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Magnetic nanoparticle-glycosyl transferase-amorphous metal organic framework composite catalytic material as well as preparation method and application thereof

A technology of metal-organic frameworks and glycosyltransferases, applied in biochemical equipment and methods, transferases, immobilized on or in inorganic carriers, etc., can solve the problems of enzymes being easily soluble in water, high catalytic costs, and affecting activity, etc. problems, to achieve the effect of low raw material cost, high catalytic activity, and mild synthesis conditions

Pending Publication Date: 2022-01-28
NANJING NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The use of amorphous metal-organic framework materials can maintain the activity of the enzyme to a greater extent and realize the repeated use of the enzyme, which solves the problem that in the glycosylation reaction, the enzyme is easily soluble in water and its activity is easily affected by the environment, and it is difficult to collect for repeated use. Utilization, the problem of high catalytic cost

Method used

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  • Magnetic nanoparticle-glycosyl transferase-amorphous metal organic framework composite catalytic material as well as preparation method and application thereof
  • Magnetic nanoparticle-glycosyl transferase-amorphous metal organic framework composite catalytic material as well as preparation method and application thereof
  • Magnetic nanoparticle-glycosyl transferase-amorphous metal organic framework composite catalytic material as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] Preparation of magnetic nanoparticles (MNPs):

[0060] FeCl 3 solution (10mL, 5mM) with FeCl 2 Put the solution (10mL, 10mM) into a double-necked flask, add a magnet, pass in nitrogen, and discharge oxygen. Place it in a water bath constant temperature magnetic stirrer at 75°C, heat and stir the reaction at a speed of 300rpm, and cool to room temperature after 1h of reaction , add ammonia water (10%) to the obtained reaction solution, the volume ratio of ammonia water to the reaction solution is 1:3, place it in a water bath constant temperature magnetic stirrer, heat and stir the reaction at 75°C, the rotation speed is 200rpm, and react for 1h; Cool to room temperature, recover magnetic nanoparticles (MNPs) under magnetic field conditions, add water after drying to prepare a certain concentration of MNP solution.

Embodiment 2

[0062] Preparation of MNPs-HS2@aZIF-90:

[0063] Get the MNPs solution that 0.6mL concentration is 1mg / mL prepared in Example 1, stir and mix and add to 1mL Zn(NO 3 ) 2 ·6H 2O (40mM) and 1mL glycosyltransferase pmHS2 (0.3mg / mL) mixed solution, then add 1mL 2-imidazole-carboxaldehyde (HICA, dissolve at high temperature, 40mM) solution and add ultrapure water to make up to 4mL. React at 37° C. and 500 rpm for 15 hours. After the reaction, centrifuge at 8,000 rpm for 4 minutes to recover the precipitate. Then, it was washed three times with ultrapure water to remove the glycosyltransferase not wrapped by the aZIF-90 material, and the magnetic nanoparticle-glycosyltransferase-amorphous metal-organic framework composite catalytic material was obtained.

Embodiment 3

[0065] The preparation method of MNPs-HS2@aZIF-90 catalyzing the glycosylation of monosaccharides to generate heparin disaccharides:

[0066] In Tris-HCl buffer (50mM pH 7.4), add uridine diphosphate-N-acetylglucosamine (final concentration 10mM), p-nitrophenyl-β-D-glucuronic acid (final concentration 10mM), magnesium chloride ( Final concentration 10mM) was shaken and mixed in a water bath at 37°C, the speed was 150rpm, and the time was 5min. After mixing, add uridine diphosphate-N-acetylglucosamine and p-nitrophenyl-β-D-glucuronic acid 5% of the MNPs-HS2@aZIF-90 composite catalyst prepared in Example 2 was shaken in a water bath at 37° C. at 150 rpm, and the reaction was completed after 12 hours of sufficient reaction. The product was centrifuged at 10,000 rpm for 5 minutes, the upper layer oil was taken, and the remaining solution was subjected to magnetic adsorption to collect the composite catalytic material, washed for reuse, and the supernatant was taken for TLC separat...

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Abstract

The invention discloses a magnetic nanoparticle-glycosyl transferase-amorphous metal organic framework composite catalytic material as well as a preparation method and application thereof. The composite catalytic material is formed by wrapping glycosyl transferase and magnetic nanoparticles in an amorphous metal organic framework as a novel immobilized enzyme material. After the composite catalytic material is prepared, the composite catalytic material is added into a mixed solution of a glycosyl donor and a glycosyl acceptor, and a heparin disaccharide product is obtained through a catalytic glycosylation reaction. The amorphous metal organic framework material has a larger aperture than a common metal organic framework, reduces mass transfer resistance, improves enzymatic efficiency, is hollow inside, and is more beneficial to maintenance of enzyme molecule activity. The composite catalytic material can be used for efficiently catalyzing the glycosylation reaction of monosaccharide, not only has good thermal stability and chemical stability, but also can improve the enzymatic reaction rate, can be recycled for multiple times, keeps higher catalytic activity and simplifies the separation step.

Description

technical field [0001] The invention relates to the technical field of catalysts, in particular to a magnetic nanoparticle-glycosyltransferase-amorphous metal-organic framework composite catalytic material and its preparation method and application. Background technique [0002] At present, heparin has been successfully used as an injectable anticoagulant drug for the prevention and treatment of thrombotic diseases. Heparin can exert anti-inflammatory activity by neutralizing inflammatory factors and selectins, and can also inhibit tumor neovascularization and tumor cell proliferation. It can also block the attachment of tumor cells to endothelial cells and inhibit the activity of tumor metastasis-related enzymes, thereby inhibiting tumor metastasis. In addition, heparin also has a variety of biological activities such as regulating blood lipids, antiviral, and antiallergic. Low-molecular-weight heparin, which is widely used at present, is mainly isolated and extracted from...

Claims

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Application Information

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IPC IPC(8): C12N9/10C12N11/089C12N11/14
CPCC12N9/1048C12N11/089C12N11/14Y02P20/584
Inventor 张幸乔梦纪元李昺之李乐林蕾
Owner NANJING NORMAL UNIVERSITY
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