Method for separating and purifying N-acetylneuraminic acid produced by microbiological fermentation

A technology of acetylneuraminic acid and microbial fermentation, which is applied in the direction of sugar derivatives, sugar derivatives, sugar derivatives preparation, etc., can solve the problems of high price and high quality demand, reduce pressure, improve purity, and improve product quality. The effect of purity

Active Publication Date: 2015-05-20
武汉中科光谷绿色生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Expensive prices and high quality requirements lead to urgent research and

Method used

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  • Method for separating and purifying N-acetylneuraminic acid produced by microbiological fermentation
  • Method for separating and purifying N-acetylneuraminic acid produced by microbiological fermentation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] In this implementation, the N-acetylneuraminic acid produced by microbial fermentation is separated and purified according to the following steps:

[0033] a. Separation of bacterial liquid: In the method disclosed in the invention patent with application number 201310600843.0, the fermented liquid containing N-acetylneuraminic acid produced by the fermentation of E. The base montmorillonite powder is used for flocculation and sedimentation of the bacteria, and then the filter cloth is used to squeeze and filter the filter cloth with a pore size of 2000 meshes. Bacterial fermentation broth, detected by high performance liquid chromatography, the content of the target product N-acetylneuraminic acid in the fermentation broth has almost no change before and after sterilization, that is, the use of montmorillonite powder to flocculate the bacteria will not cause loss of N-acetylneuraminic acid ;

[0034] b. Removal of impurity proteins: adding hydrochloric acid to the ste...

Embodiment 2

[0040] In this implementation, the N-acetylneuraminic acid produced by microbial fermentation is separated and purified according to the following steps:

[0041]a. Separation of bacterial liquid: In the method disclosed in the invention patent with application number 201310600843.0, the fermented liquid containing N-acetylneuraminic acid produced by the fermentation of E. The base montmorillonite powder is used for flocculation and sedimentation of the bacteria, and then the filter cloth is used to squeeze and filter the filter cloth with a pore size of 2000 meshes. Bacterial fermentation liquid;

[0042] b. Removal of impurity proteins: adding hydrochloric acid to the sterilizing fermentation broth, adjusting the pH to 3.8, heating at 90°C for 0.5h to precipitate the impurity proteins, centrifuging, removing the impurity protein precipitation, and obtaining the protein-removing fermentation broth;

[0043] c. Decolorization: adding activated carbon accounting for 3% of the ...

Embodiment 3

[0048] In this example, the same raw material as in Example 2 is separated and purified in the same manner as in Example 2, the only difference being that the temperature in step b is changed from 90°C to 70°C, and the purity of the final product in this implementation is 97.9% after testing. Compared with Example 2, it is worse. It shows that the effect of heat treatment at 90°C is better, it can completely denature the protein in the fermentation liquid, reduce the impurity protein in the crystallization liquid, and improve the purity of the product.

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Abstract

The invention provides a method for separating and purifying N-acetylneuraminic acid produced by microbiological fermentation. The method is characterized by comprising the following steps: by taking fermentation liquid in fermenting production of N-acetylneuraminic acid by escherichia coli as a raw material, sterilizing; removing proteins; decoloring; desalting; crystallizing and the like to obtain a high purity N-acetylneuraminic acid product. Through tests, the product purity is at least 98%, and the requirements in the fields of foods, healthcare, medicines, cosmetics and the like can be satisfied. Moreover, the method is simple and easy to operate and is particularly suitable for industrial fermentation production of N-acetylneuraminic acid.

Description

technical field [0001] The invention relates to a method for separating and purifying monomeric N-acetylneuraminic acid produced by a microbial fermentation method, and belongs to the technical field of biochemical separation engineering. Background technique [0002] The production of N-acetylneuraminic acid by microbial fermentation has been reported in many literatures. The Chinese invention patent with the application number 201310600843.0 discloses a method for producing N-acetylneuraminic acid by microbial fermentation using Escherichia coli engineering bacteria, and the method obtains a fermentation liquid containing the product. [0003] Although there are many reports on the fermentation methods and processes for the production of N-acetylneuraminic acid by microbial fermentation, there are few reports on the separation and purification methods of the downstream products, and the downstream separation and purification methods and processes directly determine the qua...

Claims

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Application Information

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IPC IPC(8): C07H13/06C07H1/06
Inventor 袁丽霞陈祥松吴金勇孙立洁王刚朱薇薇王纪姚建铭费贤春王煜
Owner 武汉中科光谷绿色生物技术有限公司
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