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Modified epoxy resin immobilized enzyme, and preparation method and application thereof

An epoxy resin, immobilized enzyme technology, applied in immobilized enzymes, biochemical equipment and methods, oxidoreductases, etc., can solve problems such as unsuitable enzyme application, and achieve the effect of reducing space inhibition and improving enzyme load

Inactive Publication Date: 2020-05-08
ASYMCHEM LIFE SCI TIANJIN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0018] For the method of immobilized enzyme reported in the prior art, the enzyme in table 1 is not suitable for industrial application, need to further improve its stability

Method used

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  • Modified epoxy resin immobilized enzyme, and preparation method and application thereof
  • Modified epoxy resin immobilized enzyme, and preparation method and application thereof
  • Modified epoxy resin immobilized enzyme, and preparation method and application thereof

Examples

Experimental program
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Effect test

Embodiment approach

[0042] According to a typical embodiment of the present invention, a preparation method of modified epoxy resin-immobilized enzyme is provided. The preparation method comprises the following steps: modifying the epoxy resin, adding polyethyleneimine to the modified epoxy resin for further modification, and then adding the enzyme to be immobilized and glutaraldehyde for immobilization to obtain the immobilized epoxy resin Catase.

[0043] Applying the technical scheme of the present invention, the epoxy resin is modified, and polyethyleneimine is added to the modified epoxy resin for further modification, and the aldehyde group in the resin is covalently bonded to the amino group in the polyethyleneimine. The enzyme is then activated by the bifunctional reagent glutaraldehyde, in this way, the steric resin arms are increased to form a network structure, and the enzyme can be more easily bound by covalent binding, because the steric inhibition is reduced, and the enzyme loading ...

Embodiment 1

[0088] Immobilization of Enzyme on Sodium Periodate Oxidized Epoxy Resin

[0089] Add 5 g of epoxy resin ECR8285 or LX1000EP into 20 mL of 1M acetic acid, stir gently at room temperature for 12 hours, wash the acetic acid-treated carrier with 20 mL of distilled water for 3 times, and suspend the treated carrier in 20 ml of 50 mM sodium periodate , stirred gently at room temperature for 2 h, filtered and washed with distilled water.

[0090] 5g modified epoxy resin was resuspended with 0.1M phosphate buffer, added PEI, the final concentration was 1%, adjusted the pH to pH7.0, after gentle stirring for 2 hours, added 50-100mM glutaraldehyde, at room temperature Stir for 1-2 hours. The carrier resin was then filtered and washed with 10 ml of water. The washed carrier was added to the enzyme solution (10ml enzyme, containing 5mg / mL cofactor, dissolved in 40mL 100mM phosphate buffer, pH7.0), stirred at 10-25°C for 4 hours, and kept in the refrigerator overnight. After standing o...

Embodiment 2

[0092] Immobilization of Enzyme on Sodium Periodate Oxidized Epoxy Resin

[0093] Add 5 g of epoxy resin beads into 50 mL of 1 M acetic acid, under gentle stirring at room temperature for 12 h, wash the acetic acid-treated support with 50 mL of distilled water for 3 times with gentle stirring for 10 min each time, suspend the treated support in 100 ml sodium iodate, stirred gently at room temperature for 2 hours, filtered and washed with distilled water.

[0094] 5 g of modified epoxy resin was resuspended with 0.1 M phosphate buffer, PEI was added to a final concentration of 1%, the pH was adjusted to pH 7.0, and stirred gently for 2 hours. The carrier was then filtered and washed with 10 ml of water. The washed carrier was added to the enzyme solution (10ml enzyme, containing 5mg / mL cofactor, dissolved in 40mL 100mM phosphate buffer, pH7.0), stirred at room temperature for 4 hours, and kept in the refrigerator overnight. After standing overnight, 20-100 mM glutaraldehyde w...

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Abstract

The present invention discloses a modified epoxy resin immobilized enzyme, and a preparation method and an application thereof. The preparation method comprises the following steps: modifying epoxy resin, adding polyethyleneimine to the modified epoxy resin for further modification, and then adding an enzyme to be immobilized and glutaraldehyde for immobilization to obtain the modified epoxy resinimmobilized enzyme. By applying the technical scheme, the epoxy resin is modified, the polyethyleneimine is added to the modified epoxy resin for further modification, aldehyde groups in the resin are covalently combined with amino groups in the polyethyleneimine to bind various enzymes, then the bifunctional reagent glutaraldehyde is used for activation, by using the method, space resin arms areincreased to form a network structure, the enzyme can be more easily bound by covalent bonding, and enzyme load can also be improved due to reduced space inhibition.

Description

technical field [0001] The invention relates to the technical field of biocatalysis, in particular to a modified epoxy resin immobilized enzyme, a preparation method and an application. Background technique [0002] Enzymes are widely used as biocatalysts in different industrial processes. Compared with chemical catalysts, enzymes have the advantages of high activity, strong selectivity and high specificity. The ability of enzymes to generate complex compounds under very mild reaction conditions has enabled the development of more sustainable processes, making enzyme-catalyzed processes an emerging platform. Since enzymes are of biological origin, they often have operating characteristics that differ from those required for industrial processes. [0003] Biocatalysts can be produced using whole living cells, dead cells, crude or purified enzymes, depending on the enzyme type and application. Scalable production of enzymes and advances in protein engineering have made comme...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/089
CPCC12N11/08C12N9/1096C12N9/0006C12N9/0073C12N9/80C12N9/001C12N9/0026C12N9/0016C12N9/0018C12N9/78C12Y114/13022C12Y104/01009C12Y104/0102C12Y305/05001
Inventor 洪浩詹姆斯·盖吉肖毅罗杰斯卡·维亚撒·威廉姆斯崔瑜霞张娜赵佳东高妍妍
Owner ASYMCHEM LIFE SCI TIANJIN
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