Application of chelating agents in inhibiting the catalytic activity and DNA cleavage of low-dimensional nanomaterials
By forming covalent bonds between chelating agents and low-dimensional nanomaterials, the stability and cost issues of natural enzymes in industrial, medical and biological fields are solved, and the catalytic activity of nanomaterials and DNA shearing are inhibited, which has important biological application potential.
Patent Information
- Application Number
- CN202311275501.6
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- Filing Date
- 2023-09-28
- Publication Date
- 2025-09-05
- Estimated Expiration
- 2043-09-28
AI Technical Summary
In the existing technology, the application of natural enzymes in industrial, medical and biological fields has problems such as high cost, low stability and difficulty in recovery, and there are no reports on the effects and interactions of chelating agents on nanomaterials.
A chelating agent is mixed with a low-dimensional nanomaterial so that the electrons of the chelating agent form covalent bonds with the dangling bonds of the material defects, thereby inhibiting the catalytic activity of the nanomaterial and the DNA shearing effect.
It effectively inhibits the catalytic activity and DNA shearing of low-dimensional nanomaterials, provides stability and cost-effectiveness, and has important biological application potential.
Smart Images
Figure CN117399034B_ABST
Abstract
Description
Technical Field
[0001] The present invention relates to the field of catalysis, and in particular to the application of a chelating agent in inhibiting the catalytic activity of low-dimensional nanomaterials and DNA shearing. Background Art
[0002] Natural enzymes, due to their high catalytic activity and substrate specificity, have been widely used in industry, medicine, biology, and other fields. Despite their promising prospects, they often suffer from inherent drawbacks, such as high cost, low operational stability, and difficulty in recycling. To overcome these shortcomings, researchers have long been committed to exploring artificial enzyme mimics.
[0003] Since the discovery of ferromagnetic nanoparticles with horseradish peroxidase-like activity in 2007, a significant amount of research on nanozymes has emerged over the following decade. Nanozymes are a class of nanomaterials with enzyme-catalytic properties. Compared to natural enzymes, nanozymes offer advantages such as low cost, high stability, and durability, and have found widespread application in industry, medicine, biology, and other fields. A deeper understanding of the potential catalytic mechanisms of nanozymes will facilitate the development of novel and efficient nanozymes, and the rational regulation of their activity is crucial. Chelating agents are often used as inhibitors in the biological field to prevent metal ions from damaging biological samples. This principle utilizes the lone electrons of the chelator to form coordination bonds with metal ions, thereby suppressing the effects of metal ions. However, the effects of chelating agents on defects in nanomaterials and the nature of their interactions have not been reported.
[0004] It should be noted that the information disclosed in the above background technology section is only used to understand the background of this application, and therefore may include information that does not constitute prior art known to ordinary technicians in this field. Summary of the Invention
[0005] The present invention provides an application of a chelating agent in inhibiting the catalytic activity of low-dimensional nanomaterials and DNA shearing.
[0006] The present invention adopts the following technical solutions:
[0007] In a first aspect, the present invention provides an application of a chelating agent in inhibiting the catalytic activity of low-dimensional nanomaterials. The chelating agent is mixed with the low-dimensional nanomaterial so that the electrons provided by the chelating agent interact with the dangling bonds of the defects in the low-dimensional nanomaterial to form covalent bonds, thereby inhibiting the catalytic activity of the low-dimensional nanomaterial.
[0008] Preferably, the chelating agent has two or more lone electrons; and the defects of the low-dimensional nanomaterial are at least one of point defects, line defects and surface defects.
[0009] Preferably, the low-dimensional nanomaterial is at least one of a two-dimensional nanomaterial, a one-dimensional nanomaterial, and a zero-dimensional nanomaterial.
[0010] Preferably, the chelating agent is ethylenediaminetetraacetic acid.
[0011] Preferably, the low-dimensional nanomaterial is molybdenum disulfide nanosheets.
[0012] Preferably, the thickness of the molybdenum disulfide nanosheets is in the range of 0.6 nm to 20 nm.
[0013] In a second aspect, the present invention also provides an application of a chelating agent in DNA shearing. The chelating agent is mixed with a low-dimensional nanomaterial and then mixed with a DNA solution, so that the electrons provided by the chelating agent interact with the dangling bonds of the defects of the low-dimensional nanomaterial to form covalent bonds, thereby inhibiting the catalytic activity of the low-dimensional nanomaterial from affecting DNA shearing.
[0014] Preferably, the application comprises the following steps:
[0015] S1, preparing a molybdenum disulfide solution by adding pure water to the molybdenum disulfide nanosheets;
[0016] S2, preparing ethylenediaminetetraacetic acid solution with pure water;
[0017] S3, mixing the molybdenum disulfide solution and the ethylenediaminetetraacetic acid solution for a predetermined time to obtain a mixed solution;
[0018] S4. Mixing the mixed solution with a DNA solution and incubating the mixture, thereby inhibiting the shearing effect of the molybdenum disulfide nanosheets on the DNA.
[0019] Preferably, the concentration of the molybdenum disulfide solution in step S3 is 100-1000 mg / L; the concentration of the ethylenediaminetetraacetic acid solution in step S3 is 1-20 mM.
[0020] Preferably, in step S3, the volume ratio of the molybdenum disulfide solution to the ethylenediaminetetraacetic acid solution is 1:1 or 1:2.
[0021] Preferably, the DNA in step S4 is a pBR322 plasmid; preferably, the concentration of the DNA solution is 100 ng / μl-500 ng / μl.
[0022] Preferably, the volume of the DNA solution is 0.5 μl-1 μl.
[0023] Preferably, in step S3, the predetermined time is 5-30 minutes; in step S4, the mixed solution and the DNA solution are mixed and incubated under the following conditions: temperature 4-85°C, pH 6-8, and mixed incubation time 5 minutes to 3 hours.
[0024] The present invention has the following advantages or beneficial effects: the application of the chelating agent provided by the present invention in inhibiting the catalytic activity of low-dimensional nanomaterials is to mix the chelating agent with the low-dimensional nanomaterial so that the electrons provided by the chelating agent interact with the dangling bonds of the defects of the low-dimensional nanomaterial to form covalent bonds, thereby inhibiting the catalytic activity of the low-dimensional nanomaterial. The present invention also provides the application of the chelating agent in DNA shearing, which is to mix the chelating agent with the low-dimensional nanomaterial and then mix it with a DNA solution so that the electrons provided by the chelating agent interact with the dangling bonds of the defects of the low-dimensional nanomaterial to form covalent bonds, thereby inhibiting the effect of the catalytic activity of the low-dimensional nanomaterial on DNA shearing. Therefore, the present invention utilizes the chelating effect of the chelating agent on the defects of the low-dimensional nanomaterial to inhibit the catalytic activity of the defects of the low-dimensional nanomaterial, and utilizes this property to successfully inhibit the shearing effect of the low-dimensional nanomaterial on DNA, which is of great significance for the later development of related biological applications of nanozymes. BRIEF DESCRIPTION OF THE DRAWINGS
[0025] Figure 1 Schematic diagram of the defect interaction between the chelating agent EDTA and molybdenum disulfide in a preferred embodiment of the present invention.
[0026] Figure 2 This is a thickness distribution diagram of molybdenum disulfide nanosheets measured using an atomic particle microscope in a preferred embodiment of the present invention.
[0027] Figure 3 It is the electrophoresis diagram of the experimental example and the comparative example of the present invention. DETAILED DESCRIPTION
[0028] The following is a detailed description of the embodiments of the present invention. It should be emphasized that the following description is merely illustrative and is not intended to limit the scope and application of the present invention. In the absence of conflict, the embodiments and features in the embodiments of this application can be combined with each other.
[0029] A specific embodiment of the present invention provides an application of a chelating agent in inhibiting the catalytic activity of low-dimensional nanomaterials. The chelating agent is mixed with the low-dimensional nanomaterial so that the electrons provided by the chelating agent interact with the dangling bonds of the defects of the low-dimensional nanomaterial to form covalent bonds, thereby inhibiting the catalytic activity of the low-dimensional nanomaterial.
[0030] The embodiments of the present invention utilize the new property of chelating agents discovered by the inventors, that is, they have a chelating effect on the defects of low-dimensional nanomaterials, and provide the application of chelating agents in inhibiting the catalytic activity of low-dimensional nanomaterials. When the chelating agent is mixed with the low-dimensional nanomaterial, the electrons provided by the chelating agent interact with the dangling bonds of the defects of the low-dimensional nanomaterial to form covalent bonds, thereby inhibiting the catalytic activity of the low-dimensional nanomaterial. The properties of the chelating agent are used to successfully inhibit the shearing effect of the low-dimensional nanomaterial on DNA.
[0031] In a preferred embodiment, the chelating agent has two or more lone electrons; and the defect of the low-dimensional nanomaterial is at least one of a point defect, a line defect, and a surface defect.
[0032] In a preferred embodiment, the chelating agent and the low-dimensional nano material are mixed in a liquid form. For example, the chelating agent and pure water can be configured to form a chelating agent solution, and the low-dimensional nano material and pure water can be configured to form a low-dimensional nano material solution, and then the two can be mixed. Alternatively, the chelating agent and the low-dimensional nano material can be added to pure water together and mixed. In addition, if the catalytic activity of the low-dimensional nano material is applied to the biological field, the chelating agent and a solvent such as acetic acid can also be configured to form a buffer solution for application.
[0033] In a preferred embodiment, the low-dimensional nanomaterial is at least one of a two-dimensional nanomaterial (such as a planar material, a heterojunction, a single-atom-doped two-dimensional nanomaterial, etc.), a one-dimensional nanomaterial (such as a tubular or linear material, a single-atom-doped one-dimensional nanomaterial), and a zero-dimensional nanomaterial (such as a point material, a single-atom-doped zero-dimensional nanomaterial, etc.).
[0034] In a preferred embodiment, low-dimensional nanomaterials can be prepared by chemical vapor deposition, lithium ion intercalation, chemical electroplating, pulsed laser deposition, mechanical exfoliation, warm ball milling, and other methods.
[0035] In a preferred embodiment, the chelating agent is ethylenediaminetetraacetic acid.
[0036] In a preferred embodiment, the low-dimensional nanomaterial is a molybdenum disulfide nanosheet; preferably, the thickness of the molybdenum disulfide nanosheet is in the range of 1-20 nm. For example, the chelating agent is ethylenediaminetetraacetic acid (EDTA) and the low-dimensional nanomaterial is a molybdenum disulfide nanosheet. Figure 1 The figure shows the interaction between the chelating agent EDTA and the Mo dangling bonds of MoS2 defects. MoS2 nanosheets are mainly composed of sulfur vacancies (a combination of point defects and line defects). The molybdenum element has excess electrons. The electrons provided by the chelating agent EDTA interact with the Mo dangling bonds of MoS2 nanosheet defects to form covalent bonds, thereby inhibiting the MoS2 nanosheet defects from exerting their catalytic activity.
[0037] The present invention also provides the application of a chelating agent in DNA shearing. The chelating agent is mixed with a low-dimensional nanomaterial and then mixed with a DNA solution, so that the electrons provided by the chelating agent interact with the dangling bonds of the defects of the low-dimensional nanomaterial to form covalent bonds, thereby inhibiting the catalytic activity of the low-dimensional nanomaterial from affecting DNA shearing.
[0038] Preferably, the chelating agent has two or more lone electrons; the defects of the low-dimensional nanomaterial are at least one of point defects, line defects, and surface defects. The low-dimensional nanomaterial is at least one of a two-dimensional nanomaterial (such as a planar material, a heterojunction, a single-atom-doped two-dimensional nanomaterial, etc.), a one-dimensional nanomaterial (such as a tube or wire material, a single-atom-doped one-dimensional nanomaterial), and a zero-dimensional nanomaterial (such as a point material, a single-atom-doped zero-dimensional nanomaterial, etc.).
[0039] In a preferred embodiment, the application comprises the following steps:
[0040] S1, preparing a molybdenum disulfide solution by adding pure water to the molybdenum disulfide nanosheets;
[0041] S2, preparing ethylenediaminetetraacetic acid solution with pure water;
[0042] S3, mixing the molybdenum disulfide solution and the ethylenediaminetetraacetic acid solution for a predetermined time to obtain a mixed solution;
[0043] S4. Mixing the mixed solution with a DNA solution (the solvent is pure water) and incubating the mixture, thereby inhibiting the shearing effect of the molybdenum disulfide nanosheets on the DNA.
[0044] In a preferred embodiment, the concentration of the molybdenum disulfide solution in step S3 is 100-1000 mg / L, for example, the concentration can be 100 mg / L, 200 mg / L, 300 mg / L, 400 mg / L, 500 mg / L, 600 mg / L, 700 mg / L, 800 mg / L, 900 mg / L, 1000 mg / L, etc.
[0045] In a preferred embodiment, the concentration of the EDTA solution in step S3 is 1-20 mM.
[0046] In a preferred embodiment, the volume ratio of the molybdenum disulfide solution to the EDTA solution in step S3 is 1:1 or 1:2.
[0047] In a preferred embodiment, the DNA in step S4 is a pBR322 plasmid.
[0048] In a preferred embodiment, the concentration of the DNA solution is 100 ng / μl-500 ng / μl, for example, the concentration of the DNA solution can be 100 ng / μl, 200 ng / μl, 300 ng / μl, 400 ng / μl, 500 ng / μl, etc.
[0049] In a preferred embodiment, the volume of the DNA solution is 0.5 μl-1 μl.
[0050] In a preferred embodiment, in step S3, the predetermined time is 5-30 minutes.
[0051] In a preferred embodiment, in step S4, the mixed solution and the DNA solution are incubated under the following conditions: a temperature of 4-85°C, a pH of 6-8, and a mixing and incubation time of 5 minutes to 3 hours, for example, a mixing and incubation time of 5 minutes, 10 minutes, 20 minutes, 30 minutes, 60 minutes, 1 hour, 2 hours, 3 hours, etc. In addition to inhibiting the catalytic activity of low-dimensional nanomaterials and thereby inhibiting the cleavage of DNA by low-dimensional nanomaterials, the chelating agents are also expected to find corresponding applications in other areas of gene editing, such as polymerase chain reaction and DNA thermal denaturation.
[0052] The following uses molybdenum disulfide nanosheets (two-dimensional materials) as an example to illustrate the inhibitory properties of EDTA on molybdenum disulfide on DNA shearing, which includes the following steps:
[0053] 1. Prepare MoS2 nanosheets by cryogenic ball milling (1-5 g of MoS2 powder is placed in a ball mill filled with liquid nitrogen to maintain low temperature, 15 cycles per minute, for a total of 150 min) to ensure that they are active and have a size within the range of 0.6 nm to 20 nm. Figure 2 Shown is the thickness distribution of MoS2 nanosheets measured by atomic force microscopy.
[0054] 2. Prepare a molybdenum disulfide nanosheet solution in pure water (PW) with a concentration of 100-1000 mg / L.
[0055] 3. Prepare EDTA solution with purified water (PW) to a concentration of 10 mM.
[0056] 4. Mix the molybdenum disulfide nanosheet solution and EDTA solution for 10 minutes, and then mix and incubate with DNA solution (the solvent is pure water, the DNA is pBR322 plasmid, and the concentration is 500 ng / μl) at room temperature (20-25°C) and pH 7 for 10 minutes.
[0057] Pure water was used instead of EDTA as a control, and the experimental examples (Experimental Example 1 and Experimental Example 2 are repeated experiments of the same experiment) and control examples (Control Example 1 and Control Example 2 are repeated experiments of the same control experiment) with the content of each substance shown in Table 1 below were used, and combined with the experimental data to demonstrate the inhibitory effect of EDTA on the catalytic activity of molybdenum disulfide nanosheets, thereby inhibiting the shearing effect of molybdenum disulfide nanosheets on DNA.
[0058] Table 1
[0059]
[0060] like Figure 3The electrophoresis diagrams of the experimental and control examples are shown in the figure. Figure 3 It can be seen that when EDTA is not added, new bands will be generated. This is because the catalytic properties of molybdenum disulfide will shear DNA. When EDTA is added, EDTA will combine with the defects of molybdenum disulfide, reducing its catalytic activity and thus inhibiting the DNA shearing effect.
[0061] The embodiment of the present invention utilizes the chelating effect of the chelating agent EDTA on the defects of low-dimensional nanomaterials to inhibit the catalytic activity of the defects of low-dimensional nanomaterials, and uses this property to successfully inhibit the shearing effect of molybdenum disulfide on DNA, which is of great significance for the later development of related biological applications of nanozymes.
[0062] In other embodiments, other chelating agents may be used in place of EDTA, and similarly, other low-dimensional nanomaterials may be used in place of molybdenum disulfide nanosheets. The above description further details the present invention in conjunction with specific / preferred embodiments, and the specific implementation of the present invention should not be construed as being limited to these descriptions. Persons skilled in the art will appreciate that, without departing from the spirit of the present invention, they may make several substitutions or modifications to the described embodiments, and these substitutions or modifications should be considered to fall within the scope of protection of the present invention. Throughout this specification, reference to terms such as "one embodiment," "some embodiments," "preferred embodiments," "examples," "specific examples," or "some examples" indicates that the specific features, structures, materials, or characteristics described in conjunction with such embodiment or example are included in at least one embodiment or example of the present invention. In this specification, the schematic representations of these terms do not necessarily refer to the same embodiment or example. Furthermore, the specific features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples. Persons skilled in the art may combine and assemble the different embodiments or examples, and features of the different embodiments or examples, described in this specification, unless they conflict with each other. Although the embodiments of the present invention and their advantages have been described in detail, it should be understood that various changes, substitutions, and alterations can be made herein without departing from the scope of protection of the patent application.
Claims
1. An application of a chelating agent in DNA shearing, characterized in that: The chelating agent is mixed with the low-dimensional nanomaterial and then mixed with the DNA solution, so that the electrons provided by the chelating agent interact with the dangling bonds of the defects of the low-dimensional nanomaterial to form covalent bonds, thereby inhibiting the catalytic activity of the low-dimensional nanomaterial on DNA shearing; the chelating agent is ethylenediaminetetraacetic acid, the DNA is pBR322 plasmid, and the low-dimensional nanomaterial is molybdenum disulfide nanosheets.
2. The use of the chelating agent as claimed in claim 1 in DNA shearing, characterized in that The application comprises the following steps: S1, preparing a molybdenum disulfide solution by adding pure water to the molybdenum disulfide nanosheets; S2, preparing ethylenediaminetetraacetic acid solution with pure water; S3, mixing the molybdenum disulfide solution and the ethylenediaminetetraacetic acid solution for a predetermined time to obtain a mixed solution; S4. Mixing the mixed solution with a DNA solution and incubating the mixture, thereby inhibiting the shearing effect of the molybdenum disulfide nanosheets on the DNA.
3. The use of the chelating agent as claimed in claim 2 in DNA shearing, characterized in that The concentration of the molybdenum disulfide solution in step S3 is 100-1000 mg / L; the concentration of the ethylenediaminetetraacetic acid solution in step S3 is 1-20 mM.
4. The use of the chelating agent as claimed in claim 3 in DNA shearing, characterized in that In step S3, the volume ratio of the molybdenum disulfide solution to the ethylenediaminetetraacetic acid solution is 1:1 or 1:
2.
5. The use of the chelating agent as claimed in claim 3 in DNA shearing, characterized in that The concentration of the DNA solution is 100 ng / μl-500 ng / μl.
6. Use of the chelating agent as claimed in claim 3 in DNA shearing, characterized in that, The volume of the DNA solution is 0.5 μl-1 μl.
7. Use of the chelating agent as claimed in claim 3 in DNA shearing, characterized in that In step S3, the predetermined time is 5-30 minutes; in step S4, the mixed solution and the DNA solution are mixed and incubated under the following conditions: temperature 4-85°C, pH 6-8, and mixed incubation time 5 minutes to 3 hours.
8. Use of the chelating agent according to claim 1 in DNA shearing, characterized in that The thickness of the molybdenum disulfide nanosheets is in the range of 0.6 nm to 20 nm.