Preparation method and application of nano-enzyme containing vanadium oxide
By preparing vanadium oxide-containing nanozymes, the problem of unstable enzyme activity of nanozymes in extreme temperatures and acidic environments was solved, and the effect of efficient degradation of chlorophenol was achieved with simple operation and low cost.
Patent Information
- Application Number
- CN202510777709.0
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-06-11
- Publication Date
- 2025-09-16
AI Technical Summary
The enzyme activity of existing nanozymes is unstable at extreme temperatures, and the degradation efficiency of chlorophenol is low in acidic environments. The preparation method is complex and costly.
P-123, sodium dodecyl sulfate, melamine and phytic acid are used as raw materials to prepare vanadium oxide nanozymes through hydrothermal reaction and calcination. The specific steps include dissolution, mixing, hydrothermal reaction, purification and calcination to form nanozymes with high peroxidase activity.
The prepared vanadium oxide nanozyme has high peroxidase activity in the acidic pH range, especially at pH=3, the degradation rate of chlorophenol reaches 99.9%. It still maintains high catalytic activity at extreme temperatures, has a long cycle life and low cost.
Smart Images
Figure CN120644190A_ABST
Abstract
Description
Technical Field
[0001] The present invention belongs to the technical field of nanozymes, and specifically relates to a preparation method and application of a nanozyme containing vanadium oxide. Background Art
[0002] Nanozymes, nanomaterials with enzymatic catalytic properties, are typically composed of inorganic nanoparticles, metal nanoparticles, or carbon-based materials. They exhibit similar properties to natural enzymes in catalytic reactions, such as specificity and efficiency. However, due to their higher stability, resistance to temperature and fouling, and wider pH adaptability, they have become an important research direction in the field of biocatalysis and have shown great potential in many applications.
[0003] The research on nanozymes began in the 1990s. In 2004, Scrimin et al. proposed the term nanozyme (Flavio et al. Nanozymes: gold-nanoparticle-based transphosphorylation catalysts. Angewandte Chemie International Edition, 43(45), 6009-6217). Since Yan et al. discovered ferromagnetic nanoparticles with enzyme-like activity in 2007 (Gao et al. Intrinsic peroxidase-like activity of ferromagnetic nanoparticles. Nature Nanotechnology, 2, 577-583), nanozymes have become the focus of attention of many scientists, and a large number of nanozymes have been developed. In the preparation of nanozymes, various nanozymes have been prepared using metal-based materials (Ti, V, Cr, Mn, Fe, Co, Ni, Cu, Zn, Zr, Mo, Ru, Rh, Pd, Ag, Cd, Sn, Sb, Ce, Hf, Pt, Au, Bi), metal-free materials (C, B, Se), and their compounds and hybrids. Nanozymes are nanomaterials with enzyme-like properties. The raw materials, including chemical composition (e.g., metal-based or non-metal-based), synthesis methods (e.g., impregnation, co-precipitation, deposition-precipitation, hydrothermal / solvothermal), and form (e.g., spherical, rod-like, ring-like, hollow structures), have been studied to mimic the antioxidant properties of natural enzymes using nanomaterials such as magnetic iron oxide nanoparticles, platinum nanoparticles, manganese oxide nanoparticles, gold nanoparticles, and vanadium pentoxide nanoparticles. Although these nanomaterials possess excellent enzymatic activity, their enzymatic properties vary.
[0004] In today's environment, climate change is extremely drastic, and the most extreme temperatures ever recorded have appeared around the world. Temperature has a strong impact on most enzymes. In this context, it is necessary to develop a material that can have efficient enzymatic activity over a wide range of temperatures. Summary of the Invention
[0005] The first technical problem to be solved by the present invention is to provide a method for preparing a nanozyme containing vanadium oxide, which is simple and easy to operate; the second technical problem to be solved by the present invention is to provide a nanozyme containing vanadium oxide, which has high peroxidase activity in the acidic pH range; the third technical problem to be solved by the present invention is to provide the application of the nanozyme containing vanadium oxide in degrading para-chlorophenol in wastewater.
[0006] In order to solve the above technical problems, the technical solutions adopted by the present invention are as follows:
[0007] A method for preparing a nanozyme containing vanadium oxide comprises the following steps:
[0008] 1) Dissolve P-123 and sodium lauryl sulfate in deionized water;
[0009] 2) adding melamine to the solution of step 1) and mixing and stirring;
[0010] 3) diluting the phytic acid solution in deionized water, stirring uniformly, and then adding the solution to the dispersed solution in step 2) to form a prepared solution;
[0011] 4) subjecting the prepared solution in step 3) to a hydrothermal reaction, cooling after the reaction, purifying, drying, grinding, and collecting the carbon precursor;
[0012] 5) The carbon precursor obtained in step 4) is mixed with the ammonium metavanadate solution, stirred, dried to form a solid, and then calcined to obtain the nanozyme.
[0013] Furthermore, in the step 1), the mass ratio of P-123 to sodium lauryl sulfate is 4-5:3-4.
[0014] Furthermore, in the step 2), the molar amount of melamine is 10 to 20 mmol.
[0015] Furthermore, in the step 3), the mass of phytic acid is 2 to 2.5 g.
[0016] Furthermore, in the step 4), the temperature of the hydrothermal reaction is 160° C., and the hydrothermal time is 12 h.
[0017] Furthermore, in the step 5), the mass ratio of the carbon precursor to ammonium metavanadate is 1:0.07.
[0018] Furthermore, in the step 5), the calcination temperature is 550° C. and the calcination time is 4 hours.
[0019] Furthermore, the method for preparing the nanozyme containing vanadium oxide prepares the nanozyme containing vanadium oxide.
[0020] Furthermore, the vanadium oxide-containing nanozyme is used to degrade p-chlorophenol in wastewater.
[0021] Furthermore, the wastewater temperature is 0-80°C, and the wastewater pH is 2-8.
[0022] Compared with the prior art, the present invention has the following advantages:
[0023] (1) The preparation method of the present invention is simple, easy to operate and low in cost.
[0024] (2) The vanadium oxide nanozyme prepared by the present invention has a long cycle life and can be used repeatedly.
[0025] (3) The vanadium oxide nanozyme prepared by the present invention has a wide range of applications and has high peroxidase activity in the acidic pH range. Its peroxidase activity is optimal at pH = 3, and the degradation rate of chlorophenol reaches 99.9%, far exceeding the common enzyme-like materials on the market. Therefore, its application value in the acidic field is extremely high.
[0026] (4) The enzyme activity of the vanadium oxide nanozyme prepared by the present invention continues to increase with the increase of temperature. In addition, it still has a peroxidase activity much higher than that of common enzyme materials on the market under low temperature conditions, and can be actually applied to wastewater treatment processes in northern China. BRIEF DESCRIPTION OF THE DRAWINGS
[0027] Figure 1 This is an appearance diagram of the carbon precursor prepared in Example 1 of the present application;
[0028] Figure 2 This is the SEM scanning result of the carbon precursor prepared in Example 1 of the present application;
[0029] Figure 3 This is the appearance of the vanadium oxidase prepared in Example 1 of the present application;
[0030] Figure 4 This is a SEM scanning result of the vanadium oxidase prepared in Example 1 of the present application;
[0031] Figure 5 This is a Fourier transform infrared result of the vanadium oxidase prepared in Example 1 of the present application;
[0032] Figure 6 This is the XPS result of the vanadium oxidase prepared in Example 1 of the present application;
[0033] Figure 7 This is a Michaelis-Menten kinetic diagram of the vanadium oxidase prepared in Example 1 of the present application at 25°C; wherein Figure a is a Michaelis-Menten plot with TMB as a substrate, Figure b is a Lineweaver-Burk plot with TMB as a substrate, Figure c is a Michaelis-Menten plot with hydrogen peroxide as a substrate, and Figure d is a Lineweaver-Burk plot with hydrogen peroxide as a substrate;
[0034] Figure 8 Comparison of the experimental graphs of the degradation of p-chlorophenol by the vanadium oxidase prepared in Example 1 of the present application at different temperatures;
[0035] Figure 9 This is a comparison chart of the degradation experiment of p-chlorophenol performed by the vanadium oxidase prepared in Example 1 of the present application at different pH values;
[0036] Figure 10 This is a comparison chart of the degradation experiment of p-chlorophenol performed by the vanadium oxidase prepared in Example 1 of the present application and natural peroxidase. DETAILED DESCRIPTION
[0037] The present invention will be further illustrated below with reference to specific examples. The examples are implemented based on the technical solutions of the present invention. It should be understood that these examples are only used to illustrate the present invention and are not used to limit the scope of the present invention.
[0038] Whether the raw materials used in the following examples are all commercially available, please see Table 1 for detailed information.
[0039] Table 1 Raw material information
[0040] drug model factory CAS Block polyether P-123 AR Aladdin 9003-11-6 Sodium lauryl sulfate AR Aladdin 151-21-3 Phytic acid solution 70％ Maclean 83-86-3 Melamine AR Maclean 108-78-1 Ammonium metavanadate AR Aladdin 7803-55-6 Horseradish peroxidase AR Aladdin 9003-99-0 sulfuric acid 98％ Aladdin 7664-93-9 Sodium hydroxide AR Aladdin 1310-73-2 phenol AR Maclean 108-95-2 p-Chlorophenol AR Aladdin 106-48-9 hydrogen peroxide 30％ Aladdin 7722-84-1
[0041] Example 1
[0042] A method for preparing a nanozyme containing vanadium oxide comprises the following steps:
[0043] (1) Weigh 40 mg of P-123 solid and 30 mg of sodium lauryl sulfate and dissolve them in 20 mL of deionized water;
[0044] (2) Weigh 10 mmol of melamine solid and add it to the solution of step (1), stirring evenly;
[0045] (3) Weigh 2.5 g of phytic acid solution, dilute it in 40 mL of deionized water, stir it evenly, and add it to the above 20 mL of the dispersed solution to form a prepared solution;
[0046] (4) The prepared solution was transferred to a hydrothermal reactor and subjected to a hydrothermal reaction at 160°C for 12 hours. After the reaction was completed, the mixture was naturally cooled to room temperature to obtain white needle-shaped crystals and black particles, which were then centrifuged at 4000 rpm for 5 minutes and washed three times with deionized water. After washing, the mixture was dried for 12 hours, ground, and the carbon precursor was collected.
[0047] (5) 1 g of the carbon precursor obtained in step (4) was mixed with 0.07 g of ammonium metavanadate solution and stirred for 24 h, followed by centrifugal drying for 12 h to form a solid; the obtained solid was placed in a muffle furnace and calcined at 550 ° C for 4 h in an air atmosphere at a heating rate of 4.4 ° C / min to obtain a dark green powdered nanozyme.
[0048] Figure 1 This is the appearance picture of the prepared carbon precursor. It can be seen that its appearance is wafer-like and is brittle after drying.
[0049] Figure 2 The SEM results of the prepared carbon precursor show that at the microscopic level, its structure is like stacked chips, and the space created can effectively adsorb metal ions.
[0050] Figure 3 This is the appearance picture of the prepared nanozyme. It can be seen that it appears as a dark green agglomerated powder and can be dispersed quickly in water.
[0051] Figure 4 The SEM results of the prepared nanozyme show that at the microscopic level, the vanadium oxide nanozyme is in the form of fine flakes, and there are uneven tiny holes on the surface of the flakes, which can produce good active sites for the catalysis of peroxides.
[0052] Figure 5 The Fourier infrared results of the prepared nanozyme show that after loading V, the material has a wavelength of 3000-3600 cm -1 The increase in vibration in the range of 1300-1800 cm -1 The increase in vibration within the range indicates that more C=O bonds and C=C bonds are generated.
[0053] Figure 6 This is the XPS result diagram of the prepared nanozyme. It can be seen that the main components of vanadium oxide nanozyme are O, V, N, C, and P, among which O element is dominant.
[0054] The nanozyme prepared in Example 1 was subjected to Michaelis-Menten kinetics. The method included analyzing the steady-state kinetics of the vanadium oxide nanozyme at different TMB concentrations (0.2, 0.52, 0.83, 1.56, 2.08, 3.12, and 4.16 mM) with a constant hydrogen peroxide concentration (1 mM), and at different hydrogen peroxide concentrations (0.0125, 0.025, 0.0375, 0.05, 0.0625, 0.075, 0.0875, 0.1, and 0.125 mM) with a constant TMB concentration (2.08 mM). The data were fitted to the Michaelis-Menten equation:
[0055]
[0056] Depend on Figure 7 It can be seen that when vanadium oxide nanozyme uses TMB as substrate, V max Up to 1.269 μM s -1 , K m When hydrogen peroxide is used as substrate, V max Up to 0.1301 μM s -1 , K m As low as 0.023, it shows that vanadium oxidase has good affinity for TMB and hydrogen peroxide, especially for hydrogen peroxide, which proves that it can efficiently catalyze hydrogen peroxide.
[0057] Example 2
[0058] A method for preparing a nanozyme containing vanadium oxide comprises the following steps:
[0059] (1) Weigh 45 mg of P-123 solid and 35 mg of sodium lauryl sulfate and dissolve them in 20 mL of deionized water;
[0060] (2) Weigh 15 mmol of melamine solid and add it to the solution of step (1), stirring evenly;
[0061] (3) Weigh 2 g of phytic acid solution, dilute it in 40 mL of deionized water, stir it evenly, and add it to the above 20 mL of the dispersed solution to form a prepared solution;
[0062] (4) The prepared solution was transferred to a hydrothermal reactor and subjected to a hydrothermal reaction at 160°C for 12 hours. After the reaction was completed, the mixture was naturally cooled to room temperature to obtain white needle-shaped crystals and black particles, which were then centrifuged at 4000 rpm for 5 minutes and washed three times with deionized water. After washing, the mixture was dried for 12 hours, ground, and the carbon precursor was collected.
[0063] (5) 1 g of the carbon precursor obtained in step (4) was mixed with 0.07 g of ammonium metavanadate solution and stirred for 24 h, followed by centrifugal drying for 12 h to form a solid; the obtained solid was placed in a muffle furnace and calcined at 550 ° C for 4 h in an air atmosphere at a heating rate of 4.4 ° C / min to obtain a dark green powdered nanozyme.
[0064] Example 3
[0065] A method for preparing a nanozyme containing vanadium oxide comprises the following steps:
[0066] (1) Weigh 50 mg of P-123 solid and 40 mg of sodium lauryl sulfate and dissolve them in 20 mL of deionized water;
[0067] (2) Weigh 20 mmol of melamine solid and add it to the solution of step (1), stirring evenly;
[0068] (3) Weigh 2.3 g of phytic acid solution, dilute it in 40 mL of deionized water, stir it evenly, and add it to the above 20 mL of the dispersed solution to form a prepared solution;
[0069] (4) The prepared solution was transferred to a hydrothermal reactor and subjected to a hydrothermal reaction at 160°C for 12 hours. After the reaction was completed, the mixture was naturally cooled to room temperature to obtain white needle-shaped crystals and black particles, which were then centrifuged at 4000 rpm for 5 minutes and washed three times with deionized water. After washing, the mixture was dried for 12 hours, ground, and the carbon precursor was collected.
[0070] (5) 1 g of the carbon precursor obtained in step (4) was mixed with a solution containing 0.07 g of ammonium metavanadate and stirred for 24 h, followed by centrifugal drying for 12 h to form a solid; the obtained solid was placed in a muffle furnace and calcined at 550 ° C for 4 h in an air atmosphere at a heating rate of 4.4 ° C / min to obtain a dark green powdered nanozyme.
[0071] Example 4
[0072] The vanadium oxide nanozymes prepared in Examples 1-3 were tested for their ability to degrade phenolic pollutants. An appropriate amount of vanadium oxide nanozyme was weighed and prepared into a 2 g / L stock solution. After thorough sonication, the solution was added to a conical flask containing 0.4 mM hydrogen peroxide and 30 μM of typical phenols (phenol and p-chlorophenol), respectively, to a final concentration of 0.2 g / L. The solution was shaken at 25°C. The degradation rates are shown in Table 2 below.
[0073] Table 2 Degradation rate of phenolic pollutants by vanadium oxide nanozymes prepared in Examples 1 to 3
[0074]
[0075] As can be seen from the results in Table 2, the vanadium oxide nanozyme in Example 1 can achieve a degradation rate of more than 99% for 30 μM phenol pollutants in about 10 minutes under the conditions of 0.2 g / L and 0.4 mM hydrogen peroxide; the vanadium oxidase in Example 2 can achieve a degradation rate of up to 97.7% for 30 μM phenol pollutants under the conditions of 0.2 g / L and 0.4 mM hydrogen peroxide; the vanadium oxidase in Example 3 can achieve a degradation rate of up to 95.1% for 30 μM phenol pollutants under the conditions of 0.2 g / L and 0.4 mM hydrogen peroxide.
[0076] Example 5
[0077] The vanadium oxide nanozyme from Example 1 was subjected to five cycles of testing. The experimental method was as follows: an appropriate amount of vanadium oxide nanozyme was weighed and prepared into a 2 g / L stock solution. After thorough sonication, the solution was added to a conical flask containing 0.4 mM hydrogen peroxide and 30 μM typical phenols (phenol and p-chlorophenol), respectively, to a final concentration of 0.2 g / L. The solution was shaken at 25°C and sampled after 30 minutes. The remaining liquid was centrifuged and evaporated to dryness. The above steps were repeated five times with the resulting solid. The degradation rates are shown in the table below.
[0078] Table 3 Degradation rate results of vanadium oxide nanozyme cycle experiment
[0079] pollutants Loop 1 Loop 2 Loop 3 Loop 4 Loop 5 phenol 99.9％ 98.2％ 94.7％ 90.1％ 80.5％ p-Chlorophenol 99.9％ 99.4％ 96.2％ 92.7％ 87.4％
[0080] As shown in Table 3, when vanadium oxide nanozyme degraded phenol, the degradation rate was still 80.5% in the fifth cycle experiment, and when degrading para-chlorophenol, the degradation rate was still 87.4% in the fifth cycle experiment, indicating that vanadium oxide nanozyme has a good cycle life.
[0081] Example 6
[0082] The degradation experiment of para-chlorophenol was carried out on the vanadium oxide nanozyme of Example 1 at 0°C, 15°C, 25°C, 60°C and 80°C. The experimental method was as follows: an appropriate amount of vanadium oxide nanozyme was weighed and prepared into a 2g / L stock solution. After sufficient ultrasound, it was added to a conical flask containing 0.4mM hydrogen peroxide and 30μM para-chlorophenol. The final concentration of the vanadium oxide nanozyme was 0.2g / L. The solution was shaken at 0°C, 15°C, 25°C, 60°C and 80°C, and samples were taken at regular intervals. The results were as follows: Figure 8 As can be seen from the figure, at 80°C, the degradation rate reached 99.9% within 5 minutes, and at 0°C, the degradation rate was 75%, indicating that vanadium oxide nanozymes still have excellent catalytic activity under high temperature conditions, and vanadium oxide nanozymes have good catalytic effects under a wide range of temperatures (0-80°C), especially good adaptability to extreme environments (80°C).
[0083] Example 7
[0084] The vanadium oxide nanozyme of Example 1 was subjected to a degradation experiment of parachlorophenol in ultrapure water with an initial pH of 2-8. The experimental method was as follows: an appropriate amount of vanadium oxide nanozyme was weighed and prepared into a 2 g / L stock solution. After sufficient sonication, it was added to a conical flask containing 0.4 mM hydrogen peroxide and 30 μM parachlorophenol solution with an initial pH of 2-8. The final concentration of the vanadium oxide nanozyme was 0.2 g / L. The solution was shaken at 25°C for 20 minutes and then sampled. The degradation results were as follows: Figure 9 As can be seen from the figure, when the pH is 2, the degradation rate reaches 96%, and when the pH is 3, the degradation rate of parachlorophenol reaches 99.9%. At pH 4-8, the degradation rate slowly decreases with the increase of pH, but at pH 8, there is still a degradation rate of nearly 70%, indicating that vanadium oxide nanozymes are active under a wide range of pH and have a good catalytic effect in highly acidic water bodies.
[0085] Example 8
[0086] The vanadium oxide nanozyme of Example 1 and natural peroxidase (HRP enzyme) were used to perform a degradation experiment of para-chlorophenol in ultrapure water with an initial pH of 3. The experimental method was as follows: appropriate amounts of vanadium oxide nanozyme and HRP enzyme were weighed and prepared into 2 g / L stock solutions, and after the vanadium oxide enzyme stock solution was fully sonicated, it was added to a conical flask containing 0.4 mM hydrogen peroxide and 30 μM para-chlorophenol solution with an initial pH of 3, and shaken at 0°C, 4°C, 15°C, 25°C, 37°C, 60°C, 80°C, and 100°C, and sampled at 20 minutes. The degradation results were as follows: Figure 10 At 0℃-37℃, HRP enzyme can achieve a degradation rate of 100%, but when the temperature is higher than 60℃, HRP enzyme loses its enzymatic activity and cannot degrade para-chlorophenol. At 0℃-15℃, vanadium oxide nanozyme can achieve degradation rates of 75%, 85%, and 90%, and at 25℃-100℃, it can reach a degradation rate of 99.9%, indicating that vanadium oxide enzyme has considerable peroxide catalytic ability at low temperatures and can maintain stable and efficient peroxide catalytic ability at high temperatures.
[0087] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.
Claims
1. A method for preparing a nanozyme containing vanadium oxide, characterized in that: The following steps are involved: 1) Dissolve P-123 and sodium lauryl sulfate in deionized water; 2) adding melamine to the solution of step 1) and mixing and stirring; 3) diluting the phytic acid solution in deionized water, stirring uniformly, and then adding the solution to the dispersed solution in step 2) to form a prepared solution; 4) subjecting the prepared solution in step 3) to a hydrothermal reaction, cooling after the reaction, purifying, drying, grinding, and collecting the carbon precursor; 5) The carbon precursor obtained in step 4) is mixed with the ammonium metavanadate solution, stirred, dried to form a solid, and then calcined to obtain the nanozyme.
2. The method for preparing a nanozyme containing vanadium oxide according to claim 1, wherein: In the step 1), the mass ratio of P-123 to sodium lauryl sulfate is 4-5:3-4.
3. The method for preparing a nanozyme containing vanadium oxide according to claim 1, wherein: In the step 2), the molar amount of melamine is 10 to 20 mmol.
4. The method for preparing a nanozyme containing vanadium oxide according to claim 1, wherein: In the step 3), the mass of phytic acid is 2 to 2.5 g.
5. The method for preparing a nanozyme containing vanadium oxide according to claim 1, wherein: In the step 4), the temperature of the hydrothermal reaction is 160° C., and the hydrothermal time is 12 h.
6. The method for preparing a nanozyme containing vanadium oxide according to claim 1, wherein: In the step 5), the mass ratio of the carbon precursor to the ammonium metavanadate is 1:0.
07.
7. The method for preparing a nanozyme containing vanadium oxide according to claim 1, wherein: In the step 5), the calcination temperature is 550° C. and the calcination time is 4 hours.
8. The method for preparing a vanadium oxide-containing nanozyme according to any one of claims 1 to 7, wherein the vanadium oxide-containing nanozyme is prepared.
9. Use of the vanadium oxide-containing nanozyme according to claim 8 to degrade p-chlorophenol in wastewater.
10. The use of the vanadium oxide-containing nanozyme according to claim 9 for degrading p-chlorophenol in wastewater, characterized in that: The wastewater temperature is 0-80°C and the wastewater pH is 2-8.