Preparation method of polysialic acid and polysialic acid product

The technology of polysialic acid and filtrate is applied in the field of bioengineering, and can solve the problems of toxicity, incompatibility, human harm and the like, and achieve the effects of obvious effect, high separation efficiency and simple preparation method.

Active Publication Date: 2021-06-22
HEFEI INSTITUTES OF PHYSICAL SCIENCE - CHINESE ACAD OF SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The complexing agents or eluents involved in the existing methods are mostly toxic and irritating reagents, which are harmful to the human body and are not suitable as reagents for extracting food raw materials. Therefore, there is still room for improvement

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Embodiment 1, a kind of preparation method of polysialic acid

[0044] A preparation method of polysialic acid, comprising the following steps:

[0045] S1. Strain fermentation: take the polysialic acid-producing bacteria frozen at -80°C, and after thawing, take a ring with an inoculation loop and spread it on a petri dish containing solid medium, and incubate at 37°C for 12 hours. Take a single colony and inoculate it in liquid LB medium, culture it at 220rpm at 37°C for 12h to form a primary seed liquid, then inoculate the primary seed liquid with 1% inoculum in the LB medium, cultivate it at 220rpm at 37°C for 6h, Form the secondary seed liquid; take the secondary seed liquid and inoculate it into a fermenter containing 50% of the liquid-filled fermentation medium according to the inoculation amount of 1%, the initial fermentation condition is 200rpm, 0.5vvm, and then adjust the ventilation during the fermentation process The amount of dissolved oxygen and the stirr...

Embodiment 2

[0052] Embodiment 2, a kind of preparation method of polysialic acid

[0053] A preparation method of polysialic acid, comprising the following steps:

[0054] S1. strain fermentation: same as embodiment 1;

[0055] S2. Passing through the membrane to remove bacteria: filter the fermented liquid prepared in step S1 through a ceramic membrane with a membrane pore size of 100 nm, intercept the bacteria, and obtain a membrane clear solution containing polysialic acid;

[0056] S3. Heat treatment: add sodium hydroxide solution to the membrane serum prepared in step S2, adjust the pH value of the membrane serum to 10, pour it into a heating tank, and heat-treat it in a water bath at 80°C for 0.5 hours to denature and precipitate the protein. heat treatment fluid;

[0057] S4. Filtration and precipitation: add 18g / L of diatomaceous earth to the heat treatment solution prepared in step S3, then press filter through a 0.22um single-layer filter membrane to remove denatured protein, ...

Embodiment 3

[0061] Embodiment 3, a kind of preparation method of polysialic acid

[0062] A preparation method of polysialic acid, comprising the following steps:

[0063] S1. strain fermentation: same as embodiment 1;

[0064] S2. Passing the membrane to remove bacteria: filter the fermentation broth prepared in step S1 through a ceramic membrane with a membrane pore size of 200 nm, intercept the bacteria, and obtain a membrane clear solution containing polysialic acid;

[0065] S3. Heat treatment: adding sodium hydroxide solution to the membrane serum prepared in step S2, adjusting the pH value of the membrane serum to 12, pouring it into a heating tank, and heat-treating it in a water bath at 90°C for 1 hour to denature and precipitate the protein. heat treatment fluid;

[0066] S4. Filtration and precipitation: add 20g / L of diatomaceous earth to the heat treatment liquid prepared in step S3, then press filter through a 0.22um single-layer filter membrane to remove denatured protein,...

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Abstract

The invention relates to the technical field of bioengineering, and particularly discloses a preparation method of polysialic acid and a polysialic acid product. The method comprises the following steps: S1, strain fermentation; S2, filtering through a membrane to remove thalli: filtering the fermentation liquid through a ceramic membrane to obtain membrane clear liquid; S3, heat treatment: adding alkali liquor into the membrane clear liquid, adjusting the pH value of the membrane clear liquid, and performing water bath heat treatment to obtain heat treatment liquid; S4, filtering and precipitating: adding diatomite into the heat treatment liquid, then carrying out filter pressing, taking filtrate I, carrying out ultrafiltration through an organic membrane, taking filtrate II, and adding absolute ethyl alcohol to obtain crude polysialic acid precipitate; S5, dissolving and decolorizing; and S6, concentrating and centrifuging. The preparation method is simple, thalli are removed more thoroughly, redundant salt and small molecule impurities are removed, the protein removal effect is obvious under the alkaline heating condition, the addition of a complexing agent is reduced, and the method is safe and effective.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular to a preparation method of polysialic acid and polysialic acid products. Background technique [0002] Polysialic acid (PSA) is a unique class of linear, uniform polyα2,8-linked sialic acid carbohydrates, which are mainly attached to neural adhesion molecules in the vertebrate nervous system through typical N-linked glycosidic bonds. Polysialic acid plays a key role in neural development by changing the adhesion of neural adhesion molecules in the nervous system to regulate nerve cell development, nerve guidance and synapse formation. [0003] Its hydrolyzed product sialic acid is a substance present in animal cells, which can prevent pathogens from attaching to cells and make cells produce immune antibodies; as a food additive, it is suitable for weaning foods for babies and nutrition for hospitalized patients; as a drug for It is effective in central or external neurologic...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/26C08B37/00C12R1/19
CPCC12P19/26C08B37/0003
Inventor 袁丽霞王煜陈祥松吴金勇朱薇薇王刚孙立洁姚建铭李翔宇王纪
Owner HEFEI INSTITUTES OF PHYSICAL SCIENCE - CHINESE ACAD OF SCI
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