A method for regulating the type of nanoszyme mimetic enzyme

Abstract

The application discloses a method for regulating the type of nano-enzyme mimetic enzyme and belongs to the field of nano-catalysis. The method comprises the following steps: calcining precursor materials containing at least one metal element and at least one element capable of interacting with the metal element by controlling the calcination temperature, and preparing nano-enzymes which can simulate different enzyme catalytic activities. The method can obtain nano-enzymes which can simulate different activities by controlling specific calcination temperatures, and can regulate the type of the prepared nano-enzymes which can simulate catalytic enzymes.

Description

Technical Field

[0001] This invention belongs to the field of nanocatalysis and provides a method for regulating the type of nanozyme to mimic enzymes. Background Technology

[0002] Natural enzymes can efficiently and specifically catalyze various biochemical reactions under mild conditions, and have important applications in medicine, agriculture, chemical industry, and food processing. However, their poor stability, high cost, and difficulty in preparation and purification greatly limit their practical applications.

[0003] Nanozymes are a class of nanomaterials with enzyme-like catalytic activity. They overcome the inherent shortcomings of natural enzymes and possess advantages such as simple structure, stable properties, and easy preparation processes. They show promising applications in the treatment of various diseases, including cancer, inflammatory diseases, and cardiovascular diseases. Despite the development of many high-performance nanozymes, their practical application still faces numerous challenges.

[0004] Nanozymes typically possess multiple catalytic active sites, often exhibiting two or more enzyme-like activities simultaneously. For example, iron oxide nanozymes possess peroxidase-like and catalase-like activities. Cerium oxide nanozymes exhibit oxidase-like, peroxidase-like, catalase-like, and superoxide dismutase-like activities. Nanozymes composed of cobalt oxides have been reported to possess peroxidase-like, oxidase-like, catalase-like, and superoxide dismutase-like activities. These enzyme activities can sometimes interfere with each other. Taking the most prevalent mimicry enzyme types in the current nanozyme field, peroxidase (POD) and oxidase (OXD), as examples, these two catalytic types often coexist within the same nanozyme. Oxidase catalyzes reactions involving oxygen, while OXD catalyzes reactions involving peroxides, represented by hydrogen peroxide. Both enzyme reactions occur in acidic environments, leading to competition and interference between them, which is detrimental to the practical application of nanozymes. Therefore, developing suitable methods to regulate the types of catalytic enzymes mimicked by nanozymes is of great significance. Summary of the Invention

[0005] I. Purpose of this invention:

[0006] The purpose of this invention is to provide a simple and highly controllable method for regulating the type of nanozyme catalytic reaction. By controlling the specific calcination temperature, nanozymes that can simulate different activities can be obtained, and the type of catalytic enzyme simulated by the prepared nanozymes can be regulated.

[0007] II. The technical solution adopted by the present invention to achieve the above objectives:

[0008] A method for regulating nanozyme mimicking enzyme types includes the following steps:

[0009] By controlling the calcination temperature T, precursor materials containing at least one metal element and at least one element capable of interacting with the metal element are calcined to prepare nanozymes that mimic the catalytic activities of different enzymes.

[0010] Furthermore, the precursor material contains metallic elements such as precious metals and transition metals.

[0011] Furthermore, the precursor material may contain non-metallic elements such as nitrogen, sulfur, and boron, or other different metallic elements.

[0012] Furthermore, the precursor material may also contain other functional materials such as the substrate material, for example, carbon materials.

[0013] Furthermore, by controlling the calcination temperature T within the range of 150–1000 °C, nanozymes simulating the activities of different catalytic enzymes were prepared.

[0014] Furthermore, by controlling the calcination temperature T within the range of 150–1000 °C, nanozymes with simulated POD and / or OXD activities were prepared.

[0015] Furthermore, the 150–1000°C range is divided into two calcination temperature ranges, with a dividing point of 350°C or 400°C. By controlling the calcination temperature T within the low calcination temperature range, nanozymes with a single POD-like activity are obtained; by controlling the calcination temperature T within the high calcination temperature range, nanozymes with both POD-like and OXD-like activities or a single OXD-like activity are obtained.

[0016] Furthermore, the precursor materials include cobalt-containing compounds, nitrogen-containing compounds, and graphylene as the base material. By controlling the calcination temperature to 150℃≤T≤400℃, nanozymes with single POD-like activity are obtained; at 400℃<T<650℃, nanozymes with both POD-like and OXD-like activities are obtained; and at 650℃≤T≤1000℃, nanozymes with single OXD-like activity are obtained.

[0017] Furthermore, the nitrogen-containing compound is melamine or cyanuric chloride.

[0018] Furthermore, the precursor materials include cobalt-containing compounds, melamine, and graphene as a substrate material. By controlling the calcination temperature to 150℃≤T≤350℃, nanozymes with single POD-like activity are obtained; and nanozymes with both POD-like and OXD-like activities are obtained at 350℃<T≤1000℃.

[0019] Furthermore, the precursor materials include manganese-containing compounds, melamine, and carbon materials as the base material. By controlling the calcination temperature to 150℃≤T≤350℃, nanozymes with single POD-like activity are obtained; and nanozymes with both POD-like and OXD-like activities are obtained at 350℃<T≤1000℃.

[0020] Furthermore, the carbon material is graphene or graphyne.

[0021] Furthermore, the precursor materials include cobalt-containing compounds and nitrogen carbide. By controlling the calcination temperature to 150℃≤T≤350℃, nanozymes with single POD-like activity are obtained; and nanozymes with both POD-like and OXD-like activities are obtained at 350℃<T≤1000℃.

[0022] III. Beneficial effects achieved by the present invention:

[0023] This invention proposes a simple and highly controllable method for regulating the catalytic reaction type of nanozymes, which enables the controllable transformation of the catalytic reaction type of nanozymes. For example, nanozymes can exhibit single POD-like activity, single OXD-like activity, or both POD-like and OXD-like activities. Nanozymes with different enzyme activities can be prepared by controlling the calcination temperature without changing the precursor materials, thereby meeting the different needs of nanozymes in practical applications. Attached Figure Description

[0024] Figure 1 The nanozymes prepared in Example 1 simulate the spectral curves of different types of enzyme catalysis;

[0025] Figure: (a) POD-like activity of nanozymes at different calcination temperatures; (b) OXD-like activity of nanozymes at different calcination temperatures.

[0026] Figure 2 The nanozymes prepared in Example 2 simulate the spectral curves of different enzyme catalytic types;

[0027] In the figure: (a) Nanozyme activity at calcination temperature of 350℃; (b) Nanozyme activity at calcination temperature of 900℃.

[0028] Figure 3 These are photographs of colorimetric reactions catalyzed by different types of enzymes, using nanozymes prepared in Example 3.

[0029] In the figure: (a) calcination temperature of 350℃; (b) nanozyme calcined at 650℃.

[0030] Figure 4 The nanozymes prepared in Example 4 simulate the spectral curves of different types of enzyme catalysis;

[0031] Figure: (a) POD-like activity of nanozymes at different calcination temperatures; (b) OXD-like activity of nanozymes at different calcination temperatures.

[0032] Figure 5 The nanozymes prepared in Example 5 simulate the spectral curves of different types of enzyme catalysis;

[0033] In the figure: (a) calcination temperature is 350℃; (b) calcination temperature is 650℃.

[0034] Figure 6 The nanozymes prepared in Example 6 simulate the spectral curves of different types of enzyme catalysis;

[0035] In the figure: (a) calcination temperature is 350℃; (b) calcination temperature is 650℃. Detailed Implementation

[0036] To make the various technical features, advantages, or effects of the present invention more apparent and understandable, a detailed description is provided below in conjunction with the accompanying drawings.

[0037] Example 1:

[0038] 1. Sample preparation: The precursor materials selected were cobalt chloride hexahydrate, melamine and graphyne. After all the precursor materials were mixed evenly, they were placed in a tube furnace and calcined at 350℃, 450℃, 500℃, 550℃, 650℃ and 900℃ for 2 hours in an inert atmosphere. After washing and drying, the sample was obtained.

[0039] 2. Enzyme-like Catalytic Performance Study: The enzyme-like catalytic performance of the samples was investigated using the chromogenic substrate TMB as a representative. In the OXD-like activity study, the samples were added to an acidic buffer solution containing TMB, and the signal of the characteristic UV-Vis absorption peak (approximately 652 nm) of TMB oxide was detected. In the POD-like activity study, the samples were added to an acidic buffer solution containing TMB and hydrogen peroxide (H₂O₂), and the signal of the characteristic UV-Vis absorption peak was detected. Experimental results are shown below. Figure 1 As shown in the figure, the sample calcined at 350℃ can only catalyze the oxidation of TMB in the presence of H2O2, indicating a single POD-like activity. Samples calcined at 350-650℃ showed a characteristic peak of 652nm in both the presence and absence of H2O2, indicating both POD-like and OXD-like activities; the calcination temperature can regulate the strength of these two enzyme-like activities. The sample calcined at 900℃ showed a characteristic peak of 652nm in the absence of H2O2, indicating a single OXD-like activity.

[0040] Example 2:

[0041] 1. Sample preparation: The precursor materials selected were cobalt chloride hexahydrate, cyanuric chloride and graphyne. After all the precursor materials were mixed evenly, they were placed in a tube furnace and calcined at 350℃ and 900℃ in an inert atmosphere for 2 hours. After washing and drying, the sample was obtained.

[0042] 2. Enzyme-like Catalytic Performance Study: The enzyme-like catalytic performance of the samples was investigated using the chromogenic substrate TMB as a representative. In the OXD-like activity study, the samples were added to an acidic buffer solution containing TMB, and the signal of the characteristic UV-Vis absorption peak (approximately 652 nm) of TMB oxide was detected. In the POD-like activity study, the samples were added to an acidic buffer solution containing TMB and hydrogen peroxide (H₂O₂), and the signal of the characteristic UV-Vis absorption peak was detected. Experimental results are shown below. Figure 2 ,from Figure 2 As shown in Figure (a), the sample calcined at 350℃ can only catalytically oxidize TMB in the presence of H2O2, indicating that it possesses a single POD-like activity. From Figure 2 As can be seen in Figure (b), the sample calcined at 900℃ can catalytically oxidize TMB and produce color in the absence of H2O2, but does not produce color in the presence of H2O2, indicating that it has a single OXD-like activity.

[0043] Example 3:

[0044] 1. Sample preparation: The precursor materials selected were cobalt chloride hexahydrate and carbon nitride C3N3. After all the precursor materials were mixed evenly, they were placed in a tube furnace and calcined at 350℃ and 650℃ in an inert atmosphere for 2 hours. After washing and drying, the sample was obtained.

[0045] 2. Enzyme-like Catalytic Performance Study: The enzyme-like catalytic performance of the samples was investigated using the chromogenic substrate TMB as a representative example. In the OXD-like activity study, the samples were added to an acidic buffer solution containing TMB, and the characteristic color of the catalytic reaction product (i.e., TMB oxide) was observed visually. In the POD-like activity study, the samples were added to an acidic buffer solution containing TMB and hydrogen peroxide (H2O2), and the characteristic color of the catalytic reaction product was observed visually. Experimental results are shown below. Figure 3 , Figure 3 (a) is a sample calcined at 350℃. It can be seen that TMB can only be catalyzed to produce color in the presence of H2O2, indicating that it has a single POD-like activity. Figure 3 (b) is the sample calcined at 650℃. It can be seen that it can catalyze the oxidation of TMB to produce color in the absence of H2O2, while the color is darker in H2O2, indicating that the sample has OXD-like and POD-like activities.

[0046] Example 4:

[0047] 1. Sample preparation: The precursor materials selected were cobalt chloride hexahydrate, melamine and graphene. After all the precursor materials were mixed evenly, they were placed in a tube furnace and calcined at 350℃, 650℃ and 900℃ for 2 hours in an inert atmosphere. After washing and drying, the samples were obtained.

[0048] 2. Enzyme-like Catalytic Performance Study: The enzyme-like catalytic performance of the samples was investigated using the chromogenic substrate TMB as a representative. In the OXD-like activity study, the samples were added to an acidic buffer solution containing TMB, and the signal of the characteristic UV-Vis absorption peak (approximately 652 nm) of TMB oxide was detected. In the POD-like activity study, the samples were added to an acidic buffer solution containing TMB and hydrogen peroxide (H₂O₂), and the signal of the characteristic UV-Vis absorption peak was detected. Experimental results are shown below. Figure 4 As can be seen from the figure, the sample calcined at 350℃ can only catalyze the oxidation of TMB to produce a characteristic peak of 652nm in the presence of H2O2, indicating that the nanozyme prepared under this condition mainly exhibits POD-like activity. The samples calcined at 650℃ and 900℃ showed OXD-like and POD-like activities, with the 650℃ sample exhibiting stronger POD-like activity.

[0049] Example 5:

[0050] 1. Sample preparation: The precursor materials selected were manganese chloride tetrahydrate, melamine and graphyne. After all the precursor materials were mixed evenly, they were placed in a tube furnace and calcined at 350℃ and 650℃ in an inert atmosphere for 2 hours. After washing and drying, the sample was obtained.

[0051] 2. Enzyme-like Catalytic Performance Study: The enzyme-like catalytic performance of the samples was investigated using the chromogenic substrate TMB as a representative. In the OXD-like activity study, the samples were added to an acidic buffer solution containing TMB, and the signal of the characteristic UV-Vis absorption peak (approximately 652 nm) of TMB oxide was detected. In the POD-like activity study, the samples were added to an acidic buffer solution containing TMB and hydrogen peroxide (H₂O₂), and the signal of the characteristic UV-Vis absorption peak was detected. Experimental results are shown below. Figure 5 As can be seen from the figure, the sample calcined at 350℃ can only catalyze the oxidation of TMB in the presence of H2O2, indicating that it has a single POD-like activity. The sample calcined at 650℃ showed a characteristic peak of 652nm in both the presence and absence of H2O2, indicating that it has both POD-like and OXD-like activities.

[0052] Example 6:

[0053] 1. Sample preparation: The precursor materials selected were manganese chloride hexahydrate, melamine and graphene. After all the precursor materials were mixed evenly, they were placed in a tube furnace and calcined at 350℃ and 650℃ in an inert atmosphere for 2 hours. After washing and drying, the sample was obtained.

[0054] 2. Enzyme-like Catalytic Performance Study: The enzyme-like catalytic performance of the samples was investigated using the chromogenic substrate TMB as a representative. In the OXD-like activity study, the samples were added to an acidic buffer solution containing TMB, and the signal of the characteristic UV-Vis absorption peak (approximately 652 nm) of TMB oxide was detected. In the POD-like activity study, the samples were added to an acidic buffer solution containing TMB and hydrogen peroxide (H₂O₂), and the signal of the characteristic UV-Vis absorption peak was detected. Experimental results are shown below. Figure 6 As can be seen from the figure, the sample calcined at 350℃ did not show a significant characteristic absorption peak at 652nm regardless of the presence of H2O2, indicating that the sample at this temperature basically does not have POD-like and OXD-like activities; the sample calcined at 650℃ showed a characteristic peak at 652nm in both the presence and absence of H2O2, indicating that it has POD-like and OXD-like activities.

[0055] Although the present invention has been disclosed above with reference to embodiments, it is not intended to limit the present invention. Appropriate modifications or equivalent substitutions made by those skilled in the art to the technical solutions of the present invention should be covered within the protection scope of the present invention, which is defined by the claims.

Claims

1. A method for regulating the type of nanozyme to mimic enzymes, characterized in that, Includes the following steps: By controlling the calcination temperature T within the range of 150~1000ºC, precursor materials containing at least one metal element and at least one element capable of interacting with the metal element are calcined to regulate the catalytic activity of nanozymes, mimicking the activity of different enzymes. Specifically, the 150~1000ºC range is divided into two calcination temperature ranges, with the dividing point being 350ºC or 400ºC. By controlling the calcination temperature T within the low calcination temperature range, nanozymes with a single POD-like activity are obtained. By controlling the calcination temperature T within the high calcination temperature range, nanozymes with both POD-like and OXD-like activities, or a single OXD-like activity, are obtained.

2. The method for regulating the type of nanozyme mimicking enzyme as described in claim 1, characterized in that, The precursor material contains at least one of noble metals and transition metals; the precursor material contains elements that can interact with metals, which are non-metallic elements or other different kinds of metallic elements, including at least one of nitrogen, sulfur, and boron.

3. The method for regulating the type of nanozyme mimicking enzyme as described in claim 2, characterized in that, The precursor materials include cobalt-containing compounds and nitrogen carbide. By controlling the calcination temperature to 150ºC≤T≤350ºC, nanozymes with single POD-like activity are obtained; and nanozymes with both POD-like and OXD-like activities are obtained at 350ºC<T≤1000ºC.

4. The method for regulating the type of nanozyme mimicking enzyme as described in claim 1 or 2, characterized in that, The precursor material also contains a substrate material.

5. The method for regulating the type of nanozyme mimicking enzyme as described in claim 4, characterized in that, The precursor materials include cobalt-containing compounds, nitrogen-containing compounds, and graphylene as the base material. The nitrogen-containing compounds are melamine or cyanuric chloride. By controlling the calcination temperature to 150ºC≤T≤400ºC, nanozymes with single POD-like activity are obtained; at 400ºC<T<650ºC, nanozymes with both POD-like and OXD-like activities are obtained; and at 650ºC≤T≤1000ºC, nanozymes with single OXD-like activity are obtained.

6. The method for regulating the type of nanozyme mimicking enzyme as described in claim 4, characterized in that, The precursor materials include cobalt-containing compounds, melamine, and graphene as a substrate material. By controlling the calcination temperature to 150ºC≤T≤350ºC, nanozymes with single POD-like activity are obtained; and nanozymes with both POD-like and OXD-like activities are obtained at 350ºC<T≤1000ºC.

7. The method for regulating the type of nanozyme mimicking enzyme as described in claim 4, characterized in that, The precursor materials include manganese-containing compounds, melamine, and carbon materials as the base material, which are graphene or graphyne. By controlling the calcination temperature to 150ºC≤T≤350ºC, nanozymes with single POD-like activity are obtained; and nanozymes with both POD-like and OXD-like activities are obtained at 350ºC<T≤1000ºC.

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