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Method for separating and purifying β-thymidine from fermentation broth

A technology for separation and purification, fermented liquid, applied in chemical instruments and methods, organic chemistry, sugar derivatives, etc., can solve the problem of lack of stereospecificity in the formation of glycosidic bonds, low removal rate of impurity proteins, high energy consumption of electrodialysis, etc. problems, to achieve good market prospects, simple process operation, and high yield

Active Publication Date: 2020-10-16
JIANGSU CHUANGUO PHARMA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

One is the production of thymidine by chemical synthesis, the process is very lengthy, and the formation of glycosidic bonds lacks stereospecificity, making the cost of the final thymidine relatively expensive
The second is the biological enzymatic method, which has the advantage of strong specificity, but the cycle is long and the product is not easy to separate
This method adopts the method of electrodialysis for desalination treatment. Although it can effectively remove pigments, inorganic salts, etc., the removal rate of impurity proteins is not high. The increase and loss of products; and the high energy consumption of electrodialysis, the high price of membranes, etc. all make the production cost very high

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0089] Stirring, the 80m 3 The pH of the β-thymidine fermentation broth was adjusted to 3.5 with 98% concentrated sulfuric acid. Steam is heated to 60°C, microfiltration is carried out with a ceramic membrane with a pore size of 200nm, the operating pressure is 0.15MPa, and when it is concentrated to 1 / 2 of the original volume, water is added for top washing, and the water volume for top washing is 160m 3 .

[0090] The obtained clarified microfiltrate was cooled to 25°C with a heat exchanger, and ultrafiltration separation was performed with a polyethersulfone roll-type ultrafiltration membrane with a molecular weight cut-off of 5000Da. The operating pressure was 1.0MPa. 3 Water top wash.

[0091] Ultrafiltrate at 10m 3 The speed of / h passes through the D001 type strongly acidic cation exchange resin column and the D201 type strongly basic anion exchange resin column in turn, and the resin loading capacity is 20m 3 .

[0092] Collect the ionized solution, use cellulose ...

Embodiment 2

[0096] With stirring, the 100m 3 The pH of the β-thymidine fermentation broth was adjusted to 3.0 with 31% hydrochloric acid. Steam is heated to 85°C, microfiltration is carried out with a ceramic membrane with a pore size of 100nm, the operating pressure is 0.2MPa, and when it is concentrated to 1 / 2 of the original volume, water is added for top washing, and the water volume for top washing is 200m 3 .

[0097] The obtained clarified microfiltrate was cooled to 20°C with a heat exchanger, and ultrafiltration separation was performed with a polyacrylonitrile roll-type ultrafiltration membrane with a molecular weight cut-off of 2500Da. The operating pressure was 1.0MPa. 3 Water top wash.

[0098] Ultrafiltrate at 10m 3 The speed of / h passes through 001X4 type strong acidic cation exchange resin and D213 type strong basic anion exchange resin column successively, and the resin loading capacity is 30m 3 .

[0099] Collect the ionized solution, use cellulose acetate membrane...

Embodiment 3

[0103] Stirring, the 50m 3 The pH of the β-thymidine fermentation broth was adjusted to 4.0 with 85% phosphoric acid. Heat the steam to 35°C, use a polysulfone membrane with a pore size of 400nm for microfiltration, and operate at a pressure of 0.3MPa. When it is concentrated to 1 / 3 of the original volume, start to add water for top washing, and the water volume for top washing is 150m 3 .

[0104] The obtained clarified microfiltrate was cooled to 30°C with a heat exchanger, and ultrafiltration separation was performed with a polyvinylidene fluoride roll-type ultrafiltration membrane with a molecular weight cut-off of 10,000 Da. The operating pressure was 1.0 MPa, and it was concentrated to 1 / 50 of the original volume. join 10m 3 Water top wash.

[0105] Ultrafiltrate at 10m 3 The speed of / h passes through the 112 type weakly acidic cation exchange resin column and the 313 type weakly basic anion exchange resin column in turn, and the resin loading capacity is 25m 3 . ...

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Abstract

The invention discloses a method for separating and purifying beta-thymidine from fermentation liquor. The method comprises the following steps: carrying out microfiltration, ultrafiltration, ion exchange resin chromatography, reverse osmosis concentration and vacuum concentration on beta-thymidine fermentation liquor in sequence to obtain a beta-thymidine crude product, and finally carrying out decoloration and recrystallization on the crude product to obtain a beta-thymidine competitive product. The extraction and purification technology disclosed by the invention is good in impurity removal effect and high in degree of automation, the chemical purity of the obtained beta-thymidine competitive product is greater than or equal to 99.5%, the content of the obtained beta-thymidine competitive product is greater than or equal to 99.5%, the total recovery of the obtained beta-thymidine competitive product is greater than or equal to 80%, and the method is extremely applicable for industrial mass production.

Description

technical field [0001] The invention relates to the field of biochemical industry, in particular to a method for separating and purifying β-thymidine from fermentation broth. Background technique [0002] β-thymidine, also known as 2'-deoxythymidine, is the precursor of anti-AIDS stavudine (3'-deoxy-2', 3'-didehydrothymidine) and zidovudine. At present, there are three main methods for the production of thymidine. One is the production of thymidine by chemical synthesis, the process is very lengthy, and the formation of glycosidic bonds lacks stereospecificity, making the cost of the final thymidine relatively expensive. The second is the biological enzymatic method, which has the advantage of strong specificity, but the cycle is long and the product is not easy to separate. The third is the biological fermentation method, which has a simple process and low subsequent separation costs, and is a very promising production method. Therefore, the research on the method of extr...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07H19/073C07H1/06
CPCC07H1/06C07H19/073
Inventor 张莹陈少雄
Owner JIANGSU CHUANGUO PHARMA CO LTD
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