Method for preparing strong magnetic iron enzyme compound under mild conditions
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A strong magnetic and complex technology, applied in the field of protein immobilization, can solve the problems of reduced apparent enzyme activity, and achieve the effect of simple preparation method, high stability, and no agglomeration
Inactive Publication Date: 2018-10-12
TIANJIN UNIV OF SCI & TECH
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CN102250869 reports a method for preparing a magnetically immobilized enzyme used to produce phytoalcohol ester; CN104560940A discloses a method for preparing a magnetically immobilized enzyme, in which the enzyme protein is fixed on polyvinylpyrrolidone-vinyltrimethoxy Silane surface; CN104844768A discloses a method for preparing temperature-sensitive magnetic protein imprinted microspheres with a core-shell structure; the above methods all belong to the preparation of magnetic particles first, and then the enzyme protein is fixed on the surface of the magnetic microspheres by adsorption or covalent bonding , there is a problem that the active site of the enzyme is masked by the carrier (steric hindrance effect) during the catalytic process, resulting in a significant decrease in the apparent enzyme activity
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Embodiment 1
[0013] (1) The enzyme protein was dissolved in 0.02M Tris-HCl buffer solution with pH 9.0, and the concentration of the enzyme protein was 0.1g / L.
[0014] (2) Add FeSO 4 To (1), make the final concentration 0.02.
[0015] (3) Add dopamine into the solution to make the final concentration 0.5g / L.
[0016] (4) Stir the above mixed solution at room temperature for 0.5 h, add a few drops of ammonia water, and separate and collect the strong magnetic iron enzyme complex under the action of a strong NdFeB magnet.
[0017] The enzyme load of the above iron enzyme complex is 600mg / g carrier, and its magnetism is 0.31emu / g, which can be collected under the action of a strong NdFeB magnet.
Embodiment 2
[0019] (1) The enzyme protein was dissolved in 0.2M Tris-HCl buffer solution with pH 6.0, and the concentration of the enzyme protein was 10g / L.
[0020] (2) Add ferrocene to (1) to make the final concentration 0.2M.
[0021] (3) Add dopamine into the solution to make the final concentration 10g / L.
[0022] (4) Stir the above mixed solution at room temperature for 10 h, add a few drops of ammonia water, and separate and collect the strong magnetic iron enzyme complex under the action of a strong neodymium-iron-boron magnet.
[0023] The enzyme load of the above iron enzyme complex is 400mg / g carrier, and its magnetism is 0.5emu / g, which can be collected under the action of strong NdFeB magnet.
Embodiment 3
[0025] (1) The enzyme protein was dissolved in 0.1M Tris-HCl buffer solution with pH 8.0, and the concentration of the enzyme protein was 2g / L.
[0026] (2) Add FeCl 3 to (1) to make the final concentration 0.1M.
[0027] (3) Add dopamine into the solution to make the final concentration 4g / L.
[0028] (4) Stir the above mixed solution at room temperature for 2 hours, add a few drops of ammonia water, and separate and collect the strong magnetic iron enzyme complex under the action of a strong NdFeB magnet.
[0029] The enzyme load of the above iron enzyme complex is 500mg / g carrier, and its magnetism is 0.4emu / g, which can be collected under the action of a strong NdFeB magnet.
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Abstract
The invention discloses a method for preparing a strong magnetic iron enzyme compound under mild conditions. The method comprises the following steps of (1) dissolving enzyme protein into a Tris-HCl buffer solution; (2) adding inorganic iron salts or inorganic ferrous salts or organic iron salts or organic ferrous salt iron precursors; (3) adding dopamine or dopamine bionic adhesives into the solution; and (4) stirring the mixed solution at normal temperature; dripping a plurality of drips of ammonia water; performing separation under the effect of a neodymium iron boron strong magnet; and colleting the strong magnetic iron enzyme compound. The method provided by the invention has the advantages that the strong magnetic iron enzyme compound simultaneously has bioactivity and strong magnetism; the compound can realize the bioactivity in the free state, and can realize the effective separation and recovery in the magnetic field; a magnetic core is generated in situ; the unit magnetic loading rate is high; no aggregation phenomenon occurs; the magnetic core stability is high; and demagnetization due to oxidization reaction cannot easily occur. The preparation method is simple; the preparation is performed under the conditions of water phase, room temperature and normal pressure; no organic solvent participates in the reaction; and the preparation conditions are green and mild.
Description
technical field [0001] The invention relates to a method for preparing a strong magnetic iron enzyme complex under mild conditions, and belongs to the technical field of protein immobilization. Background technique [0002] Due to the high efficiency and high selectivity of biocatalysts, their application in the chemical industry has become more and more attractive. They have mild reaction conditions, low energy consumption, and are easy to catalyze to obtain relatively pure products, which can reduce waste emissions and can complete tasks that traditional chemical catalysis cannot do. Among them, enzyme catalysis has the characteristics of high selectivity, strong specificity, single reaction, strong controllability, and easy purification of products. Due to the need for repeated use, cost reduction, and enhanced enzyme stability, immobilization techniques continue to develop, mainly including physical adsorption, covalent bonding, embedding, and cross-linking: adsorption ...
Claims
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Application Information
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