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Aramagnetic epoxy group mesoporous molecular sieve for immobilized biological enzymes, and preparation method thereof

A mesoporous molecular sieve, epoxy-based mesoporous technology, applied in biochemical equipment and methods, immobilized enzymes, molecular sieve compounds, etc., to reduce clogging, improve separation efficiency, and large specific surface area.

Active Publication Date: 2014-01-08
EAST CHINA UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Through a selective etching process, large-scale nanometer γ-Fe was fabricated using mesoporous silicon 2 o 3 Particle coating to obtain hollow magnetic mesoporous silicon microsphere composite material, which can be used as an adsorbent to quickly separate two antibiotics, tetracycline and sulfamethazine, in the water system. The pore size of the synthesized material is about 3nm, which is also not suitable for macromolecular enzymes Adsorption

Method used

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  • Aramagnetic epoxy group mesoporous molecular sieve for immobilized biological enzymes, and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] 0.365g FeCl 3 ·6H 2 O and 0.208g FeSO 4 ·7H 2 O(Fe 3+ / Fe 2+ The molar ratio is 1.8:1) dissolved in 10mL of water and heated to 80°C, adjusted to a pH of about 11 with concentrated ammonia water, aged at this temperature for 2 hours, cooled to room temperature and washed with water until neutral to obtain 0.15g of Fe 3 o 4 Then disperse the nanoparticles in 20mL water, adjust the pH value of the solution to 4.0-5.0 with dilute hydrochloric acid, then add 5mL of L-cysteine ​​solution with a concentration of 5g / L, react with ultrasound for 30 minutes, and finally remove the aqueous solution under an external magnetic field , to obtain L-cysteine ​​surface-modified Fe 3 o 4 Nanoparticles.

[0037] The epoxy-based mesoporous molecular sieve prepared in 0.60g comparative example and the Fe of the L-cysteine ​​surface modification of 0.15g 3 o 4 The nanoparticles were respectively dispersed in 25mL of water, and then mixed uniformly and reacted at room temperature f...

Embodiment 2

[0039] Change the weight of epoxy-based mesoporous molecular sieve into 0.70g and L-cysteine ​​surface-modified Fe in embodiment 1 3 o 4 The weight of the nanoparticles was changed to 0.30g, and other preparation processes were the same as in Example 1. The prepared paramagnetic epoxy-based mesoporous molecular sieve was used for the immobilization of penicillin acylase, and the immobilized enzyme activity obtained was 8514U / g, After 10 cycles of use, the immobilized enzyme retained 92.8% of its initial activity.

Embodiment 3

[0041] Change the weight of the epoxy-based mesoporous molecular sieve into the Fe of 0.45g and L-cysteine ​​surface modification in embodiment 1 3 o 4 The weight of the nanoparticles was changed to 0.30g, and other preparation processes were the same as in Example 1. The prepared paramagnetic epoxy-based mesoporous molecular sieve was used for the immobilization of penicillin acylase, and the activity of the immobilized enzyme obtained was 8489U / g, After 10 cycles of use, the immobilized enzyme retained 89.8% of its initial activity.

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Abstract

The invention belongs to the technical field of biology, and discloses an aramagnetic epoxy group mesoporous molecular sieve for immobilizing biological enzymes, and a preparation method thereof. According to the invention, gamma glycidoxy propyl trimethoxy silane is utilized to introduce aramagnetic epoxy groups onto the surface of the mesoporous molecular sieve, and then a covalent bond is utilized to immobilize aramagnetic Fe3O4 nano particles and biological enzymes onto the outer surface and the inner surface of the mesoporous molecular sieve respectively, wherein the aramagnetic Fe3O4 nano particles are subjected to L-cysteine surface modification and have the particle sizes larger than the pore diameter of the mesoporous molecular sieve, as a result, specially-structured aramagnetic immobilized enzymes can be prepared and are separated from the liquid phase system easily under the action of an external magnetic field, and further, the property and separation efficiency of the immobilized enzymes are improved. The aramagnetic epoxy group mesoporous molecular sieve can be used for immobilization of water-soluble biological enzymes such as penicillin acylase, glucose isomerase, glucosylase, trypsin and amylase, the activity of the prepared aramagnetic immobilized penicillin acylase is 8800 U / g, and 94.5% of the initial activity is retained after ten times of cycle use.

Description

technical field [0001] The invention relates to a paramagnetic epoxy-based mesoporous molecular sieve used for immobilizing biological enzymes and a preparation method thereof, belonging to the field of biotechnology. Background technique [0002] Penicillin acylase (EC 3.5.1.11, enzyme molecular size: ) is the most critical enzyme in the production of semi-synthetic β-lactam antibiotics. It can catalyze the hydrolysis of penicillin and its cyclic acid to remove the side chain, and produce the important intermediate 6-aminopenicillium of semi-synthetic β-lactam antibiotics Alkanoic acid (6-APA) and 7-amino-3-desacetoxycephalosporanic acid (7-ADCA) can also catalyze the condensation of 6-APA and 7-ADCA with side chains to produce a variety of semi-synthetic β-endo Amide antibiotics (such as Ampicillin, Amoxicillin, Cephalexin and Cefadroxil, etc.). [0003] There are many deficiencies in using free enzyme directly in the catalytic process, such as instability in high tempe...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N11/00C01B37/00C01B39/00
Inventor 郭杨龙杨玲杨露高振源卢冠忠詹望成王筠松郭耘张志刚
Owner EAST CHINA UNIV OF SCI & TECH
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