Preparation method of oxidation coenzyme I

An oxidized coenzyme and enzyme powder technology, which is applied in the direction of fermentation, can solve the problems of expensive products, large energy and material consumption, and restrictions on the production and development of nicotinamide adenine dinucleotide, so as to achieve high conversion rate and avoid high energy consumption , the effect of good industrial application prospects

Active Publication Date: 2012-07-25
ENZYMEWORKS
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  • Abstract
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  • Application Information

AI Technical Summary

Problems solved by technology

Although the process is mature, it consumes a lot of energy and materials, and the product is expensive, which limits the production of nicotinamide adenine dinucleotide and the development of its subsequent application process

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Add 10 mL Tris-HCl buffer solution (100 mM, pH 7.5) into a 20 mL three-necked flask, and add nicotinamide riboside ( J. Med. Chem. 2007, 50, 6458-6461 ) 50 mg, adenosine triphosphate disodium salt 55 mg, nicotinamide riboside kinase ( PLoS Biology, 2007, 5(10), 2220-2230 ) 10 mg, magnesium chloride 20 mM, stirred and reacted at 200 rpm at 37 °C, and the conversion rate of the reaction was monitored by liquid chromatography-mass spectrometry. After 6 hours of reaction, it was detected that adenosine triphosphate had been consumed, and the reaction was stopped. After further filtration, adsorption by HZ-818 macroporous resin, freeze-drying, and recrystallization with ethanol and water, the oxidized coenzyme I product can be obtained with a yield of 70%.

Embodiment 2

[0020] Add 10 mL of phosphate buffered saline (100 mM, pH 5.8) into a 20 mL three-necked flask, and add nicotinamide riboside ( J. Med. Chem. 2007, 50, 6458-6461 ) 50 mg, adenosine triphosphate disodium salt 55 mg, nicotinamide riboside kinase ( PLoS Biology, 2007, 5(10), 2220-2230 ) 5 mg, manganese chloride 20 mM, reacted at 37 °C, 200 rpm, and monitored the conversion rate of the reaction by liquid chromatography-mass spectrometry. After 10 hours of reaction, it was detected that adenosine triphosphate had been consumed, and the reaction was stopped. After further filtration, adsorption by AB-8 macroporous resin, freeze-drying, and recrystallization with ethanol and water, the oxidized coenzyme I product can be obtained with a yield of 52%.

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PUM

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Abstract

The invention relates to a preparation method of an oxidation coenzyme I. According to the method, nicotinamide ribonucleoside (NR) and adenosine disodium triphosphate (ATP-Na2) react with each other to obtain the oxidation coenzyme I (NAD+, Nicotinamide Adenine Dinucleotide+) in a buffer solution with the pH of 5.0 to 8.0 at a temperature of 30 DEG C to 40 DEG C under the catalytic action of nicotinamide ribonucleoside kinase (NRK) under the condition of the existence of divalent metal ions. According to the invention, a biocatalysis method is adopted, the oxidation coenzyme is prepared by utilizing an NRK one pot process, the reaction system is simple, the conditions are mild, and the preparation method has wide industrialization application prospect.

Description

technical field [0001] The invention relates to a preparation method of oxidized coenzyme I. Background technique [0002] Among all the enzymes discovered so far, about 30-35% are oxidoreductases. In industrial biotechnology, especially in biopharmaceutical and fine chemical industries, oxidoreductases are the most important enzymes. In the reactions catalyzed by oxidoreductases, certain coenzymes need to be consumed for the transfer of electrons and protons while producing products. According to statistics, about 80% of the reactions require nicotinamide adenine dinucleotide oxidized coenzyme I (abbreviated as NAD+, see formula I for structural formula) as a coenzyme, and about 10% of the reactions require nicotinamide adenine dinucleotide phosphate That is, oxidized coenzyme II (NADP+ for short) is used as a coenzyme, and only a small amount of reactions require flavin (FAD, FMN) and coenzyme Q (PQQ) as a coenzyme. Although the addition of a complex coenzyme regenerati...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/32
Inventor 陶军华李斌鞠鑫梁晓亮庄季昌
Owner ENZYMEWORKS
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