Method for preparing chitosan oligosaccharide by enzymolysis of supercritical carbon dioxide fluid

A carbon dioxide, supercritical technology, applied in the field of medical materials, can solve the problems of low yield of chitosan oligosaccharide, difficult separation, environmental pollution, etc., and achieve the effects of improving immunity, good water solubility, and inhibiting tumor metastasis

Inactive Publication Date: 2019-03-15
ZHEJIANG OCEAN UNIV
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  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

At present, the methods for preparing chitosan oligosaccharides from chitosan include acid hydrolysis, enzymatic hydrolysis and physical degradation. Among them, the yield of chitosan oligosaccharides in the product prepared by acid hydrolysis is low, separation is difficult, and the environment is seriously polluted. Therefore, enzymatic hydrolysis is the A green and safe method for preparing chitosan oligosaccharides

Method used

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  • Method for preparing chitosan oligosaccharide by enzymolysis of supercritical carbon dioxide fluid

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] (1) Chitosan is added in the acetic acid solution to prepare 1w / v% chitosan solution, the chitosan solution is ground for 30min, the ball-mill ratio of the grinding process is 5:1, and the rotating speed of the ball mill is 1000r / min, Spray drying to obtain pretreated chitosan powder with a particle size of 50-800 μm.

[0027] (2) According to the mass ratio of sodium alginate, glutaraldehyde and chitosanase is 1:0.6:0.2, add 2% glutaraldehyde solution in sodium alginate solution, at room temperature, with 1500r / min Stir at a rotating speed for 30 minutes, add the chitosan enzyme solution, and cross-link at 4°C for 2 hours at a low temperature of 200 r / min to obtain a sodium alginate hydrosol immobilized with chitosan enzyme. The frequency is 12 kHz and the strength is 20 W / min. cm2 ultrasonic activation treatment for 10min, the activity of 50U / g immobilized chitosanase was obtained.

[0028] (3) Add the pretreated chitosan to the ultra-high pressure reactor, add activ...

Embodiment 2

[0030] (1) Chitosan is added in the acetic acid solution to prepare 3w / v% chitosan solution, the chitosan solution is ground for 90min, the ball-milling ratio of the grinding process is 6:1, and the rotating speed of the ball mill is 2500r / min, Spray drying to obtain pretreated chitosan powder with a particle size of 50-800 μm.

[0031] (2) According to the mass ratio of sodium alginate, glutaraldehyde and chitosanase is 1:0.9:0.5, add 3% glutaraldehyde solution in sodium alginate solution, at room temperature, with 2000r / min Stir at a rotating speed for 90 minutes, add chitosan enzyme solution, and cross-link at 10°C for 4 hours at a low temperature of 500 r / min to obtain sodium alginate hydrosol immobilized chitosan enzyme. The frequency is 13 kHz and the strength is 25 W / min. cm2 ultrasonic activation treatment for 20min, to obtain the activity of 70U / g immobilized chitosanase.

[0032] (3) Add the pretreated chitosan to the ultra-high pressure reactor, add activated immob...

Embodiment 3

[0034] (1) Chitosan is added in the acetic acid solution to prepare a 1.5w / v% chitosan solution, and the chitosan solution is ground for 60 minutes, the ball-milling ratio of the grinding process is 5.5:1, and the rotating speed of the ball mill is 1500r / min , spray-dried to obtain pretreated chitosan powder with a particle size of 50-800 μm.

[0035] (2) According to the mass ratio of sodium alginate, glutaraldehyde and chitosanase is 1:0.7:0.4, add 2.5% glutaraldehyde solution in sodium alginate solution, at room temperature, with 1800r / min Stir at a rotating speed for 50 minutes, add chitosan enzyme solution, and cross-link at 7°C at a rate of 300 r / min for 3 hours to obtain sodium alginate hydrosol immobilized chitosan enzyme. The frequency is 12.5 kHz and the strength is 235 W. / cm2 of ultrasonic activation treatment for 15min, the activity was 60U / g immobilized chitosanase.

[0036] (3) Add the pretreated chitosan to the ultra-high pressure reactor, add activated immobili...

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Abstract

The invention provides a method for preparing chitosan oligosaccharide by enzymolysis of supercritical carbon dioxide fluid. The method comprises the following steps: grinding and spray-drying a chitosan solution to obtain pretreated chitosan powder; adding a sodium alginate solution in a glutaraldehyde solution, stirring at a room temperature, adding a chitosanase solution, stirring and crosslinking at a low temperature to obtain a sodium alginate hydrosol for immobilizing chitosanase, and carrying out ultrasonic activating treatment to obtain activated immobilized chitosanase; and adding pretreated chitosan in an ultra-high-pressure reaction kettle, adding the activated immobilized chitosanase, feeding carbon dioxide gas, pressurizing and heating to enable carbon dioxide to be in a supercritical state, keeping heating to an enzymolysis temperature, and carrying out enzymolysis to obtain the chitosan oligosaccharide. Micron chitosan powder and the chitosanase which is crosslinked andimmobilized to a sodium alginate hydrosol system are taken as raw materials, and with the aid of the supersolubility of the supercritical carbon dioxide fluid, the chitosan oligosaccharide which is simple and convenient to operate, high in efficiency and narrow in molecular weight is prepared.

Description

technical field [0001] The invention belongs to the technical field of medical materials, and in particular relates to a method for preparing chitosan oligosaccharide by supercritical carbon dioxide fluid enzymatic hydrolysis. Background technique [0002] Oligochitosan is a degradation product of chitosan. Oligochitosan not only has the advantages of rich sources of chitosan and good biocompatibility, but also has the advantages of reduced molecular weight and improved water solubility, which is more conducive to absorption, and also has anti-tumor, Antibacterial, immune activation and other characteristics, it has a broader application prospect than chitosan. At present, the methods for preparing chitosan oligosaccharides from chitosan include acid hydrolysis, enzymatic hydrolysis and physical degradation. Among them, the yield of chitosan oligosaccharides in the product prepared by acid hydrolysis is low, separation is difficult, and the environment is seriously polluted....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/14C12P19/26
CPCC12P19/14C12P19/26Y02P20/141Y02P20/54
Inventor 方旭波陈小娥袁高峰迟海波孙海燕
Owner ZHEJIANG OCEAN UNIV
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