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Metal organic framework material-enzyme compound as well as preparation method and application thereof

A metal-organic framework and composite technology, applied in biochemical equipment and methods, enzymes, immobilized enzymes, etc., can solve the problems of free enzyme inactivation, achieve high catalytic activity, reduce costs, good stability and dispersibility Effect

Pending Publication Date: 2021-11-05
TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, free enzymes are susceptible to various physical and chemical degradation and inactivation in vivo and in vitro, and it is difficult for negatively charged proteins to enter the cytoplasm wrapped by phospholipid bilayers to function

Method used

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  • Metal organic framework material-enzyme compound as well as preparation method and application thereof
  • Metal organic framework material-enzyme compound as well as preparation method and application thereof
  • Metal organic framework material-enzyme compound as well as preparation method and application thereof

Examples

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preparation example Construction

[0029] One aspect of the present invention provides a method for preparing a metal-organic framework material-enzyme complex, comprising the following steps:

[0030] Dissolve antioxidant enzymes, transition metal ions and organic ligands in a solvent for co-precipitation reaction;

[0031] Wherein, the antioxidant enzyme is selected from the combined enzyme of superoxide dismutase and catalase, glutathione peroxidase or horseradish peroxidase, or glutathione reductase and glutathione peroxidase Combined enzymes of oxidase, glucose dehydrogenase or alcohol dehydrogenase;

[0032] The transition metal ion is derived from a soluble metal salt, and the organic ligand is an imidazole compound.

[0033] The metal-organic framework material-enzyme complex has a pH-dependent protein release mechanism, and at the same time relies on the proton sponge effect, and can effectively escape from acidic endosomes after ingestion into cells, thereby ensuring its stability under physiological...

Embodiment 1

[0055] (1) Prepare 1.25mol / L 2-methylimidazole aqueous solution, 0.31mol / L zinc nitrate aqueous solution, 2.5mg / mL SOD solution, and 7.0mg / mL CAT solution respectively, and ultrasonicate at room temperature for 10min until completely dissolved, wherein, The molar ratio of zinc ion to 2-methylimidazole is 1:40;

[0056] (2) Add 160 μL of zinc nitrate aqueous solution, 50 μL of SOD solution, and 100 μL of CAT solution to 1.6 mL of 2-methylimidazole aqueous solution at the same time, and stir for 30 minutes at room temperature with a magnetic stirrer at a speed of 600 rpm;

[0057] (3) Centrifuge the solution obtained in step (2) at a speed of 15,000 rpm for 5 minutes, remove the supernatant, resuspend with deionized water, and repeat washing twice;

[0058] (4) The precipitate obtained in step (3) was pre-frozen at -20°C for 12 hours, and freeze-dried for 24 hours to obtain a metal-organic framework material-enzyme complex, which was designated as SC@ZIF-8.

[0059] The scannin...

Embodiment 2

[0061] The preparation method of this example is basically the same as that of Example 1, except that the concentration of the 2-methylimidazole aqueous solution is 1.875 mol / L. Specific steps are as follows:

[0062] (1) Prepare 1.875mol / L 2-methylimidazole aqueous solution, 0.31mol / L zinc nitrate aqueous solution, 2.5mg / mL SOD solution, and 7.0mg / mL CAT solution respectively, and ultrasonicate at room temperature for 10min until completely dissolved, wherein, The molar ratio of zinc ion to 2-methylimidazole is 1:60;

[0063] (2) Add 160 μL of zinc nitrate aqueous solution, 50 μL of SOD solution, and 100 μL of CAT solution to 1.6 mL of 2-methylimidazole aqueous solution at the same time, and stir for 30 minutes at room temperature with a magnetic stirrer at a speed of 600 rpm;

[0064] (3) Centrifuge the solution obtained in step (2) at a speed of 15,000 rpm for 5 minutes, remove the supernatant, resuspend with deionized water, and repeat washing twice;

[0065] (4) The pre...

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Abstract

The invention relates to the technical field of immobilized enzymes, in particular to a metal organic framework material-enzyme compound as well as a preparation method and application thereof. The preparation method of the metal organic framework material-enzyme compound comprises the following steps that antioxidant enzyme, transition metal ions and an organic ligand are dissolved in a solvent for a co-precipitation reaction, the antioxidant enzyme is selected from combined enzyme of superoxide dismutase and catalase, glutathione peroxidase or horseradish peroxidase, and the transition metal ions are selected from soluble metal salts, and the organic ligand is an imidazole compound. The metal organic framework material-enzyme compound has strong active oxygen scavenging capacity.

Description

technical field [0001] The invention relates to the technical field of immobilized enzymes, in particular to a metal organic framework material-enzyme complex and its preparation method and application. Background technique [0002] Reactive oxygen species (ROS) are normal products of metabolic cycles in the body, including superoxide ions, hydrogen peroxide, hydroxyl radicals, singlet oxygen and other electronic reduction products of oxygen, and play an important role in metabolism, immunity, and signal transduction . There is a complex antioxidant symbiotic loop in the human body to maintain the level of reactive oxygen species under normal physiological conditions, including superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), Glutathione reductase (GR) and other enzymes, as well as vitamin C, vitamin E, coenzyme Q, glutathione (GSH), flavonoids and other non-enzyme small molecules. However, when the body is exposed to a specific environment or toxi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/089C12N9/08C12N9/04C12N9/02
CPCC12N11/089C12N9/0006C12N9/0051C12N9/0065C12N9/0089C12Y115/01001C12Y111/01006C12Y111/01009C12Y111/01007C12Y108/01007C12Y101/01047C12Y101/01001
Inventor 戈钧刘雨
Owner TSINGHUA UNIV
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