CN114470149A

CN114470149A - Antibacterial solution with size-controllable protein-based nanoparticles

Info

Publication number: CN114470149A
Application number: CN202210095697.XA
Authority: CN
Inventors: 杨鹏; 李玲
Original assignee: Shaanxi Normal University
Current assignee: Shaanxi Normal University
Priority date: 2022-01-26
Filing date: 2022-01-26
Publication date: 2022-05-13
Anticipated expiration: 2042-01-26
Also published as: CN114470149B; CN117442702A

Abstract

The invention discloses an antibacterial solution with size-controllable protein-based nanoparticles, which is prepared by dissolving protein and a modifier in water, adjusting the pH value to 3-8 with NaOH, and reacting at room temperature for 5-10 hours to obtain a dispersion solution with size-controllable protein-based nanoparticles. The invention utilizes the antibacterial property of natural proteins such as lysozyme and the like, and modifies the natural proteins by a modifier, so that the antibacterial property of the natural proteins is improved. The antibacterial liquid can also inhibit the formation of bacterial biofilm and remove the formed biofilm. The antibacterial liquid can be prepared in large volume, has high biocompatibility and simple preparation method, and cannot generate drug resistance. The antibacterial agent can also be made into antibacterial coatings, gels, powder, tablets and other antibacterial agents in various forms, the use modes of the antibacterial agent are diversified, the antibacterial agent comprises oral administration, intramuscular injection, intravenous injection, patches, microneedles, atomization inhalation, spraying, gargle, toothpaste, food additives and the like, and bacterial drug resistance caused by improper use of antibiotics can be effectively avoided.

Description

Antibacterial solution with size-controllable protein-based nanoparticles

Technical Field

The invention belongs to the technical field of medical materials, and particularly relates to an antibacterial liquid with size-controllable protein-based nanoparticles, which has strong bactericidal capacity on bacteria and fungi and also has strong antiviral capacity.

Background

Bacterial infections, one of the ten leading causes of death worldwide, threaten the lives of countless people. The discovery of penicillin in the last 20 th century has created a new antibiotic antibacterial chapter, and is the most great invention in the 20 th century. Subsequent abuse of antibiotics also greatly contributed to the development of bacterial resistance. Each new antibiotic can bring new drug-resistant strains in clinical use, and even generate multi-drug-resistant 'super drug-resistant bacteria'. Antibiotic resistance is an increasing problem worldwide. According to the statistics of the world health organization, the death caused by infection in patients in the world is more than 85 percent of the death caused by the infection of drug-resistant bacteria, and the malignant result brings great burden to the life health and the economy of human beings. More than 70 million people die from drug resistant diseases each year. The threat of the "post-antibiotic age" urgently requires new antibacterial materials and methods to replace antibiotics, coping with this crisis.

Antibacterial enzymes such as lysozyme, lysostaphin and the like have gained increasing attention in recent years due to their high biocompatibility. But its development is limited by its limitations in its antimicrobial capacity. Therefore, it is a great challenge to modify it, maintain its biocompatibility, and further improve its antibacterial performance.

Disclosure of Invention

The invention aims to overcome the defect of antibiotic resistance and provide the multifunctional antibacterial liquid which is simple to prepare, is biologically safe, does not cause bacterial resistance and can effectively remove bacterial biofilms.

Aiming at the purposes, the antibacterial liquid adopted by the invention is a dispersion liquid of protein-based nanoparticles with controllable sizes, which is obtained by dissolving a modifier and proteins in water, adjusting the pH to 3-8 by NaOH, and reacting for 5-10 hours at room temperature.

The protein is one or more of lysozyme, lactalbumin, insulin, beta-lactoglobulin, bovine serum albumin, human serum albumin, alpha-lactalbumin, fibrinogen, beta-amyloid, transferrin, collagen, pepsin, keratin, myoglobin, hemoglobin, soybean protein, lactoferrin, albumin, thyrolactoglobulin, prion protein, Abeta peptide, alpha-synuclein, alpha-amylase, pepsin, horseradish peroxidase, ribonuclease A, cytochrome c, cystatin C, DNA polymerase, casein, Huntington protein and immunoglobulin light chain.

The modifier is one or more of cysteine, tris (2-carboxyethyl) phosphine hydrochloride, glutathione, mercaptoethanol, dithiothreitol, dimercaptosuccinic acid, sodium sulfite, beta-mercaptoethanol, hydrogen peroxide, ozone, sodium ferrate, trivalent cobalt salt, chlorate, potassium permanganate, persulfate, potassium dichromate, concentrated sulfuric acid, hydrochloric acid, nitric acid, hydrobromic acid, hydroiodic acid, perchloric acid, fluorine gas, chlorine gas, sodium bismuthate, periodic acid, lead dichloride, guanidine hydrochloride, urea, trifluoroethanol, hexafluoroisopropanol and trifluoroacetic acid.

The antibacterial liquid preferably has a mass ratio of protein to modifier of 1: 0.5 to 2.

The concentration of the protein dissolved in water is preferably 1 to 50mg/mL and the concentration of the modifier is preferably 1 to 50 mg/mL.

In the antibacterial liquid, the size of the protein-based nanoparticles is 10-200 nm.

And spin-coating or spray-coating the antibacterial liquid on the surfaces of different base materials to be modified to obtain the antibacterial coating. The substrate to be modified may be any material for film formation, and is not limited to any shape or material. The method specifically comprises the following steps: (1) metal material: stainless steel, titanium and its alloys, cobalt-based alloys, nickel-titanium alloys, magnesium and its alloys, zinc and its alloys, iron and its alloys, and the like; (2) inorganic materials: inorganic materials such as silicon dioxide, titanium dioxide, carbon materials, silicon, and titanium nitride; (3) high polymer material: dacron (PET), polyvinyl alcohol (PVALC), Polyethylene (PE), Polytetrafluoroethylene (PTFE), polyvinyl chloride (PVC), Polystyrene (PS), Polyurethane (PU), polypropylene (PP), polyamide, Polycarbonate (PC), polyacrylonitrile, polyacrylic acid (PAA) and its derivatives, polyetheretherketone, silicone rubber, polylactic acid, polyglycolide, polylactide, polycaprolactone, and the like; (4) natural biological material: plastic starch-based materials (PSM), sodium alginate, collagen, fibrin, sodium hyaluronate, gelatin and the like; (5) artificially synthesizing a polypeptide hydrogel material: poly-L-glutamic acid, poly-L-lysine, and the like. The type of the base material is not limited to the above materials, and may be a mixture of the above materials.

And (3) dissolving the macromolecular compound with the mass fraction of 5-30% in the antibacterial liquid to obtain the antibacterial gel. Wherein the macromolecular compound comprises one or more of starch, cellulose, gelatin, pectin, konjac glucomannan, carrageenan, Arabic gum, agar, seaweed gel, alginic acid, hyaluronic acid, chitosan, carrageenan, polysaccharide derivatives, collagen, poly-L-lysine and poly-L-glutamic acid.

Dialyzing the antibacterial liquid for 1 day, and freeze-drying to obtain an antibacterial powder preparation; and tabletting the powder preparation in a tabletting machine to obtain the antibacterial tablet preparation.

The invention has the following beneficial effects:

1. the antibacterial liquid disclosed by the invention can play an antibacterial role without adding antibiotics, can effectively avoid bacterial drug resistance caused by improper use of antibiotics, and can not cause bacteria to generate drug resistance.

2. The antibacterial liquid of the invention has net positive charge, good biocompatibility and high bactericidal performance of nano materials, and has very high antiviral performance besides good antibacterial effect on bacteria and fungi.

3. The antibacterial liquid can effectively inhibit the formation of a biological membrane, and has a good removing effect on the formed biological membrane.

4. The antibacterial liquid can inhibit the mildew of rice, traditional Chinese medicines and the like in a humid environment.

5. The antibacterial liquid has the advantages of simple preparation method, biological safety and controllable nano-particle size, can be prepared into antibacterial agents in various forms such as antibacterial coatings, gels, powder, tablets and the like, and has more diversified use modes including oral administration, intramuscular injection, intravenous injection, patches, micro-needles, atomized inhalation, spraying, gargle, toothpaste, food additives and the like.

Drawings

FIG. 1 is a size distribution diagram of nanoparticles in the antibacterial solution of example 1.

FIG. 2 is a size distribution diagram of nanoparticles in the antibacterial solution of example 2.

FIG. 3 is a graph showing the bactericidal activity against Staphylococcus aureus, Escherichia coli, and Candida tropicalis in the antibacterial liquid of example 1.

FIG. 4 is a graph showing the antiviral effect of the antibacterial solution of example 1 on adenovirus.

FIG. 5 shows the results of the hemolysis test of the antibacterial solution of example 1.

FIG. 6 shows the results of the test for the resistance of Staphylococcus aureus to the antibacterial solution of example 1.

FIG. 7 shows the results of the rice mold-proofing test using the antibacterial solution of example 1.

Detailed Description

The invention will be further described in detail with reference to the following figures and examples, but the scope of the invention is not limited to these examples.

Example 1

5mg/mL L-cysteine aqueous solution and 5mg/mL lysozyme aqueous solution were mixed in equal volume at room temperature, and then the pH was adjusted to 6.5, 7.0, and 7.5 with 1mol/L sodium hydroxide aqueous solution, respectively, and reacted at room temperature for 8 hours to obtain an antibacterial solution. As can be seen from FIG. 1, the nanoparticles of the obtained antibacterial liquid are controllable from 10nm to 100 nm.

Example 2

After mixing a 10mg/mL glutathione aqueous solution and a 10mg/mL lysozyme aqueous solution in equal volume at room temperature, adjusting the pH to 6.5, 7.0 and 7.5 respectively by using a 1mol/L sodium hydroxide aqueous solution, and reacting for 8 hours at room temperature to obtain an antibacterial solution. As can be seen from FIG. 2, the nanoparticles in the obtained antibacterial liquid can be controlled from 10nm to 200 nm.

Example 3

At room temperature, 5mg/mL bovine serum albumin aqueous solution and 5mg/mL tris (2-carboxyethyl) phosphine hydrochloride aqueous solution were mixed at a volume ratio of 2: 1, adjusting the pH to 7.0, 7.5 and 8.0 respectively by using 1mol/L sodium hydroxide aqueous solution, and reacting for 5 hours at room temperature to obtain the antibacterial solution.

Example 4

At room temperature, mixing a 5mg/mL glutathione aqueous solution and a 5mg/mL lysozyme aqueous solution according to a volume ratio of 1: 2, adjusting the pH to 7.0 by using a 1mol/L sodium hydroxide aqueous solution, and reacting at room temperature for 8 hours to obtain an antibacterial solution.

Example 5

At room temperature, mixing a 15mg/mL aqueous solution of tris (2-carboxyethyl) phosphine hydrochloride and a 15mg/mL aqueous solution of lactalbumin according to a volume ratio of 1: 2, adjusting the pH to 8.0 by using a 1mol/L sodium hydroxide aqueous solution, and reacting at room temperature for 8 hours to obtain an antibacterial solution.

Example 6

Mixing 10mg/mL L-cysteine aqueous solution and 10mg/mL lysozyme aqueous solution in equal volume at room temperature, adjusting the pH to 6.5 with 1mol/L sodium hydroxide aqueous solution, and reacting at room temperature for 8 hours to obtain the antibacterial solution. The antibacterial liquid is coated on a quartz plate in a spinning mode, and the antibacterial coating can be obtained.

Example 7

After mixing a 10mg/mL glutathione aqueous solution and a 10mg/mL lysozyme aqueous solution in equal volume at room temperature, adjusting the pH to 7.0 by using a 1mol/L sodium hydroxide aqueous solution, and reacting for 8 hours at room temperature to obtain an antibacterial solution. And spraying the antibacterial liquid on the surface of PP to obtain the antibacterial coating.

Example 8

After 5mg/mL tris (2-carboxyethyl) phosphine hydrochloride aqueous solution and 5mg/mL insulin aqueous solution were mixed in equal volumes at room temperature, the pH was adjusted to 8.0 with 1mol/L sodium hydroxide aqueous solution, and the reaction was carried out at room temperature for 8 hours to obtain an antibacterial solution. The antibacterial liquid is sprayed on the surface of the stainless steel to obtain the antibacterial coating.

Example 9

At room temperature, 5mg/mL dithiothreitol aqueous solution and 5mg/mL human serum albumin aqueous solution were mixed in a volume ratio of 2: 1, adjusting the pH to 8.0 by using a 1mol/L sodium hydroxide aqueous solution, and reacting at room temperature for 8 hours to obtain an antibacterial solution. 0.15g of gelatin is dissolved in 1mL of antibacterial solution to obtain the antibacterial gel.

Example 10

Mixing a 10mg/mL mercaptoethanol aqueous solution and a 10mg/mL lysozyme aqueous solution in equal volume at room temperature, adjusting the pH to 7.0 by using a 1mol/L sodium hydroxide aqueous solution, and reacting at room temperature for 8 hours to obtain the antibacterial liquid. 0.2g of chitosan is dissolved in 1mL of antibacterial solution to obtain the antibacterial gel.

In order to prove the beneficial effects of the invention, the inventor carries out various performance tests on the antibacterial liquid of the example 1, and the specific experiments are as follows:

1. antimicrobial Activity test

The antibacterial activity of the above antibacterial liquid was evaluated based on a colony counting method using two kinds of bacteria (staphylococcus adamantine ATCC6538 and escherichia coli ATCC25922) and one kind of fungus (candida tropicalis ATCC 1369). Before conducting the in vitro antibacterial test, the bacterial suspension was incubated overnight at 37 ℃ in a centrifuge tube of 50mL MHB medium with shaking at 70 rpm. The bacteria or fungi are allowed to grow in logarithmic phase. The cells were collected by centrifugation and washed three times with PBS buffer to remove residual medium, followed by 10⁸The concentration of CFU/mL was resuspended in PBS buffer. 100. mu.L of the resuspended suspension was added to 900. mu.L of the antibacterial solution, incubated at 37 ℃ for 8 hours, and the bacterial or fungal suspension was serially diluted and plated on MHA plates. After incubation of these MHA plates at 37 ℃ for 24 hours, the number of colonies was recorded. The antibacterial activity is represented by a bactericidal rate, which is calculated according to the following formula:

wherein C0 represents the number of colonies in blank PBS and C is the number of colonies in the antibacterial solution. The results of the experiment are shown in FIG. 3. The result shows that the antibacterial liquid has high bactericidal rate on gram-positive bacteria, gram-negative bacteria and candida tropicalis.

2. Antiviral Properties

2mL of the complete medium is used for preparing the culture medium with the density of 3-5 multiplied by 10⁴Inoculating 293T cell suspension per mL into a 6-well plate, culturing at 37 ℃ for 16-24 hours until the cell confluency reaches 30%, discarding supernatant in the well plate, adding 1mL of complete culture medium, and adding adenovirus solution (diluting adenovirus to uniform titer of 1 × 10 by using basal medium)⁸TU/mL), 10. mu.L of PBS buffer was added to three wells, and 10. mu.L of the antibacterial solution was added to the other three wells. Mixing and culturing. After 48 hours, the cells were photographed under a fluorescence microscope and the number of GFP-positive cells was counted to calculate the infection rate. The results in FIG. 4 show that the cells to which 10. mu.L of the antibacterial solution was added were not infected with viruses, indicating that the antibacterial solution had a good antiviral ability.

3. Hemolysis test

Fresh blood was centrifuged and red blood cells were collected and washed three times with PBS buffer. Red blood cells were dispersed in PBS buffer at 5% (v/v). After diluting the antibacterial solution to different concentrations, 100. mu.L of the red blood cell dispersion was added to 900. mu.L of the antibacterial solution of different concentrations, and incubated at 37 ℃ for 1 hour. A blank group was prepared by adding 100. mu.L of the red blood cell dispersion to 900. mu.L of PBS buffer, and a positive control group was prepared by adding 100. mu.L of the red blood cell dispersion to 900. mu.L of secondary water. After centrifugation, the supernatant was collected for OD measurement₅₄₀. The hemolysis rate is calculated as follows:

the results in FIG. 5 show that the hemolysis rate of the antibacterial solution is still low when the antibacterial solution reaches 10mg/mL, and thus the blood compatibility of the antibacterial solution is good.

4. Drug resistance testing

10mL of Staphylococcus aureus in logarithmic growth phase in MHB was taken and diluted to 10% with MHB⁵CFU/mL was used as the working solution. The antibacterial solution was dispersed in MHB at different concentrations, and then 100. mu.L of the mixture was mixed with 100. mu.L of the working solution and added to a 96-well plate. Placing the well plate in a microplate reader, measuring OD590 at 37 ℃, and observing bacterial growthThe situation is. Wherein the minimum concentration capable of inhibiting the growth of bacteria is the Minimum Inhibitory Concentration (MIC) of the antibacterial liquid. The bacteria were then cultured at 0.125 × MIC. The above process was repeated again with the bacteria in log phase. Fig. 6 can be obtained. The results show that the obtained antibacterial solution does not induce the bacteria to generate drug resistance.

5. Anti-mildew experiment of rice

Dividing 10g of rice into two groups on average, and soaking one group in deionized water for 0.5h to obtain a blank group; the other group was immersed in the antimicrobial solution of example 1 at pH 7.0 for 0.5h, and the test group was obtained. After being dried, the two groups of rice are placed in a constant temperature and humidity box (25 ℃, 99% RH), and the mildew condition of the two groups of rice is observed every day. As shown in FIG. 7, the rice in the blank group had significantly mildewed on day 10, with large mildew stains on day 34, and was completely covered with the mold by day 62, while the rice in the experimental group remained identical to the rice in the first day at day 62. Therefore, the antibacterial liquid can effectively inhibit the mildew of rice.

The protein in the above embodiments may be beta-lactoglobulin, alpha-lactalbumin, fibrinogen, beta-amyloid protein, transferrin, collagen, pepsin, keratin, myoglobin, hemoglobin, soy protein, lactoferrin, albumin, thyroglobulin, prion protein, Abeta peptide, alpha-synuclein, alpha-amylase, pepsin, horseradish peroxidase, ribonuclease A, cytochrome c, cystatin C, DNA polymerase, casein, Huntington protein, immunoglobulin light chain, and the modifier may be dimercaptosuccinic acid, sodium sulfite, beta-mercaptoethanol, hydrogen peroxide, ozone, sodium ferrate, cobaltous salt, chlorate, potassium permanganate, persulfate, potassium dichromate, concentrated sulfuric acid, ozone, sodium ferrate, sodium ascorbate, sodium caseinate, sodium sulfate, potassium dichromate, concentrated sulfuric acid, sodium sulfate, sodium alginate, etc., and the like, Any one or more of hydrochloric acid, nitric acid, hydrobromic acid, hydroiodic acid, perchloric acid, fluorine gas, chlorine gas, sodium bismuthate, periodic acid, lead dichloride, guanidine hydrochloride, urea, trifluoroethanol, hexafluoroisopropanol and trifluoroacetic acid.

The antibacterial liquid can be spin-coated or spray-coated on the surfaces of different base materials to obtain the antibacterial coating. The substrate may be any material for film formation that is available, and is hardly limited to shape and material. The method specifically comprises the following steps: (1) metal material: stainless steel, titanium and its alloys, cobalt-based alloys, nickel-titanium alloys, magnesium and its alloys, zinc and its alloys, iron and its alloys, and the like; (2) inorganic materials: inorganic materials such as silicon dioxide, titanium dioxide, carbon materials, silicon, and titanium nitride; (3) high polymer material: dacron (PET), polyvinyl alcohol (PVALC), Polyethylene (PE), Polytetrafluoroethylene (PTFE), polyvinyl chloride (PVC), Polystyrene (PS), Polyurethane (PU), polypropylene (PP), polyamide, Polycarbonate (PC), polyacrylonitrile, polyacrylic acid (PAA) and its derivatives, polyetheretherketone, silicone rubber, polylactic acid, polyglycolide, polylactide, polycaprolactone, and the like; (4) natural biological material: plastic starch-based materials (PSM), sodium alginate, collagen, fibrin, sodium hyaluronate, gelatin and the like; (5) artificially synthesizing a polypeptide hydrogel material: poly-L-glutamic acid, poly-L-lysine, and the like. The type of the base material is not limited to the above materials, and may be a mixture of the above materials.

The macromolecular compound with the mass fraction of 5-30% is dissolved in the antibacterial liquid to obtain the antibacterial gel. Wherein the macromolecular compound comprises one or more of starch, cellulose, gelatin, pectin, konjac glucomannan, carrageenan, Arabic gum, agar, seaweed gel, alginic acid, hyaluronic acid, chitosan, carrageenan, polysaccharide derivatives, collagen, poly-L-lysine and poly-L-glutamic acid.

Dialyzing the antibacterial solution for 1 day, and freeze-drying to obtain an antibacterial powder preparation; and tabletting the powder preparation in a tabletting machine to obtain the antibacterial tablet preparation.

Claims

1. An antiseptic solution having protein-based nanoparticles of controlled size, characterized by: the antibacterial solution is a dispersion solution of protein-based nanoparticles with controllable sizes, which is obtained by dissolving protein and a modifier in water, adjusting the pH value to 3-8 with NaOH, and reacting at room temperature for 5-10 hours.

2. The antiseptic solution with controlled size protein-based nanoparticles of claim 1, characterized by: the protein is any one or more of lysozyme, lactalbumin, insulin, beta-lactoglobulin, bovine serum albumin, human serum albumin, alpha-lactalbumin, fibrinogen, beta-amyloid, transferrin, collagen, pepsin, keratin, myoglobin, hemoglobin, soybean protein, lactoferrin, albumin, thyrolactoglobulin, prion protein, Abeta peptide, alpha-synuclein, alpha-amylase, pepsin, horseradish peroxidase, ribonuclease A, cytochrome c, cystatin C, DNA polymerase, casein, Huntington protein and immunoglobulin light chain.

3. The antiseptic solution with controlled size protein-based nanoparticles of claim 1, characterized in that: the modifier is one or more of cysteine, tris (2-carboxyethyl) phosphine hydrochloride, glutathione, mercaptoethanol, dithiothreitol, dimercaptosuccinic acid, sodium sulfite, beta-mercaptoethanol, hydrogen peroxide, ozone, sodium ferrate, trivalent cobalt salt, chlorate, potassium permanganate, persulfate, potassium dichromate, concentrated sulfuric acid, hydrochloric acid, nitric acid, hydrobromic acid, hydroiodic acid, perchloric acid, fluorine gas, chlorine gas, sodium bismuthate, periodic acid, lead dichloride, guanidine hydrochloride, urea, trifluoroethanol, hexafluoroisopropanol and trifluoroacetic acid.

4. The antiseptic solution with controllable size protein-based nanoparticles according to any one of claims 1 to 3, characterized in that: the mass ratio of the protein to the modifier is 1: 0.5 to 2.

5. The antiseptic solution with controlled size protein-based nanoparticles of claim 4, characterized by: dissolving the modifier and the protein in water to ensure that the concentration of the protein in the water is 1-50 mg/mL and the concentration of the modifier in the water is 1-50 mg/mL.

6. The antiseptic solution with controlled size protein-based nanoparticles of claim 1, characterized in that: in the antibacterial liquid, the size of the protein-based nanoparticles is 10-200 nm.

7. The antiseptic solution with controlled size protein-based nanoparticles of claim 1, characterized by: and spin-coating or spray-coating the antibacterial liquid on the surface of the substrate to be modified to obtain the antibacterial coating.

8. The antiseptic solution with controlled size protein-based nanoparticles of claim 7, characterized in that the substrate to be modified comprises any one of the following materials:

(1) metal material: stainless steel, titanium and its alloys, cobalt-based alloys, nickel-titanium alloys, magnesium and its alloys, zinc and its alloys, iron and its alloys;

(2) inorganic materials: inorganic materials such as silicon dioxide, titanium dioxide, carbon materials, silicon, and titanium nitride;

(3) high polymer material: terylene, polyvinyl alcohol, polyethylene, polytetrafluoroethylene, polyvinyl chloride, polystyrene, polyurethane, polypropylene, polyamide, polycarbonate, polyacrylonitrile, polyacrylic acid and derivatives thereof, polyether ether ketone, silicone rubber, polylactic acid, polyglycolide, polylactide and polycaprolactone;

(4) natural biological material: plastic starch-based materials, sodium alginate, collagen, fibrin, sodium hyaluronate and gelatin;

(5) artificially synthesizing a polypeptide hydrogel material: poly-L-glutamic acid, poly-L-lysine.

9. The antiseptic solution with controlled size protein-based nanoparticles of claim 1, characterized by: dissolving a macromolecular compound with the mass fraction of 5-30% in the antibacterial liquid to obtain antibacterial gel; the polymer compound comprises one or more of starch, cellulose, gelatin, pectin, konjac gum, carrageenan, arabic gum, agar, alginate jelly, alginic acid, hyaluronic acid, chitosan, carrageenan, polysaccharide derivatives, collagen, poly-L-lysine and poly-L-glutamic acid.

10. The antiseptic solution with controlled size protein-based nanoparticles of claim 1, characterized by: dialyzing the antibacterial solution and freeze-drying to obtain an antibacterial powder preparation; and tabletting the powder preparation in a tabletting machine to obtain the antibacterial tablet preparation.