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Preparation method of chitosan oligosaccharide

A technology of oligochitosan and chitosan, which is applied in the field of bioengineering, achieves the effects of high yield, high biological activity and simplified process flow

Pending Publication Date: 2021-08-24
SHANGHAI INST OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The technical problem to be solved by the present invention is: how to prepare low-cost, less-polluted and highly efficient chitosan oligosaccharides with a molecular weight range of 1-3kDa

Method used

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  • Preparation method of chitosan oligosaccharide

Examples

Experimental program
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Effect test

Embodiment 1

[0032] A kind of preparation method of chitosan oligosaccharide, comprises the steps:

[0033] (1) Preparation of chitosan solution: take 1g chitosan, dissolve in 50mL water and stir fully to make chitosan evenly dispersed in water, add hydrochloric acid to make hydrochloric acid content 0.5wt%, adjust with 2wt% sodium hydroxide pH to 5.0, stir until chitosan is completely dissolved.

[0034] (2) Enzymolysis: add chitosan enzyme to the above chitosan solution, the amount of enzyme added is 400U, the enzymolysis rotation speed is 200rpm, the temperature is 45°C, and the time is 8-10h. After the reaction is over, heat up to 85°C to inactivate the enzyme for 10-30 minutes.

[0035] (3) Separation and purification: use the microfiltration device to pass the above enzymatic hydrolysis solution through the 0.45 μm and 0.22 μm filter membranes of the water system, and then pass the above enzymatic hydrolysis solution through 5.0×10 4 Da, 1.0×10 4 Da, 3.0×10 3 Large plate ultrafil...

Embodiment 2

[0038] A kind of preparation method of chitosan oligosaccharide, comprises the steps:

[0039] (1) Preparation of chitosan solution: take 1g chitosan, dissolve in 60mL water and stir fully to make chitosan evenly dispersed in water, add acetic acid to make the acetic acid content 1wt%, adjust pH with 10wt% sodium hydroxide To 5.5, stir until the chitosan is completely dissolved.

[0040] (2) Enzymolysis: add chitosan enzyme to the above chitosan solution, the amount of enzyme added is 600U, the enzymolysis rotation speed is 150rpm, the temperature is 55°C, and the time is 4-6h. After the reaction is over, heat up to 85°C to inactivate the enzyme for 10-30 minutes.

[0041] (3) Separation and purification: use the microfiltration device to pass the above enzymatic hydrolysis solution through the 0.45 μm and 0.22 μm filter membranes of the water system, and then pass the above enzymatic hydrolysis solution through 5.0×10 4 Da, 1.0×10 4 Da, 3.0×10 3 Large plate ultrafiltratio...

Embodiment 3

[0044] A kind of preparation method of chitosan oligosaccharide, comprises the steps:

[0045] (1) Preparation of chitosan solution: take 1g chitosan, dissolve in 70mL water and stir fully to make chitosan evenly dispersed in water, add phosphoric acid to make phosphoric acid content 2wt%, adjust pH with 5wt% sodium hydroxide To 6, stir until the chitosan is completely dissolved.

[0046] (2) Enzymolysis: add chitosan enzyme to the above chitosan solution, the amount of enzyme added is 700U, the enzymolysis rotation speed is 200rpm, the temperature is 55°C, and the time is 3-5h. After the reaction is over, heat up to 85°C to inactivate the enzyme for 10-30 minutes.

[0047](3) Separation and purification: use the microfiltration device to pass the above enzymatic hydrolysis solution through the 0.45 μm and 0.22 μm filter membranes of the water system, and then pass the above enzymatic hydrolysis solution through 5.0×10 4 Da, 1.0×10 4 Da, 3.0×10 3 Large plate ultrafiltratio...

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Abstract

The invention discloses a method for preparing chitosan oligosaccharide. The preparation method comprises the following steps: dissolving chitosan to prepare a chitosan solution, adjusting the pH value, and adding chitosan for enzymolysis; carrying out microfiltration impurity removal, ultrafiltration impurity removal and concentration desalting on enzymatic hydrolysate obtained by enzymolysis in sequence, then collecting a concentrated solution, and finally freeze-drying the concentrated solution to obtain the chitosan oligosaccharide. According to the preparation method, the chitosan oligosaccharide with the molecular weight concentrated in 1-3 kDa is prepared with the yield of 60% or above, the prepared chitosan oligosaccharide is good in water solubility and high in biological activity, and the preparation method is high in yield, low in production cost and suitable for large-scale production.

Description

technical field [0001] The invention relates to a preparation method of chitosan oligosaccharide, which belongs to the technical field of bioengineering. Background technique [0002] Chitosan is a basic polysaccharide composed of D-glucosamine (GlcN) linked by β-1,4-glycosidic bonds. Oligochitosan is a product obtained from chitin or chitosan through physical methods, chemical methods, enzymatic hydrolysis and other methods. [0003] The degree of polymerization of chitosan oligosaccharide is generally 2 to 20, and it is the only positively charged basic cationic oligosaccharide in nature. Oligochitosan is easily soluble in water, low viscosity, good biocompatibility, non-toxic, low allergenicity, good cell transduction, easy gastrointestinal absorption. A large number of studies have confirmed that oligochitosan has a variety of biological activities, including antibacterial, anti-tumor, anti-oxidation, anti-inflammatory, anti-hypertensive, anti-diabetic, anti-Alzheimer'...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/26C12P19/14C08B37/08C07H3/04C07H3/06C07H1/06
CPCC12P19/26C12P19/14C08B37/003C08B37/0003C07H3/04C07H3/06C07H1/06
Inventor 李茜茜叶扬志王团团荣绍丰管世敏蔡宝国
Owner SHANGHAI INST OF TECH
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