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Method for producing coenzyme and transformant set for coenzyme production

Foreign code F180009416
File No. AF30-03WO
Posted date Apr 24, 2018
Country United States of America
Application number 201615552440
Gazette No. 20180044683
Date of filing Feb 19, 2016
Gazette Date Feb 15, 2018
International application number JP2016054872
International publication number WO2016136620
Date of international filing Feb 19, 2016
Date of international publication Sep 1, 2016
Priority data
  • P2015-033843 (Feb 24, 2015) JP
  • 2016WO-JP54872 (Feb 19, 2016) WO
Title Method for producing coenzyme and transformant set for coenzyme production
Abstract (US20180044683)
The present invention provides a method for synthesizing NAD+ or NADH in an enzyme reaction system requiring NAD+ or NADH so as to supplement NAD+ lost due to thermal decomposition in the enzyme reaction system, and a transformant set for coenzyme production used in the method.
That is, the present invention relates to a method for producing a coenzyme including performing synthesis of NAD+ or NADH in an enzyme reaction system requiring NAD+ or NADH by adding one or more thermostable enzymes required for a reaction for synthesizing NAD+ from nicotinamide, to the enzyme reaction system, and a transformant set for coenzyme production including one or more transformants into which one or more genes encoding thermostable enzymes required for a reaction for synthesizing NAD+ or NADH from nicotinamide are introduced by using a non-thermostable microorganism as a host.
Scope of claims [claim1]
1. A method for producing a coenzyme, the method comprising: performing synthesis of NAD+ in an enzyme reaction system requiring NAD+ by adding one or more thermostable enzymes required for a reaction for synthesizing NAD+ from ADP-ribose and nicotinamide, to the enzyme reaction system.
[claim2]
2. The method for producing a coenzyme according to claim 1, wherein the thermostable enzymes required for a reaction for synthesizing NAD+ are nicotinamidase, nicotinate phosphoribosyltransferase, nicotinate-nucleotide adenylyltransferase, NAD+ synthase, ADP-ribose pyrophosphatase, and ribose-phosphate pyrophosphokinase.
[claim3]
3. A method for producing a coenzyme, the method comprising: performing synthesis of NADH in an enzyme reaction system requiring NADH by adding one or more thermostable enzymes required for a reaction for synthesizing NADH from ADP-ribose and nicotinamide, to the enzyme reaction system.
[claim4]
4. The method for producing a coenzyme according to claim 3, wherein the thermostable enzymes required for a reaction for synthesizing NADH are nicotinamidase, nicotinate phosphoribosyltransferase, nicotinate-nucleotide adenylyltransferase, NAD+ synthase, ADP-ribose pyrophosphatase, ribose-phosphate pyrophosphokinase, and oxidoreductase that catalyzes a reaction for synthesizing NADH from NAD+.
[claim5]
5. The method for producing a coenzyme according to claim 4, wherein the oxidoreductase is dehydrogenase with sugar, alcohol, or organic acid as a substrate.
[claim6]
6. The method for producing a coenzyme according to claim 1, further comprising: performing synthesis of ATP in the enzyme reaction system by adding one or more thermostable enzymes required for a reaction for synthesizing ATP from AMP or ADP, to the enzyme reaction system.
[claim7]
7. The method for producing a coenzyme according to claim 6, wherein the thermostable enzymes required for a reaction for synthesizing ATP are adenylate kinase and polyphosphate kinase.
[claim8]
8. The method for producing a coenzyme according to claim 1, wherein the thermostable enzyme is synthesized by an expression system of a non-thermostable microorganism.
[claim9]
9. The method for producing a coenzyme according to claim 1, wherein the thermostable enzyme is synthesized by a transformant into which a gene encoding the thermostable enzyme is introduced by using a non-thermostable microorganism as a host.
[claim10]
10. The method for producing a coenzyme according to claim 9, wherein a heat treated product of microbial cells obtained by culturing the transformant is added to the enzyme reaction system.
[claim11]
11. The method for producing a coenzyme according to claim 8, wherein the non-thermostable microorganism is E. coli.
[claim12]
12. A transformant set for coenzyme production, comprising: one or more transformants into which one or more genes encoding thermostable enzymes required for a reaction for synthesizing NAD+ or NADH from ADP-ribose and nicotinamide are introduced by using a non-thermostable microorganism as a host.
[claim13]
13. The transformant set for coenzyme production according to claim 12, wherein the thermostable enzymes are nicotinamidase, nicotinate phosphoribosyltransferase, nicotinate-nucleotide adenylyltransferase, NAD+ synthase, ADP-ribose pyrophosphatase, and ribose-phosphate pyrophosphokinase.
[claim14]
14. The transformant set for coenzyme production according to claim 12, wherein the thermostable enzymes are nicotinamidase, nicotinate phosphoribosyltransferase, nicotinate-nucleotide adenylyhransferase, NAD+ synthase, ADP-ribose pyrophosphatase, ribose-phosphate pyrophosphokinase, and oxidoreductase that catalyzes a reaction for synthesizing NADH from NAD+.
[claim15]
15. The transformant set for coenzyme production according to claim 12, further comprising: one or more transformants into which one or more genes encoding thermostable enzymes required for a reaction for synthesizing ATP from AMP or ADP are introduced by using a non-thermostable microorganism as a host.
[claim16]
16. The transformant set for coenzyme production according to claim 12, wherein one type of genes encoding the thermostable enzymes is introduced into one transformant.
[claim17]
17. The transformant set for coenzyme production according to claim 12, wherein the non-thermostable microorganism is E. coli.
[claim18]
18. The transformant set for coenzyme production according to claim 12, wherein the transformant is heat treated.
[claim19]
19. The method for producing a coenzyme according to claim 3, further comprising: performing synthesis of ATP in the enzyme reaction system by adding one or more thermostable enzymes required for a reaction for synthesizing ATP from AMP or ADP, to the enzyme reaction system.
[claim20]
20. The method for producing a coenzyme according to claim 19, wherein the thermostable enzymes required for a reaction for synthesizing ATP are adenylate kinase and polyphosphate kinase.
  • Inventor, and Inventor/Applicant
  • HONDA KOHSUKE
  • ATOMI HARUYUKI
  • JAPAN SCIENCE AND TECHNOLOGY AGENCY
IPC(International Patent Classification)
Reference ( R and D project ) CREST Creation of Basic Technology for Improved Bioenergy Production through Functional Analysis and Regulation of Algae and Other Aquatic Microorganisms AREA
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