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Preparation method of nano-magnetic chitin immobilized enzyme

A nano-magnetic, immobilized enzyme technology, applied in the direction of immobilization on/in an organic carrier, can solve the problems of limited use range, low catalytic activity, and inaccessibility, and achieve simple recovery, high protein loading, and improved The effect of utilization

Inactive Publication Date: 2007-01-31
SOUTH CHINA UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The existing immobilized enzyme has large particle size, inaccessibility and low catalytic activity, which limit its application range

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0013] Disperse 0.1 g of nano-magnetic chitosan particles containing acyl chloride functional groups on the surface in 20 mL of a solution containing 0.01% trypsin, stir at 20 °C for 2 hours, separate from solid-liquid in a magnetic field, and wash the magnetic precipitate with phosphate buffer solution. After three times, the desired nano-magnetic chitosan-immobilized trypsin was obtained. Using N-benzoyl-L-arginine ethyl ester hydrochloride as the substrate, the activity of the immobilized enzyme was determined. The protein loading of the magnetically immobilized enzyme was 5.8 mg / g, and the activity recovery rate was 74.2%.

Embodiment 2

[0015] Disperse 0.1 g of nano-magnetic chitosan particles containing acyl chloride functional groups on the surface in 10 mL of a 0.05% lipase solution, stir and react at 23 °C for 6 hours, separate solid-liquid in a magnetic field, and wash the magnetic precipitate with phosphate buffer solution. After three times, the desired nano-magnetic chitosan-immobilized lipase was obtained. Using triglyceride as the substrate, the activity of the immobilized enzyme was determined. The protein loading of the magnetically immobilized enzyme was 6.1 mg / g, and the activity recovery rate was 67.2%.

Embodiment 3

[0017] Disperse 0.1 g of nano-magnetic chitosan particles containing acyl chloride functional groups on the surface in 5 mL of a solution containing 0.1% aprotinin, and stir the reaction at 25 °C for 8 hours. After washing three times, the desired nano-magnetic chitosan-immobilized aprotinin was obtained. Using trypsin and N-benzoyl-L-arginine ethyl ester hydrochloride as substrates, the immobilized enzyme activity was determined. The protein loading of the magnetically immobilized enzyme was 6.8 mg / g, and the activity recovery rate was 75.3%.

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Abstract

The present invention discloses a preparation method of nano magnetic chitosan immobilized enzyme. Said method includes the following steps: making nano magnetic chitosan particles whose surface contains acylchloro functional group be dispersed in enzyme-bearing solution, stirring them to make reaction for 2-8 hr at 20-25 deg.C, making solid-liquid separation in magnetic field, using phosphoric acid buffer solution to wash the magnetic precipitate for three times so as to obtain the required nano magnetic chitosan immobilized enzyme, the concentration of enzyme in enzyme-bearing solution is 0.01-0.1%. and the solid-liquid ratio of nano magnetic chitosan whose surface contains acylchloro functional group and enzyme-bearing solution is 1:50-1:200 g / ml.

Description

technical field [0001] The invention belongs to the field of magnetic chitosan immobilized enzymes, and particularly relates to a preparation method of nano-magnetic chitosan immobilized enzymes. Background technique [0002] Magnetic polymer particles are a new type of functional polymer materials, which can be used as immobilization carriers of biomolecules. It can introduce various reactive functional groups on the surface of magnetic particles through chemical reactions such as copolymerization and surface modification, such as hydroxyl and carboxyl groups. , aldehyde groups, amino groups, etc., can be combined with biologically active substances such as enzymes, cells, antibodies, etc. through covalent bonds, and because of their magnetic properties, they can move or separate rapidly under the action of an external magnetic field. Therefore, magnetic polymer particles are used in biomedicine. , cytology, bioengineering and other fields have a wide range of applications....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/10
Inventor 安小宁
Owner SOUTH CHINA UNIV OF TECH
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