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Preparation method of ZIF-8/enzyme composite material

A technology of ZIF-8 and composite materials, applied in the direction of fixing on/in the organic carrier, fixing on or in the inorganic carrier, etc., can solve the problems of restricting the diffusion of large-size substrates and hindering the application, and achieve excellent Catalytic ability, low price, high efficiency effect

Active Publication Date: 2020-07-14
FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] However, this approach has significant limitations
Due to the small pore size of ZIF-8 (approximately ), which severely limits the diffusion of large-sized substrates, resulting in only a small number of small-substrate enzymes (such as catalase, cytochrome c, glucose oxidase, etc.) applicable to this strategy, which greatly hinders the application of this strategy in industry. application

Method used

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  • Preparation method of ZIF-8/enzyme composite material
  • Preparation method of ZIF-8/enzyme composite material
  • Preparation method of ZIF-8/enzyme composite material

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Screen and optimize the conditions for the preparation of HSAMS@ZIF-8. Its method steps are:

[0037] (1) Dissolve 0.65 mg, 1.3 mg, 1.9 mg, 2.5 mg, 5 mg, and 10 mg of zinc nitrate hexahydrate in 420 μL of water, respectively, to obtain zinc nitrate solutions 1, 2, 3, 4, 5, and 6;

[0038] (2) Dissolve 0.7mg, 1.4mg, 2.1mg, 2.8mg, 5.5mg, and 11mg of 2-methylimidazole in 420μL of water, respectively, and then add 20μL of HSAMS (concentration: 6mg / mL), and stir for 5min to obtain 2 - Methylimidazole / enzyme solutions 1, 2, 3, 4, 5, 6;

[0039] (3) Mix the 2-methylimidazole / enzyme solutions 1, 2, 3, 4, 5, and 6 with the corresponding zinc nitrate solutions, stir at room temperature for 4 hours, then centrifuge (10000rpm, 5min), and use a concentration of 0.1mol / L, Tris-HCl buffer solution with pH=8.0 washed the solid 3 times to obtain HSAMS@ZIF-8 samples 1, 2, 3, 4, 5, 6;

[0040] (4) Prepare the HSAMS reaction substrate mixed solution, numbered as solution a. The prepar...

Embodiment 2

[0044] It was proved that HSAMS@ZIF-8 has good catalytic ability, and the reactivity of HSAMS@ZIF-8 was compared with that of free HSAMS. Its method steps are:

[0045] (1) Dissolve 1.3 mg of zinc nitrate hexahydrate in 420 μL of water to obtain a zinc nitrate solution;

[0046] (2) Dissolve 1.4 mg of 2-methylimidazole in 420 μL of water, then add 20 μL of HSAMS (concentration: 6 mg / mL), and stir for 5 minutes to obtain 2-methylimidazole / enzyme solution;

[0047] (3) Mix the 2-methylimidazole / enzyme solution with the zinc nitrate solution, stir at room temperature for 4h, centrifuge (10000rpm, 5min), and wash the solid with Tris-HCl buffer solution with a concentration of 0.1mol / L and pH=8.0 3 times, get HSAMS@ZIF-8;

[0048] (4) Add 20 μL of solution a to HSAMS@ZIF-8, 180 μL of Tris-HCl buffer with a concentration of 0.1mol / L and pH=8.0; add 20 μL of solution a to 20 μL of HSAMS (concentration of 6 mg / mL), 160 μL of 0.1mol / L, Tris-HCl buffer solution with pH=8.0. The two we...

Embodiment 3

[0050] It was proved that HSAMS@ZIF-8 has better reaction stability than free HSAMS, and the reactivity of HSAMS@ZIF-8 and free HSAMS was compared under the conditions of high temperature, organic solvent and trypsin solution. Its method steps are:

[0051] (1) Prepare 5 copies of HSAMS@ZIF-8, numbered a1, a2, a3, a4, a5 respectively. For the experimental operation method, refer to the steps (1) (2) (3) of Example 2. Take 5 copies of 20μL HSAMS (concentration: 6mg / mL), numbered b1, b2, b3, b4, b5 respectively;

[0052] (2) Add 20 μL solution a, 80 μL Tris-HCl buffer solution with a concentration of 0.1 mol / L, pH=8.0, and 100 μL methanol to a1; add 20 μL solution a, 30 μL concentration of 0.1 mol / L, pH= 8.0 Tris-HCl buffer solution, 150 μL dimethylsulfoxide (DMSO); add 20 μL solution a to a3, 180 μL Tris-HCl buffer solution with a concentration of 0.1mol / L, pH=8.0, 0.01 mg trypsin ; Add 20 μL of solution a, 180 μL of Tris-HCl buffer solution with a concentration of 0.1 mol / L...

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Abstract

The invention relates to a preparation method of a ZIF-8 / enzyme composite material. The preparation method comprises the steps of regulating the concentration of a precursor to enable a ZIF-8 crystalto grow on the surface of an enzyme in a lamellar structure, mixing a metal salt water solution with a mixed solution of a ligand water solution and the enzyme, performing stirring at room temperaturefor a period of time, and separating out a solid product, and thereby obtaining the ZIF-8 / enzyme composite material. Different from a composite material obtained through a traditional method, the method does not depend on ducts of ZIF-8 to diffuse substrate molecules, and a large-size substrate makes contact with the active center of the enzyme through gaps between ZIF-8 lamellar structures, so that a catalytic reaction is completed. ZIF-8 nanoflowers growing on the surface of the enzyme can well protect the enzyme from reacting under harsh conditions, and meanwhile, the ZIF-8 / enzyme nanoflower composite material also has good circularity and storage capacity.

Description

technical field [0001] The invention relates to the technical field of composite material preparation, in particular to a preparation method of a ZIF-8 / enzyme composite material. Background technique [0002] Enzyme is a very important biological macromolecule, which has a catalytic efficiency far exceeding that of artificial catalysts in catalyzing biological reactions. Therefore, the application of enzymes in industrial production has great value and prospects. However, enzymes are difficult to apply in industry due to their low thermal stability, narrow applicable pH range, poor tolerance to organic solvents and metal ions, and difficult separation after reaction. To address this issue, immobilizing enzymes on metal-organic frameworks (MOFs) is a viable strategy. This strategy can effectively improve the stability of enzyme storage and reaction, and the enzyme can also be recycled. [0003] MOFs are a class of hybrid materials composed of metal nodes and organic ligand...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/02C12N11/14
CPCC12N11/02C12N11/14
Inventor 张琪
Owner FUDAN UNIV
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