Vector containing a DNA coding for a novel glucose dehydrogenase and method

• US 9,587,228 B2
• Filed: 02/24/2016
• Issued: 03/07/2017
• 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:
  US15/051,813
• Publication Number:
  US 20160168544A1
• Time in Patent Office: 377 Days
• 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 a NADP-specific glucose dehydrogenase showing a NADP/NAD activity ratio of no lower than 300, derived from Lactobacillus plantarum, which is selected from the group consisting of:

(a) a DNA comprising SEQ ID NO;

2;

(b) a DNA coding for a polypeptide having the amino acid sequence of SEQ ID NO;

1; and

(c) a DNA coding for a polypeptide at least 90% sequence identical to the polypeptide of SEQ ID NO;

1.

2. A transformant obtained by transforming a host cell with the vector of 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 according to 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. A method for detecting NADPH and/or NADP and/or measuring the concentration of NADPH and/or NADP in a sample which comprises contacting the sample with the biosensor of claim 5.

7. A method for detecting D-glucose or 2-deoxy-D-glucose and/or measuring the concentration of D-glucose or 2-deoxy-D-glucose in a sample which comprises contacting the sample with the biosensor of claim 5.