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Method for preparing beta-glucosidase fixed with magnetic nano-particle

A technology of magnetic nanoparticles and glucosidase, which is applied in the direction of fixing on/in the organic carrier, can solve the problems of short service life and storage period, expensive price and limitation of β-glucosidase, and achieve low cost , uniform shape, and high recovery rate of enzyme activity

Inactive Publication Date: 2008-09-24
DONGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, β-glucosidase is expensive, and its service life and storage period are short, so it is limited in actual production and application.

Method used

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  • Method for preparing beta-glucosidase fixed with magnetic nano-particle
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  • Method for preparing beta-glucosidase fixed with magnetic nano-particle

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] (1) Add 0.1MFe 2+ and Fe 3+ After mixing the chloride solution with a ratio of 1:2, use 4M NaOH to stir at a high speed at 80°C and pH=10.0 for precipitation reaction, then wash, filter, and dry the precipitate to obtain Fe with a size of 8-10 nm. 3 o 4 particles, in this chemical co-precipitation method to prepare nano-Fe 3 o 4 Due to the different chemical characteristics of the dispersion system used in the process of particle processing, it will have a certain impact on the performance of magnetic particles.

[0030] (2) Dissolving 1 g of chitosan in 100 mL of acetic acid solution with a mass fraction of 10% forms a uniform and transparent colloidal solution.

[0031] (3) the above-prepared Fe 3 o 4 The particles were poured into the chitosan solution, stirred for 20 minutes, washed repeatedly with distilled water until neutral, and dried for later use.

[0032] (4) Weigh 0.1 g of the absolute dry magnetic nanoparticles prepared above, add 1 mL of β-glucosida...

Embodiment 2

[0035] (1) Fe 2+ and Fe 3+ After the chloride solution is mixed in a ratio of 1:2, excess NaOH is used to stir at a high speed at a certain temperature and pH value for precipitation reaction, and then the precipitate is washed, filtered, and dried to obtain Fe with a size of 8-10 nm. 3 o 4 particles.

[0036] (2) Dissolving 1 g of chitosan in 100 mL of acetic acid solution with a mass fraction of 10% forms a uniform and transparent colloidal solution.

[0037] (3) the above-prepared Fe 3 o 4 The particles were poured into the chitosan solution, stirred for 20 minutes, washed repeatedly with distilled water until neutral, and dried for later use.

[0038] (4) Add 0.1M citric acid buffer solution with a pH value of 4.0, 5.0, 6.0, 7.0, 8.0 to the immobilized β-glucosidase solution and the free β-glucosidase solution respectively, and react at 45°C for 10 minutes and measure the activity of the enzyme.

Embodiment 3

[0040] (1) Fe 2+ and Fe 3+ After the chloride solution is mixed in a ratio of 1:2, excess NaOH is used to stir at a high speed at a certain temperature and pH value for precipitation reaction, and then the precipitate is washed, filtered, and dried to obtain Fe with a size of 8-10 nm. 3 o 4 particles.

[0041] (2) Dissolving 1 g of chitosan in 100 mL of acetic acid solution with a mass fraction of 10% forms a uniform and transparent colloidal solution.

[0042] (3) the above-prepared Fe 3 o 4 The particles were poured into the chitosan solution, stirred for 20 minutes, washed repeatedly with distilled water until neutral, and dried for later use.

[0043] (4) Add 0.1 M citric acid buffer solution with pH=5.0 to the immobilized β-glucosidase solution and the free β-glucosidase solution respectively, react at 45° C. for 10 minutes, and measure the enzyme activity every other week.

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Abstract

The invention relates to a preparation method for magnetic nanoparticle fixed Beta-glucosaccharase, which comprises the steps that (1) precipitation reaction occurs between chloride solution of Fe<2+> and Fe<3+> and NaOH solution to prepare Fe3O4 particles; (2) chitosan is dissolved in acetum to form homogeneous transparent colloidal solution, and Fe3O4 particles are mixed with the homogeneous transparent colloidal solution, and the mixture is beaten, thus getting magnetic nanoparticles; (3) the magnetic nanoparticles are added to Beta-glucosaccharase solution, thus getting chitosan magnetic nanoparticle immobilized enzyme through absorption and cross linkage of glutaric dialdehyde solution; (4) citric acid buffer solution is added to immobilized Beta-glucosaccharase and free Beta-glucosaccharase solution, respectively, and reaction occurs after the temperature is adjusted; and the enzyme activity is examined. The method is quick, simple, convenient and is low in cost. The product got has the characteristics of uniform shape, large specific area, high coefficient of recovery of enzyme activity and so on.

Description

technical field [0001] The invention belongs to the technical field of preparing immobilized enzymes, in particular to a preparation method for immobilizing beta-glucosidase with magnetic nanoparticles. Background technique [0002] Nano-ultrafine particles usually refer to ultra-fine particles with a particle size of 1-100nm. When the particle size of a substance enters the nanometer level, it has unusual small size effects, surface effects, quantum size effects, and macroscopic quantum tunneling effects. The unique optical, electrical, thermal, magnetic, mechanical and chemical properties of nanomaterials make nanomaterials play an irreplaceable role in the fields of new energy materials, ecological environment materials, high-performance electronic materials and new rare earth materials. With the wide application of nano-magnetic materials in various fields, as an important magnetic material-nano-Fe 3 o 4 Research and development has received widespread attention. Nano...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/04
Inventor 朱利民薛勇聂华丽周毓婷
Owner DONGHUA UNIV
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