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Preparation and application of dopamine-coated urchin-shaped manganese dioxide hollow microspheres

A technology of manganese dioxide and dopamine, which is applied to the application of the immobilized acetylcholinesterase in the screening of acetylcholinesterase inhibitors in traditional Chinese medicine, and the immobilized enzyme carrier is used in the field of preparing immobilized acetylcholinesterase, which can solve the problem of enzyme immobilization efficiency Low cost, poor stability of free enzyme, etc., to achieve the effect of less sample consumption, rich active sites, and short diffusion path

Pending Publication Date: 2022-03-29
LANZHOU INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, sea urchin-like manganese dioxide hollow microspheres are used as an enzyme immobilization carrier for the immobilization of acetylcholinesterase, the stability of the free enzyme is poor, and the enzyme immobilization efficiency is low. Therefore, it is necessary to modify the manganese dioxide hollow microspheres , in order to be used as an immobilized acetylcholinesterase carrier for screening acetylcholinesterase inhibitors

Method used

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  • Preparation and application of dopamine-coated urchin-shaped manganese dioxide hollow microspheres
  • Preparation and application of dopamine-coated urchin-shaped manganese dioxide hollow microspheres
  • Preparation and application of dopamine-coated urchin-shaped manganese dioxide hollow microspheres

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Experimental program
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Effect test

Embodiment 1

[0043] 1. Accurately weigh 0.4g of potassium permanganate, fully dissolve it in 20mL of ultrapure water; then add 2mL of concentrated hydrochloric acid drop by drop, and stir continuously at room temperature for 30 min; transfer the obtained mixed solution to polytetrafluoroethylene In an autoclave of ethylene, react at 120°C for 4 h; after the autoclave is cooled to room temperature, the product is collected by centrifugation, washed several times with ethanol and ultrapure water, and dried to obtain manganese dioxide hollow microspheres h-MnO 2 .

[0044] 2. Take 10mgh-MnO 2 Dissolve completely in 10 mL of Tris-HCl buffer solution (pH value 8.5), sonicate for 15 min; add 10 mg of dopamine hydrochloride, sonicate for 5 min, collect the product by centrifugation, wash several times with ethanol and ultrapure water, freeze-dry, Obtained dopamine-coated manganese dioxide hollow microspheres h-MnO 2 @PDA.

[0045] 3. Take 4mgh-MnO 2 @PDA, dispersed in 4mL phosphate buffer sol...

Embodiment 2

[0052] 1. Accurately weigh 0.4g potassium permanganate, fully dissolve it in 25mL ultrapure water; then add 4mL concentrated hydrochloric acid drop by drop, and stir continuously at room temperature for 40 min; transfer the obtained mixed solution to polytetrafluoroethylene In an autoclave of ethylene, react at 140°C for 3 h; after the autoclave is cooled to room temperature, the product is collected by centrifugation, washed several times with ethanol and ultrapure water, and dried to obtain manganese dioxide hollow microspheres h-MnO 2 ;

[0053] 2. Weigh 10mgh-MnO 2 , completely dissolved in 20 mL of Tris-HCl buffer solution (pH value 9.5), sonicated for 25 minutes; added 20 mg of dopamine hydrochloride, sonicated for 15 minutes, collected by centrifugation, washed several times with ethanol and ultrapure water, and dried. Obtained dopamine-coated manganese dioxide hollow microspheres h-MnO 2 @PDA;

[0054] 3. Weigh 4mgh-MnO 2 @PDA, dispersed in 8mL of phosphate buffer ...

Embodiment 3

[0057] 1. Accurately weigh 0.4g potassium permanganate, fully dissolve it in 30mL ultrapure water; then add 5mL concentrated hydrochloric acid drop by drop, and stir continuously at room temperature for 35 min; transfer the obtained mixed solution to polytetrafluoroethylene In a high-pressure reactor, the reaction was carried out at 130 °C for 4 h. After the autoclave was cooled to room temperature, the product was collected by centrifugation, washed several times with ethanol and ultrapure water, and dried to obtain manganese dioxide hollow microspheres h-MnO 2 ;

[0058] 2. Take 10mgh-MnO 2 , completely dissolved in 30 mL of Tris-HCl buffer (pH value 8), sonicated for 15 min; added 30 mg of dopamine hydrochloride, sonicated for 25 min, centrifuged to collect the product, washed several times with ethanol and ultrapure water, and dried. Obtained dopamine-coated manganese dioxide hollow microspheres h-MnO 2 @PDA;

[0059] 3. Take 4mgh-MnO 2 @PDA, dispersed in 10mL of phosph...

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Abstract

The invention provides preparation of dopamine-coated sea urchin-shaped manganese dioxide hollow microspheres, which comprises the following steps: completely dissolving manganese dioxide hollow microspheres in a tris (hydroxymethyl) aminomethane hydrochloride solution, then adding dopamine hydrochloride, carrying out ultrasonic treatment for 10-25 minutes, centrifuging, washing and drying to obtain the dopamine-coated manganese dioxide hollow microspheres. The hollow microspheres have the advantages of thermal stability, easiness in synthesis, porous structure, low density, short diffusion path, high specific surface area, rich active sites and the like, and moreover, the formed aminated manganese dioxide hollow microspheres are beneficial to immobilization of acetylcholin esterase, so that the aminated manganese dioxide hollow microspheres serving as a novel enzyme immobilization carrier have a good application prospect. After the acetylcholin esterase is immobilized, the capillary electrophoresis technology is combined, and the acetylcholin esterase inhibitor is screened from traditional Chinese medicines, so that the method has the characteristics of high separation efficiency, short analysis time, low sample consumption, low operation cost and the like, and a technical support is provided for discovery of new medicines for treating Alzheimer's disease.

Description

technical field [0001] The present invention relates to a method for preparing dopamine-coated manganese dioxide hollow microspheres, and the present invention also relates to a method for preparing immobilized acetylcholinesterase using the dopamine-coated manganese dioxide hollow microspheres as an immobilized enzyme carrier. The invention also relates to the application of the immobilized acetylcholinesterase in screening acetylcholinesterase inhibitors in traditional Chinese medicines. Background technique [0002] Alzheimer's disease (AD) is the most common neurodegenerative disease in the elderly that is associated with a decrease in the neurotransmitter acetylcholine (ACh) in the brain. The pathogenesis of AD has not yet been fully elucidated. At present, the main theories are Aβ (Amyloid β-protein) theory, cholinergic theory, Tau (Microtubule-associated protein tau) protein abnormal phosphorylation theory, oxidative stress theory and inflammatory response theory. . ...

Claims

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Application Information

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IPC IPC(8): C12N11/14C12N9/18C01G45/02C12Q1/46B01J13/02
CPCC12N11/14C12N9/18C01G45/02B01J13/02C12Y301/01007C12Q1/46C01P2004/34C01P2004/03C01P2004/04G01N2333/918
Inventor 刘佳师彦平哈伟
Owner LANZHOU INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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