Method for preparing N-acetvlneuraminic acid through integrated polysialic acid separation and purification
A technology of acetylneuraminic acid and polysialic acid, which is applied in the preparation of sugar derivatives, chemical instruments and methods, sugar compounds with non-glycosyl groups, etc., can solve problems such as high price, achieve low cost, simplify production steps, The effect of reducing production costs
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Embodiment 1
[0028] The method for the separation and purification of polysialic acid integrated in this example to prepare N-acetylneuraminic acid is as follows:
[0029] 1. Pretreatment of polysialic feed liquid: polysialic acid derived from microbial fermentation is used as raw material, and the feed liquid containing polysialic acid is obtained through membrane filtration and sterilization steps;
[0030] 2. Hydrolysis of polysialic acid: add 0.1M oxalic acid aqueous solution to the pretreated fermentation broth containing polysialic acid, heat the temperature to 80°C, and heat for 4 hours to hydrolyze polysialic acid into N-acetylneuramine acid;
[0031] 3. Feed liquid neutralization: Add calcium hydroxide aqueous solution to the above hydrolyzed aqueous solution, and neutralize the pH of the solution to 7.0.
[0032] 4. Precipitate protein and salt: Add 2 times the volume of ethanol to the above neutralized solution to precipitate impurity proteins and some salt impurities that are ...
Embodiment 2
[0036] The method for the separation and purification of polysialic acid integrated in this example to prepare N-acetylneuraminic acid is as follows:
[0037] 1. Pretreatment of polysialic feed liquid: polysialic acid derived from microbial fermentation is used as raw material, and the feed liquid containing polysialic acid is obtained through membrane filtration and sterilization steps;
[0038] 2. Hydrolysis of polysialic acid: Add 0.5M oxalic acid aqueous solution to the pretreated fermentation broth containing polysialic acid, heat the temperature to 80°C for 4 hours, and hydrolyze polysialic acid into N-acetylneuramine acid;
[0039] 3. Feed liquid neutralization: Add calcium bicarbonate aqueous solution to the above hydrolyzed aqueous solution, and neutralize the pH of the solution to 7.0.
[0040] 4. Precipitate protein and salt: Add 3 times the volume of ethanol to the above neutralized solution to precipitate impurity protein and some salt impurities insoluble in eth...
Embodiment 3
[0044] The method for the separation and purification of polysialic acid integrated in this example to prepare N-acetylneuraminic acid is as follows:
[0045] 1. Pretreatment of polysialic feed liquid: polysialic acid derived from microbial fermentation is used as raw material, and the feed liquid containing polysialic acid is obtained through membrane filtration and sterilization steps;
[0046] 2. Hydrolysis of polysialic acid: add 0.1M oxalic acid aqueous solution to the pretreated fermentation broth containing polysialic acid, heat the temperature to 70°C for 5 hours, and hydrolyze polysialic acid into N-acetylneuramine acid;
[0047] 3. Feed liquid neutralization: Add calcium hydroxide aqueous solution to the above hydrolyzed aqueous solution, and neutralize the pH of the solution to 7.0.
[0048] 4. Precipitate protein and salt: Add 4 times the volume of ethanol to the above neutralized solution to precipitate impurity proteins and some salt impurities that are insolubl...
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