CN115463151A - Nano enzyme, preparation method and application thereof, and bacteriostatic agent - Google Patents
Nano enzyme, preparation method and application thereof, and bacteriostatic agent Download PDFInfo
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- CN115463151A CN115463151A CN202211002839.XA CN202211002839A CN115463151A CN 115463151 A CN115463151 A CN 115463151A CN 202211002839 A CN202211002839 A CN 202211002839A CN 115463151 A CN115463151 A CN 115463151A
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K33/00—Medicinal preparations containing inorganic active ingredients
- A61K33/24—Heavy metals; Compounds thereof
- A61K33/30—Zinc; Compounds thereof
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K33/00—Medicinal preparations containing inorganic active ingredients
- A61K33/24—Heavy metals; Compounds thereof
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
- A61P17/02—Drugs for dermatological disorders for treating wounds, ulcers, burns, scars, keloids, or the like
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y40/00—Manufacture or treatment of nanostructures
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y5/00—Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Abstract
The invention discloses a nano enzyme, a preparation method, an application and a bacteriostatic agent thereof, wherein the nano enzyme is of a cubic structure, and C, zn and Co are uniformly dispersed in the cubic structure. The invention aims to provide a nano enzyme bacteriostatic agent with good bacteriostatic performance and good effect of promoting wound healing.
Description
Technical Field
The invention relates to the technical field of biological and medical sterilization, and particularly relates to a nano enzyme, a preparation method and application thereof, and a bacteriostatic agent.
Background
The nano enzyme is a nano material with enzyme catalytic activity and mainly comprises two types: one is that natural enzyme or group with enzyme catalytic activity is modified on nano material, and the nano material plays the role of carrier; another is that the nanomaterial itself has catalytic activity similar to that of enzymes. Compared with natural enzymes, the nano-enzyme has the characteristics of low price, simple preparation process, good stability, high recycling efficiency and the like, and has application prospects in the fields of detection, sewage treatment, sterilization, inflammation diminishing, cancer treatment and the like.
Diseases caused by bacteria have become one of the biggest global health problems, and affect tens of thousands of people every year. The traditional antibacterial drugs are mainly antibiotics, and in addition, inorganic reagents of metal inorganic salts and organic reagents of vanillin, quaternary ammonium salts and the like can be used as antibacterial materials. However, these organic and inorganic antibacterial agents often have certain defects, such as complex preparation and synthesis process, high cost, easy generation of drug resistance, pollution to the surrounding environment, and the like. For this reason, new materials are being sought for antibacterial therapy.
The process of wound tissue repair and regeneration is one of the most complex biological processes, mainly involving cell proliferation, angiogenesis, and tissue remodeling. And the repair process may be delayed due to bacterial infection and slow differentiation of fibroblasts, resulting in local pain, swelling, and even life-threatening complications of infection. Therefore, there is an urgent need to find ways to accelerate tissue repair at a wound site while performing antimicrobial therapy.
Reactive Oxygen Species (ROS) are oxygen-containing, highly reactive chemical species with unpaired electrons, including superoxide anion (O2) .- ) Hydroxy radical(s) (iii) . OH)、H 2 O 2 In addition, ROS can cause damage to chemical substances such as microbial DNA, proteins and the like, and finallyResulting in cell death. The nanometer enzyme can adjust ROS level, and is used for developing novel antibacterial drugs. MSN-AuNPs can produce excessive ROS, thereby producing strong antibacterial performance. Has obvious inhibition effect on the proliferation of gram-positive staphylococcus aureus and gram-negative escherichia coli. The nano enzyme is used as a mimic of peroxidase, and can be used for inhibiting wound infection caused by bacteria and promoting wound healing. For example, the magnetic nanoparticles and the graphene quantum dots have peroxidase mimic enzyme activity, so that the magnetic nanoparticles and the graphene quantum dots have excellent antibacterial performance in wound infection. At present, the drug resistance of bacteria is a worldwide problem, and the nano enzyme can be developed into a novel antibacterial agent, so that a new idea is provided for solving the problem.
Disclosure of Invention
The invention mainly aims to provide a nano enzyme, a preparation method and application thereof, and a bacteriostatic agent, and aims to provide a nano enzyme bacteriostatic agent with good bacteriostatic performance and wound healing promotion effect.
In order to achieve the purpose, the invention provides a nano enzyme which is of a cubic structure, wherein C, zn and Co are uniformly dispersed in the cubic structure.
The invention further provides a preparation method of the nano enzyme, which comprises the following steps:
adding zinc salt and 2-methylimidazole into water, mixing and stirring uniformly, and then carrying out centrifugal treatment to obtain a precipitate;
adding cobalt salt into a methanol solution, carrying out ultrasonic treatment, then adding the first precipitate, and stirring to obtain a first solution;
adding 2-methylimidazole into a methanol solution, and performing ultrasonic treatment to obtain a second solution;
and adding the second solution into the first solution, standing, centrifuging, drying and calcining to obtain the nano enzyme.
Optionally, adding the zinc salt and the 2-methylimidazole into water, mixing and uniformly stirring, and then performing centrifugation treatment, wherein in the step of taking out the precipitate, the centrifugation treatment parameters are as follows:
centrifuging at 9000-10000 rpm for 10-15 min.
Optionally, the second solution is added into the first solution, the first solution is left to stand and centrifuged, and a precipitate is taken out to be dried and calcined to obtain the nano-enzyme, wherein the drying and calcining comprises the following steps:
drying at 70-75 deg.c for 8-12 hr, and calcining at 400-410 deg.c for 2-3 hr at 2-3 deg.c/min.
Alternatively, the zinc salt and 2-methylimidazole are added to water and mixed and stirred uniformly, and then the precipitate is washed with deionized water in the step of centrifuging and taking the precipitate.
Optionally, the second solution is added to the first solution, and the mixture is subjected to standing centrifugation, drying and calcination to obtain the nano-enzyme, wherein the nano-enzyme is washed with methanol after the standing centrifugation.
Optionally, the second solution is added into the first solution, standing and centrifuging are carried out, and the precipitate is taken out, dried and calcined to obtain the nano enzyme, wherein the standing time is 8-24 hours.
The invention also provides a bacteriostatic agent which comprises the nano enzyme or pharmaceutically acceptable salt thereof.
Optionally, the bacteriostatic agent further comprises a compound solvent of nano-enzyme, and in the bacteriostatic agent, the concentration of the nano-enzyme is 1 mg/mL-3 mg/mL.
The invention also provides application of the nano enzyme or the salt thereof in preparing the bacteriostatic agent.
The nano enzyme provided by the invention has a large number of active sites on the surface, is high in catalytic activity, and achieves the effects of high-efficiency broad-spectrum antibacterial performance and promotion of wound healing by combining the synergistic effect of bimetal. The antibacterial agent has multiple mimic enzyme activities of oxidase and catalase, and pharmaceutically acceptable salts and solvates thereof have good antibacterial performance and effect of promoting wound healing in preparation of the antibacterial agent.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only some embodiments of the present invention, and for those skilled in the art, other related drawings can be obtained according to the drawings without creative efforts.
FIG. 1 is an X-ray diffraction spectrum of the nano-enzyme provided by the invention;
FIG. 2 shows ZnO-Co provided by the present invention 3 A bacteriostatic performance schematic diagram of O nanoenzyme;
FIG. 3 is a graph showing the results of treatment of wounds in mice in the control group and groups 1 to 3 provided by the present invention;
FIG. 4 is a flow chart of a method for preparing nanoenzyme according to an embodiment of the present invention;
FIG. 5 is a microscopic schematic view of a nanoenzyme provided by the present invention.
The implementation, functional features and advantages of the objects of the present invention will be further explained with reference to the accompanying drawings.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. It is to be understood that the described embodiments are merely a few embodiments of the invention, and not all embodiments.
It should be noted that those whose specific conditions are not specified in the examples were performed according to the conventional conditions or the conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products available commercially. In addition, the meaning of "and/or" appearing throughout includes three juxtapositions, exemplified by "A and/or B" including either A or B or both A and B. In addition, technical solutions between various embodiments may be combined with each other, but must be realized by a person skilled in the art, and when the technical solutions are contradictory or cannot be realized, such a combination should not be considered to exist, and is not within the protection scope of the present invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Reactive Oxygen Species (ROS) are oxygen-containing, highly reactive chemical species with unpaired electrons, including superoxide anion (O2) .- ) Hydroxy radical(s) (iii) . OH)、H 2 O 2 And the ROS can cause damage to chemical substances such as microbial DNA, proteins and the like, and finally cause cell death. The nanometer enzyme can adjust the ROS level and is used for developing novel antibacterial drugs. MSN-AuNPs can produce excessive ROS, thereby producing strong antibacterial performance. Has obvious inhibition effect on the proliferation of gram-positive staphylococcus aureus and gram-negative escherichia coli. The nano enzyme is used as a mimic of peroxidase, and can be used for inhibiting wound infection caused by bacteria and promoting wound healing. For example, the magnetic nanoparticles and the graphene quantum dots have peroxidase mimic enzyme activity, so that the magnetic nanoparticles and the graphene quantum dots have excellent antibacterial performance in wound infection. At present, the drug resistance of bacteria is a worldwide problem, and the nano enzyme can be developed into a novel antibacterial agent, so that a new idea is provided for solving the problem. In view of the above, the present invention provides a ZnO-Co 3 O 4 A heterostructure nanoenzyme.
Referring to fig. 5, the present invention provides a nanoenzyme, which has a cubic structure, wherein C, zn and Co are uniformly dispersed in the cubic structure. That is, C, zn and Co are uniformly distributed to constitute the nanoenzyme. Referring to parts A and B of FIG. 5, the nanoenzymes are shown in the form of isolated cubes under a Scanning Electron Microscope (SEM). The C part is a Transmission Electron Microscope (TEM) image of the nanoenzyme, from which ZnO (100), znO (101) and Co can be clearly observed 3 O 4 (511) D is ZnO-Co 3 O 4 Element Mapping (Mapping) of (1), which demonstrates ZnO-Co 3 O 4 The elements of (A) O, zn and Co are uniformly dispersed in the cubic structure. In conclusion, the successful preparation of the nanoenzyme is demonstrated.
Further, the invention also provides a preparation method of the nano enzyme, which comprises the following steps:
s10, adding zinc salt and 2-methylimidazole into water, mixing and uniformly stirring, and then carrying out centrifugal treatment to obtain a precipitate;
in particular, the zinc salt is preferably Zn (NO) 3 ) 2 ·6H 2 O, the parameters of the centrifugal treatment are as follows: centrifuging at 9000-10000 rpm for 10-15 min.
Further, before the step S10 and the step S20, the method further includes washing the precipitate with deionized water.
S20, adding cobalt salt into a methanol solution, carrying out ultrasonic treatment, adding the first precipitate, and stirring to obtain a first solution;
specifically, the cobalt salt is preferably Co (NO) 3 ) 2 ·6H 2 O。
S30, adding 2-methylimidazole into the methanol solution, and performing ultrasonic treatment to obtain a second solution;
and S40, adding the second solution into the first solution, standing, centrifuging, drying and calcining to obtain the heterostructure nano enzyme.
Specifically, the drying and calcining comprises drying for 8-12 h at 70-75 ℃, and calcining for 2-3 h at 400-410 ℃ at a heating rate of 2-3 ℃/min. After standing for 8-24h, centrifuging, and washing with methanol.
Furthermore, the invention also provides a bacteriostatic agent which comprises the nano enzyme or pharmaceutically acceptable salt thereof. The nano-enzyme has high catalytic activity, and achieves the effects of high-efficiency broad-spectrum bacteriostasis and wound healing promotion by combining the synergistic effect of bimetal. The bacteriostatic agent has multiple mimic enzyme activities of oxidase and peroxidase, and pharmaceutically acceptable salts and solvates thereof have good bacteriostatic performance and the effect of promoting wound healing in the preparation of bacteriostatic agents.
Specifically, the bacteriostatic agent can be introduced into the body such as muscle, subcutaneous, intradermal, intravenous and mucosal tissues by injection, spray, nasal drop, eye drop, penetration, absorption, physical or chemical mediated method, or can be mixed or coated with other substances and then introduced into the body with good effect.
Specifically, the bacteriostatic agent also comprises a compound solvent of nano-enzyme, and in the bacteriostatic agent, the concentration of the nano-enzyme is 1 mg/mL-3 mg/mL. Wherein the compound solvent is preferably hydrogen peroxide (H) 2 O 2 )。
Furthermore, the invention also provides application of various nano enzymes or salts thereof in preparing bacteriostatic agents, wherein the bacteriostatic agent is preferably the bacteriostatic agent. In particular to application of the nanoenzyme or pharmaceutically acceptable salt thereof in preparing medicaments for inhibiting gram-positive bacteria and gram-negative bacteria.
The technical solutions of the present invention are further described in detail below with reference to specific examples and drawings, it should be understood that the following examples are merely illustrative of the present invention and are not intended to limit the present invention.
Example 1: application of nano enzyme serving as bacteriostatic agent
1. Preparation of nanoenzyme
(1) Adding Zn (NO) 3 ) 2 ·6H 2 Adding O and 2-methylimidazole into water, mixing and uniformly stirring, centrifuging for 10min at a rotation speed of 9000rpm to obtain a precipitate, and washing the precipitate with deionized water;
(2) Mixing Co (NO) 3 ) 2 ·6H 2 Adding O into a methanol solution, carrying out ultrasonic treatment, adding the first precipitate, and stirring to obtain a first solution;
(3) And adding the second solution into the first solution, standing for 24h, centrifuging, washing with methanol, drying at 70 ℃ for 12h, and calcining at 400 ℃ for 2h at the heating rate of 2 ℃/min to obtain the nano-enzyme.
2. Plate count.
Continuously diluting activated Staphylococcus aureus, pseudomonas aeruginosa, escherichia coli, and Bacillus subtilis to 10 times 6 CFU/mL. 2mg/mL of ZnO-Co 3 O 4 Heterojunction nanoenzyme, 5mmol/L H 2 O 2 Mixing with the bacterial liquid, incubating at 37 deg.C for 2h, spreading on solid culture medium with spreader which has been burned with alcohol and cooled to warm, and incubating overnight (about 18-24 h) in 37 deg.C incubator. To pairColonies on the plate were counted.
3. Group culture
Control group: 100 mul of culture medium and 100 mul of bacterial liquid;
group 1:50 μ L of culture medium, 100 μ L of bacterial liquid and 50 μ L of ZnO-Co 3 O 4 (2mg/mL)；
Group 2:50 μ L Medium +50 μ L H 2 O 2 (5 mmol/L) + 100. Mu.L of the bacterial solution;
group 3:50 μ LH 2 O 2 (5 mmol/L) + 100. Mu.L of bacterial solution + 50. Mu.L of ZnO-Co 3 O 4 (2mg/mL)。
4. Sterilization rate = (number of control colony-number of experimental colony)/number of control colony 100%
5. And the wound healing of the mouse is promoted.
Constructing wound with diameter of 6mm on the skin of the back of mice after adaptive feeding for 7 days and infecting staphylococcus aureus, and treating with nano enzyme and H at certain concentration 2 O 2 The administration, daily monitoring of mouse body weight, spirit, food intake, observation of mouse wound healing and recording of photographs.
The invention is further described below with reference to the accompanying drawings.
(1) Please refer to fig. 1, which shows ZnO-Co 3 O 4 In the X-ray diffraction spectrum (XRD) of (1), the diffraction peaks (100), (002), (101), (102), (110), (103), (112), (201) and (202) are assigned to ZnO, and the diffraction peaks (111), (400), (511) and (440) are assigned to Co 3 O 4 XRD spectrum shows that ZnO-Co 3 O 4 Has good crystal structure.
(2) Referring to FIG. 2, it shows a nano-enzyme bacteriostatic agent ZnO-Co 3 O 4 The results of the plate counting method for four strains of staphylococcus aureus, pseudomonas aeruginosa, escherichia coli and bacillus subtilis. Wherein, the sterilization rates of the group 2, the group 3 and the group 4 to gram-negative bacteria Escherichia coli are respectively 74.06%, 83.33% and 98.34% by calculation, which shows that 50 mu L of ZnO-Co 3 O 4 (2 mg/mL) for 5mmol/L H 2 O 2 And 50. Mu.L of 5mmol/L H 2 O 2 Can kill part of Escherichia coli, and 50 μ L of nanometer enzyme inhibitorMicrobial inoculum ZnO-Co 3 O 4 (2 mg/mL) and 50. Mu.L, 5mmol/L of H 2 O 2 Almost complete killing of E.coli is possible. The same bactericidal effect is also reflected in staphylococcus aureus, pseudomonas aeruginosa and bacillus subtilis.
(3) Wound model is created on mouse back by using Staphylococcus aureus as infection strain, see FIG. 3, for nano enzyme bacteriostatic agent ZnO-Co 3 O 4 A treatment picture for inhibiting bacteria and promoting wound healing. Wherein, when compared with the control group, the physiological saline and H are obviously used 2 O 2 The wound is treated, the capability of promoting wound healing and repair is not obvious, but the nano enzyme bacteriostatic agent ZnO-Co is used for treating the wound 3 O 4 And H 2 O 2 After treatment, the staphylococcus aureus can be effectively inhibited, and the healing of wounds can be accelerated.
In conclusion, the invention provides the heterojunction nano-enzyme bacteriostatic agent prepared by the method, and the nano-enzyme bacteriostatic agent can efficiently catalyze H 2 O 2 Decomposing to generate ROS. At low concentration of H 2 O 2 Good bacteriostatic activity against gram-negative and gram-positive bacteria is achieved in the presence. The nano enzyme bacteriostatic agent is a nano enzyme material prepared by carrying out low-temperature molten salt method and high-temperature calcination on bimetallic oxide, and can catalyze H 2 O 2 Decomposing into ROS with super strong sterilization capability, and combining the synergistic catalytic effect between the bimetallic oxides, the catalytic action of the nano enzyme is greatly enhanced, and the purpose of high-efficiency sterilization is achieved.
Example 2: preparation of nanoenzyme
(1) Zn (NO) 3 ) 2 ·6H 2 Adding O and 2-methylimidazole into water, mixing and uniformly stirring, centrifuging for 15min at a rotation speed of 9000rpm to obtain a precipitate, and washing the precipitate with deionized water;
(2) Mixing Co (NO) 3 ) 2 ·6H 2 Adding O into a methanol solution, carrying out ultrasonic treatment, adding the first precipitate, and stirring to obtain a first solution;
(3) And adding the second solution into the first solution, standing for 12h, centrifuging, washing with methanol, drying at 75 ℃ for 8h, and calcining at 400 ℃ at the temperature rise rate of 2 ℃/min for 2h to obtain the nano-enzyme.
Example 3: preparation of nanoenzyme
(1) Adding Zn (NO) 3 ) 2 ·6H 2 Adding O and 2-methylimidazole into water, mixing and uniformly stirring, centrifuging for 10min at the rotation speed of 10000rpm to obtain a precipitate, and washing the precipitate with deionized water;
(2) Mixing Co (NO) 3 ) 2 ·6H 2 Adding O into a methanol solution, carrying out ultrasonic treatment, adding the first precipitate, and stirring to obtain a first solution;
(3) And adding the second solution into the first solution, standing for 12h, centrifuging, washing with methanol, drying at 70 ℃ for 10h, and calcining at 410 ℃ at a heating rate of 3 ℃/min for 2h to obtain the nano-enzyme.
The above is only a preferred embodiment of the present invention, and it is not intended to limit the scope of the invention, and various modifications and changes will occur to those skilled in the art. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention shall be included in the scope of the present invention.
Claims (10)
1. The nanoenzyme is characterized in that the nanoenzyme is in a cubic structure, wherein C, zn and Co are uniformly dispersed in the cubic structure.
2. A preparation method of nano enzyme is characterized by comprising the following steps:
adding zinc salt and 2-methylimidazole into water, mixing and stirring uniformly, and then carrying out centrifugal treatment to obtain a precipitate;
adding cobalt salt into a methanol solution, carrying out ultrasonic treatment, then adding the first precipitate, and stirring to obtain a first solution;
adding 2-methylimidazole into a methanol solution, and performing ultrasonic treatment to obtain a second solution;
and adding the second solution into the first solution, standing, centrifuging, drying and calcining to obtain the nano enzyme.
3. The process for preparing nanoenzyme according to claim 2, wherein the zinc salt and 2-methylimidazole are added to water, mixed and stirred uniformly, and then centrifuged, and the precipitate is taken out, wherein the centrifugation parameters are as follows:
centrifuging for 10min to 15min at a rotation speed of 9000rpm to 10000 rpm.
4. The method for preparing nanoenzyme according to claim 2, wherein the step of adding the second solution to the first solution, standing, centrifuging, and drying and calcining the precipitate to obtain nanoenzyme comprises:
drying at 70-75 deg.c for 8-12 hr, and calcining at 400-410 deg.c for 2-3 hr at 2-3 deg.c/min.
5. The method for preparing nanoenzyme according to claim 2, wherein the zinc salt and 2-methylimidazole are added to water and mixed and stirred uniformly, and then the precipitate is washed with deionized water in the step of centrifuging and taking the precipitate.
6. The method for preparing nanoenzyme according to claim 2, wherein the step of adding the second solution to the first solution, standing for centrifugation, drying and calcining to obtain nanoenzyme, wherein the step of standing for centrifugation is followed by washing with methanol.
7. The method for preparing nanoenzyme according to claim 2, wherein the step of adding the second solution to the first solution, standing for centrifugation, taking out the precipitate, drying and calcining to obtain nanoenzyme, wherein the standing time is 8-24 hours.
8. A bacteriostatic agent, which is characterized by comprising nano enzyme or pharmaceutically acceptable salt thereof.
9. The bacteriostatic agent according to claim 8, wherein the bacteriostatic agent further comprises a compound solvent of nano-enzyme, and the concentration of the nano-enzyme in the bacteriostatic agent is 1 mg/mL-3 mg/mL.
10. An application of nano enzyme or its salt in preparing antibacterial agent.
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