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Magnetic nano-particle immobilized basic protease and preparation method and application thereof

A magnetic nanoparticle, nanoparticle technology, applied in the direction of being fixed on or in an inorganic carrier, chemical industry, climate sustainability, etc., can solve the problems of poor stability, easy leakage of enzymes, inactivation, etc. Fast recycling, good stability, not easy to leak

Inactive Publication Date: 2011-06-29
HUAZHONG UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the existing immobilization technology, the immobilized enzyme is easy to leak and inactivate during storage or reaction, the stability is not good, and the recycling process is cumbersome.

Method used

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  • Magnetic nano-particle immobilized basic protease and preparation method and application thereof
  • Magnetic nano-particle immobilized basic protease and preparation method and application thereof
  • Magnetic nano-particle immobilized basic protease and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] The implementation of the preparation of alkaline protease immobilized on magnetic nanoparticles in the present invention will be specifically described below by taking 1 ml of alkaline protease (0.6 U / μl) immobilized on 1 ml of magnetic nanoparticles (3.6 mg / ml) as an example.

[0022] (1) Preparation of FeSO 4 (0.06M) and FeCl 3 (0.1M) mixed solution, adjust the pH value to 1.5 with hydrochloric acid, under argon protection, add 25% ammonia water drop by drop and stir vigorously until the pH rises to 9, continue to feed argon, stir for 0.5h, and then use Magnetic separation of Fe 3 o 4 The granules were washed 5 times with water and 2 times with ethanol, and the finally obtained granules were dispersed in absolute ethanol with a particle concentration of about 3.45mg / ml (0.015M).

[0023] (2) Take out 25ml of the above-mentioned Fe 3 o 4 Disperse in 50ml of ethanol and 75ml of water, pass through argon to exclude the air, then sonicate for 0.5h, add an appropriat...

Embodiment 2

[0026] The implementation of the application of the magnetic nanoparticle-immobilized alkaline protease in biomass green conversion will be specifically described below by taking the hydrolysis of casein and rapeseed meal catalyzed by the magnetic nanoparticle-immobilized alkaline protease of the present invention as an example.

[0027] (1) Detection of the catalytic activity of alkaline protease immobilized on magnetic nanoparticles to casein

[0028] The catalytic activity of alkaline protease immobilized on magnetic nanoparticles was detected by Folin reagent chromogenic method. Take out 1ml of alkaline protease solution immobilized with magnetic nanoparticles, preheat at 40°C for 2min, add 1ml of casein (10mg / ml), react in water bath at 40°C for 10min, add 2ml of trichloroacetic acid (0.4mol / L), mix well, 5000rad Centrifuge for 10min, take out 1ml supernatant, add 5ml Na 2 CO 3solution (0.4mol / L), then add 1ml of Folin’s reagent (diluted twice), develop color in a water...

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Abstract

The invention provides a magnetic nano-particle immobilized basic protease, the magnetic nano-particle immobilized basic protease is obtained by utilizing a silylating reagent of 3-aminopropyl-triethoxysilane to modify Fe3O4 nano-particles so as to improve water solubility and stability of the Fe3O4 nano-particles, introducing a free amino terminal onto the surface of the Fe3O4 nano-particles, and then using a bifunctional reagent of p-phenylene diisothiocyanate to fix the basic protease into the Fe3O4 nano-particles modified by the silylating reagent, and the magnetic nano-particles with surface functionalization are taken as a vector for immobilizing the basic protease so as to be applied in green transformation of biomass. The preparation method is simple and easy to operate, and a biological catalyst can be rapidly and conveniently recycled through an externally applied magnetic field, thereby being conductive to realization of continuous large-scale production through biological catalytic reaction.

Description

technical field [0001] The invention belongs to the field of biocatalysis and nanotechnology, and in particular relates to a new method for preparing magnetic nanoparticle-immobilized alkaline protease, which is used to efficiently and greenly convert biomass raw materials such as animal and vegetable proteins, animal and vegetable oils, and plant cellulose into polypeptides , fatty acid esters, sugar and other products have a wide range of applications. Background technique [0002] Enzymes are natural biocatalysts with extremely high catalytic efficiency and strong reaction specificity, and are widely used in food processing, pharmaceutical and other industries. However, due to the poor stability of natural enzymes, it is not easy to recycle and reuse, which hinders its application in industrial production. In recent years, the preparation of immobilized biological enzymes based on enzyme immobilization technology has attracted more and more attention in the field of biom...

Claims

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Application Information

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IPC IPC(8): C12N11/14C12P21/06C01G49/08
CPCY02P20/50
Inventor 赵元弟金欣董绪燕黄平英姜达海李菊芳陈洪
Owner HUAZHONG UNIV OF SCI & TECH
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