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Medium immobilized enzyme and preparation method and application thereof

A technology for immobilizing enzymes and media, applied to biochemical equipment and methods, immobilized on or in inorganic carriers, fixed on/in organic carriers, etc., can solve the problem of low enzyme loading and achieve loading High enzyme content, high stability, and high enzyme activity

Active Publication Date: 2019-09-24
INST OF PROCESS ENG CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Although the polydopamine coating can expand the application range of immobilized enzymes, its loading capacity is low. Therefore, it is expected to develop a new medium with higher enzyme loading capacity to immobilize enzymes.

Method used

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  • Medium immobilized enzyme and preparation method and application thereof
  • Medium immobilized enzyme and preparation method and application thereof
  • Medium immobilized enzyme and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] This embodiment provides a medium-immobilized enzyme, such as figure 1 As shown, it consists of substrate 1, immobilized coating 2 and enzyme molecule 4.

[0062] Wherein, the matrix is ​​sodium alginate microspheres, chitosan microspheres and montmorillonite microspheres with an average particle size of 1-3mm, and the raw materials for the fixed coating include tannic acid buffer solution and 3-aminopropyltriethoxysilane ethanol solution, the enzyme molecule is horseradish peroxidase.

[0063] The preparation method is as follows:

[0064] (1) Dissolve tannic acid in 25mL10mM Tris-HCl buffer solution (pH=8.5) to obtain a tannic acid buffer solution with a concentration of 2g / L, and dissolve 3-aminopropyltriethoxysilane in anhydrous In ethanol (analytical pure), obtain the 3-aminopropyltriethoxysilane solution that concentration is 10g / L; 5:1 mixed to obtain a mixed solution;

[0065] (2) Sodium alginate microspheres, chitosan microspheres and montmorillonite micros...

Embodiment 2-7

[0068] The difference with Example 1 is that, in the present example, the volume ratio of tannic acid buffer solution and 3-aminopropyltriethoxysilane solution is 0.2:1 (Example 2), 1:1 (Example 2). 3), 2:1 (Example 4), 8:1 (Example 5), 0.1:1 (Example 6), 10:1 (Example 7).

Embodiment 8

[0070] This embodiment provides a medium immobilized enzyme, which consists of a matrix, an immobilized coating and enzyme molecules.

[0071] Wherein, the substrate is a nylon film, the raw material of the fixed coating includes a tannic acid buffer solution and a 3-aminopropyltriethoxysilane ethanol solution with a solution volume ratio of 8:1, and the enzyme molecule is sucrase.

[0072] The preparation method is as follows:

[0073] (1) With reference to Example 1, the difference is that the solution volume ratio of tannic acid and 3-aminopropyltriethoxysilane is 8:1;

[0074] (2) Place the nylon membrane in the mixed solution of step (1), rinse it with deionized water after stirring at 150rpm / min at room temperature for 18h, to obtain a substrate with a fixed coating;

[0075] (3) Place the substrate with a fixed coating obtained in step (2) in 0.15g / L sucrase solution (pH=5.010mM acetic acid buffer solution), stir at 150rpm / min at room temperature for 2h, then rinse wit...

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Abstract

The invention provides a medium immobilized enzyme and a preparation method and application thereof. The medium immobilized enzyme comprises a substrate, a fixed coating and enzyme molecules; wherein the raw materials of the fixed coating include tannic acid and 3-aminopropyl triethoxysilane. By adopting the fixed coating of the medium immobilized enzyme, it can be achieved that enzyme molecules can be fixed while the coating does not easily shed, and the stability is high. The medium immobilized enzyme has high enzyme carrying capacity and high enzyme activity.

Description

technical field [0001] The invention belongs to the technical field of immobilized enzymes, and relates to a medium-immobilized enzyme and its preparation method and application. Background technique [0002] As a biocatalyst, enzymes are sensitive to the environment and are easily inactivated. The stability and reusability of enzymes can be improved by immobilizing biological enzymes on the surface of substrates such as membranes or microspheres. Conventional enzyme immobilization methods include physical adsorption, entrapment, covalent bonding, and chemical cross-linking. Srivastava P K etc. covalently immobilized acid phosphatase on glutaraldehyde-activated chitosan beads (Srivastava P K, Anand A. Immobilization of acid phosphatase from Vignaaconitifolia seeds on chitosan beads and its characterization[J].International Journal of Biological Macromolecules , 2014,64:150-154.), studies have shown that the immobilized enzyme has enhanced tolerance to pH and temperature, an...

Claims

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Application Information

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IPC IPC(8): C12N11/14C12N11/10C12N11/08C12N11/06
CPCC12N9/0065C12N9/1051C12N11/06C12N11/08C12N11/10C12N11/14C12Y111/01007
Inventor 罗建泉周芳芳万印华
Owner INST OF PROCESS ENG CHINESE ACAD OF SCI
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