Preparation method of magnetic immobilized enzyme

A technology of immobilized enzymes and immobilized phospholipases, which is applied to the direction of immobilization on or in inorganic carriers, or in or on organic carriers, which can solve the problems of free enzyme reactions being sensitive to the environment and unable to recover production costs, etc. , to achieve the effect of reducing the operation

Inactive Publication Date: 2015-04-29
NORTHEAST AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The purpose of the present invention is to solve the problems that the free enzyme is sensitive to the reaction environment, cannot be recycled and the production cost is high, so the free enzyme is immobilized on the nano-scale magnetic carrier PVP-DB-171 / SiO 2 / Fe 3 o 4

Method used

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Effect test

specific Embodiment approach 1

[0013] Embodiment 1: The method for preparing the magnetically immobilized enzyme carrier of this embodiment. Carry out as follows: one, prepare Fe 3 o 4 Magnetic microspheres: make a solution of ferric salt and ferrous salt in a certain proportion, and select an alkaline precipitant (such as NH 3 ·H 2 O, NaOH, etc.) for co-precipitation, and select excellent process conditions to obtain magnetic particles with excellent performance; 2. Preparation of SiO 2 / Fe 3 o 4 Core-shell particles; 3. Preparation of magnetic carrier PVP-DB-171 / SiO 2 / Fe 3 o 4 , the specific method is: add 0.14g DB-171, 3.98g NVP and 0.20g AIBN to 80mL ethanol, and reflux at 75°C for 6h. After rotary evaporation, vacuum-dry at 60°C for 24 hours to obtain PVP-DB-171. Add 2g of SiO to a 100mL three-necked bottle 2 / Fe 3 o 4 200mL water solution and 200mL water, after mixing evenly, add 20mL ethanol solution of 50mg PVP-DB-171 dropwise, add 0.3g triethylamine, reflux at 70℃ for 10h, use NdFeB pe...

specific Embodiment approach 2

[0014] Embodiment 2: This embodiment differs from Embodiment 1 in that the rotary evaporation temperature for preparing the magnetic carrier PVP-DB-171 in step 3 is 30-80°C. Other steps are the same as in the first embodiment.

specific Embodiment approach 3

[0015] Embodiment 3: This embodiment differs from Embodiment 1 in that the vacuum drying time for preparing the magnetic carrier PVP-DB-171 in step 3 is 12-36 hours. Other steps are the same as in the first embodiment.

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Abstract

The invention provides a preparation method of a magnetic immobilized enzyme, relating to a preparation method of a novel carrier PVP-DB-171 (polyvinylpyrrolidone-vinyltrimethoxysilane)/SiO2/Fe3O4. The invention aims to solve the problem that the free enzyme is sensitive to the reaction environment and can not be recovered and the problems of high production cost and the like, and thus, the free enzyme is immobilized in the nano magnetic carrier PVP-DB-171/SiO2/Fe3O4. The magnetic immobilized enzyme is an enzyme with magnetic response characteristic and has operational stability; and the nano immobilized enzyme is uniformly dispersed in the liquid phase. The magnetic response performance can be utilized to achieve the goal of completely separating the immobilized enzyme from the reaction medium. The active recovery rate of the finally obtained immobilized enzyme is 85%; and the enzyme activity after 10 times of repeated use is kept at 70% above.

Description

technical field [0001] The invention belongs to the technical field of chemical materials, and specifically relates to a method for preparing magnetically immobilized enzymes, that is, a carrier is PVP-DB-171 (polyvinylpyrrolidone-vinyltrimethoxysilane) / SiO 2 / Fe 3 o 4 The preparation method of the immobilized enzyme. Background technique [0002] Immobilized enzymes bind enzyme molecules to carriers through physical or chemical methods, while maintaining their high-efficiency, specificity and mild enzyme-catalyzed reaction characteristics, they also overcome the shortcomings of free enzymes and present storage stability High, easy to separate and recover, can be reused many times, continuous and controllable operation and so on. Immobilized enzymes are widely used in the fields of environment, energy, materials, chemistry, and pharmaceuticals. [0003] The existing immobilization methods include adsorption method, embedding method, cross-linking method, covalent bindin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/14C12N11/08
Inventor 江连洲于殿宇赵清霞王俊国王立琦赵丹丹杜华楠刘飞
Owner NORTHEAST AGRICULTURAL UNIVERSITY
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