Application of nano-copper selenide as a peroxide mimetic enzyme in the detection of hydrogen peroxide and l-cysteine

A peroxide and cysteine ​​technology, applied in the field of nano-biochemical detection, can solve the problems of high storage and use costs, expensive natural enzymes, damaged enzyme molecules, etc., to meet the requirements of production and application, high stability, The effect of simple process

Active Publication Date: 2022-03-29
QINGDAO UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the catalysis of natural enzymes requires strict environmental conditions. Any factors that can denature proteins or destroy their structures, such as strong acid, strong alkali, high temperature, etc., can damage enzyme molecules and make them inactive, and natural enzymes are expensive. Purification is difficult, storage and use costs are high, so the study of mimic enzymes is of great significance

Method used

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  • Application of nano-copper selenide as a peroxide mimetic enzyme in the detection of hydrogen peroxide and l-cysteine
  • Application of nano-copper selenide as a peroxide mimetic enzyme in the detection of hydrogen peroxide and l-cysteine
  • Application of nano-copper selenide as a peroxide mimetic enzyme in the detection of hydrogen peroxide and l-cysteine

Examples

Experimental program
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Effect test

Embodiment 1

[0030] Synthesis of nano-copper selenide.

[0031] Mix 0.01mol selenium powder and 10mL NaOH (13mol / L) solution, and stir until the selenium powder is completely dissolved. 10mL0.05mol / LCuSO 4 Mix 10mL of 0.05mol / L EDTA solution, stir, then add selenium-NaOH solution and stir evenly, then use NaOH solution to adjust the pH of the mixed reactant to 12, then put the mixture solution into a 100mL stainless steel autoclave, 180 ℃ hydrothermal reaction for 36h. When the reaction mixture was cooled to room temperature, the precipitate was centrifuged and washed several times with absolute ethanol. Then the sample was vacuum-dried at 60° C. for 6 hours to obtain nano-copper selenide.

[0032] figure 1 The scanning electron micrograph of the nano-copper selenide prepared for the present invention.

[0033] figure 2 The XRD characterization figure of the nano-copper selenide prepared for the present invention.

Embodiment 2

[0035] Nano-copper selenide was used as peroxide mimetic enzyme to detect hydrogen peroxide content.

[0036] In a series of 5mL colorimetric tubes, add 0.2mL 1mmol / L TMB solution, 0.2mL 1mg / L CuSe solution, and different amounts of hydrogen peroxide solution in sequence, and mix well. Dilute to volume with 0.1 mol / L phosphate buffer solution (pH=4.5). Measure the absorbance with a UV-visible spectrophotometer, draw the absorbance-hydrogen peroxide concentration standard curve, and deduce H from the absorbance value of the solution. 2 o 2 concentration.

[0037] image 3 This is the standard curve for detecting hydrogen peroxide with TMB as the chromogenic agent and nano-copper selenide.

Embodiment 3

[0039] Nano-copper selenide is used as a peroxide mimetic enzyme to detect the content of L-cysteine.

[0040] Mix 200 μL TMB (1 mM), 200 μL CuSe turbid solution, 200 μL H 2 o 2 Added to 1400 μL of PBS buffer and then measured the absorbance with a UV-Vis spectrophotometer. Add different concentrations of L-cysteine ​​into the above CuSe reaction system, react for 5 minutes, and measure the absorbance of the solution. The difference between the absorbance of the solution without L-cysteine ​​and the absorbance of the solutions containing L-cysteine ​​with different concentrations is the ordinate, and the concentration of the L-cysteine ​​solution is the abscissa to draw a standard curve, The concentration of L-cysteine ​​can be extrapolated from the standard curve.

[0041] Figure 4 In order to use TMB as the chromogen, in the presence of hydrogen peroxide, the standard curve of the detection of L-cysteine ​​by nano-copper selenide.

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Abstract

The invention discloses an application of nano copper selenide (CuSe) as a peroxide mimetic enzyme for measuring hydrogen peroxide and L-cysteine. Nano-copper selenide acts as a peroxide mimetic enzyme and has peroxidase activity. The present invention finds that nano-copper selenide has the function of peroxide imitating enzyme for the first time, and can be used to detect the content of hydrogen peroxide; 2 o 2 The catalytic oxidation reaction can be used to detect the concentration of L-cysteine; the nano-copper selenide prepared by the present invention has high stability, reusability, and efficient peroxide mimetic enzyme catalysis as a catalyst compared with HRP Activity; the present invention finds that in the absence of hydrogen peroxide, nano-copper selenide can also catalyze the chromogenic agent and be used as a simulated enzyme; the preparation method in the present invention is simple in process and strong in operability, and can further meet production and application requirements.

Description

technical field [0001] The invention relates to an application of nanometer copper selenide (CuSe) as a peroxide mimetic enzyme for measuring hydrogen peroxide and L-cysteine. The invention belongs to the field of nanometer biochemical detection. Background technique [0002] Biological enzymes are high-efficiency catalysts with high catalytic efficiency, strong specificity and mild reaction conditions. However, the catalysis of natural enzymes requires strict environmental conditions. Any factors that can denature proteins or destroy their structures, such as strong acid, strong alkali, high temperature, etc., can damage enzyme molecules and make them inactive, and natural enzymes are expensive. Purification is difficult, and the cost of storage and use is high, so the study of mimic enzymes is of great significance. Since the first discovery of magnetic ferric oxide nanoparticles (Fe 3 o 4 NPs) have the catalytic activity of peroxidase, that is, Fe 3 o 4 NPs can cata...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/78G01N21/31
CPCG01N21/78G01N21/31G01N2021/775
Inventor 万均白卯娟徐成成王卫
Owner QINGDAO UNIV OF SCI & TECH
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