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Preparation method of fluorogold nano-clusters by enzyme synthesis

A technology of fluorescent gold nanometers and gold nanoclusters, which is applied in metal processing equipment, chemical instruments and methods, luminescent materials, etc., can solve the problems of complex response process and high dosage of acetylcholinesterase, and achieve simple production steps and low concentration requirements , the effect of high fluorescence intensity

Active Publication Date: 2021-05-11
SOUTH CHINA UNIV OF TECH
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  • Summary
  • 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

Method used

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  • Preparation method of fluorogold nano-clusters by enzyme synthesis
  • Preparation method of fluorogold nano-clusters by enzyme synthesis
  • Preparation method of fluorogold nano-clusters by enzyme synthesis

Examples

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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 of gold nano-clusters by acetylcholinesterase synthesis. The method comprises the following steps: uniformly mixing acetylcholinesterase with chloroauric acid, adjusting the pH value to 8.0-14.0, and carrying out reacting for 8-24 hours at a certain temperature to obtain an acetylcholinesterase gold nano-cluster solution, wherein the concentration ratio of acetylcholinesterase to chloroauric acid in the solution is (1-20) mU: 1 mmol / L. The gold nano-clusters are synthesized by taking chloroauric acid and acetylcholinesterase as raw materials in one step, the prepared acetylcholinesterase gold nano-clusters emit blue light, the maximum excitation wavelength and emission wavelength are 315 nm and 398 nm respectively, and the application potential of simplifying the pollutant fluorescence detection response process participated by acetylcholinesterase is achieved; and the particle size distribution is between 0.8 nm and 2.6 nm, the average particle size is 1.6 nm, the small nanometer size realizes a unique electronic structure, the fluorescence intensity is high, and the fluorescence intensity and maximum emission peak position are kept stable within one week during storage at 4 DEG C.

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 Applications(China)
IPC IPC(8): B22F9/24B22F1/00C09K11/58
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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