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Method for preparing monatomic copper artificial mimic peroxidase

A peroxidase and artificial simulation technology, which is applied in the field of preparing single-atom copper artificially simulated peroxidase, can solve the difficulty of functionalization of artificial simulated peroxidase, is not suitable for large-scale industrial synthesis, and restricts mass production and application. and other problems, to achieve the effect of reducing domestic pollution, inhibiting secondary pollution, and overcoming the large amount of metal consumption

Active Publication Date: 2020-02-18
CHINA UNIV OF GEOSCIENCES (BEIJING)
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Problems solved by technology

[0007] (1) The macromolecules of synthetic artificial enzymes are expensive, the structure is not stable enough and the storage conditions are harsh, so they are not suitable for large-scale industrial synthesis;
[0008] (2) The catalytic activity of artificial simulated peroxidase is still significantly lower than that of biological enzymes, and cannot completely replace the position of biological enzymes;
[0009] (3) It is difficult to artificially simulate the functionalization of peroxidase, and it is easy to cause pollution during production, which restricts its mass production and application
[0010] The above defects make it difficult to obtain peroxide mimetic enzymes with stable properties, low price, green environmental protection and comparable catalytic activity to natural enzymes with the existing technology, which limits its further application in the biological field

Method used

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  • Method for preparing monatomic copper artificial mimic peroxidase
  • Method for preparing monatomic copper artificial mimic peroxidase

Examples

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no. 1 example

[0034] Collect 10L of fresh human urine, let it stand for 10 hours, add 2kg of basic anion exchange resin, stir and adsorb the amino acid ligand compound in the urine, then desorb the amino acid ligand compound with high concentration salt solution, and use Concentration method to increase the concentration of amino acid ligands in the aqueous solution, add copper nitrate solution with a volume ratio of 0.5% and a concentration of 10 g / L to the ligands, stir for 24 hours, and let it stand for 2 to 3 hours to precipitate, and obtain a blue-green solid precipitate , discard the supernatant, add an equal volume of glacial acetic acid to the precipitate, stir and heat to boiling, then add a 40 g / L aqueous sodium hydroxide solution that accounts for 20% of the volume of glacial acetic acid, and continue stirring for 5 minutes to obtain a blue-black precipitate. The blue-black precipitate obtained by the reaction was centrifuged, washed three times with absolute ethanol, and dried un...

no. 2 example

[0036] Add 5 mL of sodium dihydrogen phosphate-disodium hydrogen phosphate buffer (pH 4.5) with a concentration of 20 mmol / L into the container, then add 0.5 mL of hydrogen peroxide and 0.2 mL of 3,3',5, with a concentration of 20 mmol / L 5'-Tetramethylbenzidine hydrochloride, finally add 0.1mg monoatomic copper artificial simulated peroxidase, stir vigorously, mix evenly and react at room temperature for 10min, measure the absorbance at 652nm wavelength. When the concentration of hydrogen peroxide is in the range of 0.02-1.0 mmol / L, the absorbance of the chromogenic product is linearly correlated with the concentration of hydrogen peroxide, and the conversion rate of hydrogen peroxide of different concentrations is greater than 95%. figure 2 The absorbance at 652nm corresponding to the hydrogen peroxide reaction solution of different concentrations catalyzed by peroxidase is artificially simulated for single atom copper, which proves that the conversion efficiency of hydrogen ...

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Abstract

The invention discloses a method for preparing monatomic copper artificial mimic peroxidase. According to the preparation of the artificial mimic peroxidase, human urine which is cheap, easy to obtainand rich in resource is directly used as a ligand source, and a monatomic catalyst is used as a mimic enzyme active center, so that the use amount of copper can be reduced, and meanwhile, the functional activity of the artificial peroxidase is improved. The monatomic copper artificial mimic peroxidase has excellent peroxidase activity, can catalyze hydrogen peroxide to efficiently oxidize a substrate 3,3',5,5'-tetramethylbenzidine hydrochloride, and shows the activity characteristic similar to peroxidase; the preparation method has the advantages of simpleness, wide source and low cost, provides the human waste treatment effect to a certain extent, and has high industrial application and environmental application potentials.

Description

technical field [0001] The invention relates to the technical fields of coordination chemistry and enzyme catalysis, in particular to a method for preparing single-atom copper artificial simulated peroxidase. Background technique [0002] Enzymes are a class of proteins that can perform biological catalysis. In a biological reaction system, all metabolic molecules can be considered as substrates. Enzymes are involved in almost all intracellular reactions to improve the working efficiency of biological systems. The essence of enzyme-catalyzed biological reactions is to lower the activation energy barrier of chemical reactions. Generally speaking, enzymes can increase the reaction rate by up to several million times without any change before and after the reaction, and the chemical balance of the reaction is not affected. Due to the specificity, specificity and high efficiency of enzyme catalysis, its mass production has extremely high industrial value. [0003] Natural en...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B01J31/22B01J35/00
CPCB01J31/003B01J31/2217B01J2531/16B01J35/391
Inventor 代云容段存戌王思宇朱繁刘富明
Owner CHINA UNIV OF GEOSCIENCES (BEIJING)
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