Method for preparing chitosan oligosaccharide by using lipase

Abstract

The invention belongs to the technical field of oligosaccharide preparation, and particularly relates to a method for preparing chitosan oligosaccharide by using lipase. The method for preparing the chitosan oligosaccharide by using the lipase comprises the steps that chitosan is dissolved in an acetic acid-sodium acetate buffer solution and sucrose fatty acid ester is added so as to prepare a chitosan solution, lipase liquid is added into the chitosan solution for enzymatic hydrolysis, an enzymatic hydrolysate is dialyzed after the enzymatic hydrolysis is completed, and the obtained dialysateis concentrated and then dried. The method for preparing the chitosan oligosaccharide by using the lipase is easy and rapid to operate, the required preparation time is short, and the obtained degradation product, namely the chitosan oligosaccharide has a high yield, uniform distribution of molecular weight and consistent color formation.

Description

technical field [0001] The invention belongs to the technical field of oligosaccharide preparation, and in particular relates to a method for preparing chitosan oligosaccharide by using lipase. Background technique [0002] Chitosan, also known as deacetylated chitin, is a long-chain macromolecule. It has poor water solubility under physiological conditions and high solution viscosity. Its application as a biological effector is very limited. Oligochitosan is a low-molecular-weight polymer with a low degree of polymerization (2-20) obtained by degrading chitosan. Its segment molecules are connected with a large number of reactive groups hydroxyl and amino groups, so it has a low molecular weight. , high solubility, good moisture absorption and moisturizing properties, strong absorption capacity and good biocompatibility, etc., and also has anti-inflammatory, enhanced immunity, lower cholesterol, lower blood fat, anti-tumor, antibacterial, anti-virus, clear Physiological ac...

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Problems solved by technology

[0004] Chinese patent document CN 1320123 C discloses a method for producing low-molecular-weight chitosan by using lipase. The enzymolysis reaction is carried out under the conditions of a substrate concentration of 10-100 g / L and a reaction time of 5-6 hours, and the filtrate is vacuum-concentrated and filtered. Gel chromatographic column to obtain low molecular weight chitosan, this method effectively reduces the production cost, but the required reaction time is long, the enzyme liquid and the air contact time are long and easy to be oxidized, which inevitably affects the degradation products in the later stage. color makes a big difference

Tan Peiyi and others studied the orthogonal experiment method to optimize the process of preparing chitosan oligosaccharides by lipase. The optimal conditions determined were: the amount of lipase added was 1.2g / 100mL, the enzymatic hydrolysis temperature was 50°C, the pH of the solution was 4.5, and the enzymatic hydrolysis The time is 6h, but the time required for the reaction is long and the yield is low

Chinese patent document CN 102492664 B discloses a method for preparing chitosan oligosaccharides using compound enzymes, wherein the compound enzymes include cellulase, lysozyme, amylase, lipase, glucose and papain. rate, the obtained chitosan oligosaccharide products are evenly distributed, but there are many types of enzymes required, and the steps are complicated

Therefore, it is necessary to overcome the problems of low yield, long reaction time and complicated steps caused by the high viscosity of the current chitosan substrate, so as to provide people with a new type of simple operation and short reaction time using lipase to prepare chitosan. sugar method

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