Enzymatic synthesis process for 5'-cytidine monophosphate

A synthesis process, the technology of cytidylic acid, which is applied in the field of enzyme-catalyzed synthesis of 5′-cytidylic acid, can solve the problems of unfavorable human health and industrial production, multiple sources of RNA, and complicated separation operations, and achieve a short process cycle , a wide range of sources, and the effect of improving the preparation efficiency

Inactive Publication Date: 2017-11-21
NANTONG SANE BIOLOGICAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the first method can obtain four products at one time, except for cytidylic acid, the other three become by-products, and the source of RNA is more restricted, and the separation operation in the production process is complicated and the yield is low; At pre

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] a. Synthesis of cytidine: take a 1000ml four-necked flask, add 111.10g cytosine, 150.13g D-ribose, 600g pure water and 0.11g PEG400, place the four-necked flask on a constant temperature stirrer, start stirring and adjust the constant temperature stirring Adjust the temperature of the tank to 37-39°C, adjust the pH of the mixture in the four-necked flask to 6.7, and then add 55.6 g of synthetase I to the four-necked flask. Synthetase I includes cytidine ligase and NAD+, and the content of NAD+ is 0.01%-0.02% of the content of glycoside ligase, keep warm and stir, take out samples from the four-necked flask every one hour for liquid spectrum analysis, when the liquid spectrum detects that cytosine is completely reacted, the reaction is terminated, and the reaction in the four-necked flask The obtained solution is filtered, and the filter cake is rinsed with 200 g of pure water, and the obtained filtrate and the rinsing liquid are combined together, which is the cytidine f...

Embodiment 2

[0026] a. Synthesis of cytidine: take a 1000ml four-necked flask, add 111.10g cytosine, 150.13g D-ribose, 600g pure water and 0.11g PEG400, place the four-necked flask on a constant temperature stirrer, start stirring and adjust the constant temperature stirring Adjust the temperature of the tank to 37-39°C, adjust the pH of the mixture in the four-necked flask to 7.0, and then add 55.6 g of synthetase I to the four-necked flask. Synthetase I includes cytidine ligase and NAD+, and the content of NAD+ is 0.01%-0.02% of the content of glycoside ligase, keep warm and stir, take out samples from the four-necked flask every one hour for liquid spectrum analysis, when the liquid spectrum detects that cytosine is completely reacted, the reaction is terminated, and the reaction in the four-necked flask The obtained solution is filtered, and the filter cake is rinsed with 200 g of pure water, and the obtained filtrate and the rinsing liquid are combined together, which is the cytidine f...

Embodiment 3

[0031] a. Synthesis of cytidine: take a 1000ml four-necked flask, add 111.10g cytosine, 150.13g D-ribose, 600g pure water and 0.11g PEG400, place the four-necked flask on a constant temperature stirrer, start stirring and adjust the constant temperature stirring Adjust the temperature of the tank to 37-39°C, adjust the pH of the mixture in the four-necked flask to 6.7, and then add 111.1 g of synthetase I to the four-necked flask. Synthetase I includes cytidine ligase and NAD+, and the content of NAD+ is 0.01%-0.02% of the content of glycoside ligase, keep warm and stir, take out samples from the four-necked flask every one hour for liquid chromatography analysis, when the liquid chromatography detects that cytosine is completely reacted, the reaction is terminated, and the The obtained solution is filtered, and the filter cake is rinsed with 200 g of pure water, and the obtained filtrate and the rinsing liquid are combined together, which is the cytidine filtrate;

[0032] b....

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PUM

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Abstract

The invention discloses an enzymatic synthesis process for 5'-cytidine monophosphate. The enzymatic synthesis process for 5'-cytidine monophosphate comprises the following steps: synthesis of cytidine: synthesizing the cytidine by using cytosine and D-ribose under the catalytic action of synthetase I, and filtering a solution obtained from reaction to obtain cytidine filtrate; preparation of 5'-cytidine monophosphate mother liquor: synthetizing 5'-cytidine monophosphate from cytidine filtrate and sodium dihydrogen phosphate under catalytic action of synthetase II, stirring obtained reaction liquid for a period of time under the condition of insulating and filtering to obtain 5'-cytidine monophosphate mother liquid; and refined extraction of the 5'-cytidine monophosphate: regulating pH of the 5'-cytidine monophosphate mother liquid, then filtering to obtain a 5'-cytidine monophosphate crude product, and refining the 5'-cytidine monophosphate crude product to obtain the product 5'-cytidine monophosphate. The enzymatic synthesis process for 5'-cytidine monophosphate has the advantages that the production cost of the 5'-cytidine monophosphate can be reduced; the process period is short, and the preparation efficiency of the 5'-cytidine monophosphate can be improved; the total yield of the 5'-cytidine monophosphate can be 90% or above, and the content is greater than 99%; and the 5'-cytidine monophosphate is harmless to the environment, equipment and operators.

Description

technical field [0001] The invention relates to the field of biochemical industry, in particular to an enzyme-catalyzed synthesis process of 5'-cytidylic acid. Background technique [0002] 5'-cytidylic acid is an important biochemical substance in biological cells and participates in various physiological and biochemical reactions. In industry, 5'-cytidylic acid is mainly used in the manufacture of citicoline, cytidine triphosphate, cytarabine, polyinosin and other drugs. In recent years, it can also be added to milk powder in combination with other nucleotides to enhance the baby's immunity. [0003] There are two main production methods for 5'-cytidylic acid, 1) degraded from RNA (ribonucleic acid), and then separated; 2) starting from cytidine, chemically attaching a phosphate group to its 5' position to form 5'-cytidine '-cytidylic acid. Although the first method can obtain four products at one time, except for cytidylic acid, the other three become by-products, and ...

Claims

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Application Information

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IPC IPC(8): C12P19/30
CPCC12P19/305
Inventor 张剑刘拥军于广张玮琪张军
Owner NANTONG SANE BIOLOGICAL
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