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Preparation of low-molecular weight chitoglycan or chitooligose

A low molecular weight, chitosan technology, applied in the field of chemical biology of renewable resources, can solve the problems of difficult to obtain in large quantities, harsh reaction conditions, modification, etc., and achieve the effect of improving immobilization rate, good mechanical properties, and increasing affinity

Inactive Publication Date: 2007-06-06
WUHAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the methods for preparing low molecular weight chitosan and chitosan oligosaccharide mainly include chemical method and biological method. The reaction process of chemical method is violent, it is not easy to obtain low molecular weight chitosan and the product structure is modified.
Enzymatic degradation only breaks the β(14) glycosidic bonds of chitosan, and generally does not change the residue structure of chitosan molecules, but specific enzymes are expensive and difficult to obtain in large quantities, so they are not suitable for low molecular weight chitosan and chitosan. Industrialized production of chitosan oligosaccharides; use free enzyme hydrolysis to prepare low molecular weight chitosan and chitosan oligosaccharides, the enzyme cannot be reused after the reaction, it is uneconomical, the reaction conditions are harsh, and the product contains 0.1% (w / w) sugar and enzyme protein Compounds, this product cannot be used in food, medicine and other fields because it can cause pyrogen reactions

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] Embodiment 1: N-succinyl chitosan is made into 0.25g / ml aqueous solution, squeezes into 30ml 0.3g / ml calcium chloride and 70ml dehydrated alcohol to form with syringe (3# needle) dropwise under magnetic stirring In the mixed solution, stirring was continued at room temperature for 6 hours to obtain N-succinyl chitosan hydrogel balls. Weigh 4g (wet weight) of this hydrogel ball, place it in 12ml of citric acid-disodium hydrogen phosphate buffer solution (0.05M, pH3.0) with 0.5% glutaraldehyde dissolved in the volume percentage, shake at room temperature for 12 hours . The cross-linked N-succinyl chitosan hydrogel balls were collected by filtration, and washed repeatedly with water until the washing solution had no absorption at 245 nm. Dissolve neutral protease with a ratio of 0.2 to the immobilized carrier weight (wet weight) in 0.05M citric acid-disodium hydrogen phosphate buffer solution (pH3.0), add cross-linked immobilized carrier to the enzyme solution, Shake con...

Embodiment 2

[0016] Embodiment 2: N-succinyl chitosan is made into 0.30g / ml aqueous solution, squeezes into 40ml 0.15g / ml calcium chloride and 60ml dehydrated alcohol to form with syringe (4# needle) dropwise under magnetic stirring In the mixed solution, stirring was continued at room temperature for 3 hours to obtain N-succinyl chitosan hydrogel balls. Weigh 4 g (wet weight) of this hydrogel ball, place it in 16 ml of citric acid-disodium hydrogen phosphate buffer solution (0.2 M, pH 3.5) with a volume percentage of 0.8% glutaraldehyde, shake at room temperature for 10 hours . The cross-linked N-succinyl chitosan hydrogel balls were collected by filtration, and washed repeatedly with water until the washing solution had no absorption at 245 nm. Dissolve neutral protease with a ratio of 0.45 to the immobilized carrier weight (wet weight) in 0.2M citric acid-disodium hydrogen phosphate buffer solution (pH3.5), add cross-linked immobilized carrier to this enzyme solution, Shake continuous...

Embodiment 3

[0017] Embodiment 3: N-succinyl chitosan is made into 0.30g / ml aqueous solution, squeezes into 50ml 0.2g / ml calcium chloride and 50ml dehydrated alcohol to form with syringe (5# needle) dropwise under magnetic stirring In the mixed solution, stirring was continued at room temperature for 4 hours to obtain N-succinyl chitosan hydrogel balls. Weigh 4g (wet weight) of this hydrogel ball, place it in 20ml of citric acid-disodium hydrogen phosphate buffer solution (0.3M, pH4.0) with 1.0% glutaraldehyde dissolved in the volume percentage, shake at room temperature for 8 hours . The cross-linked N-succinyl chitosan hydrogel balls were collected by filtration, and washed repeatedly with 0.3M citric acid-disodium hydrogen phosphate buffer solution (pH4.0) until the washing solution had no absorption at 245nm. Dissolve neutral protease with a ratio of 0.6 to the immobilized carrier weight (wet weight) in 0.3M citric acid-disodium hydrogen phosphate buffer solution (pH4.0), add cross-li...

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Abstract

Production of low-molecular weight chitoglycan or chitooligose is carried out by fixing dispase on N-succinchitoglycan aquogel ball from cross-linking method, degrading chitoglycan from fixed enzyme, controlling enzymolysis time, regulating degraded product pH to 8-9 from sodium hydroxide, depositing by alcohol, washing and vacuum drying to obtain low-molecular weight chitoglycan or chitooligose with free amidogen. It achieves much yield, less molecular weight distribution and continuous industrial production. It can be used for medicine and food.

Description

technical field [0001] The invention relates to a preparation method of low molecular weight chitosan or chitosan oligosaccharide, which belongs to the field of chemical biology of renewable resources. Background technique [0002] Low molecular weight chitosan and chitosan oligosaccharide are the degradation products of chitin deacetylation, which have better solubility and biocompatibility than high molecular weight chitosan. Chitosan with different molecular weights has different biological and physiological activities, so the preparation of chitosan with different molecular weights is of great significance for the application of chitosan in food and medicine. At present, the methods for preparing low molecular weight chitosan and chitosan oligosaccharide mainly include chemical method and biological method. The reaction process of chemical method is violent, it is not easy to obtain low molecular weight chitosan and the product structure is modified. Enzymatic degradati...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P19/04
Inventor 杜予民李瑾
Owner WUHAN UNIV
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