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Preparation method and application of gold nanoparticle based nano-enzyme

A gold nanoparticle and nanozyme technology, which is applied in the preparation of artificial nanozymes and the field of artificial enzyme preparation, can solve the problems of bulkiness, hinder the binding of substrates and key amino acids, and the exposure of active sites is not obvious, and achieve stable secondary structure, excellent catalytic activity, and the effect of excellent catalytic activity

Active Publication Date: 2019-08-30
SHANGHAI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Or use a polymer material as the skeleton, but the space structure of the polymer material is complex and huge, and the exposure of the active site is not obvious, which will hinder the combination of the substrate and the key amino acid to a certain extent, thereby affecting the proton exchange between the key amino acids
Or design polypeptide polymers, relying on random combinations of key amino acids to build active pockets, without a specific conformation, and the catalytic triplets formed have low catalytic efficiency

Method used

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  • Preparation method and application of gold nanoparticle based nano-enzyme
  • Preparation method and application of gold nanoparticle based nano-enzyme
  • Preparation method and application of gold nanoparticle based nano-enzyme

Examples

Experimental program
Comparison scheme
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Embodiment 1

[0029] In the present embodiment, a method for preparing a nanozyme based on gold nanoparticles, the steps are as follows:

[0030] a. A polypeptide consisting of twelve amino acids is used, and the sequence of the polypeptide is NCLDALHSCLSY;

[0031] b. Add 40 μL of sodium citrate solution with a concentration of 0.2M to 24 mL of citrate-coated gold nanoparticles solution, and mix well to obtain gold nanoparticles solution; then add 4 mL of alkaline peptide solution containing NaOH with a concentration of 1.6 mM dropwise Add it into the nano-gold solution that is being stirred, and at room temperature, the control speed is 800r / min to stir the mixed solution, and perform a preliminary reaction for 1 hour; then place the preliminary product solution in a shaker, and at room temperature, control The rotating speed is 100r / min to stir the mixed solution, and continue to react for 12 hours; when the reaction is completed, it is concentrated by ultrafiltration to obtain a concent...

Embodiment 2

[0040] This embodiment is basically the same as Embodiment 1, especially in that:

[0041] In the present embodiment, a method for preparing a nanozyme based on gold nanoparticles, the steps are as follows:

[0042] a. A polypeptide consisting of twelve amino acids is used, and the sequence of the polypeptide is NCLDALHSCLAY;

[0043] b. Add 40 μL of sodium citrate solution with a concentration of 0.2M to 24 mL of citrate-coated gold nanoparticles solution, and mix well to obtain gold nanoparticles solution; then add 4 mL of alkaline peptide solution containing NaOH with a concentration of 1.6 mM dropwise Add it into the nano-gold solution that is being stirred, and at room temperature, the control speed is 800r / min to stir the mixed solution, and perform a preliminary reaction for 1 hour; then place the preliminary product solution in a shaker, and at room temperature, control The rotating speed is 100r / min to stir the mixed solution, and continue to react for 12 hours; when...

Embodiment 3

[0046] This embodiment is basically the same as the previous embodiment, and the special features are:

[0047] In the present embodiment, a method for preparing a nanozyme based on gold nanoparticles, the steps are as follows:

[0048] a. A polypeptide consisting of twelve amino acids is used, and the sequence of the polypeptide is NCLDALHACLSY;

[0049] b. Add 40 μL of sodium citrate solution with a concentration of 0.2M to 24 mL of citrate-coated gold nanoparticles solution, and mix well to obtain gold nanoparticles solution; then add 4 mL of alkaline peptide solution containing NaOH with a concentration of 1.6 mM dropwise Add it into the nano-gold solution that is being stirred, and at room temperature, the control speed is 800r / min to stir the mixed solution, and perform a preliminary reaction for 1 hour; then place the preliminary product solution in a shaker, and at room temperature, control The rotating speed is 100r / min to stir the mixed solution, and continue to rea...

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Abstract

The invention discloses a preparation method and application of a gold nanoparticle based nano-enzyme. According to the preparation method, a polypeptide is modified to the surface of a gold nanoparticle through an Au-S bond and is induced to represent an alpha-helical structure on the gold nanoparticle with trifluoroethanol, so that a triplet is catalyzed to be positioned in a corresponding spatial position to form a charge relay network, and advanced simulation of a natural hydrolase from the molecular level. The nano-enzyme has excellent performance of difficult degeneration due to the stability of the Au-S bond; after the trifluoroethanol is removed, the nano-enzyme still as a stable secondary structure and excellent catalytic activity. The nano-enzyme has a simple synthesizing methodand high catalyzing efficiency. The method selects the gold nanoparticle as a rigid frame and can greatly improve the stability of the nano-enzyme, and the surface of the nano-enzyme is formed by conformation adjustable polypeptides and has a flexible dynamic secondary structure. The conformation adjustment of the nano-enzyme enables a group without a function to have a function and is an advancedsimulation of natural enzyme.

Description

technical field [0001] The present invention relates to a preparation method and application of an artificial enzyme, in particular to a preparation method and application of an artificial nano-enzyme simulating the structure of a natural enzyme, which is applied to inorganic metal-organic hybrid nanomaterials and biochemical and enzyme catalysts technology field. Background technique [0002] Natural enzymes are a kind of biocatalysts produced by living organisms in the process of continuous evolution. They have significant advantages such as high catalytic efficiency, strong reaction selectivity, and mild reaction conditions. However, natural enzymes can only be catalyzed in a mild water environment, have poor stability, and their sources are limited, making extraction difficult. [0003] Mimicking the structure of natural enzymes to prepare artificial enzymes and reproduce their natural activities is a major strategy for the design of artificial enzymes. The preparation...

Claims

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Application Information

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IPC IPC(8): C12N11/14
CPCC12N11/14
Inventor 曹傲能曹奕王海芳
Owner SHANGHAI UNIV
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