Preparation method of nicotinamide adenine dinucleotide

A technology of nicotinamide adenine and nicotinamide nucleotide adenosine, which is applied in the field of preparation of nicotinamide adenine dinucleotide, can solve the problems of high concentration and energy consumption, low conversion rate of chemical synthesis, and huge equipment. To achieve the effect of simple and optimized process

Inactive Publication Date: 2013-01-16
SYNCOZYMES SHANGHAI
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Problems solved by technology

[0005] Extraction from yeast is the main source of NAD widely used in industry at present. Although this method has a mature process, the equipment is huge, the concentration energy consumption is high, and silver-containing salts or Hg(COOCH 3 ) 2 Reagents such as heavy metal ions, so the production efficiency is low and the cost is high
[0006] Due to the low conversion rate, the reagents used are expensive, and there are a large number of operations through silica gel columns and gel chromatography columns, the chemical synthesis method is still in the laboratory research stage and can only be prepared at milligram or gram levels.
[0007] In summary, the existing methods for preparing NAD are still difficult to meet the requirements of commercial production

Method used

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  • Preparation method of nicotinamide adenine dinucleotide
  • Preparation method of nicotinamide adenine dinucleotide

Examples

Experimental program
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Embodiment 1

[0031] see figure 1 The preparation and synthesis process roadmap of

[0032] Using 1,2,3,5-tetraacetyl-β-D-ribofuranose as the starting material;

[0033] Step 1: condensation reaction, synthesis such as figure 1 Intermediate 1 shown:

[0034] In a 20L glass reactor, set the stirring and thermometer (range 0-100°C), add 12L of dichloromethane with a triangular funnel, start stirring, and set the stirring speed to 42.5rpm. Throw in 1.2kg (3.77mol) of 1,2,3,5-tetraacetyl-β-D-ribofuranose and stir to dissolve. Open the jacketed circulating water of the reaction kettle (set at 12°C), and when the internal temperature reaches 12°C, add 0.84kg (3.77mol) trimethylsilicone trifluoromethanesulfonate to the dropping funnel, and start to add it slowly, Add 0.84kg (5.65mol) nicotinyl ethyl ester after dropping. The temperature of the circulating water is raised to 50°C, and the internal temperature reaches 48°C, and heat preservation and reflux are started. After reflux for 4h, tak...

Embodiment 2

[0042] Steps one to three are the same as in Example 1.

[0043] Step four:

[0044] Set stirring and a thermometer with a range of 0-100°C on the 20L glass reactor, add a buffer solution made of 0.33kg of sodium dihydrogen phosphate, 0.48kg of disodium hydrogen phosphate and 16L of water from the mouth of the kettle with a triangular funnel, and open the reactor. Circulating water (set at 30°C) and stirring, the stirring speed is set at 42.5rpm. Throw 0.78kg (2.32mol) of intermediate 3 nicotinamide mononucleotide and 1.09kg (2.78mol) of adenosine triphosphate. After the materials are completely dissolved, add 0.3kg of whole cells of NMNAT and 0.6kg of whole cells of PPase, and react 10h, sampling. If HPLC shows that the remaining raw materials are ≤ 3% in 4.3 minutes, start to lower the temperature; if HPLC shows that the remaining raw materials are > 3%, continue the reaction until the remaining raw materials are ≤ 3% and then stop the reaction. Blowing, ultrafiltration, ...

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Abstract

The invention relates to a preparation method of nicotinamide adenine dinucleotide, which comprises the following steps of: with 1,2,3,5-tetraacetyl-beta-D-ribofuranose as a starting raw material, preparing nicotinamide mononucleotide through sequentially a condensation reaction, an ammonolysis reaction and a phosphorylation reaction; and then mixing the prepared nicotinamide mononucleotide with adenosine triphosphate, generating an enzymic catalytic reaction in the presence of nicotinamide adenosine nucleotide transferase and pyrophosphatase to prepare the nicotinamide adenine dinucleotide. Compared with the prior art, the technical scheme provided by the invention has the advantage that the whole process is simplified and optimized by combination of a chemical method and an enzymic method.

Description

technical field [0001] The invention relates to a preparation method of nicotinamide adenine dinucleotide. Background technique [0002] Nicotinamide adenine dinucleotide (hereinafter referred to as NAD) is a very common coenzyme of dehydrogenase in organisms. Dehydrogenase plays a decisive role in the metabolism of organisms. Some basic metabolic movements of organisms, such as protein decomposition, carbohydrate decomposition, and fat decomposition, cannot proceed normally without dehydrogenase, and organisms will lose their lives. signs. Since dehydrogenase must be combined with NAD to promote metabolic operation, it is an indispensable substance in the metabolism of NAD organisms. [0003] NAD is one of the essential coenzymes in modern biocatalytic reactions. Modern biocatalysis technology simulates the reaction completed by enzymes in organisms. The simulated conditions of this reaction are strictly based on the metabolic characteristics of organisms, so like common...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/36
Inventor 王波孙勇文军
Owner SYNCOZYMES SHANGHAI
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