Protein having fructosyl valyl histidine oxidase activity, modified protein, and use of the protein or the modified protein

• US 8,993,255 B2
• Filed: 10/06/2009
• Issued: 03/31/2015
• Est. Priority Date: 10/10/2008
• Status: Active Grant

• Current Assignee: Toyobo Company Limited
• Original Assignee: Gekkeikan Sake Co., Ltd., Toyoda Gosei Company Limited (Toyota Motor Corporation)
• Inventors: Hirao, Rie, Kitabayashi, Masao, Nishiya, Yoshiaki, Ishida, Hiroki, Hata, Yoji
• Primary Examiner(s): HOBBS, LISA JOE
• Application Number:
  US13/123,052
• Publication Number:
  US 20110195444A1
• Time in Patent Office: 2,002 Days
• Field of Search: None
• US Class Current: 435/25
• CPC Class Codes: C12N 9/0022 with oxygen as acceptor (1.4.3)
• CPC Class Codes: C12Q 1/26 involving oxidoreductase
• CPC Class Codes: G01N 33/723 Glycosylated haemoglobin

Abstract

The present invention provides: a protein having a fructosyl amino acid oxidase activity which protein is useful for measurement of a glycosylated protein (particularly, glycosylated hemoglobin); a modified protein thereof; and use of the protein or the modified protein. The protein of the present invention is, for example, a fructosyl valyl histidine oxidase derived from Phaeosphaeria nodorum, the fructosyl valyl histidine oxidase having excellent thermal stability and substrate specificity and also having a small Km value to fructosyl valyl histidine. This allows a glycosylated protein measuring reagent to be stored in a long time and measurement accuracy of the glycosylated protein measuring reagent to be improved.

Claims

1. A protein having a fructosyl valyl histidine oxidase activity, the protein being any one of proteins (I) through (III), below:

(I) a protein comprising the amino acid sequence shown in SEQ ID NO;

1;

(II) a protein comprising the amino acid sequence shown in SEQ ID NO;

1 in which 10 or less amino acid residues are replaced, deleted, inserted and/or added, the protein having the fructosyl valyl histidine oxidase activity; and

(III) a protein comprising an amino acid sequence having 90% or greater homology with the amino acid sequence shown in SEQ ID NO;

1, the protein having the fructosyl valyl histidine oxidase activity,

wherein, in the proteins of (II) and (III), glycine at 110th position from the amino terminal is replaced with an amino acid selected from the group consisting of methionine, threonine, cysteine, histidine, asparagine, arginine, serine, valine, leucine, asparagine acid, isoleucine, tyrosine, phenylalanine, glutamine, lysine, glutamic acid and alanine.

2. The protein as set forth in claim 1, wherein:

the protein is the protein described in (II) or (III), and isoleucine at 58th position from the amino terminal is replaced with another amino acid.

3. The protein as set forth in claim 1, wherein:

the protein is the protein described in (II) or (III), and phenylalanine at 282th position from the amino terminal is replaced with another amino acid.

4. The protein as set forth in claim 2, wherein:

the isoleucine at 58th position from the amino terminal is replaced with methionine, threonine, alanine, asparagine, serine, valine or leucine.

5. The protein as set forth in claim 3, wherein:

the phenylalanine at 282th position from the amino terminal is replaced with tyrosine.

6. A method for measuring a glycosylated protein, comprising the step of causing a protein as set forth in claim 1 to act on a glycosylated amine.

7. A method for measuring glycosylated hemoglobin included in a sample, comprising the steps of:

preparing a glycosylated amine by use of the glycosylated hemoglobin included in the sample; and

causing a protein as set forth in claim 1 to act on the sample.