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Preparation of recyclable magnetic nanometer immobilized enzyme

A technology of immobilized enzymes and magnetic nanometers, applied to biochemical equipment and methods, and enzymes immobilized on or in inorganic carriers, can solve the problems of loss of enzyme activity, achieve high protein loading, and improve enzyme utilization The effect of the rate and method is simple and easy to implement

Inactive Publication Date: 2008-10-08
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Using ferric oxide nanoparticles as a carrier to prepare immobilized enzymes can effectively solve the problem of enzyme product recycling, but the existing immobilization methods mostly involve surface modification by introducing silyl groups and carboxyl groups on the surface of ferric oxide. Furthermore, cross-linking agents such as glutaraldehyde and carbodiimide are added to cross-link the enzyme molecules, but due to the introduction of various chemical reagents in the surface modification process, the enzyme activity is seriously lost

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Disperse 1.5 grams of nanometer iron ferric oxide powder in 10 milliliters of distilled water, take 1.5 milliliters and add 50 microliters of 25% glutaraldehyde, then add 1 milliliter of 1 percent lipase (derived from porcine pancreas) solution at room temperature (about 25°C) Crosslinked for 0.5 hours. The solid-liquid is separated under the action of an external magnetic field, and the cross-linked product is washed three times with distilled water to obtain the required magnetic nano-immobilized enzyme. The prepared immobilized enzyme is further freeze-dried to obtain the immobilized enzyme dry powder which is reusable and easy to store and transport.

[0021] Using p-nitrophenol palmitate as substrate, the activity of immobilized enzyme was determined. After the reaction, solid-liquid separation was performed under the action of an external magnetic field, the cross-linked product was washed with distilled water three times, and the substrate was added to repeat th...

Embodiment 2

[0023] Disperse 3 grams of nanometer iron ferric oxide powder in 10 milliliters of distilled water, take 0.6 milliliters and add 600 microliters of 25% glutaraldehyde, then add 0.75 milliliters of 0.01 percent lipase (derived from Pseudomonas) solution, cross-link at 30 ° C 1 hour. The solid-liquid is separated under the action of an external magnetic field, and the cross-linked product is washed three times with distilled water to obtain the required magnetic nano-immobilized enzyme.

[0024] Using p-nitrophenol palmitate as substrate, the activity of immobilized enzyme was determined. After the reaction, solid-liquid separation was performed under the action of an external magnetic field, the cross-linked product was washed with distilled water three times, and the substrate was added to repeat the measurement ten times.

Embodiment 3

[0026] Disperse 5 grams of nanometer iron ferric oxide powder in 10 milliliters of distilled water, take 1.5 milliliters and add 1000 microliters of 25% glutaraldehyde, then add 3 milliliters of 10 percent lipase (from Candida) solution, and cross-link at 30 ° C for 1.5 Hour. The solid-liquid is separated under the action of an external magnetic field, and the cross-linked product is washed three times with distilled water to obtain the required magnetic nano-immobilized enzyme.

[0027] Using p-nitrophenol palmitate as substrate, the activity of immobilized enzyme was determined. After the reaction, solid-liquid separation was performed under the action of an external magnetic field, the cross-linked product was washed with distilled water three times, and the substrate was added to repeat the measurement ten times.

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PUM

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Abstract

The invention discloses a method for preparing reusable magnetic nanometer immobilized enzyme. The method comprises the following steps: nanometer Fe3O4 powder with surface on for amino group adsorption is prepared with a coprecipitation method; by using the surface groups, the nanometer Fe3O4 powder reacts with enzyme molecule amino group under the action of cross linking agent, and accomplishes cross linking process, then solid-liquid separation is performed under the action of external magnetic field, immobilized enzyme is recycled, and the magnetic nanometer immobilized enzyme which can be reused can be obtained. The magnetic nanometer immobilized enzyme of the invention has the advantages of high protein load capacity, high enzyme activity, convenient recycling and stable chemical property, and is suitable for the catalytic field, the degeneration of environment pollutant and the synthesizing process of energy material.

Description

technical field [0001] The invention relates to a preparation method of magnetic nanometer immobilized enzyme, in particular to a preparation method of reusable magnetic nanometer immobilized enzyme with iron ferric oxide nano particles as the carrier. Background technique [0002] As an efficient and specific biocatalyst, enzymes are widely used in brewing, food, medicine and other fields. However, due to the protein properties of enzymes, they are unstable in heat, strong acids, strong alkalis, and organic solvents, so enzymes are usually used in aqueous solutions, making their recovery and reuse a problem. [0003] Using ferric oxide nanoparticles as a carrier to prepare immobilized enzymes can effectively solve the problem of enzyme product recycling, but the existing immobilization methods mostly involve surface modification by introducing silyl groups and carboxyl groups on the surface of ferric oxide. Furthermore, cross-linking agents such as glutaraldehyde and carbo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/14C12N13/00
Inventor 王霞许平窦培培赵鹏赵传明
Owner SHANDONG UNIV
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