Chitosan nano particles embedded with alpha-galactosidase and preparation method thereof

A technology of galactosidase and chitosan nanometer, which is applied in the molding or processing of animal feed, animal husbandry, additional food elements, etc. The effect of roundness, good spheroidization and simple preparation and operation

Inactive Publication Date: 2012-05-23
ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There is no report on the preparation of composite nanoparticles by embedding α-galactosidase in chitosan as a carrier. The composite nanoparticles release α-galactosidase rapidly under acidic (such as pH 4.0) or alkaline (such as pH 10.0) conditions. - The characterization of galactosidase has also not been reported
At the same time, the commonly used dosage form of α-galactosidase for feeding is enzyme powder or liquid enzyme dosage form, which has problems such as easy inactivation of α-galactosidase, heat resistance, and storage resistance.

Method used

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  • Chitosan nano particles embedded with alpha-galactosidase and preparation method thereof
  • Chitosan nano particles embedded with alpha-galactosidase and preparation method thereof
  • Chitosan nano particles embedded with alpha-galactosidase and preparation method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0034] Dissolve 50mg chitosan (50kd) in 50mL 1% acetic acid solution, adjust the pH to 4.0; slowly add an equal volume of α-galactosidase crude enzyme solution (0.1mg / mL) under magnetic stirring; Add 25mL sodium tripolyphosphate solution (1mg / mL) dropwise to the α-galactosidase; stir at room temperature for 30 minutes, the α-galactosidase mixture presents a uniform blue-white color, that is, the embedded α-galactosidase has been formed. Chitosan nanoparticles of lactosylase. The encapsulation efficiency of α-galactosidase was 63.0%; under the condition of pH 5.5, the tolerance was increased by 35.5% after warming at 60°C for 30 minutes; under the condition of pH 5.5, the enzyme activity was lost by 6.9% when stored at room temperature for 90 days; Under the environment of pH 3.0, the release rate within 10 minutes was 93.2%.

Embodiment 2

[0036] Dissolve 1000mg chitosan (1800kd) in 50mL 1% acetic acid solution, adjust the pH to 6.0; slowly add an equal volume of α-galactosidase crude enzyme solution (5.0mg / mL) under magnetic stirring; Add 250mL sodium tripolyphosphate solution (1mg / mL) dropwise to the α-galactosidase mixture; stir at room temperature for 30 minutes, the α-galactosidase mixture appears blue-white with a small amount of flocculent precipitate, namely Chitosan nanoparticles entrapping alpha-galactosidase have been formed. The encapsulation efficiency of α-galactosidase was 72.5%; under the condition of pH 5.5, the tolerance was increased by 50.2% after warming at 60°C for 30 minutes; under the condition of pH 5.5, the enzyme activity was lost by 4.6% when stored at room temperature for 90 days; The release rate was 82.4% within 10 minutes under the environment of pH 10.0.

Embodiment 3

[0038] Dissolve 475mg chitosan (900kd) in 50mL 1% acetic acid solution, adjust the pH to 5.0; slowly add an equal volume of α-galactosidase crude enzyme solution (2.45mg / mL) under magnetic stirring; Add 120mL sodium tripolyphosphate solution (1mg / mL) dropwise to the α-galactosidase mixture; stir at room temperature for 30 minutes, the α-galactosidase mixture appears uniform blue-white, that is, the embedded α - Chitosan nanoparticles of galactosidase. The encapsulation efficiency of α-galactosidase was 87.3%; under the condition of pH 5.5, the tolerance was increased by 60.3% after warming at 60°C for 30 minutes; under the condition of pH 5.5, the enzyme activity was lost by 2.7% when stored at room temperature for 90 days; The release rate was 35.2% within 10 minutes under the environment of pH 7.0.

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Abstract

The invention relates to a preparation method of chitosan nano particles embedded with alpha-galactosidase in the technical field of feeding enzymic preparations, which comprises the steps: preparing the chitosan stock solution; adding alpha-galactosidase crude enzyme solution with the same volume into the chitosan stock solution, and evenly mixing by adopting the magnetic stirring method to prepare alpha-galactosidase mixed solution; adding sodium tripolyphosphate water solution drop by drop into the alpha-galactosidase mixed solution, and magnetic stirring at room temperature to prepare thechitosan nano particles embedded with alpha-galactosidase. The invention has simple preparation operation and can obtain chitosan nano particles embedded with alpha-galactosidase by the simple mixingof chitosan / alpha-galactosidase / sodium tripolyphosphate; the particles are not adhered, have good ballability and round surfaces, can release enzymic preparations fast at the acid or alkaline condition consistent with the acidity and alkalinity of the stomach and intestines of the pig, and is beneficial to the better efficacy exertion of the feeding alpha-galactosidase.

Description

technical field [0001] The invention relates to a chitosan nanoparticle in the technical field of feed additives and a preparation method thereof, in particular to a chitosan nanoparticle embedding alpha-galactosidase and a preparation method thereof. Background technique [0002] α-galactosidase (α-D-galactoside galactohydrolases E.C.3.2.1.22) is one of the feed enzyme preparations approved in my country. α-Galactosidase has a wide range of sources and has a variety of enzyme gene sequences. Its activity can be divided into two types: exonuclease activity and endonuclease activity. α-galactosidase is unstable to heat and sensitive to heavy metal ions, and is easily inactivated during storage and feed pelleting. Proper embedding of α-galactosidase can effectively improve its thermal stability and storage period. The production of feed enzyme preparations in our country is gradually developing from the production of solid enzyme powder by solid fermentation to the productio...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A23K1/165A23K20/163A23K20/189A23K40/30
Inventor 许尧兴柳永李艳丽许少春孙燕
Owner ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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