Method for preparing chitosan oligosaccharide from papain through freeze drying

A papain and chitosan oligosaccharide technology, which is applied in the field of oligosaccharide preparation, can solve the problems of low molecular weight of chitosan oligosaccharide, difficult reaction control, complicated preparation steps, etc., and achieves convenient drug storage, good industrial promotion practicability and value , the effect of uniform molecular weight distribution

Inactive Publication Date: 2018-02-27
GUANGDONG PHARMA UNIV
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Problems solved by technology

The research on the preparation of chitosan oligosaccharides by chemical degradation method has made great progress, but the steps are more complicated, and the chemical degradation method requires a large amount of chemical reagents, which pollutes the environment more, and the reaction is not easy to control
Chinese patent application CN105884932A discloses a preparation method of low-molecular-weight chitosan, including: step 1: coordinate chitosan with metal ions to convert chitosan metal complexes into chitosan metal complexes, and step 2: chitosan metal complexes Oxidative degradation of the substance, step three: the product obtained in step two is separated and purified, the molecular weight of chitosan oligosaccharide prepared by this method is relatively low, but the preparation steps are relatively complicated
Ultrasonic degradation method is easy to operate, but 95% of the ultrasonic energy is used for system heating instead of product degradation, so the degradation efficiency is low, and it has the problem of wide molecular weight distribution and high molecular weight of the product

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  • Method for preparing chitosan oligosaccharide from papain through freeze drying
  • Method for preparing chitosan oligosaccharide from papain through freeze drying
  • Method for preparing chitosan oligosaccharide from papain through freeze drying

Examples

Experimental program
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Effect test

Embodiment 1

[0029] Embodiment 1, preparation of chitosan oligosaccharide and reducing sugar content determination experiment

[0030] 1. Preparation of Chitooligosaccharide by Enzymatic Hydrolysis of Papain

[0031] In the 25 milliliter colorimetric tube, add the mass concentration of 10mLpH4.5 and be 1.0% chitosan solution, add the papain solution that mass concentration is 1.0%, make the enzyme base mass ratio be 5%, in the water bath environment of 45 ℃ , water bath for 2h, after 2h, take out the boiling water bath for 10min to inactivate, cool to room temperature with running water and filter, discard the filter residue, take 0.5mL filtrate (or less, determined according to absorbance) in a 10mL colorimetric tube, add distilled water to 1mL, Then add 1.0mL DNS reagent, develop color in a boiling water bath for 5min, take it out, cool it to room temperature with running water, and set the volume to 10mL, use the blank tube as a reference, and measure the absorbance value at the maximum...

Embodiment 2

[0037] Embodiment 2, papain enzymatic hydrolysis prepares chitosan oligosaccharide condition optimization experiment

[0038] 1. Substrate concentration single factor experiment

[0039] Under the condition that the mass ratio of fixed enzyme substrate is 5%, the enzymolysis time is 2h, and pH4.5, the selected substrate concentration (chitosan mass concentration) is respectively 0, 0.2%, 0.5%, 0.8%, 1.0%, 1.2%, 1.5%, by measuring the chitosan oligosaccharide content after the reaction as an evaluation index, the impact of the chitosan concentration on the chitosan oligosaccharide content was investigated. The experimental results of the impact of chitosan concentration on chitosan oligosaccharide content are as follows: image 3 shown. Depend on image 3 It can be seen that under the same enzymolysis conditions, the mass ratio of the enzyme substrate is 5%, the enzymolysis time is 2h, pH4.5, and the chitosan oligosaccharide content first increases with the concentration of ...

Embodiment 3

[0051] Embodiment 3, freeze-drying process optimization experiment

[0052] In the investigation of the liquid loading area of ​​freeze-drying, it is known that the diameter of the tray is 180mm, and the thickness of the liquid loading should not exceed 10mm. Prepare a 5% chitosan oligosaccharide solution, take a certain volume in 5 identical trays, make the thickness of the carrier liquid be 1mm, 3mm, 5mm, 7mm, 9mm, and then put it into a freeze dryer for slow freezing. To freeze the material, wait until the temperature of the cold hydrazine and the temperature of the material remain unchanged, and the naked eye judges that the cold hydrazine is completed before vacuum sublimation. The relationship between the thickness of the carrier liquid and the freeze-drying time is as follows: Image 6 shown.

[0053] Depend on Image 6 It can be obtained that the smaller the thickness of the carrier liquid, the faster the temperature will drop in the same low-temperature atmosphere,...

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Abstract

The invention belongs to the field of an oligosaccharide preparation method, and particularly relates to a method for preparing chitosan oligosaccharide from papain through freeze drying. The method for preparing the chitosan oligosaccharide from the papain through freeze drying provided by the invention comprises the following steps of dissolving chitosan into an acetic acid-sodium acetate buffersolution for preparing a chitosan solution; adding the papain into the chitosan solution for enzymolysis; after the enzymolysis is completed, dialyzing filter liquid by a dialyzing method; then, performing heating concentration on the collected dialyzing liquid; next, performing freeze drying and the like. The method for preparing the chitosan oligosaccharide from the papain through freeze dryingprovided by the invention has the advantages that simplicity, convenience, high speed and high efficiency are realized; the method is suitable for industrialized production; good industrial popularization practicability and values can be realized.

Description

technical field [0001] The invention belongs to the field of preparation methods of oligosaccharides, in particular to a method for preparing chitosan oligosaccharides by freeze-drying with papain. Background technique [0002] Chitosan is a natural high-molecular polymer, which belongs to aminopolysaccharide, and is known as the sixth life element after protein, fat, sugar, vitamin and inorganic salt. Chitosan is also known as deacetylated chitin, and its chemical name is polyglucosamine (1-4)-2-amino-B-D-glucose, which is composed of 2-acetylglucosamine and 2-glucosamine through β-1,4-glucoside The relative molecular mass ranges from thousands to millions due to different raw materials and different preparation methods. In recent years, researchers have found that chitosan has unique effects in regulating blood pressure and blood lipids, balancing blood cholesterol levels, and losing weight. And safer, so chitosan has become a new type of food and medicine for weight los...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08B37/08
CPCC08B37/003
Inventor 苏政权
Owner GUANGDONG PHARMA UNIV
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