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A kind of preparation method of covalent organic framework encapsulated enzyme

A technology of covalent organic framework and enzyme solution, which is applied in the field of nanomaterials, can solve the problems of unstable fixation recovery rate and low enzyme activity, achieve improved recovery rate and recycling rate, simple operation, and enhanced environmental tolerance Effect

Active Publication Date: 2022-07-29
LUDONG UNIVERSITY
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Problems solved by technology

The traditional enzyme immobilization technology uses physical or chemical methods such as adsorption, embedding, covalent bonding and cross-linking to improve the environmental stability, recovery rate and recycling rate of free enzymes to a certain extent, but there are still problems that cannot be ignored. Disadvantages, such as low recovery rate due to poor enzyme immobilization; low enzyme activity due to incomplete retention of the active site of the enzyme, etc.

Method used

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  • A kind of preparation method of covalent organic framework encapsulated enzyme
  • A kind of preparation method of covalent organic framework encapsulated enzyme

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Experimental program
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Effect test

Embodiment 1

[0043] 1. Prepare covalent organic framework encapsulating enzyme (organic framework encapsulating single enzyme), the steps are as follows:

[0044] (1) Under normal temperature, glucose oxidase was dissolved in 50 mL of PBS buffer (pH7.4) to obtain an enzyme solution;

[0045] (2) at normal temperature, add polyvinyl polypyrrolidone to the enzyme solution obtained in step (1), stir evenly, and obtain mixed solution A;

[0046] (3) Under normal temperature, take a 250mL round-bottomed flask, dissolve 1,3,5-trialdehyde-based pyrogallol in 25mL of absolute ethanol to obtain solution A;

[0047] (4) at normal temperature, add the mixed solution A obtained in step (2) into the solution A obtained in step (3), and stir at room temperature for 1 h to obtain mixed solution B;

[0048] (5) at normal temperature, benzidine diamine is dissolved in 25mL absolute ethanol to obtain solution B;

[0049] (6) adding the solution B obtained in step (5) to the mixed solution B obtained in st...

Embodiment 2

[0055] 1. To prepare covalent organic skeleton encapsulation enzyme (organic skeleton encapsulation double enzyme), the steps are as follows:

[0056] (1) Under normal temperature, glucose oxidase and horseradish peroxidase were dissolved in 50 mL of PBS buffer (pH7.4) to obtain an enzyme solution;

[0057] (2) at normal temperature, add polyvinyl polypyrrolidone to the enzyme solution obtained in step (1), stir evenly, and obtain mixed solution A;

[0058] (3) Under normal temperature, take a 250mL round-bottomed flask, dissolve 1.3.5-trialdehyde-based pyrogallol in 25mL of absolute ethanol to obtain solution A;

[0059] (4) at normal temperature, add the mixed solution A obtained in step (2) into the solution A obtained in step (3), and stir at room temperature for 1 h to obtain mixed solution B;

[0060] (5) at normal temperature, benzidine diamine is dissolved in 25mL absolute ethanol to obtain solution B;

[0061] (6) adding solution B obtained in step (5) to mixed solu...

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Abstract

The invention discloses a preparation method of a covalent organic skeleton encapsulating enzyme. The enzyme solution protected by polyvinyl polypyrrolidone is first mixed with an aldehyde-based ligand solution, then mixed with an amino ligand solution, and centrifuged, and the obtained precipitate is a valent organic skeleton. Encapsulated enzymes. The aldehyde-based ligand is 1,3,5-trialdehyde-based pyrogallol, and the amino ligand is biphenylenediamine. The method of the invention adopts a one-step synthesis method to prepare the covalent organic framework encapsulated enzyme, and has the advantages of simple operation, good repeatability and high stability. Compared with the traditional enzyme immobilization method, the crystalline porous covalent organic framework prepared by the present invention can overcome the defect of incomplete enzyme active site retention caused by traditional enzyme immobilization. The covalent organic framework encapsulating enzyme of the present invention can enhance the environmental tolerance of the free enzyme, and improve the recovery rate and the recycling rate of the free enzyme.

Description

technical field [0001] The invention belongs to the field of nanomaterials, and in particular relates to a preparation method of a covalent organic framework encapsulating enzyme. Background technique [0002] Enzymes are a kind of natural biocatalysts with high catalytic efficiency, good selectivity, environmental protection and economy, most of which are proteins, and have a wide range of applications in the fields of food, medicine, chemical industry and environment. Environmental conditions affect the stability and activity of enzymes, and enzymes can exhibit high catalytic activity under mild conditions such as room temperature, moderate pH, and aqueous media. The traditional enzyme immobilization technology improves the environmental stability, recovery rate and recycling rate of free enzymes to a certain extent through physical or chemical methods such as adsorption, embedding, covalent bonding and cross-linking, but there are still problems that cannot be ignored. D...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N11/082C12N9/08C12N9/04
CPCC12N11/082C12N9/0006C12N9/0065C12Y101/03004C12Y111/01007
Inventor 王鲁良杨晓帅孙舒扬柳全文刘玉申刘芳洁胡振华
Owner LUDONG UNIVERSITY
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