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Preparation method of integrase

A technology of integrating enzymes and skeleton materials, applied in biochemical equipment and methods, chemical instruments and methods, medical preparations containing active ingredients, etc. To achieve the effect of no toxic effect, sensitive results and simple operation process

Pending Publication Date: 2022-06-28
SOUTHEAST UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this detection process involves glucose oxidase and horseradish peroxidase. The natural enzymes are unstable, easily inactivated in the catalytic system, and difficult to recover, so they cannot be recycled.
In addition, glucose oxidase and horseradish peroxidase need to achieve their optimal catalytic efficiency under different pH conditions (Small.2018, 14, 1803256), so the detection requires a two-step reaction, which is cumbersome and time-consuming

Method used

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  • Preparation method of integrase
  • Preparation method of integrase
  • Preparation method of integrase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] (1) Preparation of zeolite imidazolate framework material: 9.5197g of zinc nitrate hexahydrate and 3.0769g of 2-methylimidazole were dissolved in 500mL and 400mL of methanol, respectively, after the dissolution was complete, mixed, and stirred at room temperature at 1000 rpm for 2 hours . Centrifuge at 13,000 rpm, wash with methanol three times, and vacuum dry at 60°C overnight.

[0034] (2) Preparation of mesoporous silicon-wrapped zeolite imidazolate framework material: 2.6 g of zeolite imidazolate framework material was dispersed in 24 mL of methanol. 216 mL of distilled water was added (total volume 240 mL, 10 vol% methanol). The pH was adjusted to 11 by adding 1M sodium hydroxide. 0.2016 g of cetyltrimethylammonium bromide was added and stirred for 30 minutes. 1.2 mL of tetraethyl orthosilicate was added under stirring at 1000 rpm and stirred for 30 minutes. Centrifuge at 13,000 rpm and wash three times with methanol. Dry under vacuum at 60°C overnight.

[0035...

Embodiment 2

[0040] (1) Preparation of zeolite imidazolate framework material: 9.5197g of zinc nitrate hexahydrate and 3.0769g of 2-methylimidazole were dissolved in 500mL and 400mL of methanol, respectively, after the dissolution was complete, mixed, and stirred at room temperature at 1000 rpm for 2 hours . Centrifuge at 13,000 rpm, wash with methanol three times, and vacuum dry at 60°C overnight.

[0041] (2) Preparation of mesoporous silicon-wrapped zeolite imidazolate framework material: 2.6 g of zeolite imidazolate framework material was dispersed in 24 mL of methanol. 216 mL of distilled water was added (total volume 240 mL, 10 vol% methanol). The pH was adjusted to 11 by adding 1M sodium hydroxide. 0.2016 g of cetyltrimethylammonium bromide was added and stirred for 30 minutes. 1.2 mL of tetraethyl orthosilicate was added at 1000 rpm and stirred for 30 minutes. Centrifuge at 13,000 rpm and wash three times with methanol. Dry under vacuum at 60°C overnight.

[0042] (3) Prepara...

Embodiment 3

[0046] (1) Preparation of zeolite imidazolate framework material: 9.5197g of zinc nitrate hexahydrate and 3.0769g of 2-methylimidazole were dissolved in 500mL and 400mL of methanol, respectively, after the dissolution was complete, mixed, and stirred at room temperature at 1000 rpm for 2 hours . Centrifuge at 13,000 rpm, wash with methanol three times, and vacuum dry at 60°C overnight.

[0047] (2) Preparation of mesoporous silicon-wrapped zeolite imidazolate framework material: 2.6 g of zeolite imidazolate framework material was dispersed in 24 mL of methanol. 216 mL of distilled water was added (total volume 240 mL, 10 vol% methanol). The pH was adjusted to 11 by adding 1M sodium hydroxide. 0.2016 g of cetyltrimethylammonium bromide was added and stirred for 30 minutes. 1.2 mL of tetraethyl orthosilicate was added at 1000 rpm and stirred for 30 minutes. Centrifuge at 13,000 rpm and wash three times with methanol. Dry under vacuum at 60°C overnight.

[0048] (3) Prepara...

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Abstract

The invention discloses a preparation method of integrase, which comprises the following steps: pyrolyzing a zinc-based zeolite imidazate framework material coated with mesoporous silicon, taking a generated porous metal nitrogen carbon material as a substrate, and loading gold nanoparticles in situ by utilizing a chemical reduction method to construct nano-enzyme; and finally, coupling the natural glucose oxidase with the nano-enzyme through electrostatic adsorption to obtain the integrase. According to the method, the catalytic activity of the nano enzyme can be reserved, and the stability and the catalytic activity of the natural enzyme can be improved. Besides, the prepared integrase can be used for further detecting glucose, the operation process is simple and rapid, the result is sensitive, the selectivity is high, the detection limit is low (0.77 mu M), the detection range is wide (1-300 mu M), and the integrase has a good application prospect in sensitive blood glucose detection. In addition, the integrase also has a three-in-one tumor treatment effect of hunger, chemical kinetics and photo-thermal.

Description

technical field [0001] The present invention relates to the preparation method of analytical chemistry and nanometer medicine, in particular to a preparation method of integrase. Background technique [0002] At present, diabetes has become the third chronic disease that seriously endangers human health after tumor and cardiovascular and cerebrovascular diseases due to its wide range of diseases and serious complications. In order to prevent and treat diabetes, it is of great practical significance to accurately measure the content of glucose in blood and body fluids. In living organisms, glucose can be catalyzed by glucose oxidase in the presence of oxygen to produce gluconic acid and hydrogen peroxide. Therefore, the most common method to detect glucose is to use horseradish peroxidase to quantitatively detect hydrogen peroxide, the metabolite of glucose. However, this detection process involves glucose oxidase and horseradish peroxidase, and the natural enzymes are unst...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B01J27/24B01J31/00C12N9/04C12N9/08C12Q1/54C12Q1/26C12Q1/28A61K33/242A61K38/44A61P35/00B82Y5/00
CPCB01J27/24B01J31/003C12N9/0006C12N9/0065C12Y101/03004C12Y111/01C12Q1/54C12Q1/26C12Q1/28A61K33/242A61K38/443A61P35/00B82Y5/00C12Q2326/12A61K2300/00
Inventor 李颖陈雨洁李淑娟刘松琴
Owner SOUTHEAST UNIV
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