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Immobilized enzyme and preparation method thereof

An immobilized enzyme, amination technology, applied in the direction of being immobilized on or in an inorganic carrier, immobilized on/in an organic carrier, etc., can solve the problems of inability to realize recycling, instability, and small immobilization amount, etc. Achieve the effect of increasing fixed amount, improving efficiency and avoiding pollution

Active Publication Date: 2019-09-17
OCEAN UNIV OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The above methods all use glutaraldehyde cross-linking to immobilize the corresponding enzymes, the immobilization amount is small, unstable, and cannot be recycled.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0066] This embodiment provides a preparation method of immobilized enzyme, comprising the following specific steps:

[0067] (1) After the mass concentration is 8g / L chitin, the molar concentration is 0.5mM TEMPO, the molar concentration is 5mM sodium bromide and the molar concentration is 5mM sodium hypochlorite, after mixing, stir the reaction, and adjust the pH during the reaction to keep 9;

[0068] (2) Centrifuge the reacted mixture at 6000rpm for 15min, collect the precipitate, and perform ultrasonic crushing with a power of 200W for 40min after purification;

[0069] (3) Activate the crushed chitin nanofibers at a temperature of 15° C. for 3 hours by passing them through EDC with a mass concentration of 5 mg / mL and NHS with a mass concentration of 40 mg / mL;

[0070] Wherein, the length of the chitin nanofiber is 30-100nm;

[0071] (4) Aminating the magnetic nanomaterial, adding aminated ferric oxide to activated chitin nanofibers for magnetic modification;

[0072] W...

Embodiment 2

[0075] This embodiment provides a preparation method of immobilized enzyme, comprising the following specific steps:

[0076] (1) after the mass concentration is 15g / L chitin, the molar concentration is 3mM TEMPO, the molar concentration is 15mM sodium bromide and the molar concentration is 15mM sodium hypochlorite, stir the reaction, and adjust the pH during the reaction to keep 11;

[0077] (2) Centrifuge the reacted mixture at a speed of 15000rpm for 5min, collect the precipitate, and perform ultrasonic crushing with a power of 500W for 20min after purification;

[0078] (3) Activate the crushed chitin nanofibers at a temperature of 25° C. for 3 hours by passing them through EDC with a mass concentration of 15 mg / mL and NHS with a mass concentration of 60 mg / mL;

[0079] Wherein, the length of the chitin nanofiber is 150-300nm;

[0080] (4) Aminating the magnetic nanomaterial, adding aminated ferric oxide to activated chitin nanofibers for magnetic modification;

[0081] ...

Embodiment 3

[0085] This embodiment provides a preparation method of immobilized enzyme, comprising the following specific steps:

[0086] (1) After the mass concentration is 9g / L chitin, the molar concentration is 0.8mM TEMPO, the molar concentration is 8mM sodium bromide and the molar concentration is 12mM sodium hypochlorite, stir and react, and adjust the pH to keep 10 during the reaction;

[0087] (2) Centrifuge the reacted mixture at 12000rpm for 10min, collect the precipitate, and perform ultrasonic crushing with a power of 300W for 25min after purification;

[0088] (3) Activate the crushed chitin nanofibers at a temperature of 20° C. for 3 hours by passing them through EDC with a mass concentration of 12 mg / mL and NHS with a mass concentration of 50 mg / mL;

[0089] Wherein, the length of the chitin nanofiber is 50-200nm;

[0090] (4) Aminating the magnetic nanomaterial, adding aminated ferric oxide to activated chitin nanofibers for magnetic modification;

[0091] Wherein, the a...

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Abstract

The invention relates to an immobilized enzyme and a preparation method thereof. Chitin nanofibers are used as a carrier, wherein the enzyme is fixed to the carrier in a covalent connection manner, and the hydroxyl of the chitin nanofibers is oxidized into a carboxylic acid group for covalent connection of the enzyme and the chitin nanofibers. The hydroxyl of the chitin nanofibers is oxidized into the carboxylic acid group, so that the enzyme can be in covalent connection through the carboxylic acid group to be fixed to the carrier, and a cross-linking agent is used as assistance for further fixing the enzyme; and through cooperation of the methods, the fixing quantity of the enzyme is further increased, the stability of the enzyme is improved, and the catalyzing efficiency of the enzyme is further improved.

Description

technical field [0001] The invention belongs to the field of nanomaterials and relates to an immobilized enzyme and a preparation method thereof, in particular to an immobilized enzyme with chitin as a carrier and a preparation method thereof. Background technique [0002] As a biocatalyst, enzymes have broad research and application prospects in the fields of medicine, food, chemical industry, etc. However, enzymes are unstable in the catalytic process, but natural enzymes have poor stability, are easily inactivated, cannot be recycled, and are mixed with products after the reaction And other problems limit the application of enzymes. Enzyme immobilization is a technology that uses solid materials to limit enzyme molecules to a certain area. The immobilized enzyme technology improves the stability of the enzyme, realizes the repeated use of the enzyme, and reduces the cost. [0003] Enzyme immobilization methods can be mainly divided into adsorption method, covalent coupli...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/10C12N11/14
CPCC12N11/10C12N11/14Y02P20/50
Inventor 毛相朝黄文灿薛长湖
Owner OCEAN UNIV OF CHINA
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