Preparation and purification method of chitosan oligosaccharide

Abstract

The invention discloses a preparation and purification method of chitosan oligosaccharide. The method comprises the following steps of: taking chitosan dried to constant weight, adding a HCl solution,and adjusting a pH value; adding a chitosanase solution, performing insulation for 2h-6h, and then heating the material in a boiling water bath and performing enzyme deactivation; performing rotary evaporation and condensation at 50 DEG C or lower, and then placing the material in a freeze dryer to obtain a crude chitosan oligosaccharide extract; dissolving the crude chitosan oligosaccharide extract in a NH4HCO3 solution, and after completely dissolved, filtering the material through a filter membrane to obtain a chitosan oligosaccharide solution; dumping resin into a chromatography column, using NH4HCO3 for flushing resin until the resin is compacted, slowly dumping the chitosan oligosaccharide solution into the chromatography column, collecting an eluate, and performing rotary evaporation and condensation to obtain chitosan oligosaccharide. The method has the advantages that the chitosanase is used to decompose chitosan to prepare chitosan oligosaccharide, and the chitosan oligosaccharide with a single polymerization degree is prepared by gel column chromatographic separation and purifying, the purity can achieve more than 97%, the prepared chitosan oligosaccharide has good hypolipidemic activity, protects the liver, and enhances the antioxidant activity of the body.

Description

technical field [0001] The invention relates to a preparation and purification method of chitosan oligosaccharide. Background technique [0002] Chitosan is derived from shrimp and crab shells. It is obtained by removing the acetyl group on N from chitin under the action of concentrated alkali or deacetylase. (1,4) Linked by glycosidic bonds, it is a natural, non-toxic preservative and flocculant in the food industry, and can absorb heavy metal ions such as cadmium, mercury, and copper in water, but chitosan has a large molecular weight and is insoluble Therefore, it is very necessary to degrade chitosan to obtain chitosan oligosaccharides with small molecular weight, good water solubility and easy absorption by human body. However, it is difficult to prepare high-purity chitosan monomers. However, chitosan oligosaccharide mixtures are often used for biological activity evaluation, which limits the in-depth study of the biological activity mechanism of chitosan oligosacchar...

AI Technical Summary

Problems solved by technology

[0002] Chitosan is derived from shrimp and crab shells. It is obtained by removing the acetyl group on N from chitin under the action of concentrated alkali or deacetylase. (1,4) Linked by glycosidic bonds, it is a natural, non-toxic preservative and flocculant in the food industry, and can absorb heavy metal ions such as cadmium, mercury, and copper in water, but chitosan has a large molecular weight and is insoluble Therefore, it is very necessary to degrade chitosan to obtain chitosan oligosaccharides with small molecular weight, good water solubility and easy absorption by human body. However, it is difficult to prepare high-purity chitosan monomers. , the biological activity evaluation of chitosan oligosaccharide mixture is often used, which limits the in-depth study of the biological activity mechanism of chitosan oligosaccharides