Preparation method of covalent organic framework encapsulated enzyme
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A kind of technology of covalent organic framework, enzyme solution
Active Publication Date: 2021-08-27
LUDONG UNIVERSITY
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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.
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Embodiment 1
[0043] 1. Prepare covalent organic framework encapsulation enzyme (organic framework encapsulation single enzyme), the steps are as follows:
[0044] (1) At room temperature, dissolve glucose oxidase in 50 mL of PBS buffer solution (pH 7.4) to obtain an enzyme solution;
[0045] (2) At room temperature, add polyvinylpolypyrrolidone to the enzyme solution obtained in step (1), and stir evenly to obtain a mixed solution A;
[0046] (3) At room temperature, take a 250mL round bottom flask, and dissolve 1,3,5-trialdehydepyroglucinol in 25mL of absolute ethanol to obtain solution A;
[0047] (4) At room temperature, add the mixed solution A obtained in step (2) to the solution A obtained in step (3), and stir at room temperature for 1 hour to obtain mixed solution B;
[0048] (5) At room temperature, dissolve benzidine in 25 mL of absolute ethanol to obtain solution B;
[0049] (6) Add the solution B obtained in step (5) to the mixed solution B obtained in step (4), and stir at r...
Embodiment 2
[0055] 1. Prepare covalent organic framework encapsulation enzyme (organic framework encapsulation double enzyme), the steps are as follows:
[0056] (1) At room temperature, dissolve glucose oxidase and horseradish peroxidase in 50 mL of PBS buffer (pH7.4) to obtain an enzyme solution;
[0057] (2) At room temperature, add polyvinylpolypyrrolidone to the enzyme solution obtained in step (1), and stir evenly to obtain a mixed solution A;
[0058] (3) At room temperature, take a 250mL round bottom flask, and dissolve 1.3.5-trialdehydepyroglucinol in 25mL of absolute ethanol to obtain solution A;
[0059] (4) At room temperature, add the mixed solution A obtained in step (2) to the solution A obtained in step (3), and stir at room temperature for 1 hour to obtain mixed solution B;
[0060] (5) At room temperature, dissolve benzidine in 25 mL of absolute ethanol to obtain solution B;
[0061] (6) Add the solution B obtained in step (5) to the mixed solution B obtained in step (...
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Abstract
The invention discloses a preparation method of a covalent organic framework encapsulated enzyme, which comprises the following steps: firstly mixing an enzyme solution protected by polyvinyl polypyrrolidone with an aldehyde ligand solution, then conducting mixing with an amino ligand solution, and conducting centrifuging to obtain a precipitate which is the covalent organic framework encapsulated enzyme. The aldehyde group ligand is 1,3,5-triformylphloroglucinol, and the amino ligand is biphenyl diamine. The covalent organic framework encapsulated enzyme is prepared by adopting a one-step synthesis method, and the method is simple to operate, good in repeatability and high in stability. Compared with a traditional enzyme immobilization method, the crystalline porous covalent organic framework prepared by the method can overcome the defect of incomplete retention of enzyme active sites caused by traditional enzyme immobilization. The covalent organic framework encapsulated enzyme can enhance the environmental tolerance of the free enzyme and improve the recovery rate and cyclic utilization rate of the free enzyme.
Description
technical field [0001] The invention belongs to the field of nanometer materials, and in particular relates to a preparation method of covalent organic framework encapsulation enzyme. Background technique [0002] Enzymes are a class of natural biocatalysts with high catalytic efficiency, good selectivity, environmental protection and economy. Most of them are proteins, and they are widely used in food, medicine, chemical industry and environmental fields. Environmental conditions affect the stability and activity of enzymes. Enzymes can exhibit high catalytic activity under mild conditions such as room temperature, moderate pH, and aqueous media. 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 ...
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