A preparation method for enzymatically synthesizing fluorescent gold nanoclusters

A technology of fluorescent gold nanometers and gold nanoclusters, which is applied in nanotechnology, nanotechnology, nanomedicine, etc., can solve the problems of high dosage of acetylcholinesterase and complex response process, and achieve low concentration requirements, simple production steps, and high fluorescence intensity. big effect

Active Publication Date: 2022-04-22
SOUTH CHINA UNIV OF TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Acetylcholinesterase is a key enzyme in biological nerve transmission. It can selectively hydrolyze acetylcholine and specifically bind to organophosphorus substances. In recent years, it has been widely used as an intermediate participating substance in the fluorescence of hydrogen peroxide, acetylcholine and organophosphorus pesticides. In the detection, the response process is more complicated, and the amount of acetylcholinesterase is more

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  • A preparation method for enzymatically synthesizing fluorescent gold nanoclusters
  • A preparation method for enzymatically synthesizing fluorescent gold nanoclusters
  • A preparation method for enzymatically synthesizing fluorescent gold nanoclusters

Examples

Experimental program
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Effect test

Embodiment 1

[0033] Preparation of gold nanocluster fluorescent material: 10mmol / L chloroauric acid 1mL mixed with 0.2U / mL acetylcholinesterase (purchased from McLean) 100μL, preheated at 80°C for 5 minutes, then vigorously shaken and mixed, and added 1.0mol / L tris-HCl buffer solution (pH=7.4) and 1mol / L sodium hydroxide solution adjust the pH to 12.3, and make the final reaction volume of the mixed solution reach 2mL, at this moment acetylcholinesterase and chloroauric acid concentration ratio (mU:mmol / L) was 4:1, and the solution color turned dark green. React at 80°C for 18 hours, filter large particles of gold with a 0.45 μm filter head, and obtain a light white solution after ultrafiltration with a 10kDa ultrafiltration tube for 13 minutes, which has obvious blue fluorescence under the irradiation of a 365nm ultraviolet light source, that is, gold nanoparticles cluster solution. The obtained gold nanocluster maximum excitation wavelength and emission wavelength are 315nm and 398nm ...

Embodiment 2

[0035] Preparation of gold nanocluster fluorescent material: Mix 10mmol / L chloroauric acid 1mL with 0.2U / mL acetylcholinesterase (50ul~500ul), preheat at 40°C for 5 minutes, shake vigorously, add 1.0mol / L tris-HCl buffer solution (pH=7.4) and 1mol / L sodium hydroxide solution adjust the pH to 14.0, and make the final reaction volume of the mixed solution reach 2mL, at this time the acetylcholinesterase and chloroauric acid concentration ratio (mU:mmol / L ) is 2:1 to 20:1, and the color of the solution turns dark green. React at 40° C. for 24 hours, filter large particles of gold with a 0.45 μm filter head, and obtain a gold nanocluster solution after ultrafiltration with a 10 kDa ultrafiltration tube for 13 minutes. Such as image 3 As shown, after 12 hours of reaction, the fluorescence intensity value (at 315nm) of the gold nanoclusters obtained when the concentration ratio of acetylcholinesterase and chloroauric acid (mU:mmol / L) is 4:1 is the strongest.

Embodiment 3

[0037] Preparation of gold nanocluster fluorescent material: Mix 10mmol / L chloroauric acid 1mL with 0.2U / mL acetylcholinesterase 100ul, preheat at 40°C for 5 minutes, shake vigorously, add 1.0mol / L tris-HCl buffer Solution (pH=7.4) and 1mol / L sodium hydroxide solution adjust pH to 4.0~14.0, and make mixed liquor final reaction volume reach 2mL, this moment, acetylcholinesterase and chloroauric acid concentration ratio (mU:mmol / L) are 4:1, the color of the solution turns dark green. React at 40° C. for 24 hours, filter large particles of gold with a 0.45 μm filter head, and obtain a gold nanocluster solution after ultrafiltration with a 10 kDa ultrafiltration tube for 13 minutes. Such as Figure 4 As shown, after reacting for 8 hours, the fluorescence intensity value (at 315 nm) of the gold nanoclusters obtained when the pH of the reaction solution is 12.3 is the strongest.

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Abstract

The invention discloses a preparation method for synthesizing gold nanoclusters by acetylcholinesterase. The acetylcholinesterase and chloroauric acid are uniformly mixed, adjusted to pH 8.0-14.0, and reacted at a certain temperature for 8-24 hours to obtain acetylcholinesterase gold Nano-cluster solution; the concentration ratio of acetylcholinesterase and chloroauric acid in the solution is 1-20mU:1mmol / L. The present invention uses chloroauric acid and acetylcholinesterase as raw materials to synthesize gold nano-clusters in one step, and the prepared acetylcholinesterase gold nano-clusters emit blue light, and the maximum excitation wavelength and emission wavelength are 315nm and 398nm respectively, and have simplified acetylcholinesterase participating in pollutant fluorescence The application potential of the detection response process; the particle size distribution is between 0.8nm-2.6nm, the average particle size is 1.6nm, the smaller nanometer size has a unique electronic structure, and the fluorescence intensity is large. Stored at 4°C, the fluorescence intensity and the maximum The position of the emission peak remained stable for one week.

Description

technical field [0001] The invention belongs to the technical field of fluorescent nanometer material preparation, and in particular relates to a method for preparing gold nanoclusters synthesized by acetylcholinesterase. Background technique [0002] Gold nanoclusters are composed of dozens or hundreds of gold atoms, and their diameters are highly stable at 1-2nm. Since its diameter is close to the electron Fermi wavelength, the discontinuous energy level distribution leads to its unique electrical, optical and chemical properties. nature. It has attracted much attention because of its ultrafine, non-toxic, unique core / surface structure, molecular-like optical properties, high fluorescence, good biocompatibility, and high stability in saline solution. [0003] Protein-stabilized gold nanoclusters are a unique class of bionanomaterials with strong luminescence and highly specific chemical recognition properties, suitable for fluorescence detection of ions, inorganic substan...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): B22F9/24B22F1/054B82Y5/00B82Y15/00B82Y40/00C09K11/58
CPCB22F9/24C09K11/58B22F1/07B22F1/054
Inventor 朱能武刘煌睿黄熙贤李敏婷吴平霄
Owner SOUTH CHINA UNIV OF TECH
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