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Immobilized enzyme taking bacterial cellulose as carrier and preparation method of immobilized enzyme

A technology of bacterial cellulose and bacterial cellulose membrane, which is applied in the field of immobilized enzymes and its preparation, can solve the problems of severe carrier activation and immobilization reaction conditions, complex carrier activation and immobilization operations, single carrier shape and size, etc. Achieve the effects of mild activation and immobilization conditions, good shape and size controllability, and simple operation

Inactive Publication Date: 2017-01-04
SHENZHEN INST OF ADVANCED TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] However, based on the existing technologies of covalently immobilized enzymes, it is found that these methods have the following disadvantages: (1), the activation and immobilization of the carrier are complicated and the process is cumbersome; (2), the activation and immobilization of the carrier are under severe reaction conditions , can easily affect the activity of the carrier and enzyme; (3), the shape and size of the carrier are too single, which is not conducive to the preparation of immobilized enzymes with diverse shapes and sizes

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  • Immobilized enzyme taking bacterial cellulose as carrier and preparation method of immobilized enzyme
  • Immobilized enzyme taking bacterial cellulose as carrier and preparation method of immobilized enzyme
  • Immobilized enzyme taking bacterial cellulose as carrier and preparation method of immobilized enzyme

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preparation example Construction

[0041] The second aspect of the present invention provides a kind of preparation method of the immobilized enzyme that takes bacterial cellulose as carrier, comprises the following steps:

[0042] (1) Bacterial cellulose is provided, the bacterial cellulose is placed in an oxidant solution, and stirred in the dark at room temperature for 8-12 hours, the ortho-dihydroxyl group in the bacterial cellulose structure is oxidized into an aldehyde group, and after washing, the activated Bacterial cellulose; the oxidizing agent is selected from at least one of sodium periodate, potassium metaperiodate, sodium metaperiodate and lead tetraacetate; the mass ratio of oxidizing agent and bacterial cellulose is 10:1-135:1;

[0043] (2) Put the activated bacterial cellulose in the enzyme solution, and stir in the dark for 2-6 hours at room temperature, the mass ratio of the activated bacterial cellulose to the enzyme is 0.72:1-4.28:1; then add the reducing agent Sodium cyanoborohydride, cont...

Embodiment 1

[0068] A kind of preparation method taking bacterial cellulose as the immobilized enzyme of carrier, comprises the following steps:

[0069] (1) Get 10.6950g (wet weight, water content is 99.2%) the bacterial cellulose ball that diameter is 1.0cm, join in the sodium periodate solution of 50ml 0.1mol / L, under room temperature avoid light and stir for 8 hours, bacterium The ortho-dihydroxyl groups in the cellulose structure are oxidized to form aldehyde groups. After the reaction, they are washed successively with 0.2mol / L ethylene glycol solution and double distilled water to obtain activated bacterial cellulose;

[0070] (2) Get the lipase solution of 10ml 2mg / ml, add the bacterial cellulose after the above-mentioned activation, under room temperature avoid light and fully stir for 2 hours, then add 0.1258g sodium cyanoborohydride, continue under room temperature avoid light and stir for 8 hours, Fully washed with distilled water and freeze-dried to obtain 0.9056 g of immobili...

Embodiment 2

[0074] A kind of preparation method taking bacterial cellulose as the immobilized enzyme of carrier, comprises the following steps:

[0075] (1) get 10.3711g (wet weight, water content is 99.2%) side length is 2.5cm * 2.5cm, the bacterial cellulose film that thickness is 0.2mm, joins in the sodium periodate solution of 50ml 0.3mol / L, Stir in the dark at room temperature for 12 hours, the o-dihydroxyl group in the bacterial cellulose structure is oxidized into aldehyde groups, and after the reaction, wash with 0.2mol / L ethylene glycol solution and double distilled water respectively to obtain activated bacterial cellulose .

[0076] (2) Get 10ml of 4mg / ml lipase solution, add the above-mentioned activated bacterial cellulose, fully stir in the dark at room temperature for 6 hours, add 0.1258g sodium cyanoborohydride, continue to stir in the dark at room temperature for 10 hours, distilled water Fully washed and freeze-dried to obtain 0.8936 g of immobilized lipase supported by...

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Abstract

The invention provides an immobilized enzyme taking bacterial cellulose as a carrier. The immobilized enzyme comprises the bacterial cellulose and enzyme, wherein the bacterial cellulose and the enzyme are subjected to covalent binding through C-N bonds. The size and shape of the bacterial cellulose are controllable, and thus the immobilized enzymes with diversified shapes and sizes can be obtained. The invention further provides a preparation method of the immobilized enzyme taking thebacterial cellulose as the carrier. The preparation method comprises the following steps: carrying out one-step activation on the bacterial cellulose under the room temperature, thus obtaining the activated bacterial cellulose, reacting the activated bacterial cellulose and the enzyme at the room temperature, and adding with a reducing agent, thus obtaining the immobilized enzyme taking the bacterial cellulose as the carrier. In the preparation process, the activation and immobilization conditions are mild, the process is simple, and the prepared immobilized enzyme taking the bacterial cellulose as the carrier is controllable in shape and size, and can be flexibly designed.

Description

technical field [0001] The invention relates to the technical field of immobilized enzymes, in particular to an immobilized enzyme with bacterial cellulose as a carrier and a preparation method thereof. Background technique [0002] Immobilized enzyme (immobilized enzyme) is a technology that binds the enzyme to a water-insoluble carrier or in a certain space by physical or chemical methods, thereby restricting the free flow of enzyme molecules, but enabling the enzyme to fully play its catalytic role. After the reaction, the enzyme can be separated from the substrate and the product, and the enzyme can be reused. The preparation methods of immobilized enzymes can be divided into the following categories: adsorption, covalent binding, embedding and cross-linking. The covalent binding method is to link the enzyme and the carrier through a covalent bond. In order to achieve effective covalent linkage, relevant functional groups on the carrier or enzyme must be activated firs...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/12
Inventor 阮长顺蔡青青王青山胡阳潘浩波
Owner SHENZHEN INST OF ADVANCED TECH
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