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Preparation method of copper phosphate-enzyme mineralized material as well as product and application thereof

A technology of copper phosphate and phosphate buffer, applied in the field of nanomaterials, can solve the problems of toxicity, easy to cause rejection, poor stability, etc., and achieve the effect of improving stability, simple preparation method, and small size

Pending Publication Date: 2020-04-17
HUNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the conventional generation of singlet oxygen is mainly produced by photosensitization of dyes, but the stability of such dyes in some organic chemical catalytic applications is relatively poor. In vivo, because dyes may themselves be toxic, it is easy to cause Rejection

Method used

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  • Preparation method of copper phosphate-enzyme mineralized material as well as product and application thereof
  • Preparation method of copper phosphate-enzyme mineralized material as well as product and application thereof
  • Preparation method of copper phosphate-enzyme mineralized material as well as product and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] The preparation of copper phosphate-glucose oxidase mineralization material specifically comprises the following steps:

[0032] (1) Prepare 100 U / mg of glucose oxidase (GOD) into a 10 mg / mL aqueous solution, and dihydrate copper chloride (CuCl 2 2H 2 O) It is configured as a 200mM aqueous solution, and disodium hydrogen phosphate and sodium dihydrogen phosphate are formulated as a buffer solution with a pH of 7.4.

[0033] (2) Add 100 μL of GOD aqueous solution to 1.0 mL of buffer solution, mix well, and then add 30 μL of CuCl 2 2H 2 O aqueous solution, after mixing thoroughly, let it stand for 5 minutes.

[0034] (3) centrifugation+water cleaning and purification of the mixed solution obtained in step (2) 3 times, wherein: the centrifugal speed is 8000r / min, and each centrifugation time is 10min to obtain Cu-GOD mineralized material, and the obtained Cu-GOD The mineralized material was dispersed in 0.5 mL of water and stored at 4°C.

[0035] (4) The Cu-GOD disper...

Embodiment 2

[0039] The preparation method of copper phosphate and catalase mineralization material specifically comprises the following steps:

[0040] (1) 200U / mg of catalase (CAT) was configured into a 100mg / mL aqueous solution, and copper chloride dihydrate (CuCl 2 2H 2 O) It is configured as a 200mM aqueous solution, and disodium hydrogen phosphate and sodium dihydrogen phosphate are formulated as a buffer solution with a pH of 7.4.

[0041] (2) Add 100 μL of CAT aqueous solution to 0.3 mL of buffer solution, then add 0.7 mL of water, mix well, and then add 30 μL of CuCl 2 2H 2 O aqueous solution, after fully mixing, let it stand for 5 minutes;

[0042] (3) Centrifugation+water cleaning and purification of the mixed solution obtained in step (2) 5 times, wherein: the centrifugal speed is 8000r / min, and each centrifugation time is 10min to obtain Cu-CAT mineralized material, and the obtained Cu-CAT The mineralized material was dispersed in 0.5 mL and stored at 4°C.

[0043] (4) Th...

Embodiment 3

[0046] 1. GOD consumption of glucose to produce hydrogen peroxide ability test

[0047] Method: Mix 10 μL of the Cu-GOD dispersion prepared in Example 1, 100 μL of TMB (1.5 mM, dissolved in DMSO), and 100 μL of glucose (200 mM aqueous solution), and incubate at room temperature for 15 min, 45 min, 90 min and 120 min respectively UV measurements were carried out at the time points, and the absorbance at 650 nm was recorded.

[0048] The result is as Figure 5 As shown, it can be known from the figure that GOD consumes glucose to produce hydrogen peroxide, and then hydrogen peroxide reacts with copper phosphate to produce hydroxyl radicals, which can be detected through the ultraviolet absorption of TMB. As time goes by, the content of hydrogen peroxide produced increases. The oxidation absorption peak gradually increases.

[0049] 2. Cu-GOD consumption glutathione ability test

[0050] Method: Different volumes of Cu-GOD were mixed with 100 μL of glutathione (5 mM) to a fina...

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Abstract

The invention discloses a preparation method of a copper phosphate-enzyme mineralized material as well as a product and application thereof. The preparation method comprises the following steps: 1) respectively preparing an aqueous solution of a copper salt, an aqueous solution of an enzyme, a buffer solution composed of disodium hydrogen phosphate and sodium dihydrogen phosphate; 2) adding a buffer solution into the aqueous solution of the enzyme, carrying out uniform mixing under stirring, adding the aqueous solution of the copper salt, carrying out full and uniform mixing, performing standing, then carrying out centrifugation and rinsing and purification with water to obtain the copper phosphate-enzyme mineralized material, dispersing the copper phosphate-enzyme mineralized material inwater, and carrying out storing. The preparation method is simple; the generated copper phosphate-enzyme mineralized material can improve the stability of the enzyme to a great extent; singlet oxygencan be generated under the condition that photon excitation is not needed; and the copper phosphate-enzyme mineralized material has good application prospects in the fields of organic synthesis and biology. The size of the prepared copper phosphate-enzyme mineralized material is 100 nm or below, and therefore, the copper phosphate-enzyme mineralized material is large in specific surface area, goodin catalytic activity, small in size and easier to enter organisms.

Description

technical field [0001] The invention belongs to the technical field of nanomaterials, and in particular relates to a preparation method of a copper phosphate-enzyme mineralization material and its product and application. Background technique [0002] Singlet oxygen, that is, excited oxygen molecules. After the ground state oxygen atom (triplet oxygen molecule) is excited, the two spin-parallel electrons in the original two 2pπ* orbitals can occupy a 2pπ* orbital at the same time, with opposite spins, or occupy two 2pπ* orbitals respectively , with the opposite spin. In the past people's cognition, singlet oxygen is a highly active oxygen free radical with cytotoxicity, and the cell membrane, mitochondria and other parts are most sensitive to it, and can interact with a variety of biological macromolecules in cells. Combining with molecules can cause damage to the cell membrane system, and studies have proved that it has a great relationship with blood diseases, so many pe...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/14B01J27/18B01J35/10B01J35/02B01J32/00C01B15/022C01B13/02A61K41/00A61P35/00B82Y5/00B82Y30/00B01J35/00
CPCC12N11/14C12N9/0006C12N9/0065C12Y101/03004C12Y111/01006B01J27/1817C01B15/022C01B13/0214A61K41/0057A61K41/0052A61P35/00B82Y5/00B82Y30/00B01J35/61B01J35/40A61K2300/00
Inventor 张晓兵赵岩宋国胜孔维恒
Owner HUNAN UNIV
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