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Method for producing Chitosan oligosaccharide by using enzyme method to degrade chitosan

A technology of enzymatic degradation and chitosan oligosaccharides, which is applied in the field of chitosan oligosaccharides production, can solve the problems of low substrate concentration, high energy consumption, and low enzyme activity, and achieve low pollution, low energy consumption, and high yield. Effect

Inactive Publication Date: 2008-08-13
WUHAN EAST ANGEL BIOENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, it generally has problems such as low substrate concentration, low enzyme activity, many side reactions, long enzymatic hydrolysis time, and high energy consumption.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] 1. Utilize Bacillus TQK, which can highly express chitosanase, remove impurities through fermentation, separation and purification, and obtain TQK enzyme liquid with a measured enzyme activity of 718u / ml, and store it at a low temperature (0-4°C) for later use.

[0021] 2. Preparation of chitosan solution: prepare the substrate concentration at 15%, add 2% acetic acid solution, adjust pH 4, add TQK enzyme solution according to the enzyme-substrate ratio of 1:10.

[0022] 3. Enzyme hydrolysis reaction.

[0023] Stir at 200-300 rpm, pH 4, temperature 45°C, time 6 hours; monitor with GPC instrument, take samples to determine the viscosity of the reactant is lower than 10.0mpa.s, stop the enzymatic hydrolysis reaction.

[0024] 4. Centrifugal separation.

[0025] Centrifuge the reacted enzymolysis sugar solution at a speed of 14000r / min for 10 minutes to separate slag and other insoluble matter.

[0026] 5. Flocculation, clarification and filtration.

[0027] Heat the ce...

Embodiment 2

[0037] 1, utilize the TQK enzyme liquid of embodiment 1;

[0038] 2. Prepare chitosan solution: prepare substrate concentration at 18%, add 2% acetic acid solution, adjust pH to 4.6, add TQK enzyme solution according to the enzyme-to-substrate ratio of 1:10;

[0039] 3. Enzymolysis reaction, stirring 200-300 rpm, pH 4.6, temperature 49°C, time 5 hours; monitor with GPC instrument, take samples to determine that the viscosity of the reactant is lower than 10.0mpa.s, stop the enzymolysis reaction;

[0040] Following centrifugation; flocculation, clarification, filtration treatment; ion exchange treatment; ultrafiltration classification treatment; vacuum concentration treatment; spray drying treatment;

Embodiment 3

[0042] 1, utilize the TQK enzyme liquid of embodiment 1;

[0043] 2. Preparation of chitosan substrate solution: substrate concentration is 20% preparation, by adding 2% acetic acid solution, adjusting pH5.2, adding TQK enzyme solution according to the ratio of enzyme bottom to 1:10;

[0044] 3. Enzymolysis reaction, stirring 200-300 rpm, pH 5.2, temperature 52°C, time 5.5 hours; monitor with GPC instrument, take samples to determine that the viscosity of the reactant is lower than 10.0mpa.s, stop the enzymolysis reaction;

[0045] Following centrifugation; flocculation, clarification, filtration treatment; ion exchange treatment; ultrafiltration classification treatment; vacuum concentration treatment; spray drying treatment;

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Abstract

The invention discloses a method for preparing chitosan oligosaccharide by degrading chitose comprising steps removing extraneous constituents through fermentation culturing and separation purifying by utilizing shell dextranase bacillus TQK to obtain TQK enzyme liquor which enzyme activation is up to 718u / ml, putting TQK enzyme liquor into 10-20poncentration degrading liquor which uses chitose as bottom liquor, the ratio of TQK enzyme liquor to bottom object is 1:10, and the enzymolysis reaction condition is: mixing speed is 200-300 r / min, Ph is 4-5.5, temperature is 45-52 DEG C and time is 4-6 hours, then centrifugal separating, flocculating, fining, filtrating, ion exchanging, hyperfiltration stage treating, vacuum concentration disposing, sponging drying disposing to prepare chitosan oligosaccharide. The enzymolysis bubatrate concentration can be up to 15-20, enzymolysis time is only 4-6 hours, receiving rate is high, energy consumption is low, pollution is small, also the invention can prepare narrow molecular weight distribution chitosan oligosaccharide with average molecular weight D1500 in quack speed, low consumption and large mass.

Description

technical field [0001] The invention relates to a method for producing chitosan oligosaccharides, in particular to a method for enzymatically degrading chitosan to produce chitosan oligosaccharides with an average molecular weight less than D1500. Background technique [0002] The content of chitin in nature is second only to cellulose, and it is mainly divided into the following three levels of derivative products based on its different processing methods: 1. Chitin (chitin), with a molecular weight of more than 1 million, suitable for industry and environmental protection Should be shipped, insoluble in water, it is difficult to be absorbed by the body. 2. Chitosan (chitosan), with a molecular weight of about 100,000, is generally only soluble in organic weak acids, insoluble in water, can be absorbed by the human body, and can be used as food additives and raw materials for health products. 3. Chitosan oligosaccharide is a kind of oligosaccharide with 2-10 glucosamines b...

Claims

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Application Information

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IPC IPC(8): C12P19/04
Inventor 黄代勇黄芳王光荣
Owner WUHAN EAST ANGEL BIOENG
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