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A kind of preparation method and application of biomimetic laccase based on dipeptide

A biomimetic lacquer and laccase technology, applied in the field of materials, can solve the problems of poor stability, low catalytic efficiency, and difficult preparation, and achieve the effects of good stability, high catalytic efficiency, and non-toxic metabolic mechanism.

Active Publication Date: 2021-03-26
INST OF CHEM CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Aiming at the problems that the existing bionic enzyme (bionic laccase) is difficult to prepare, has low catalytic efficiency and poor stability, the present invention provides a dipeptide-based bionic laccase and its preparation method and application

Method used

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  • A kind of preparation method and application of biomimetic laccase based on dipeptide
  • A kind of preparation method and application of biomimetic laccase based on dipeptide
  • A kind of preparation method and application of biomimetic laccase based on dipeptide

Examples

Experimental program
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Effect test

Embodiment 1

[0073] Example 1. Preparation of cationic dipeptide-glutaraldehyde-copper assembly biomimetic laccase

[0074] (1) Weigh 10 mg of lyophilized powder of cationic phenylalanine dipeptide (CDP), dissolve CDP with 1,1,1,3,3,3-hexafluoroisopropanol (referred to as HFIP) as solvent, and ultrasonically oscillate 5min, so that the powder is completely dissolved, the CDP concentration is 10mM;

[0075] (2) Use a pipette gun to draw a certain amount of CDP solution and add it to a glutaraldehyde (GA) aqueous solution with a concentration of 5 mM, wherein the volume ratio of CDP solution to GA solution is 1:10, and after shaking, incubate in the dark for 20 hours. Obtain a light yellow suspension, which is the CDP-GA nanosuspension, centrifuge and wash 3 times, add water to disperse, and obtain an aqueous dispersion of nanoparticles (the concentration of the dipeptide-crosslinking agent assembly is 2 mM);

[0076] (3) Add the 20mM copper sulfate aqueous solution to the CDP-GA nanopartic...

Embodiment 2

[0088] Example 2. Preparation of cationic dipeptide-glutaraldehyde-copper assembly biomimetic laccase

[0089] (1) Weigh 10 mg of lyophilized powder of cationic phenylalanine dipeptide (CDP), dissolve CDP with 1,1,1,3,3,3-hexafluoroisopropanol (HFIP for short), and ultrasonically shake 5min, the powder is completely dissolved, and the CDP concentration is 20mM;

[0090] (2) Use a pipette gun to absorb a certain amount of CDP solvent and add it to a 5mM glutaraldehyde (GA) aqueous solution, wherein the volume ratio of the CDP solution to the GA solution is 1:20. After shaking and incubating in the dark for 30 hours, the obtained The light yellow suspension, which is the CDP-GA nanosuspension, was centrifuged and washed 3 times to obtain an aqueous dispersion of nanoparticles, and the concentration of the dispersion was 4 mM;

[0091] (3) Add the copper sulfate aqueous solution that concentration is 50mM in step (2) gained CDP-GA nanoparticle dispersion liquid, make CDP and GA ...

Embodiment 3

[0097] Example 3. Preparation of cationic dipeptide-glutaraldehyde-copper assembly biomimetic laccase (directly mix three components to prepare biomimetic laccase)

[0098] (1) Weigh 10 mg of lyophilized powder of cationic phenylalanine dipeptide (CDP), dissolve CDP with 1,1,1,3,3,3-hexafluoroisopropanol (HFIP for short), and ultrasonically shake 5min, so that the powder is completely dissolved, the CDP concentration is 10mM;

[0099] (2) Dilute the glutaraldehyde (GA) aqueous solution with an initial concentration of 0.1M, and the dilution factor is 20 times to obtain a GA aqueous solution with a concentration of 5mM;

[0100](3) Weigh 0.2 g of copper sulfate pentahydrate powder, dissolve it in deionized water, and configure it into an aqueous solution of copper sulfate with a concentration of 40 mM;

[0101] (4) Mix the solution configured in steps (1), (2), and (3) with a pipette gun, and the volume ratio of the CDP solution, GA solution and copper sulfate solution is 1:10...

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Abstract

The invention relates to a preparation method and an application for bionic laccase based on dipeptide. The bionic laccase based on the dipeptide takes metal ions as a catalytic active center and dipeptide as an organic framework, and has laccase-like activity; and the bionic laccase based on the dipeptide is a dipeptide-crosslinking agent-metal compound. The bionic laccase prepared by using the method provided by the invention has the following advantages: the preparation process is simple, and the preparation conditions are mild; the prepared bionic laccase has the catalytic activity of simulating natural laccase; the catalytic efficiency of the bionic laccase is far higher than the catalytic efficiency of natural laccase; and the bionic laccase has high stability and good biocompatibility. The preparation method for the series of bionic laccase based on the dipeptide has potential application in development of industrial applications of wastewater treatment, textile bleaching, foodsafety and the like.

Description

technical field [0001] The invention belongs to the field of materials, and in particular relates to a preparation method and application of a dipeptide-based biomimetic laccase. Background technique [0002] Biomimetic enzymes are defined as artificially constructed materials with enzyme functions. The construction of biomimetic enzymes based on peptide molecules has attracted widespread attention in basic research and practical applications. Compared with natural enzymes, biomimetic enzymes have the advantages of higher catalytic activity and higher stability, while compared with biomimetic enzymes constructed of metal oxides or carbon-based materials, peptidyl biomimetic enzymes have good biocompatibility And the advantages of mild preparation conditions, it has a wide range of applications in biosensing, immunoassay, cancer diagnosis and treatment, etc. [0003] Peptidyl covalently assembled nanozymes can achieve high affinity and strong stability for substrates on the ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): B01J31/04C07C46/06C07C50/04
CPCB01J31/04B01J2231/70C07C46/06C07C50/04Y02P20/55
Inventor 李峻柏王晨蕾费进波
Owner INST OF CHEM CHINESE ACAD OF SCI