Method for extracting polysialic acid

A technology for polysialic acid and purpose, applied in the field of extracting polysialic acid

Inactive Publication Date: 2008-06-11
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Moreover, there is no company in the world that can carry out large-scale production of polysialic acid, so it is

Method used

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  • Method for extracting polysialic acid
  • Method for extracting polysialic acid

Examples

Experimental program
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Embodiment 1

[0024] Embodiment 1. Preparation of polysialic acid fermentation broth

[0025] Strain Escherichia coli K235-JY II-74.

[0026] Fermentation Medium (g / L) Sorbitol 20, Ammonium Chloride 2.0, Dipotassium Hydrogen Phosphate 1.5, MgSO 4 0.9, tryptone 1.5, pH 7.8.

[0027] Utilize 30L fermentation medium, adopt pH control method to ferment on 50L B.Braum mechanical stirring tank, inoculum size 4%, add a small amount of polyoxyethylene polyoxypropylene glyceryl ether (bubble enemy) to control the inoculum in the fermentation process before sterilization For foaming, the rotation speed of the fermenter is 300-500rpm, the ventilation rate is 30 L / min, the gauge pressure is 0.05Mpa, and the fermentation temperature is 37°C. The ammonia water to adjust the pH value is concentrated ammonia water, and the concentration of hydrochloric acid is 3M. The pH in the growth stage of the bacteria naturally drops to 6.4, and then the pH value is controlled at about 6.4 with ammonia water and h...

Embodiment 2

[0028] Example 2. Extraction of polysialic acid

[0029]Take 5L of fermented liquid, remove bacteria through plate and frame filtration, add 100-250g of sodium chloride (NaCl) to the resulting filtered clear liquid and stir to dissolve it, then add 2 to 4 times the volume of 95% of the clear liquid containing sodium chloride Mass concentration of ethanol, precipitate polysialic acid, and let it stand overnight, obtain polysialic acid precipitate by decantation or centrifugation, then wash with a small amount of 75% mass concentration ethanol for 2 to 4 times, and then dissolve the polysialic acid precipitate in an appropriate amount In deionized water, the obtained turbid liquid was filtered to remove part of the protein, and the obtained filtrate was adjusted to pH 7-8, and about 50 mL of alkaline protease (enzyme activity 2-50,000 U / mL) was added, and it was heated in a constant temperature water bath at 30-70 °C. Treat for 3-7 hours under the same conditions, then add exces...

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Abstract

The invention relates to a method for extracting polysialic acid, belonging to biological engineering technical field. The invention uses the fermentation liquor of one Escherichia coli generating polysialic acid as material, filters and removes thallus, adds some solvents as sodium chloride and alcohol into supernatant to deposit polysialic acid, dissolves the polysialic acid deposits via deionized water, filters to remove modified protein, uses alkali prolease to treat filtrate, complexes deposits via cetyl pyridinium chloride (CPC) or cetyltrimethylammonium bromide (CTAB), and dissociates, uses over alcohol to deposit polysialic acid, dissolves the deposits via ultra-pure water, grades and purifies via chromatography column, collects and dialyzes eluent, cools and dries to obtain polysialic acid. The invention provides an effective polysialic acid separation purification method, which an obtain polysialic acid product with more than 97% purity, to be used as materials of drug and cosmetic product.

Description

technical field [0001] A method for extracting polysialic acid belongs to the technical field of bioengineering. Background technique [0002] Sialic acid is a derivative of neuraminic acid. Its chemical structure is an acidic amino sugar with 9 carbon atoms and a pyranose structure. It is named 5-amino-3,5-dideoxy-D-glycerol -D-Galacnonylose. Different sialic acid derivatives are formed according to different linking groups on the 5th carbon position. The two most important sialic acid derivatives are 5-acetylamino-3,5-dideoxy-D-glycerol-D-galactonanone Sugars (NANA, Neu5Ac, N-acetylneuraminic acid) and 3-deoxy-D-glycerol-D-galactononulose (KDN), from which the rest of the sialic acids were derived. Among them, N-acetylneuraminic acid is more common in vertebrates, such as fish, amphibians, reptiles, birds and other animals. Sialic acid is ubiquitous in mammals and is relatively abundant in cerebrospinal fluid and mucus. Babies have higher levels of sialic acid in infan...

Claims

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Application Information

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IPC IPC(8): C08B37/00
Inventor 詹晓北吴剑荣郑志永朱莉
Owner JIANGNAN UNIV
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