Immobilized enzyme, immobilized enzyme carrier and preparation method of immobilized enzyme carrier

A technology of immobilizing enzyme carriers and fossils, which is applied in the field of enzyme immobilization, can solve the problems of unfavorable enzyme activity, operational stability, and poor immobilization of free enzymes, and achieve good reusable performance

Active Publication Date: 2017-03-29
INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to overcome above-mentioned technical deficiencies, propose a kind of immobilized enzyme, immobilized enzyme carrier and preparation method thereof, solve the problem that immobilized enzyme carrier has poor immobilization to free enzyme in the prior art, is unfavorable for improving the activity and operation of enzyme technical issues with stability

Method used

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  • Immobilized enzyme, immobilized enzyme carrier and preparation method of immobilized enzyme carrier
  • Immobilized enzyme, immobilized enzyme carrier and preparation method of immobilized enzyme carrier
  • Immobilized enzyme, immobilized enzyme carrier and preparation method of immobilized enzyme carrier

Examples

Experimental program
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Effect test

Embodiment 1

[0024] Embodiment 1 of the present invention provides an immobilized enzyme carrier, and its specific preparation steps are as follows:

[0025] Take a certain amount of graphene oxide (replaced by GO below) and disperse it in deionized water, add an appropriate amount of zinc acetate, wherein the amount of zinc acetate added is to ensure that the mass percentage of zinc oxide in the formed immobilized enzyme carrier is 30%, and ultrasonic treatment for 1 hour; under the situation of stirring, add the mixed solution of NaOH and ethylene glycol, wherein the amount of NaOH is the same molar amount as zinc acetate, and the mass part of NaOH and the volume part of ethylene glycol are 1:10, specifically when When NaOH is 1g, ethylene glycol is 10ml; then continue to stir, add sodium citrate 10 times the mass of GO, stir for 1 hour, put the mixture in the reaction kettle, and react at 100°C for 12 hours; the reaction is over After that, wash and filter with ethanol and water, and dr...

Embodiment 2

[0032] Embodiment 2 of the present invention provides an immobilized enzyme carrier, and its specific preparation steps are as follows:

[0033] Weigh a certain amount of GO and disperse it in isopropanol, then add a certain amount of MnCl 2 4H 2 O, where MnCl was added 2 4H 2The amount of O is 30% to ensure that the mass percentage of manganese dioxide in the immobilized enzyme carrier that forms is 30%, ultrasonic treatment 1 hour; Add the ammoniacal solution of 10ml 2mol / L in the above-mentioned mixed solution again, continue ultrasonic stirring 30min; Then mix The solution was poured into the reaction kettle and heated at 120°C for 12 hours; after the reaction was completed, it was repeatedly washed with deionized water until neutral, then dried at 80°C, ground and burned to obtain GO / MnO 2 carrier material.

[0034] The method for preparing the immobilized enzyme in this example is basically the same as that in Example 1, so details will not be repeated here.

Embodiment 3

[0036] Embodiment 2 of the present invention provides an immobilized enzyme carrier, and its specific preparation steps are as follows:

[0037] Get 10g of surfactant sodium dodecylbenzenesulfonate and dissolve it in 100ml of deionized water, weigh 1g of GO to disperse it and ultrasonically form a uniform GO suspension; weigh 1.025g of Mg(NO 3 ) 2 ·6H 2 O, 0.375g Al(NO 3 ) 3 9H 2 O and urea are added in the there-necked flask that holds 200ml hydrogen peroxide solution, wherein the mass concentration of hydrogen peroxide solution is 20%, the solution in the three-necked flask is mechanically stirred for 15min and then added to the GO suspension, continuously stirred, and the stirred The solution was heated to 150° C. under the protection of nitrogen in an oil bath, kept stirring vigorously, and condensed and refluxed at 150° C. for 8 hours. After the reaction was completed, the reaction product was filtered and washed 4 times with ultrapure water. The GO / LDH carrier mater...

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Abstract

The invention discloses an immobilized enzyme, an immobilized enzyme carrier and a preparation method of the immobilized enzyme carrier. The immobilized enzyme carrier comprises oxidized graphene and inorganic nano-particles, wherein the inorganic nano-particles modify the surface of oxidized graphene and contain multiple electron groups. The immobilized enzyme carrier is prepared by the steps of enabling a modifying agent in a solvent to adequately react with a surfactant to form the inorganic nano-particles, and modifying the surface of oxidized graphene with the inorganic nano-particles. The immobilized enzyme comprises the immobilized enzyme carrier and lipase immobilized on the immobilized enzyme carrier. The immobilized enzyme carrier has the beneficial effects that on one hand, the surface contains a large number of oxygen-containing functional groups, so that the binding rate with the enzyme is relatively high, the enzyme can be firmly immobilized on the surface of the carrier, and the operational stability of the enzyme is improved; on the other hand, the specific surface area is relatively large, so that a sufficient catalytic interface environment and mass transfer are provided for the catalytic process of the immobilized enzyme; and furthermore, the agglomeration of oxidized graphene in repeated use is inhibited by the immobilized enzyme carrier, so that the coverage of the enzyme by a graphite layer is reduced, and the repeated usability of the enzyme is improved.

Description

technical field [0001] The invention relates to an enzyme immobilization technology, in particular to an immobilized enzyme, an immobilized enzyme carrier and a preparation method thereof. Background technique [0002] As a biocatalyst with special catalytic functions, enzymes are widely used in bioengineering, food industry, medicine, fine chemical industry and other fields due to their high catalytic efficiency, high specificity and mild reaction conditions. Lipase is a ubiquitous enzyme in various biological enzyme reactions including: enantioselective hydrolysis and esterification, chiral resolution, enantiomeric monomer preparation and macromolecular polymerization. However, there are still many problems in the application of free enzymes, such as easy inactivation or denaturation of enzymes during storage and reaction, resulting in poor operational stability and reusability; in addition, enzymes are not easy to separate from substrates and products after enzymatic reac...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C01G9/02C12N9/20C12N11/14
CPCC12N9/20C12N11/14C12Y301/01003C01G9/02C01G45/02C01P2004/80C01F7/785
Inventor 黄凤洪时杰张珊郑明明邓乾春
Owner INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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