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A method for embedding and immobilizing enzymes in polymer microcapsules

A technology for immobilizing enzymes and polymers, which is applied in the direction of immobilizing enzymes, biochemical equipment and methods, and immobilizing on or in inorganic carriers. It can solve the problems of thin polymer microcapsules and poor stability. Controllable, stable structure, uniform mesh size effect

Active Publication Date: 2021-03-26
BEIJING UNIV OF CHEM TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

On the surface of the calcium carbonate microspheres embedding enzymes, the immobilized enzymes are embedded by layer-by-layer self-assembly method. Although the method is simple and fast, the thickness of the polymer microcapsules is thin and the stability is poor.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Take 10 mL of a mixed solution containing 0.6 mol / L calcium chloride and 3 mg / mL glucose oxidase, stir evenly, add an equal volume of 0.6 mol / L sodium carbonate solution, stir for 5 minutes with magnetic force, centrifuge and wash for later use. Take 0.5 g of calcium carbonate microspheres embedding the enzyme and add it to a polyethyleneimine solution with a mass fraction of 1%. The adsorption reaction was carried out for 20 minutes, and it was washed by centrifugation before use. Take 0.5 g of calcium carbonate microspheres embedded with enzyme after absorbing polyethyleneimine, and 0.05 g of photoinitiator thioxanthone catechol-O,O'-diacetic acid, and add them to polyethyleneimine containing 10% mass fraction. In the alcohol diacrylate (PEGDA molecular weight 575) solution, the volume of the solution is 50 mL, and the solvent is a phosphate buffer solution with pH=7. After sealing the device, pass nitrogen gas, and move into the visible light lamp irradiation device...

Embodiment 2

[0022] Take 10 mL of a mixed solution containing 0.6 mol / L calcium chloride and 3 mg / mL glucose oxidase, stir evenly, add an equal volume of 0.6 mol / L sodium carbonate solution, stir for 5 minutes with magnetic force, centrifuge and wash for later use. Take 0.25 g of enzyme-embedded calcium carbonate microspheres and add it to a polyethyleneimine solution with a mass fraction of 1%. The volume of the solution is 50 mL. The solvent in the polyethyleneimine solution is a phosphate buffer solution with a pH of 7. Magnetic stirring The adsorption reaction was carried out for 20 minutes, and it was washed by centrifugation before use. Take 0.25 g of calcium carbonate microspheres embedded with enzyme after absorbing polyethyleneimine, and 0.05 g of photoinitiator thioxanthone catechol-O,O'-diacetic acid, and add them to polyethyleneimine containing 10% mass fraction. In the alcohol diacrylate (PEGDA molecular weight 575) solution, the volume of the solution is 50 mL, and the solven...

Embodiment 3

[0026] Take 10 mL of a mixed solution containing 0.4 mol / L calcium chloride and 1 mg / mL glucose oxidase, stir evenly, add an equal volume of 0.4 mol / L sodium carbonate solution, stir for 5 minutes with magnetic force, centrifuge and wash for later use. Take 0.5 g of calcium carbonate microspheres embedding the enzyme and add it to a polyethyleneimine solution with a mass fraction of 1%. The adsorption reaction was carried out for 10 minutes, and it was washed by centrifugation before use. Take 0.25 g of calcium carbonate microspheres embedded with enzyme after absorbing polyethyleneimine, and 0.03 g of photoinitiator thioxanthone catechol-O, O′-diacetic acid, and add them to polyethyleneimine containing 10% by mass. Alcohol diacrylate (PEGDA molecular weight 575) and 0.01% by mass of methylenebisacrylamide solution, the volume of the solution is 50 mL, and the solvent is phosphate buffer with pH=7. After the device is sealed, nitrogen gas is passed, and it is moved into a vis...

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Abstract

The invention belongs to the field of immobilized enzyme preparation and discloses a method for embedding immobilization of enzymes by polymer microcapsules. The method includes steps: step one, preparing calcium carbonate microspheres for embedding immobilization of the enzymes; step two, performing adsorption of a layer of a positive-charge polymer on the surfaces of the enzyme-embedding calciumcarbonate microspheres; step three, under visible light irradiation, initiating polymerization reaction by a photoinitiator, and performing surface grafting of cross-linked polymer layers on the surfaces of the enzyme-embedding calcium carbonate microspheres after polymer adsorption; step four, removing calcium carbonate, and preparing enzyme coating microcapsules. Enzyme immobilization at the room temperature under visible light is realized, reaction conditions are mild, and enzyme activity keeping is benefited; enzyme stability is improved by immobilization, and enzyme recycling is realized.

Description

technical field [0001] The invention belongs to the technical field of preparation of immobilized enzymes, and relates to performing visible light graft cross-linking polymerization on the surface of calcium carbonate microspheres embedding enzymes, and then dissolving calcium carbonate to form polymer microcapsules to immobilize enzymes. Background technique [0002] Because the biological enzyme reaction is milder than the chemical reaction, the selectivity is better, and the side reaction products of the reaction are less, and many reactions that are difficult to synthesize and products with special isomerism can be completed by one or more enzymes. Therefore, enzyme preparation has huge market potential and is one of the important technical means to realize green chemistry and sustainable development. However, enzyme preparations also have certain defects. Enzymes are easily denatured and inactivated under special conditions, and their activity will be affected under con...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N11/14C12N11/089C12N11/096C12N11/087
Inventor 赵长稳邵广俊张凯马育红杨万泰
Owner BEIJING UNIV OF CHEM TECH