Glucose dehydrogenase

Patent metadata

Field	Value
Publication number	US-9796963-B2
Application number	US-201314426679-A
Country	US
Kind code	B2
Filing date	Sep 6, 2013
Priority date	Sep 10, 2012
Publication date	Oct 24, 2017
Grant date	Oct 24, 2017

Abstract

The present invention provides a novel glucose dehydrogenase that has excellent substrate specificity and that is suitable for use in SMBG. The present invention provides a flavin-bound glucose dehydrogenase having the following characteristics (1) to (5): (1) Temperature stability: stable at a temperature of 45° C. or lower; (2) stable at a pH range of 4.5 to 7.5; (3) substrate specificity: the reactivity to D-xylose, maltose, or D-galactose is 2% or less, based on the reactivity to D-glucose taken as 100%; (4) optimal activity temperature: 34 to 47° C.; and (5) optimal activity pH: 6.3 to 6.7.

First claim

The invention claimed is:

1. An electrode comprising a flavin-bound glucose dehydrogenase having the following characteristics (1) to (6):
(1) temperature stability: stable at a temperature of 45° C. or lower;
(2) stable at a pH range of 4.5 to 7.5;
(3) substrate specificity: the reactivity to D-xylose is 0.5% or less, based on the reactivity to D-glucose taken as 100%;
(4) optimal activity temperature: 34 to 47° C.;
(5) optimal activity pH: 6.3 to 6.7; and
(6) a molecular weight of about 70 kDa when measured by SDS-PAGE;
wherein the flavin-bound glucose dehydrogenase is immobilized onto the electrode.

2. The electrode according to claim 1 , wherein the flavin-bound glucose dehydrogenase further has the following characteristic (7):
(7) derived from microorganisms of the genus Arthrinium and the genus Apiospora , which is the sexual stage of Arthrinium.

3. The electrode according to claim 1 , wherein the electrode is selected from the group consisting of a platinum electrode, a carbon electrode, and a gold electrode.

4. A glucose sensor comprising the electrode of claim 1 .

5. A method of measuring glucose concentration in a sample comprising:
(i) providing a solution comprising a flavin-bound glucose dehydrogenase having the following characteristics (1) to (6):
(1) temperature stability: stable at a temperature of 45° C. or lower;
(2) stable at a pH range of 4.5 to 7.5;
(3) substrate specificity: the reactivity to D-xylose is 0.5% or less, based on the reactivity to D-glucose taken as 100%;
(4) optimal activity temperature: 34 to 47° C.;
(5) optimal activity pH: 6.3 to 6.7; and
(6) a molecular weight of about 70 kDa when measured by SDS-PAGE;

and a mediator,
(ii) contacting the solution with the sample,
(iii) measuring the transmission of an electron that is generated when the flavin-bound glucose dehydrogenase dehydrolzyes glucose in the sample, and
(iv) correlating the measurement from step (iii) with a concentration of glucose in the sample.

6. The method according to claim 5 , wherein the mediator is at least one selected from the group consisting of potassium ferricyanide and phenazine methosulfate.

7. The method according to claim 5 , wherein the flavin-bound glucose dehydrogenase further has the following characteristic (7):
(7) derived from microorganisms of the genus Arthrinium and the genus Apiospora , which is the sexual stage of Arthrinium.

8. The method according to claim 7 , wherein the mediator is at least one selected from the group consisting of potassium ferricyanide and phenazine methosulfate.

Assignees

• Toyo Boseki

Inventors

• Yamazaki Yasuhiro
• Sumida Yosuke
• Hirao Rie
• Utashima Yuu
• Aiba Hiroshi
• Kishimoto Takahide
• Yanagidani Shusaku

Classifications

• C12Q1/54

  involving glucose or galactose · CPC title

• C12N9/0006 Primary

  acting on CH-OH groups as donors (1.1) · CPC title