CN-121975133-A - Preparation method and application of metal polyphenol-based nano-enzyme with multienzyme activity

CN121975133ACN 121975133 ACN121975133 ACN 121975133ACN-121975133-A

Abstract

The invention discloses a preparation method and application of metal polyphenol-based nano-enzyme with multienzyme activity. The invention prepares the metal polyphenol-based nano-enzyme with multi-enzyme activity by using the compound rich in phenolic hydroxyl as a ligand through a coordination self-assembly reaction, and realizes multifunctional integrated application. The phenolic hydroxyl groups in the compounds are helpful for the formation of hydrogen bonds and pi-pi stacking interactions, can stabilize the self-assembly process of a two-dimensional structure, and can enhance the affinity of nano-enzymes to catalytic substrates through ligand engineering. Meanwhile, the phenolic hydroxyl is taken as a strong coordination site to form a stable coordination bond with metal ions, and the specific coordination mode regulates and controls the electron cloud density and valence state of the metal center, thereby creating an electrophilic or nucleophilic microenvironment which is favorable for substrate combination, and further endowing the nano-enzyme with various enzyme-like activities. The metal polyphenol-based nano-enzyme can be widely applied to the fields of detection of related substances in foods, medicines and environment, removal of pollutants and diagnosis and treatment of diseases.

Inventors

JI SHUNLI
CHANG SHAN
Wan Shuoshuo
LUO CHENXI
TIAN MENG
XIA QIAN

Assignees

中国药科大学

Dates

Publication Date: 20260505
Application Date: 20260116

Claims (10)

1. The metal polyphenol-based nano-enzyme is characterized by being prepared by the following steps: Step 1, after ultrasonic treatment of the ligand, dissolving the ligand in ultrapure water, filtering the obtained solution to separate soluble components, and then diluting to obtain a solution A; and 2, uniformly mixing the solution A, the metal salt solution, the buffer solution and the ultrapure water, standing, and centrifugally collecting the precipitate to obtain the metal polyphenol-based nano-enzyme.
2. The metal polyphenol based nanoenzyme of claim 1, wherein said ligand is a phenolic hydroxyl group rich compound including, but not limited to, one or more of phloroglucinol, resorcinol, hydroquinone, gallic acid, pyrogallic acid, anthocyanidin, flavonols.
3. The metal polyphenol based nanoenzyme of claim 1, wherein said metal salt is a metal compound having a Fenton/Fenton-like reaction including, but not limited to, one or more of ferrous, ferric, cobalt, cadmium, manganese, silver, copper, nickel salts.
4. The metal polyphenol based nanoenzyme of claim 1, wherein said buffer is selected from one or more of Tris-HCl buffer, PBS buffer, MES buffer or HEPES buffer.
5. The metal polyphenol-based nano-enzyme according to claim 4, wherein the buffer solution has a pH of 6.5-10.5 and contains substances in an amount concentration of 5-150mmol/L.
6. The metal polyphenol based nanoenzyme of claim 1, wherein in step 2, the molar ratio of buffer, ligand and metal salt solution is 0.5-1.5:1-15:1-20.
7. The metal polyphenol based nanoenzyme of claim 1, wherein in step 2, the reaction time is 10s-30min and the reaction temperature is room temperature.
8. The metal polyphenol based nanoenzyme of claim 1, wherein the final system concentration of the ligand in step 1 is 5-200mmol/L.
9. The metal polyphenol based nanoenzyme of claim 1, wherein in step2, the concentration of the metal salt solution is 20-200mmol/L.
10. The use of the metal polyphenol-based nano-enzyme according to any one of claims 1 to 9 in the preparation of an enzyme-like preparation, wherein the enzyme-like preparation is an oxidase-like preparation, laccase-like preparation, superoxide dismutase-like preparation or catalase-like preparation, and the enzyme-like preparation is an antibacterial agent, an antitumor agent, a wastewater treatment agent, a tissue repair agent or a detection reagent.

Description

Preparation method and application of metal polyphenol-based nano-enzyme with multienzyme activity Technical Field The invention belongs to the technical field of chemical materials, and particularly relates to preparation and application of metal polyphenol-based nano-enzyme with multienzyme activity. Background After graphene is successfully mechanically peeled from graphite in 2004, the two-dimensional material brings about a great deal of attention to researchers by virtue of unique mechanical, electrical, photonics and magnetic properties compared with bulk lamellar materials. Meanwhile, nanoenzymes have been developed to have the potential to replace natural enzymes by virtue of their versatility and excellent catalytic efficiency in various biochemical reactions. The unique performance of the two-dimensional material is combined with the bionic activity of the nano enzyme, so that a hybrid system with better performance can be designed, and the hybrid system has wide application prospects in various fields of catalysis, sensing, electronic devices, environmental remediation and the like. The length, geometry, functional group and other properties of the organic ligand directly determine the crystal structure, topology, pore size and specific surface area of the metal organic nano enzyme. Meanwhile, the method plays a key role in natural enzyme simulation by constructing a binding pocket and providing an optimized catalytic microenvironment. Therefore, the reasonable selection of the ligand can obviously improve the actual application efficiency of the metal organic nano enzyme. The controllable preparation of existing metal-organic nanoenzymes, particularly two-dimensional lamellar structures, still faces significant challenges. The core is how to precisely limit crystal growth to the vertical (i.e., thickness) to the nanometer scale without affecting the other two lateral dimensions. When the metal organic nano enzyme is prepared by the current mainstream solvothermal method, the method needs to be carried out under the high-temperature condition, so that the method has the advantages of long reaction time, limited yield, high cost and difficult precise regulation and control of crystal growth. In enzyme activity design, the nano-enzyme is often limited by single enzyme activity, and introduction of multi-metal sites can try to expand catalytic functions, but inevitably increases preparation cost and complexity, and can possibly cause adsorption competition of different intermediates, thereby affecting catalytic efficiency. Therefore, the novel ligand which is rich in source, low in price and capable of realizing controllable assembly of a two-dimensional structure and integration of multi-enzyme activity through a simple and mild process is developed, and has important significance for promoting practical application of high-performance metal organic nano enzyme. In view of the above, the invention prepares the metal polyphenol-based nano-enzyme with multi-enzyme activity by simulating the Cu-O coordination environment of the natural enzyme and self-assembling coordination of the medicine rich in phenolic hydroxyl and metal ions at room temperature in one step, which is inspired by the active site and electron transfer path of the natural laccase and the self-assembling coordination of the peptide and copper, and has practical significance for promoting the application of the high-performance metal organic nano-enzyme in the fields of biological detection, environmental repair, disease diagnosis and treatment and the like. Disclosure of Invention The invention aims to provide a metal polyphenol-based nano-enzyme with multi-enzyme activity, which is prepared by the following steps: step 1, dissolving a ligand in ultrapure water after ultrasonic treatment, filtering the obtained solution to separate soluble components, and then diluting to obtain a solution A; and 2, uniformly mixing the solution A, the metal salt solution, the buffer solution and the ultrapure water, standing, and centrifugally collecting the precipitate to obtain the metal polyphenol-based nano-enzyme. Further, in step 1, the ligand is a compound rich in phenolic hydroxyl groups, including but not limited to one or more of phloroglucinol, resorcinol, hydroquinone, gallic acid, pyrogallic acid, anthocyanin, flavonols, and the like. Further, in step 2, the metal salt is a metal compound having a Fenton/Fenton-like reaction, including but not limited to one or more of ferrous salt, ferric salt, cobalt salt, cadmium salt, manganese salt, silver salt, copper salt, nickel salt, and the like. Further, in step 2, the buffer is selected from one or more of Tris-HCl buffer, PBS buffer, MES buffer and HEPES buffer. Further, in step 2, the pH of the buffer is 6.5-10.5, and the concentration of the contained substance is 5-150mmol/L. Further, in step 2, the molar ratio of buffer solution, ligand and metal salt solution is 0.5-1.5:1-15:1-20. Fur