Process for preparing chitoligose by coupling enzyme degradation of chitosan with membrane separation

A technology of enzymatic degradation and chitosan, applied in oligosaccharide and other directions, can solve the problems of affecting large-scale production, serious pollution, complicated operation, etc., and achieve the effect of preventing inhibition, reducing pollution, and simplifying the process.

Inactive Publication Date: 2003-11-05
DALIAN GLYCOBIO
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although these methods can realize the effective separation of active chitosan oligosaccharides, they affect large-scale production due to complicated operations, serious pollution and high cost.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] The enzymatic degradation reaction of embodiment 1 chitosan

[0022] Dissolve 0.5-3.0% chitosan in acetic acid buffer (pH5.6, 0.05M), add a certain amount of enzyme, enzyme: substrate = 1:20, react at 15-60°C, use the reaction solution The percentage decrease in viscosity characterizes the degradation activity of various enzymes. Use Ping's viscometer to measure the drop time t of the reaction solution passing through the measuring section at the initial stage and after 12 hours of degradation 0 , t, then the percentage of chitosan viscosity decrease is t 0 -t / t 0 .

[0023] Used

Embodiment 2

[0024] Example 2 Coupling experiment of enzymatic degradation reaction and membrane separation

[0025] In a stainless steel reaction kettle, dissolve 1% chitosan in 20L of 1% acetic acid solution, add 1% (to chitosan) chitosanase into the reaction system, stir at 40°C for 1 hour, the viscosity Decrease to 40-60% to start ultrafiltration, the ultrafilter has a membrane area of ​​2m 2 Hollow fiber membrane filter with cutting molecules of 2000-10000, fed intermittently, fed 6 batches in total, ultrafiltration filtrate 20L. The 14L ultrafiltrate was treated with a roll-type flat membrane nanofilter at 2 o'clock, and the membrane was treated with MgSO 4 The rejection rate is 80-95%, and the membrane area is 0.5m 2 , to get concentrated solution 2L, filtrate 12L, the degree of polymerization of concentrated liquid chitosan oligosaccharides is mainly 3-8 sugars, and only contains trace amounts above 10 sugars.

Embodiment 3

[0026] Embodiment 3 intermittent feed and continuous feed experiment

[0027] The total reaction volume is 20L, and the test conditions are the same as in Example 2. After 1 hour of feeding, the ultrafiltration starts, and at the same time, the raw material and acetic acid are continuously added at the same speed, and the ultrafiltration filtrate is 60L, and 1.8L of concentrated solution is obtained through nanofiltration. In feeding and continuous feeding, the proportions of chitosan oligosaccharides with different degrees of polymerization are shown in the table below. Continuous feeding reduces the relative amount of 3-4 sugars, while the relative amount of 6-8 sugars increases.

[0028] Polymerization

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PUM

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Abstract

A method for producing chitosan oligosaccharides coupled with enzymatic degradation of chitosan and membrane separation is characterized in that the production process is as follows: (1) preparing chitosan oligosaccharides by enzymatic degradation of chitosan; (2) adopting hollow fiber or flat ultra- Combine the filter with the degradation reaction to separate the active oligosaccharides in situ; (3) filter the ultrafiltrate with a nanofilter to separate high-concentration active chitosan oligosaccharides. The invention realizes the integration of reaction and separation, and prevents secondary degradation of active oligosaccharides. Since the process realizes continuous operation, the degrading enzyme can be used continuously, the yield is improved, the cost is reduced, and it is beneficial to realize industrialization.

Description

technical field [0001] The invention is a technology for producing chitosan with high physiological activity by combining enzymatic degradation of chitosan and membrane separation. Background technique [0002] Oligochitosan refers to a kind of oligosaccharides with 2-10 glucosamines bound to the β-1 and 4 positions. It has the ability to inhibit tumor growth, enhance the body’s immunity, and reduce blood cholesterol; it can promote bifidity. Bacillus proliferation, inhibit the growth of intestinal harmful bacteria; it also has the ability to hinder the growth and reproduction of pathogenic bacteria, and enhance the defense ability of plants against diseases and insect pests, so it is widely used in medicine, health food and agriculture. In addition, it is also used in the chemical industry as a flocculant, separation membrane, enzyme immobilization carrier, etc. Chitosan oligosaccharides are usually obtained by degrading chitosan. At present, acid degradation, oxidative de...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07H3/06
Inventor 杜昱光张铭俊王毓福曲天明张虎白雪芳虞星炬
Owner DALIAN GLYCOBIO
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