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Method for preparing chitooligosaccharide with snailase and application thereof

A technology of snail enzyme and chitosan oligosaccharide, which is applied in the direction of medical preparations, applications, and pharmaceutical formulas containing active ingredients, and can solve the problems of limited source of specific enzymes, many steps in chemical degradation methods, and low efficiency of single enzyme degradation, etc. problems, achieve good industrial promotion practicability and value, improve idiopathic pulmonary fibrosis, and improve the effect of lung fibroblast activation

Active Publication Date: 2019-01-04
GUANGDONG PHARMA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The purpose of the present invention is to provide a method for preparing chitooligosaccharides with helicase, which mainly aims at the problems in the prior art: the cost of physical degradation method is high and the yield is low, the steps of chemical degradation method are various, and the obtained product monosaccharide is more , Chitosan oligosaccharide content is low and a large number of chemical reagents will pollute the environment, and when the commonly used enzymatic hydrolysis method is used to prepare chitosan oligosaccharides, the degradation efficiency of each single enzyme is not high and the source of some specific enzymes is limited and expensive. When using several non-specific enzymes, the problem of interaction degradation must be considered, which leads to limitations in the selection of enzymes

Method used

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  • Method for preparing chitooligosaccharide with snailase and application thereof
  • Method for preparing chitooligosaccharide with snailase and application thereof
  • Method for preparing chitooligosaccharide with snailase and application thereof

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Embodiment 1

[0036] Embodiment 1, a kind of method that prepares chitosan oligosaccharide with helicase

[0037] S1 Take the chitosan raw material and dry it to constant weight, dissolve it in 0.2mol / L acetic acid-sodium acetate buffer solution with a pH value of 4.5, stir and dissolve, and prepare a chitosan solution with a mass concentration of 1.0%;

[0038]S2 Adding a helicase solution with a mass concentration of 1.0% to the chitosan solution obtained in step S1, the mass ratio of the helicase to the chitosan substrate is 1:10, after enzymolysis in a water bath at 40°C for 2.5h, Inactivate in a water bath at 100°C for 10 minutes, cool to room temperature, stir continuously, add dropwise NaOH solution with a mass concentration of 5%, adjust the pH value to 4.5, filter, and collect the filtrate;

[0039] S3 put the filtrate obtained in step S2 into a dialysis device with a regenerated cellulose dialysis bag with a molecular weight cut-off of 3000 Da in the inner layer and a regenerated ...

Embodiment 2

[0041] Embodiment 2, a kind of method that prepares chitosan oligosaccharide with helicase

[0042] S1 Take the chitosan raw material and dry it to constant weight, dissolve it in 0.2mol / L acetic acid-sodium acetate buffer solution with a pH value of 4.5, stir and dissolve, and prepare a chitosan solution with a mass concentration of 1.0%;

[0043] S2 adds the helicase solution that mass concentration is 1.0% to the chitosan solution obtained in step S1, and described helicase and chitosan substrate mass ratio are 1:10, after enzymolysis 2h in 40 ℃ of water bath boxes, 100 Inactivate in a water bath at ℃ for 15 minutes, cool to room temperature, stir continuously, add NaOH solution with a mass concentration of 5% dropwise, adjust the pH value to 6, filter, and collect the filtrate;

[0044] S3 put the filtrate obtained in step S2 into a dialysis device with a regenerated cellulose dialysis bag with a molecular weight cut-off of 3000 Da in the inner layer and a regenerated cell...

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Abstract

The invention belongs to the technical field of oligosaccharide and particularly relates to a method for preparing chitooligosaccharide with snailase and an application thereof. The method for preparing the chitooligosaccharide with the snailase has the advantages of exploring and optimizing the specific process conditions such as enzymatic hydrolysis time, enzymatic hydrolysis temperature, substrate concentration and enzyme bottom mass ratio of enzymatic hydrolysis of chitosan by snailase to improve enzymatic efficiency, maintaining the properties of the chitooligosaccharide in combination with a freeze-drying method to facilitate the preservation and further use of products, and providing a basis for the large-scale production of the oligosaccharide with the snailase. The prepared oligosaccharide has the advantages of significantly improving fibroblast activation in a rat model of idiopathic pulmonary fibrosis, having a good effect on improving the idiopathic pulmonary fibrosis, andbeing beneficial to the rehabilitation of patients with the idiopathic pulmonary fibrosis.

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 helicase and its application. Background technique [0002] Chitosan, also known as acetyl chitin, is a compound that removes N-acetyl groups from chitin, and has biodegradability, safety, and good biocompatibility. However, chitosan is a long-chain macromolecule with poor water solubility and high solution viscosity under physiological conditions, so its application as a biological effector is greatly limited. Chitosan oligosaccharide is its degradation product, which refers to a polymer of glucosamine with a degree of polymerization less than 10. It has a small molecular weight, good water solubility, is easy to be dispersed and absorbed, and has many biological properties similar to chitosan, especially Oligochitosan with a degree of polymerization of 5 and 6 has anti-oxidation, anti-tumo...

Claims

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Application Information

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IPC IPC(8): C12P19/26C12P19/14A61K31/722A23L33/125A61P11/00
CPCA23V2002/00A61K31/722A23L33/125A61P11/00C12P19/14C12P19/26A23V2200/314A23V2250/28
Inventor 苏政权曹华
Owner GUANGDONG PHARMA UNIV
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