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Method for decoloring digest of poly sialic acid

A technology of polysialic acid and hydrolyzate, applied in the field of bioengineering, can solve problems such as low selectivity, limited pigment adsorption capacity, and inability to extract and remove organic solvents.

Inactive Publication Date: 2007-02-21
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] 1. Sialic acid contains carboxyl ions, and cationic resins have poor adsorption to sialic acid, which can ensure the recovery rate of sialic acid, but the adsorption capacity of pigment is quite limited and the selectivity is not high
[0006] 2. Sialic acid is insoluble in organic solvents such as ether and chloroform, and organic solvents can remove some pigments, but there are still many water-soluble pigments in polysialic acid hydrolyzate, which cannot be extracted and removed by organic solvents, so it is necessary to find some more effective pigments. decolorization method

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1. Preparation of polysialic acid fermentation broth

[0022] The strain Escherichia coli WXJYL-11 is preserved and provided by the Biochemical Engineering Laboratory, School of Bioengineering, Jiangnan University, and the strain has been published in "Journal of Wuxi University of Light Industry", 2002, 9(5): 456-459.

[0023] The medium used was: sorbitol 40 g / L, ammonium chloride 4 g / L, dipotassium hydrogen phosphate 20 g / L, anhydrous magnesium sulfate 0.9 g / L, tryptone 1.5 g / L, pH 7.8.

[0024] Using 9L fermentation medium, fermentation was carried out on a 15L Biostat C10-3 mechanical stirring tank, the inoculum amount was 4%, the stirring speed of the fermentation tank was 500r / min, the ventilation rate was 10L / min, the tank pressure was maintained at 0.01MPa, and the fermentation temperature was 37°C. After 60 hours of fermentation, polysialic acid fermentation broth was obtained, and the polysialic acid content was determined to be 2.7 g / L.

Embodiment 2

[0025] Example 2. Preparation of polysialic acid hydrolyzate

[0026] The 9L polysialic acid fermentation broth obtained in Example 1 was centrifuged at 8000 r / min for 10 min to remove the bacterial cells, and 3 times of ethanol was added to the supernatant for precipitation. A polysialic acid solution with a mass concentration of 2% was obtained. Adjust the pH of the polysialic acid solution to 1.0-2.0 with a 5%-10% (v / v) mineral acid solution (such as sulfuric acid or hydrochloric acid), heat it to 75-85°C in a water bath, and maintain the temperature for 2-4 hours Hydrolysis yields sialic acid monomers. Add an alkaline solution (such as barium hydroxide or sodium hydroxide) to the pH of the solution to 7.0, and centrifuge at 7000 r / min for 15 minutes to remove the precipitate to obtain a sialic acid supernatant.

Embodiment 3

[0027] Example 3. Decolorization of sialic acid hydrolyzate and product preparation

[0028] 50 grams of attapulgite fillers were infiltrated and suspended with water and then loaded into a Φ40×250mm chromatography column, and the fillers were acid washed (soaked in 0.5-1.5% hydrochloric acid for 6 hours) and washed with water (washed with water to neutral pH) to obtain a pre-treatment solution. Treated attapulgite filler. Take 1-2 liters of the neutralized and centrifuged sialic acid supernatant, pump it into a chromatography column at a rate of 0.01-0.2 column volume per minute for decolorization, and collect the effluent. After the sample is processed, pump 100-300 ml of deionized water to rinse the chromatography column and collect the outflowing chromatographic liquid. The sialic acid product was obtained after combining the chromatographic fluids and freeze-drying. The optimal loading rate of polysialic acid hydrolyzate is 0.01-0.1 packed bed volume per minute. The pi...

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Abstract

This invention discloses a method for decoloring polysialic acid hydrolysate. The method comprises: (1) performing aerobic fermentation on Escherichia coli strain to obtain fermented liquid containing polysialic acid, removing bacteria, precipitating with ethanol, and dissolving again in deionized water to prepare 2 wt.% polysialic acid solution; (2) hydrolyzing with an acid, neutralizing with an alkali, and centrifuging; (3) passing through an adsorption chromatographic column filled with attapulgite as the filler, eluting, collecting the eluant, and freeze-drying to obtain sialic acid product. Containing deep brown and red substances, the polysialic acid hydrolysate is treated by adsorption chromatography to obtain white powdery sialic acid product.

Description

technical field [0001] A decolorization method of polysialic acid hydrolyzate belongs to the technical field of bioengineering. Background technique [0002] Sialic acid, also known as bird's nest acid, chemically named N-acetylneuraminic acid, is named after it is the main component of bird's nest. Sialic acid is widely distributed in nature. It is mainly connected to the end of glycoproteins, glycolipids and oligosaccharides through α-glycosidic bonds through the hydroxyl group of the 2-anomeric carbon. It has important biological functions. Sialic acid is also an important component of breast milk. one. Sialic acid can effectively inhibit the invasion of red blood cells by cold viruses and some pathogenic microorganisms. The sialic acid analogs Zanamivir (N-acetyl-2,3-dideoxy-4-guanidinosialic acid) and Tamiflu have been approved by the U.S. Food and Drug Administration (FDA) as new drugs for the treatment of influenza, and has been listed. SialylLewis, a derivative o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07H7/02C07H5/06
Inventor 詹晓北郑志永吴剑荣朱莉
Owner JIANGNAN UNIV
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