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Method for extracting sodium hyaluronate from fermentation liquor containing hyaluronic acid

A technology of sodium hyaluronate and hyaluronic acid, applied in the field of biochemical industry, can solve the problems of low molecular weight, difficult to dissolve, and hard form of sodium hyaluronate

Active Publication Date: 2011-04-20
ANHUI BBCA FERMENTATION TECH ENG RES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The present invention aims to overcome the deficiencies in the prior art that the obtained sodium hyaluronate product is hard in shape, difficult to dissolve, high in protein content and contains other crude polysaccharide impurities, and low in molecular weight, and provides a low-cost, fast and efficient Method for preparing high-purity sodium hyaluronate

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] 1) Take 500ml of fermentation broth, containing 2g of HA, with a molecular weight of 2.38Mda. The fermentation broth was diluted 2 times with deionized water.

[0025] 2) Add 3 g / L of diatomaceous earth and 2 g / L of activated carbon into the solution, stir for two hours, and then filter to obtain a filtrate.

[0026] 3) Add 10% CTAB to the filtrate in an amount of HA:CTAB=1:2.0 (g / g), stir and complex, and filter to obtain the complex.

[0027] 4) Add 0.6M sodium chloride solution to the volume of the original fermentation broth, stir and dissociate at 200 rpm for 24 hours.

[0028] 5) Pass the dissociated solution through an anion exchange column DEAE-52 chromatography column at 1.5 BV / h, and collect the column liquid.

[0029] 6) 95% ethanol 1000ml is flowed into the filtrate, and the addition is HA solution: ethanol=1: 2 (v / v), while stirring at 150rpm, flow adds 15~20% (mass percentage) sodium chloride solution, until there is Small white flocs appeared, and powd...

Embodiment 2

[0032] 1) Take 500ml of fermented broth, containing 2g of HA, with a molecular weight of 2.38Mda, and dilute the fermented broth by 2.5 times with deionized water.

[0033] 2) Add 3 g / L of diatomaceous earth and 2 g / L of activated carbon into the solution, stir for two hours, and then filter to obtain a filtrate.

[0034] 3) Add 10% CTAB to the filtrate in an amount of HA:CTAB=1:2.5 (g / g), stir and complex, and filter to obtain the complex.

[0035] 4) Add 0.8M sodium chloride solution to the volume of the original fermentation broth, stir and dissociate at 200 rpm for 24 hours.

[0036] 5) After passing the dissociation solution through the chromatographic column at 2.0 BV / h, collect the column liquid.

[0037] 6) 95% ethanol 1500ml is flowed into the filtrate, and the addition is HA solution: ethanol=1: 3 (v / v), while stirring at 150rpm, flow adds 15~20% (mass percentage) sodium chloride solution, until there is Small white flocs appeared, and powdered sodium hyaluronate s...

Embodiment 3

[0040] 1) Take 500ml of fermentation broth, containing 2g of HA, with a molecular weight of 2.38Mda, and dilute the fermentation broth 3 times with deionized water.

[0041] 2) Add 3 g / L of diatomaceous earth and 2 g / L of activated carbon into the solution, stir for two hours, and then filter to obtain a filtrate.

[0042] 3) Add 10% CTAB to the filtrate in an amount of HA:CTAB=1:3.0 (g / g), stir and complex, and filter to obtain the complex.

[0043] 4) Add 0.8M sodium chloride solution to the volume of the original fermentation broth, stir and dissociate at 200 rpm for 24 hours.

[0044] 5) After passing the dissociation solution through the chromatographic column at 2.5 BV / h, collect the column liquid.

[0045] 6) 95% ethanol 1500ml is flowed into the filtrate, and the addition is HA solution: ethanol=1: 3 (v / v), while stirring at 150rpm, flow adds 15~20% (mass percentage) sodium chloride solution, until there is Small white flocs appeared, and powdered sodium hyaluronate ...

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Abstract

The invention provides a method for extracting sodium hyaluronate from a fermentation liquor containing hyaluronic acid, which comprises the steps of fading and degerming the diluted fermentation liquor which contains hyaluronic acid, filtering, adding CTAB into the filtrate, centrifugally separating, adding sodium chloride solution into the sediment and dissociating, pouring the dissociated solution onto an anion exchange column DEAE-52 chromatographic column, depositing the solution passed through the column with ethanol, and centrifugally drying so as to obtain the powder sodium hyaluronate solid. The sodium hyaluronate extracted by using the method of the invention has the advantages of high yield, high product purity, low protein content, simple process and convenient operation.

Description

technical field [0001] The invention relates to the field of biochemical industry, in particular to a method for extracting sodium hyaluronate from a fermented liquid containing hyaluronic acid. Background technique [0002] Hyaluronic acid (HA for short) is a kind of acidic mucopolysaccharide substance, a linear polymer polysaccharide composed of glucuronic acid-N-acetylglucosamine as disaccharide units, and the number of disaccharide units is 300 -1100 pairs, molecular weight range 10 5 -10 7 Da. [0003] HA is soluble in water, but insoluble in organic solvents such as alcohol, ketone, and ether. Because its solution has super water holding capacity, viscoelasticity and bioabsorbability, it is widely used in ophthalmic surgery, prevention of adhesion in abdominal surgery, arthritis and cosmetics and other fields. Hyaluronic acid is usually found in its sodium salt form. The properties, biological activity, production process, and clinical application of HA have been ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08B37/08
Inventor 张雪锋魏生
Owner ANHUI BBCA FERMENTATION TECH ENG RES
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