Amyloid protein-based photosensitive coating material as well as preparation method and application thereof
By using amyloid-based photosensitive coating materials, the problems of unsustainable antibacterial effects, insufficient adhesion and poor weather resistance of traditional antibacterial coatings are solved, and the continuous production of bactericidal reactive oxygen groups under light conditions is achieved, providing a broad spectrum, efficient and durable antibacterial effect.
Patent Information
- Application Number
- CN202510194099.1
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-02-21
- Publication Date
- 2025-05-13
AI Technical Summary
Traditional antibacterial coatings have problems such as unsustainable antibacterial effects, insufficient adhesion, and poor weather resistance, which limits its long-term effectiveness and wide applicability in the food and medical fields.
An amyloid-based photosensitive coating material is used, which forms a coating that can continuously produce bactericidal reactive oxygen groups under light conditions by mixing an aqueous protein solution with a disulfide bond modifier and a photosensitizer after ultrasonic reaction.
The coating material can be firmly adhered to the surfaces of various substrates for a long time, has stable and long-lasting bactericidal properties, and provides a broad spectrum, efficient and durable antibacterial effect.
Smart Images
Figure CN119979005A_ABST
Abstract
Description
Technical Field
[0001] The present invention relates to the technical field of photosensitive coating materials, and in particular to an amyloid protein-based photosensitive coating material and a preparation method and application thereof. Background Art
[0002] The widespread spread of pathogenic bacteria is an important factor endangering food safety and public health. Especially during food processing, storage and transportation, cross-contamination of pathogenic microorganisms is often the key cause of food contamination and foodborne diseases. Therefore, giving materials that come into contact with pathogens a sustained and effective bactericidal property can effectively reduce the spread and cross-contamination of pathogens and protect public health and food safety. At present, traditional antimicrobial coating technology usually relies on mixing consumable antimicrobial agents (such as silver, antimicrobial essential oils, antibiotics, etc.) with different polymer solutions, and then preparing the coating by dipping, spraying, etc. This type of coating prevents the attachment and reproduction of pathogens by providing antibacterial function on the surface of the material.
[0003] However, these traditional antibacterial coatings have certain limitations. First, the consumable nature of antimicrobial agents means that the antibacterial effect will gradually weaken during use, resulting in unsustainable antibacterial properties of the coating. These antimicrobial agents usually undergo irreversible consumption after contact with bacteria, gradually reducing their effectiveness. Secondly, due to the difference in affinity between the polymer and different substrates, the coating may have weak adhesion on some substrates, causing the coating to easily fall off or peel off, thereby affecting its long-term stability. In addition, traditional antibacterial coatings have poor weather resistance and are easily affected by environmental factors (such as temperature, humidity, ultraviolet radiation, etc.), thereby reducing the durability of their antibacterial effects. These problems limit the long-term effectiveness and wide applicability of traditional antibacterial coatings in practical applications, especially in demanding environments such as food and medical fields. Therefore, the development of coating materials that are long-lasting, weather-resistant, stable and can continuously exert antibacterial effects has become an urgent need to solve these problems. Summary of the invention
[0004] The purpose of the present invention is to provide an amyloid protein-based photosensitive coating material and its preparation method and application, so as to solve the problems existing in the above-mentioned prior art. The present invention provides a preparation and application of an amyloid protein-based photosensitive antibacterial coating material, which can firmly adhere to the surface of various substrates for a long time, and can continuously generate bactericidal active oxygen groups under light conditions, and has stable and lasting bactericidal performance. Existing antibacterial coatings have the problems of poor application versatility, insufficient adhesion, and short-lasting antibacterial effect.
[0005] To achieve the above object, the present invention provides the following solutions:
[0006] One of the technical solutions of the present invention is a method for preparing an amyloid-based photosensitive coating material, comprising the following steps:
[0007] The protein aqueous solution is mixed with a disulfide bond modifier and the pH is adjusted. A photosensitizer is added and subjected to ultrasonic reaction to obtain a mixed solution, which is the amyloid protein-based photosensitive coating material.
[0008] The second technical solution of the present invention is the amyloid-based photosensitive coating material prepared by the preparation method.
[0009] The third technical solution of the present invention is the application of the amyloid protein-based photosensitive coating material in antibacterial materials.
[0010] A fourth technical solution of the present invention is an antibacterial material, which includes the amyloid-based photosensitive coating material and a substrate.
[0011] Based on the above technical solution, the present invention has the following technical effects:
[0012] The amyloid-based photosensitive antibacterial coating material provided by the present invention can firmly and persistently adhere to the surface of various substrates, and has the advantage of wide application. The photosensitive antibacterial coating prepared by the present invention can continuously and efficiently generate ROS groups under visible light, providing broad-spectrum, efficient and durable antibacterial properties. BRIEF DESCRIPTION OF THE DRAWINGS
[0013] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.
[0014] Figure 1 This is a morphological diagram of the amyloid-based photosensitive antibacterial coating on the glass surface of the present invention.
[0015] Figure 2 This is a ThT fluorescence staining image of the amyloid protein-based photosensitive antibacterial coating on the glass surface of the present invention.
[0016] Figure 3 The figure is a quantitative determination result of hydroxyl radicals of the amyloid-based photosensitive antibacterial coating on the surfaces of glass, stainless steel and polystyrene under simulated sunlight.
[0017] Figure 4 The figure is a quantitative determination result of hydrogen peroxide of the amyloid-based photosensitive antibacterial coating on the surfaces of glass, stainless steel and polystyrene under simulated sunlight.
[0018] Figure 5The figure is a quantitative measurement result of singlet oxygen of the amyloid-based photosensitive antibacterial coating on the surface of glass, stainless steel and polystyrene under simulated sunlight.
[0019] Figure 6 This is a diagram showing the sterilization effect of the amyloid protein-based photosensitive antibacterial coating on the glass surface of the present invention under simulated sunlight.
[0020] Figure 7 This is a diagram showing the sterilization effect of the amyloid protein-based photosensitive antibacterial coating on the surface of stainless steel under simulated sunlight.
[0021] Figure 8 This is a diagram showing the sterilization effect of the amyloid-based photosensitive antibacterial coating on the surface of polystyrene under simulated sunlight. DETAILED DESCRIPTION
[0022] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but should be understood as a more detailed description of certain aspects, features, and embodiments of the present invention.
[0023] It should be understood that the terms described in the present invention are only for describing special embodiments and are not intended to limit the present invention. In addition, for the numerical range in the present invention, it should be understood that each intermediate value between the upper and lower limits of the scope is also specifically disclosed. Each smaller range between the intermediate value in any stated value or stated range and any other stated value or intermediate value in the described range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.
[0024] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of a conflict with any incorporated document, the content of this specification shall prevail.
[0025] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments of the present invention description without departing from the scope or spirit of the present invention. Other embodiments derived from the present invention description will be apparent to those skilled in the art. The present application description and examples are exemplary only.
[0026] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.
[0027] The technical solutions described in the present invention, unless otherwise specified, are all conventional solutions in the art, and the reagents or raw materials used, unless otherwise specified, are purchased from commercial channels or have been disclosed.
[0028] An embodiment of the present invention provides a method for preparing an amyloid-based photosensitive coating material, comprising the following steps:
[0029] The protein aqueous solution is mixed with a disulfide bond modifier and the pH is adjusted. A photosensitizer is added and subjected to ultrasonic reaction to obtain a mixed solution, which is the amyloid protein-based photosensitive coating material.
[0030] In some specific embodiments, the concentration of the aqueous protein solution is 1-50 mg / mL; the final concentration of the disulfide bond modifier is 10-100 mM; and the final concentration of the photosensitizer is 1-10 mg / mL.
[0031] In some specific embodiments, the protein is selected from at least one of bovine serum albumin, lactoferrin, lysozyme, insulin, α-lactalbumin, human serum albumin, fibrinogen, β-amyloid protein, Aβ peptide, prion protein, α-synuclein, cystatin C, huntingtin protein, immunoglobulin, or soy protein isolate;
[0032] The disulfide bond modifier is selected from at least one of cysteine, tris(2-carboxyethyl)phosphine hydrochloride, glutathione, mercaptoethanol, dithiothreitol, dimercaptosuccinic acid, sodium sulfite, β-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, chlorine, sodium bismuthate, periodic acid, lead dichloride, guanidine hydrochloride, urea, trifluoroethanol, hexafluoroisopropanol or trifluoroacetic acid;
[0033] The photosensitizer is selected from menadione and its derivatives, riboflavin and its derivatives, porphyrins, phthalocyanines, anthraquinones, metal complexes such as platinum, palladium, rhodium, TiO 2 , ZnO, CdS, ZnS, graphene, carbon nanotubes, graphene oxide or Mxene.
[0034] In some specific embodiments, the method for adjusting pH is: adjusting pH to 1-14 with NaOH;
[0035] The ultrasonic reaction is carried out at 40 kHz for 10 to 60 minutes.
[0036] The embodiment of the present invention also provides an amyloid protein-based photosensitive coating material prepared by the preparation method.
[0037] The embodiment of the present invention also provides the use of the amyloid protein-based photosensitive coating material in antibacterial materials.
[0038] An embodiment of the present invention further provides an antibacterial material, which includes the amyloid-based photosensitive coating material and a substrate.
[0039] In some specific embodiments, the substrate is selected from an alloy of at least one of magnesium, aluminum, gold, silver, platinum, nickel, copper and titanium.
[0040] Or at least one selected from silicon, glass, quartz, mica, porcelain, polymethyl methacrylate, polypropylene, polyethylene, polyethylene terephthalate, polypropylene, polycarbonate, photosensitive polyimide, cellulose film, polytetrafluoroethylene film, polyvinylidene fluoride film, nylon film, wood and paper.
[0041] In some specific embodiments, the thickness of the amyloid-based photosensitive coating material on the substrate is 10 to 200 nm.
[0042] The embodiment of the present invention also provides the amyloid protein-based photosensitive coating material for controlling pathogenic bacteria on food contact surfaces and medical device surfaces.
[0043] The photosensitive coating provided by the present invention is formed by blending a photosensitizer and an amyloid protein, wherein the amyloid protein is an oligomer induced by a disulfide bond reducing agent. The present invention can form a stable coating on the surface of a common solid material by simple dipping or spraying. Compared with traditional antibacterial coatings, the preparation process of the amyloid protein-based photosensitive coating material of the present invention is convenient and simple, the reaction conditions are mild and environmentally friendly, the biosafety is high, the selectivity of the base material is wide, the photostability is good and the antibacterial efficiency is high. It can be widely used in the antibacterial modification of food contact surfaces and medical devices.
[0044] Example 1
[0045] (1) At room temperature, 500 mg of bovine albumin (BSA) was dissolved in 50 mL of 50 mM tris(2-carboxyethyl)phosphine (TECP) solution, the pH was adjusted to 5 with NaOH, 500 mg of menaquinone sodium bisulfite (MSB) photosensitizer was added, and water bath ultrasound (40 kHz) was applied for 10 min to obtain a mixed solution.
[0046] (2) A clean glass disc with a diameter of 2 cm was immersed in an excess of the mixed solution obtained in step (1), and was incubated at room temperature in the dark for 16 hours to allow the BSA protein to undergo amyloid-like degeneration and to be composite-modified with the MSB photosensitizer on the surface of the glass disc. After ultrasonic washing for 10 minutes, a protein-based photosensitive coating with strong adhesion properties was formed on the glass disc, and the coating thickness was about 20 μm. The scanning electron microscope image of its surface morphology is shown in FIG. Figure 1As shown, the protein complex can be seen adhering to the surface of the glass slide in the form of nanoparticles.
[0047] (3) The protein-based photosensitive coated glass sheet obtained in step (2) was immersed in an excess of 1 mM thioflavin T (ThT) fluorescent dye and incubated in the dark for 30 minutes, then the glass sheet was rinsed with excess water and naturally dried. The ThT fluorescent staining microscope image of the surface was as follows: Figure 2 As shown, obvious green fluorescence signals can be seen, indicating that most of the proteins undergo phase transition and form a large amount of β-sheet secondary structure.
[0048] Example 2
[0049] (1) At room temperature, 500 mg of bovine albumin (BSA) was dissolved in 50 mL of 50 mM TECP solution, the pH was adjusted to 5 with NaOH, 500 mg of menaquinone sodium bisulfite (MSB) photosensitizer was added, and water bath ultrasound (40 kHz) was applied for 10 min to obtain a mixed solution.
[0050] (2) A clean stainless steel disc with a diameter of 2 cm was immersed in an excess of the mixed solution obtained in step 1, and incubated at room temperature in the dark for 16 hours to allow the BSA protein to undergo a phase transition and be composite-modified with the MSB photosensitizer on the surface of the stainless steel disc. After ultrasonic washing for 10 minutes, a protein-based photosensitive coating with strong adhesion properties was formed on the stainless steel disc, and the coating thickness was about 20 μm.
[0051] Example 3
[0052] (1) At room temperature, 500 mg of bovine albumin (BSA) was dissolved in 50 mL of 50 mM TECP solution, the pH was adjusted to 5 with NaOH, 500 mg of menaquinone sodium bisulfite (MSB) photosensitizer was added, and water bath ultrasound (40 kHz) was applied for 10 min to obtain a mixed solution.
[0053] (2) A clean polystyrene plastic disc with a diameter of 2 cm was immersed in an excess of the mixed solution obtained in step 1, and incubated at room temperature in the dark for 16 hours to allow the BSA protein to undergo a phase transition and be composite-modified with the MSB photosensitizer on the surface of the stainless steel sheet. After ultrasonic washing for 10 minutes, a protein-based photosensitive coating with strong adhesion properties was formed on the polystyrene plastic sheet, and the coating thickness was about 20 μm.
[0054] Example 4
[0055] Figure 3 , 45 is a comparison chart of the production of hydroxyl radicals, hydrogen peroxide and singlet oxygen by the protein-based photosensitive coatings prepared in Examples 1 to 3 under simulated sunlight (D65) illumination conditions. It can be seen from the figure that the three typical ROS are only produced under illumination conditions and not under dark conditions. The production rates of hydroxyl radicals in Examples 1, 2 and 3 are 7.35, 11.33 and 13.00 μg / h / piece, respectively, the production rates of hydrogen peroxide are 4.08, 4.96 and 5.53 μg / h / piece, respectively, and the production rates of singlet oxygen are 5.05, 2.20 and 2.58 μg / h / piece, respectively. This shows that the MSB photosensitizer can firmly adhere to the surfaces of various substrates through proteins and maintain its photosensitivity, and can continuously produce a variety of ROS groups under illumination conditions.
[0056] Example 5
[0057] Figure 6 , 7 8 are bactericidal kinetics diagrams of the protein-based photosensitive coatings prepared in Examples 1 and 2 against typical foodborne pathogenic bacteria Escherichia coli (ATCC) under simulated sunlight (D65) illumination conditions. The experimental group has a photosensitive coating, and the control group is the same as the experimental group, but without the addition of MSB. As can be seen from the figure, Example 2 can achieve a bactericidal effect of about 6log CFU / piece (99.9999%) after 15 minutes of D65 illumination. While Example 1 requires 45 minutes of D65 illumination to achieve the same amount of bactericidal activity. This indicates that the bactericidal activity of hydroxyl radicals may be higher than that of singlet oxygen.
[0058] Obviously, the above embodiments of the present invention are merely examples for clearly illustrating the present invention, and are not intended to limit the implementation methods of the present invention. For ordinary technical users in the relevant field, other different forms of changes or modifications can be made on the basis of the above description. It is not necessary and impossible to list all the implementation methods here. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the present invention should be included in the protection scope of the claims of the present invention.
Claims
1. A method for preparing an amyloid-based photosensitive coating material, characterized in that: The following steps are involved: The protein aqueous solution is mixed with a disulfide bond modifier and the pH is adjusted. A photosensitizer is added and subjected to ultrasonic reaction to obtain a mixed solution, which is the amyloid protein-based photosensitive coating material.
2. The preparation method according to claim 1, characterized in that: The concentration of the protein aqueous solution is 1-50 mg / mL; the final concentration of the disulfide bond modifier is 10-100 mM; and the final concentration of the photosensitizer is 1-10 mg / mL.
3. The preparation method according to claim 1, characterized in that: The protein is selected from at least one of bovine serum albumin, lactoferrin, lysozyme, insulin, α-lactalbumin, human serum albumin, fibrinogen, β-amyloid protein, Aβ peptide, prion protein, α-synuclein, cystatin C, huntingtin protein, immunoglobulin or soy protein isolate; The disulfide bond modifier is selected from at least one of cysteine, tris(2-carboxyethyl)phosphine hydrochloride, glutathione, mercaptoethanol, dithiothreitol, dimercaptosuccinic acid, sodium sulfite, β-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, chlorine, sodium bismuthate, periodic acid, lead dichloride, guanidine hydrochloride, urea, trifluoroethanol, hexafluoroisopropanol or trifluoroacetic acid; The photosensitizer is selected from at least one of menadione and its derivatives, riboflavin and its derivatives, porphyrins, phthalocyanines, anthraquinones, metal complexes such as platinum, palladium, rhodium, TiO2, ZnO, CdS, ZnS, graphene, carbon nanotubes, graphene oxide or Mxene.
4. The preparation method according to claim 1, characterized in that: The method for adjusting pH is: adjusting pH to 1-14 with NaOH; The ultrasonic reaction is carried out at 40 kHz for 10 to 60 minutes.
5. The amyloid-based photosensitive coating material prepared by the preparation method according to any one of claims 1 to 4.
6. Use of the amyloid-based photosensitive coating material as claimed in claim 5 in antibacterial materials.
7. An antibacterial material, characterized in that: The antibacterial material comprises the amyloid-based photosensitive coating material according to claim 5 and a substrate.
8. The antibacterial material according to claim 7, characterized in that: The substrate is selected from at least one alloy of magnesium, aluminum, gold, silver, platinum, nickel, copper and titanium, Or at least one selected from silicon, glass, quartz, mica, porcelain, polymethyl methacrylate, polypropylene, polyethylene, polyethylene terephthalate, polypropylene, polycarbonate, photosensitive polyimide, cellulose film, polytetrafluoroethylene film, polyvinylidene fluoride film, nylon film, wood and paper.