DNA coding for a novel glucose dehydrogenase

• US 9,303,280 B2
• Filed: 09/24/2008
• Issued: 04/05/2016
• Est. Priority Date: 09/26/2007
• Status: Active Grant

• Current Assignee: Kaneka Corporation
• Original Assignee: Kaneka Corporation
• Inventors: Kawano, Shigeru, Yasohara, Yoshihiko
• Primary Examiner(s): Ramirez, Delia
• Application Number:
  US12/680,050
• Publication Number:
  US 20130183716A1
• Time in Patent Office: 2,750 Days
• Field of Search: 435/190, 435/69.1, 435/320.1, 435/252.3, 435/325, 536/23.2
• US Class Current: 1/1
• CPC Class Codes: C12N 9/0006 acting on CH-OH groups as donors (1.1)
• CPC Class Codes: C12Q 1/006 for glucose
• CPC Class Codes: C12Q 1/32 involving dehydrogenase
• CPC Class Codes: C12Q 1/54 involving glucose or galactose
• CPC Class Codes: C12Y 101/01047 Glucose 1-dehydrogenase (1.1.1.47)

Abstract

The invention relates to a glucose dehydrogenase showing an NADP/NAD activity ratio, namely the value obtained by dividing the enzyme activity value obtained by using NADP as a coenzyme by the enzyme activity value obtained by using NAD as a coenzyme, of not lower than 300, to a gene coding therefor, and to a transformant harboring that gene. The enzyme of the invention is very high in NADP specificity and can be suitably used, for example, for NADPH production, for coenzyme regeneration in enzymatic reduction reactions, and in biosensors for glucose concentration measurements.

Claims

1. A recombinant vector comprising a DNA coding for an NADP-specific glucose dehydrogenase showing an NADP/NAD activity ratio of no lower than 300 derived from bacteria belonging to the genus Lactobacillus, which is selected from the group consisting of:

(a) a polypeptide having the amino acid sequence of SEQ ID NO;

3;

(b) a polypeptide having an amino acid sequence which differs from SEQ ID NO;

3 solely by substitution, deletion, insertion and/or deletion of no more than 25 amino acid residues, wherein the polypeptide is capable of acting on glucose and NADP to form gluconolactone and NADPH; and

(c) a polypeptide having an amino acid sequence identity of at least 90% with the polypeptide of SEQ ID NO;

3, wherein the polypeptide is capable of acting on glucose and NADP to form gluconolactone and NADPH.

2. A transformant obtained by transforming a host cell with the vector according to claim 1.

3. A reduction reaction system for an enzymatic reduction reaction using a reductase source requiring NADPH as a coenzyme, wherein the reduction reaction system comprises the transformant of claim 2.

4. The reduction reaction system of claim 3, wherein the transformant further comprises a DNA coding for a reductase requiring NADPH as a coenzyme.

5. A biosensor comprising the transformant of claim 2.

6. The biosensor according to claim 5 which is used for (a) detecting NADPH and/or NADP and/or (b) measuring the concentration of NADPH and/or NADP.

7. The biosensor according to claim 5 which is used for (a) detecting D-glucose or 2-deoxy-D-glucose and/or (b) measuring the concentration of D-glucose or 2-deoxy-D-glucose.

8. A method of producing NADPH which comprises culturing the transformant of claim 2 to generate NADPH from NADP and glucose.