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Protein and amorphous metal organic framework compound and preparation method thereof

A metal-organic framework, amorphous technology, applied in biochemical equipment and methods, immobilized on/in organic carriers, immobilized on or in inorganic carriers, etc., can solve the problem of inability to use biologically active proteins and Amorphous metal-organic framework complexes, low activity of complexes, cumbersome synthesis steps, etc., to achieve mild conditions, high protein entrapment rate, and good protein stability.

Active Publication Date: 2020-11-10
TSINGHUA UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The first type of preparation method uses crystalline metal-organic framework compounds, usually with a microporous structure (pore size between 0 and 2 nm). On the one hand, the too small pore structure limits the conformational change of the internal protein during the catalytic process. On the other hand, It also limits the mass transfer process of the substrate, so the activity of the prepared complex is much lower than that of the natural enzyme
In the second type of method, mesoporous metal-organic frameworks matching the size of proteins need to first synthesize complex organic ligands, and then coordinate with metal ions to form mesoporous materials, and complex organic ligands usually require multi-step organic synthesis reactions. The steps are cumbersome, the yield is low, and the energy consumption is high
And the Gpa-level high-pressure preparation conditions also cannot make the enzyme molecules retain their activity well.
[0006] Therefore, the one-step preparation of biologically active protein-amorphous metal-organic framework complexes cannot be carried out by the above physical methods.

Method used

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  • Protein and amorphous metal organic framework compound and preparation method thereof
  • Protein and amorphous metal organic framework compound and preparation method thereof
  • Protein and amorphous metal organic framework compound and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0082] Embodiment 1, the synthesis of metal organic framework compound

[0083] 1. Prepare an aqueous solution of zinc acetate hydrate with a concentration of 20mmol / L and an aqueous solution of 2-methylimidazole with a concentration of 80mmol / L, respectively, and ultrasonicate the resulting solutions for 15 minutes.

[0084] 2. Mix 1 mL each of the zinc ion solution and 2-methylimidazole solution obtained in step 1, and react at 25° C. for 1 hour.

[0085] 4. The product obtained in step 3 was separated by centrifugation, repeatedly washed with deionized water for 3 times, and the washed product was freeze-dried for 24 hours to obtain a metal organic framework compound.

[0086] The scanning electron micrographs and transmission electron micrographs of the metal-organic framework compound prepared in this embodiment are as follows: figure 1 and figure 2 shown by figure 1 and figure 2 It can be seen that the main body shape of the obtained compound is spherical, and the ...

Embodiment 2

[0094] Embodiment 2, the synthesis of protein and amorphous metal-organic framework complex

[0095] 1. Prepare an aqueous solution containing 10 mg / mL of glucose oxidase.

[0096] 2. An aqueous solution of zinc acetate hydrate with a concentration of 20mmol / L and an aqueous solution of 2-methylimidazole with a concentration of 80mmol / L were respectively prepared, and the obtained solutions were ultrasonically treated for 15 minutes.

[0097] 3. Mix 100 μL of the solution obtained in step 1 with 1 mL each of the zinc ion solution and 2-methylimidazole solution in step 2, and react at 25° C. for 1 hour.

[0098] 4. The product obtained in step 3 was separated by centrifugation, repeatedly washed with deionized water for 3 times, and the washed product was freeze-dried for 8 hours to obtain a protein embedding rate of 100% in the product.

[0099] The scanning electron micrographs and transmission electron micrographs of the compound prepared in this embodiment are as follows: ...

Embodiment 3

[0107] Embodiment 3, the synthesis of compound alcohol dehydrogenase-metal organic framework compound

[0108] 1. Prepare 10mg / mL alcohol dehydrogenase solution.

[0109] 2. Prepare an aqueous solution of zinc acetate hydrate with a concentration of 40mmol / L and an aqueous solution of 2-methylimidazole with a concentration of 160mmol / L, respectively.

[0110] 3. Take 1 mL each of the zinc ion solution and 2-methylimidazole solution obtained in step 1 and 100 μL of the protein solution obtained in step 2, and react at 25°C for 30 min.

[0111] 4. The product obtained in step 3 was separated by centrifugation, repeatedly washed with deionized water for 3 times, and the washed product was freeze-dried for 8 hours to obtain a metal organic framework compound.

[0112] The transmission electron microscope figure of the complex of the compound alcohol dehydrogenase prepared by the present embodiment is as follows Figure 5 As shown, the scanning electron microscope image is shown ...

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Abstract

The invention discloses a protein and amorphous metal organic framework compound and a preparation method thereof. The organic framework compound has a 2 nm-50 nm mesoporous structure. The preparationmethod comprises the following step of enabling a protein, zinc ions and an organic ligand to react in a solvent, wherein the organic ligand is a compound containing an imidazole group; the protein is one or a combination of several kinds of cytochrome C, cytochrome P450, horse radish peroxidase, alcohol dehydrogenase, lipase, acetylcholin esterase, laccase, a green fluorescent protein, glucose dehydrogenase, glucose oxidase, trypsin, bacillus subtilis protease, carbonic anhydrase, aldehyde ketone reductase, amylase, saccharase, superoxide dismutase, urease and catalase. The preparation method of the protein and amorphous metal organic framework compound provided by the invention is simple to operate and mild in condition, the obtained product is high in protein embedding rate and good inprotein stability, and the biologic al activity of the protein is reserved to a greater extent.

Description

technical field [0001] The invention relates to a compound of protein and amorphous metal-organic framework and a preparation method thereof, belonging to the technical field of biological material preparation. Background technique [0002] Proteins have abundant physiological and catalytic functions. Metal-organic framework compounds are complexes synthesized by the coordination of metal ions and organic ligands. It has the advantages of rich pore structure, large specific surface area, good chemical stability, and adjustable structure. It is widely used in the fields of catalysis and separation. All have good application prospects. Combining proteins and metal-organic framework compounds to construct protein-metal-organic framework complexes has attracted widespread attention in recent years, and is expected to play an important role in the fields of biocatalysis, drug delivery, and analysis and detection. [0003] In previous patents and literature reports, the combinat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/14C12N11/02C07K17/14C07K17/02
CPCC12N11/14C12N11/02C12N9/0065C12N9/0006C12N9/20C12N9/18C12N9/0061C12N9/6427C12N9/54C12N9/88C12N9/2402C12N9/0089C12N9/80C07K17/14C07K17/02C12Y111/01007C12Y101/01001C12Y301/01003C12Y301/01007C12Y110/03002C12Y101/9901C12Y101/03004C12Y304/21004C12Y304/21062C12Y402/01001C12Y302/0101C12Y115/01001C12Y305/01005C12Y111/01006
Inventor 戈钧吴晓玲侯淼
Owner TSINGHUA UNIV
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