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Method for separating and preparing 5'-nucleotide using emulated moving bed

A technology for simulating moving bed and nucleotides, applied in the field of biological product processing, can solve the problems of difficult scale, cumbersome operation, intermittent operation, etc., and achieve the effect of improving separation efficiency, ensuring high quality and good effect.

Inactive Publication Date: 2006-12-27
NANJING UNIV OF TECH
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  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to overcome the defects of cumbersome operation, multi-step separation, batch type and difficulty in scale-up in the current 5'-nucleotide separation process, and provide a combination of simulated moving bed ion exchange chromatography and nanofiltration membrane separation Method for the separation and preparation of 5'-nucleotides

Method used

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  • Method for separating and preparing 5'-nucleotide using emulated moving bed

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036]The sample concentration of the nucleotide enzymatic hydrolysis solution was 30 g / L. Pack 24 ion-exchange columns with the wet packing method, each ion-exchange resin column is filled with 200 grams of 001×6 type resin, the resin particle size is 100-150 mesh, and the diameter-to-height ratio of the column is 1:5. The following process is used for separation, 2 ion-exchange columns in the first zone, 7 ion-exchange columns in the second zone, 7 ion-exchange columns in the third zone, and 7 ion-exchange columns in the fourth zone. Then, the clarified nucleotide solution is loaded onto the column for adsorption from the adsorption area, and the sample loading flow rate is 10ml / min. Each zone was eluted with pH 5.0 deionized water, and the flow rates were: 10ml / min for the first zone, 18ml / min for the second zone, 17ml / min for the third zone, and 35ml / min for the fourth zone. The three outlets of the desorption zone collect UMP, GMP, CMP and AMP solutions respectively, and...

Embodiment 2

[0038] The sample concentration of nucleotidase hydrolysis liquid was 25g / L. Pack 24 ion-exchange columns by wet column packing method, each ion-exchange resin column is filled with 200 grams of 001×7 type resin, the resin particle size is 100-150 mesh, and the diameter-to-height ratio of the column is 1:5, and adopt The following process is used for separation, 3 ion-exchange columns in the first zone, 6 ion-exchange columns in the second zone, 7 ion-exchange columns in the third zone, and 8 ion-exchange columns in the fourth zone. Then, the clarified nucleotide solution is loaded onto the column for adsorption from the adsorption area, and the sample loading flow rate is 10ml / min. Each zone was eluted with pH 5.0 deionized water, and the flow rates were: 15ml / min for the first zone, 23ml / min for the second zone, 22ml / min for the third zone, and 43ml / min for the fourth zone. The three outlets of the desorption zone collect UMP, GMP, CMP and AMP solutions respectively, and th...

Embodiment 3

[0040] The sample concentration of the nucleotide enzymatic hydrolysis solution was 31 g / L. Pack 24 ion-exchange columns with the wet packing method, each ion-exchange resin column is filled with 200 grams of 001×5 type resin, the resin particle size is 120-150 mesh, and the diameter-to-height ratio of the column is 1:5. The following process is used for separation, 2 ion-exchange columns in the first zone, 7 ion-exchange columns in the second zone, 7 ion-exchange columns in the third zone, and 8 ion-exchange columns in the fourth zone. Then, the clarified nucleotide solution is loaded onto the column for adsorption from the adsorption area, and the sample loading flow rate is 10ml / min. Each zone was eluted with pH 5.0 deionized water, and the flow rates were: 13ml / min for the first zone, 22ml / min for the second zone, 21ml / min for the third zone, and 41ml / min for the fourth zone. The three outlets of the desorption zone collect UMP, GMP, CMP and AMP solutions respectively, an...

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Abstract

The invention discloses a method for separating and preparing 5'-nucleic acid with simulated moving bed. The method adopts simulated moving bed system and nanofiltration combining technique to separate four mixed 5'-nucleic acid, namely that pumping enzyme-hydrolysis cleaning fluid containing four mixed 5'-nucleic acid into simulated moving bed system filled with cationic ion-exchange resin to adsorb, eluting with deionized water, controlling drift velocity of nucleic acid in every domain to make corresponding domain exit individually drop off four mononucleotide solution; switching import and export position of every domain of simulated moving bed to switch every domain according to liquid flow direction and original line sequence, repeating the said steps to make corresponding domain exit individually drop off four mononucleotide solution, circulating up-and-down in this way; synchronously thickening and desalting the ion exchange elute of the obtained four mononucleotide solution with nanofiltration film, and obtaining the products by crystallization and dehumidification. The invention can perform continuous segregation, the product fineness is high, the investment is small, the cost is low, and so it can be produced with full scale.

Description

technical field [0001] The invention relates to a method for separating and preparing 5'-nucleotides by using a simulated moving bed, and belongs to the field of biological product processing. Background technique [0002] Nucleotides and their derivatives are important biochemical raw materials, which can be used as food additives, health food and pharmaceutical intermediates. Generally, yeast can be obtained by fermentation, and then nucleotides are extracted from them, and nuclease P 1 Separation after enzymatic hydrolysis, some separation methods have been reported, such as: [0003] The research results of the Institute of Microbiology, Chinese Academy of Sciences (Production and Application of Nucleotides, 1971) showed that nucleotides can be separated by combining cation-anion exchange resins, but 5'-adenylic acid, cytidylic acid And the separation effect of uridine acid is not ideal. At the same time, this document also reported another separation method, which dir...

Claims

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Application Information

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IPC IPC(8): C12N15/10
Inventor 应汉杰吕浩赵谷林
Owner NANJING UNIV OF TECH
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