis KDK1004 (FERM BP-7041). The α-GARE is contained in the culture supernatant of this strain and α-GA can be released from a glycated peptide by using the same, as shown in FIG. 1.

Claims

1-25. -25. cancelled

26. An isolated enzyme that releases an amino acid having a glycated α

-amino group from a glycated protein or a glycated peptide, wherein the enzyme is derived from Sphingomonas parapaucimobilis.

27. An enzyme according to claim 26, wherein Sphingomonas parapaucimobilis is Sphingomonas parapaucimobilis KDK 1004 (FERM BP-7041).

28. An enzyme according to claim 26, wherein the amino acid having a glycated α

-amino group to be released is valine having a glycated α

-amino group.

29. A method for determining an amount of glycated protein or a glycated peptide in a sample, comprising:

(a) degrading a glycated protein or peptide with an enzyme according to claim 26 to give a degradation product;

(b) performing a redox reaction between the degradation product and a fructosyl amino acid oxidase; and

(c) measuring a product or reactant of the redox reaction to determine the amount of glycated protein or glycated peptide.

30. A method according to claim 29, wherein the product is hydrogen peroxide and an amount of hydrogen peroxide produced by the redox reaction is measured.

31. A method according to claim 30, wherein the amount of hydrogen peroxide produced by the redox reaction is determined by measuring oxidation of a substrate by a peroxidase.

32. A method according to claim 31, wherein the substrate is chromogenic.

33. A method according to claim 29, wherein the reactant is oxygen and an amount of oxygen consumed by the redox reaction is measured.

34. A method according to claim 29, wherein the glycated protein or glycated peptide is glycated hemoglobin.

35. A kit for determining an amount of glycated protein or glycated peptide, comprising:

(a) an enzyme according to claim 26;

(b) a fructosyl amino acid oxidase;

(c) a peroxidase; and

(d) a substrate that is oxidized by the peroxidase.