Method for preparing strong magnetic iron enzyme compound under mild conditions

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 ...

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Problems solved by technology

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