A long-acting amyloid-like protein-mediated metal-organic framework antifouling agent, and a preparation method and application thereof

Abstract

The application discloses a kind of amyloid-like protein-mediated long-acting metal organic framework antifouling agent and preparation method and application thereof, and belongs to the field of marine antifouling agent material preparation.The antifouling agent is based on MOF with antifouling function, and amyloid-like protein self-assembles to form a dense protective shell layer on the surface of MOF.In the preparation method, metal ions and organic ligands are connected by coordination bond to form MOF structure, and MOF particles are obtained by centrifugation, collection, precipitation, washing and drying;under the action of water-soluble organic reducing substance, the disulfide bond of protein is completely opened to form unfolded protein chain, and amyloid-like protein solution is obtained by self-assembly;MOF particles are incubated in amyloid-like protein solution by simple one-step method to obtain amyloid-like protein-mediated long-acting metal organic framework antifouling agent.The application uses a new method of green, high stability, fast and efficient mild modification to prepare antifouling agent, which has stability, durability, universality and high efficiency.

Description

Technical Field

[0001] This invention belongs to the field of marine antifouling material preparation, specifically relating to a long-acting metal-organic framework antifouling agent mediated by amyloid protein, its preparation method, and its application. Background Technology

[0002] Marine biofouling has long been a concern for coastal nations worldwide, representing a persistent global challenge since the dawn of human marine activities. In the marine industry, secretions from fouling organisms such as plants, animals, and microorganisms trigger microbial corrosion, leading to decreased efficiency and shortened service life of marine equipment. The attachment of fouling organisms also increases drag on ships, reducing maneuverability, increasing fuel consumption, and causing significant economic losses. Furthermore, it introduces problems such as sonar interference, posing safety hazards, and, more seriously, damaging marine ecosystems. Marine biofouling often causes immense harm to coastal industries, maritime transport, marine pipelines, and fisheries.

[0003] Currently, antifouling agents are the most technically and economically feasible and effective method for solving biofouling problems. The use of marine antifouling agents began with early applications of highly toxic substances such as arsenic and mercury, but their antifouling effects were limited. The advent of highly efficient organotin antifouling agents in the 20th century brought a revolutionary breakthrough, significantly reducing ship drag. However, with increasing environmental awareness, it was discovered that toxic antifouling agents such as tributyltin accumulate in marine organisms, ultimately harming the marine ecosystem, leading to their ban. Subsequently, the industry shifted to antifouling agents primarily based on cuprous oxide. While this reduced environmental risks, the release and high toxicity of copper ions still threaten near-shore ecosystems. In recent years, marine antifouling agents have focused on biomimetic design and smart responsive materials. However, emerging environmentally friendly technologies lack stability in complex marine environments, resulting in poor antifouling effects and high costs that hinder industrialization. The current core challenge lies in developing new materials that are effective in preventing fouling, uniform and stable, environmentally friendly, and cost-effective, to balance the needs of marine industry and ecological protection.

[0004] Metal-organic frameworks (MOFs), composed of metal ions and organic ligands, possess abundant active sites due to their tunable porous structure and high specific surface area, demonstrating great potential in the field of antifouling: they can achieve highly efficient antifouling by releasing metal ions / antifouling ligands or loading antifouling active substances. However, most MOFs are prone to structural decomposition in complex marine environments such as seawater and acidic / alkaline conditions, exhibiting two major drawbacks: poor stability and excessively rapid release of antifouling components, severely restricting practical applications. To overcome the short-acting bottleneck of MOFs, constructing a protective shell to achieve sustained release is a key strategy. However, existing modification techniques face significant challenges: polymer brush grafting: low grafting density, large solvent consumption (such as DMF / THF), and catalyst residues leading to biotoxicity; encapsulation: MOF structures are prone to collapse, encapsulation efficiency is low, and interfacial bonding is unstable; coordination crosslinking: highly dependent on specific metal sites on the MOF surface, limiting its universality. Therefore, developing green, highly stable, rapid, and efficient mild modification methods has become the core direction for the functionalization modification of MOF antifouling agents.

[0005] Amyloid aggregates, formed by biomacromolecules such as peptides and proteins with β-sheet structures as their core, are an important type of biopolymer assembly structure found in nature. Researchers have proposed that amyloid protein assembly systems can modify the surfaces of micro / nano particles. These systems are characterized by mild and controllable assembly conditions, rapid assembly speed, and significantly reduced material and process costs, enabling successful adhesion to micro / nano particles from organic, inorganic, metallic, and living cell types. Summary of the Invention

[0006] To address the problems of existing technologies, this invention aims to propose an innovative strategy for functionalizing MOF surfaces using an amyloid-like protein-mediated surface modification system. It provides an amyloid-like protein-mediated long-lasting metal-organic framework antifouling agent, its preparation method, and its applications. This method prepares low-cost, biocompatible nanoscale protein oligomers, which spontaneously assemble into dense amyloid-like protein aggregates on the MOF surface through hydrophobic interactions, thus achieving successful modification of MOF particles. This results in a simple, controllable, long-lasting, stable, and environmentally friendly MOF antifouling agent. Furthermore, this preparation method can greenly, mildly, and universally modify various MOF materials, significantly improving stability and enhancing environmental friendliness while maintaining high antifouling activity. It overcomes the limitations of traditional MOF antifouling agents, such as short-lasting effects and high environmental costs, providing a new paradigm for developing long-lasting green marine antifouling materials.

[0007] The objective of this invention is achieved through the following technical solution:

[0008] In a first aspect, the present invention provides a long-lasting metal-organic framework antifouling agent mediated by amyloid protein, based on an antifouling MOF, wherein amyloid protein self-assembles on the surface of the MOF to form a dense protective shell.

[0009] Furthermore, MOFs with antifouling functions include MOF-5, MOF-74, HKUST-1, Ag-MOF, ZIF-8, UiO-66, PCN-222, or MIL-100, etc.

[0010] In a second aspect, the present invention also provides a method for preparing a long-acting metal-organic framework antifouling agent mediated by amyloid protein, comprising the following steps:

[0011] Preparation of MOFs:

[0012] Metal ions and organic ligands were added separately to a polar solvent and stirred to form a homogeneous solution. Then, the metal ion solution was added to the organic ligand solution and stirred until homogeneous. The metal ions and organic ligands were linked by coordinate bonds to form a MOF structure. After the reaction was completed, the MOF particles were obtained by centrifugation, collection, precipitation, washing and drying.

[0013] Preparation of amyloid-like protein (ALP):

[0014] The protein was dissolved in a buffer solution and stirred to form a homogeneous solution. Then, an excess of water-soluble organic reducing agent was added. After the two were mixed, the natural protein completely opened its disulfide bonds under the action of the reducing agent, forming unfolded protein chains, exposing abundant chemical groups, and self-assembling into nanoscale oligomers with interfacial activity. Subsequently, the amyloid protein solution was obtained through self-assembly.

[0015] Surface modification of MOF:

[0016] MOF particles were incubated in an amyloid protein solution. The unfolded nanoscale protein oligomers bound to the surface of the MOF particles through strong chemical bonds, thereby modifying the MOF surface. After the reaction, the MOF particles were centrifuged, collected, precipitated, washed, and dried to obtain an amyloid protein-mediated long-lasting metal-organic framework antifouling agent, hereinafter referred to as MOF@ALP.

[0017] Furthermore, the metal ion used in the preparation of MOF is Zn. 2+ Cu 2+ Ag + Fe 3+ or Zr 4+ ;

[0018] The organic ligands are 2-methylimidazolium (2-MI), terephthalic acid (H2BDC), porphyrin tetracarboxylic acid (H4TCPP), 2,5-dihydroxyterephthalic acid (H4DHBDC), or 1,3,5-benzenetricarboxylic acid (H3BDC).

[0019] The solvent is H2O, ethanol, or N,N-dimethylformamide (DMF).

[0020] Furthermore, in the preparation of MOF, the molar ratio of metal ions to organic ligands is 1:(4-12), the reaction time is 4h-36h, the reaction temperature is 20℃-120℃, the centrifugation conditions are 8000rpm-12000rpm for 5min-20min, and the drying temperature is 60℃-120℃.

[0021] Furthermore, the proteins used in the preparation of amyloid protein include serum albumin, lysozyme, α-lactalbumin, insulin, β-lactoglobulin, oat protein, or soy protein, etc.

[0022] The buffer solution is either Hepes buffer or Tris buffer;

[0023] The water-soluble organic reducing agent is tris(2-carboxyethyl) phosphate hydrochloride (TCEP) or L-cysteine ​​(L-Cysteine).

[0024] Furthermore, the protein concentration in the preparation of amyloid protein is 0.1 mg / mL-40 mg / mL;

[0025] The buffer concentration is 10mM-50mM.

[0026] Furthermore, a simple one-step method was used for incubation in the surface modification of MOFs;

[0027] The mass-to-volume ratio of MOF particles to amyloid protein (mg:mL) is 10:(1-10);

[0028] The incubation reaction time for MOF surface modification is 10 min-120 min;

[0029] Centrifugation conditions: 8000rpm-12000rpm for 5min-20min;

[0030] The drying temperature is 60℃-120℃.

[0031] Thirdly, the present invention also provides an application of the amyloid-like protein-mediated long-acting metal-organic framework antifouling agent in the field of marine antifouling.

[0032] Furthermore, when the amount of amyloid-mediated long-acting metal-organic framework antifouling agent added is ≥150mg / L, the bacterial survival rate against Bacillus vivax and Pseudomonas aeruginosa is <9%.

[0033] Compared with the prior art, the invention has the following beneficial effects:

[0034] 1. This invention selects MOF with inherent antifouling properties. After synthesizing MOF, amyloid protein is selected to modify its surface. The modified amyloid protein provides a natural armor for MOF, which greatly improves the stability of the antifouling agent in complex marine environments.

[0035] 2. This invention uses an amyloid protein-like system to modify MOFs, and develops a universal modification method applicable to MOFs such as MOF-5, MOF-74, HKUST-1, Ag-MOF, ZIF-8, UiO-66, PCN-222, and MIL-100, breaking through crystal form limitations.

[0036] 3. This invention selects amyloid-like proteins to modify the surface of MOFs. The amyloid-like proteins spontaneously assemble into a dense protective shell on the MOF surface. Its advantages are that the assembly conditions are mild and controllable, the speed is fast, and the material and process costs are greatly reduced.

[0037] 4. The amyloid-like protein-mediated long-acting metal-organic framework antifouling agent provided by this invention adopts a green, highly stable, rapid and efficient mild modification method, and has stability, durability, versatility and high efficiency, providing a way for the development of efficient, stable and green antifouling agents in marine engineering. Attached Figure Description

[0038] Figure 1 A scanning electron microscope (SEM) image of the ZIF-8 particles prepared in Example 1;

[0039] Figure 2 The image shows a scanning electron microscope (SEM) image of ZIF-8@ALP prepared in Example 1.

[0040] Figure 3 Transmission electron microscope (TEM) image of ZIF-8 particles prepared in Example 1;

[0041] Figure 4 Transmission electron microscope (TEM) image of ZIF-8@ALP prepared in Example 1;

[0042] Figure 5 The elemental distribution (TEM-EDS) of the ZIF-8 particles prepared in Example 1 is shown.

[0043] Figure 6 The elemental distribution (TEM-EDS) of ZIF-8@ALP prepared in Example 1 is shown.

[0044] Figure 7 X-ray diffraction (XRD) patterns of ZIF-8 particles and ZIF-8@ALP prepared in Example 1;

[0045] Figure 8Infrared spectra of ZIF-8 particles and ZIF-8@ALP prepared in Example 1;

[0046] Figure 9 This is a comparison chart of the sustained-release effects of ZIF-8 particles and ZIF-8@ALP prepared in Example 1;

[0047] Figure 10 The images are scanning electron microscope (SEM) images of the stability test of ZIF-8 particles prepared in Example 1, where: (a) is the SEM image at pH=5, (b) is the SEM image at pH=7, and (c) is the SEM image at pH=13.

[0048] Figure 11 The images are scanning electron microscope (SEM) images of the stability test of ZIF-8@ALP prepared in Example 1, where: (a) is the SEM image at pH=5, (b) is the SEM image at pH=7, and (c) is the SEM image at pH=13.

[0049] Figure 12 The graph shows the change in bacterial survival rate when ZIF-8 particles and ZIF-8@ALP prepared in Example 1 were tested for antibacterial efficacy against Bacillus vinifera.

[0050] Figure 13 The graph shows the changes in bacterial survival rate during the antibacterial effect test of ZIF-8 particles and ZIF-8@ALP prepared in Example 1 against Pseudomonas aeruginosa. Detailed Implementation

[0051] The specific embodiments of the present invention will be described in further detail below with reference to the accompanying drawings and examples.

[0052] A long-lasting metal-organic framework antifouling agent mediated by amyloid protein, based on MOFs with antifouling function, including MOF-5, MOF-74, HKUST-1, Ag-MOF, ZIF-8, UiO-66, PCN-222 or MIL-100, wherein amyloid protein self-assembles on the surface of MOF to form a dense protective shell.

[0053] A method for preparing a long-acting metal-organic framework antifouling agent mediated by amyloid protein includes the following steps:

[0054] Preparation of MOFs:

[0055] Metal ions and organic ligands were added separately to a polar solvent and stirred to form a homogeneous solution. Then, the metal ion solution was added to the organic ligand solution at a molar ratio of 1:(4-12). The mixture was stirred until homogeneous and reacted at 20℃-120℃ for 4-36 hours. The metal ions and organic ligands were linked by coordinate bonds to form a MOF structure. After the reaction was completed, the mixture was centrifuged at 8000rpm-12000rpm for 5-20 minutes. The particles were then collected, precipitated, washed, and dried at 60℃-120℃ to obtain MOF particles.

[0056] Preparation of amyloid proteins:

[0057] Proteins (0.1 mg / mL-40 mg / mL) are dissolved in Hepes buffer (10 mM-50 mM) or Tris buffer (10 mM-50 mM). The proteins include serum albumin, lysozyme, α-lactalbumin, insulin, β-lactoglobulin, oat protein, or soy protein. The mixture is stirred to form a homogeneous solution. Then, an excess of water-soluble organic reducing agent tris(2-carboxyethyl) phosphate hydrochloride or L-cysteine ​​is added. After mixing, the natural protein completely opens its disulfide bonds under the action of the reducing agent, forming unfolded protein chains, exposing abundant chemical groups, and self-assembling into interfacially active nano-sized oligomers. Subsequently, the amyloid protein solution is obtained through self-assembly.

[0058] Surface modification of MOF:

[0059] MOF particles were placed in an amyloid protein solution and incubated for 10-120 minutes using a simple one-step method. The mass-to-volume ratio of MOF particles to amyloid protein was 10:(1-10 mg / mL). The unfolded nanoscale protein oligomers were bound to the surface of the MOF particles through strong chemical bonds, thereby modifying the MOF surface. After the reaction, the mixture was centrifuged at 8000-12000 rpm for 5-20 minutes, collected, precipitated, washed, and dried at 60-120℃ to obtain an amyloid protein-mediated long-lasting metal-organic framework antifouling agent, hereinafter referred to as MOF@ALP.

[0060] The application of the aforementioned amyloid-like protein-mediated long-acting metal-organic framework antifouling agent in the field of marine antifouling shows that, when the addition amount is ≥150 mg / L, the bacterial survival rate against Bacillus vivax and Pseudomonas aeruginosa is <9%.

[0061] This invention provides a MOF-based marine antifouling agent that can select different metal ions and organic ligands. By exploring the reaction conditions and reactants, the morphology and size of the MOF can be designed, thereby achieving controllable regulation of the MOF. Amyloid-like proteins are selected to modify the surface of the MOF. The amyloid-like proteins (ALPs) are strongly bonded to the surface of the MOF particles through various chemical bonds, providing a natural armor for the MOF and greatly improving the stability of the antifouling agent in complex marine environments.

[0062] Based on this, the structure and function of the MOF itself are designed by selecting metal ions or organic ligands with bactericidal properties. The release of metals and ligands has an antifouling effect. Amyloid-like proteins are used to modify the surface of the MOF. The unfolded protein exposes a large number of functional groups, which bind to the MOF particles through strong chemical bonds, spontaneously assembling into dense amyloid-like protein aggregates on the MOF surface. The modified amyloid-like proteins provide a dense and uniform protective shell for the MOF.

[0063] Example 1

[0064] A long-lasting metal-organic framework antifouling agent mediated by amyloid protein, wherein amyloid protein self-assembles on the surface of ZIF-8 to form a dense protective shell.

[0065] A method for preparing a long-acting metal-organic framework antifouling agent mediated by amyloid protein specifically includes the following steps:

[0066] Preparation of MOFs:

[0067] Zn 2+ Zn and 2-methylimidazole were added separately to deionized water and stirred to form a homogeneous solution. 2+ The molar ratio of Zn to 2-methylimidazole is 1:8. 2+ The solution was added to a 2-methylimidazole solution and magnetically stirred until homogeneous. The mixture was reacted at 20°C for 4 hours. After the reaction was completed, the mixture was centrifuged at 8000 rpm for 5 minutes. The precipitate was collected and washed three times with deionized water. The precipitate was then dried at 60°C to obtain ZIF-8 particles.

[0068] Preparation of amyloid proteins:

[0069] 0.1 mg / mL bovine serum albumin was dissolved in 10 mM Hepes buffer and magnetically stirred to form a homogeneous solution. 50 mM of the reducing agent tris(2-carboxyethyl) phosphate hydrochloride was added to completely open the disulfide bonds of the protein. The two were then mixed to obtain ALP.

[0070] Surface modification of MOF:

[0071] 10 mg of ZIF-8 particles were placed in 10 mL of amyloid protein solution and incubated in a simple one-step method. The amyloid protein binds to the surface of ZIF-8 particles through strong chemical bonds. After reacting for 120 min, the mixture was centrifuged at 8000 rpm for 5 min, the precipitate was collected and washed three times with deionized water, and dried at 60 °C to obtain ZIF-8@ALP.

[0072] Characterization tests:

[0073] The ZIF-8 particles and ZIF-8@ALP prepared in Example 1 were tested by scanning electron microscopy, as follows: Figure 1 As shown, ZIF-8 with clear morphology and uniform particle size was successfully synthesized; Figure 2 As shown, ZIF-8@ALP is completely encapsulated by nanoscale particles and is firmly bound, with amyloid proteins spontaneously assembling on the ZIF-8 surface.

[0074] Transmission electron microscopy was performed on the ZIF-8 particles and ZIF-8@ALP prepared in Example 1, as follows: Figure 3 As shown, the synthesized ZIF-8 morphology is a rhombic dodecahedron with clear boundaries; Figure 4 As shown, the TEM image of ZIF-8@ALP and Figure 3 Compared to the ZIF-8 TEM image, it can be observed that amyloid proteins form a stable shell on the ZIF-8 surface.

[0075] Elemental distribution tests were performed on the ZIF-8 particles and ZIF-8@ALP prepared in Example 1, such as... Figure 5 As shown, the ZIF-8 elemental distribution is uniform; as Figure 6 As shown, the ZIF-8@ALP has a uniform elemental distribution, indicating that amyloid proteins are uniformly distributed on the ZIF-8 surface.

[0076] X-ray diffraction tests were performed on the ZIF-8 particles and ZIF-8@ALP prepared in Example 1, as follows: Figure 7 As shown, a comparison with the diffraction peaks on the ZIF-8 standard XRD diffraction card reveals that both ZIF-8 and ZIF-8@ALP exhibit characteristic diffraction peaks at 7.5°, 10.5°, and 12°, proving that the modification of amyloid protein did not alter the crystal form of ZIF-8.

[0077] Infrared spectroscopy tests were performed on the ZIF-8 particles, ALP, and ZIF-8@ALP prepared in Example 1. Figure 8 As shown, ZIF-8 particles and ZIF-8@ALP at 3133 cm⁻¹ -1 and 2921cm -1The absorption peaks at 424 cm⁻¹ are attributed to the stretching vibrations of the CH bonds in the methyl and imidazole rings, respectively. -1 The absorption peak at 1640 cm⁻¹ belongs to the Zn-N stretching band, and ALP and ZIF-8@ALP have absorption peaks at 1640 cm⁻¹. -1 The absorption peak at this location is a characteristic peak of the β-sheet of amyloid protein, indicating that ZIF-8 and ZIF-8@ALP were successfully synthesized.

[0078] Performance testing:

[0079] The sustained-release performance of ZIF-8@ALP prepared in Example 1 was evaluated, and the Zn content was investigated using the dialysis bag method. 2+ The release rate of Zn was determined using the following experimental steps: ZIF-8 and ZIF-8@ALP were dispersed separately in dialysis bags containing 20 mL of H2O with a molecular weight cutoff of 3500, and immediately transferred to clean beakers. 480 mL of H2O was added to submerge the dialysis bags. The mixture was magnetically stirred at room temperature. Three parallel experiments were conducted (the average value of the test results was taken). At fixed time intervals, 10 mL of the solution was pipetted and stored for later testing. Inductively coupled plasma mass spectrometry (ICP-MS) was used to measure the Zn content. 2+ Release concentration, such as Figure 9 As shown, the results indicate that ZIF-8@ALP has a significantly improved sustained-release effect compared to ZIF-8.

[0080] The stability of ZIF-8@ALP prepared in Example 1 was evaluated. The specific experimental steps are as follows: ZIF-8 and ZIF-8@ALP were dispersed in solutions with pH values ​​of 5, 7, and 13, respectively, and soaked. After reacting at 100 rpm for 24 h in a shaker, the mixture was centrifuged at 8000 rpm for 5 min, washed three times with ultrapure water, and the precipitate was collected and observed under a scanning electron microscope. Figure 10 , Figure 11 As shown, under harsh conditions, ZIF-8@ALP can still maintain morphological integrity and stability compared to ZIF-8, exhibiting superior stability.

[0081] The antibacterial properties of ZIF-8@ALP prepared in Example 1 were evaluated. The antibacterial performance of ZIF-8@ALP was assessed by examining bacterial survival rate. Bacillus vietnamese and Pseudomonas aeruginosa, typical Gram-positive bacteria from the marine environment, were selected as experimental strains. Single colonies were isolated using the streak plate method and cultured at 37°C for 12-16 hours in 2216 E or Luria-Bertani liquid medium until the bacteria reached the logarithmic growth phase. The specific steps of the antibacterial experiment are as follows: Bacteria (initial concentration 10) were... 6ZIF-8 and ZIF-8@ALP at different concentrations (25 mg / L, 50 mg / L, 100 mg / L, 125 mg / L, and 150 mg / L) were co-cultured for 24 h in simulated seawater medium containing yeast extract (1 g / L) and tryptone (1 g / L), respectively. A control group without ZIF-8 and ZIF-8@ALP was used. During the entire culture period, the optical density (OD) at 600 nm was measured. 600 Characterize bacterial growth and explore bacterial survival rate based on the following expression:

[0082] Bacterial survival rate ;

[0083] in OD of the sample processing group 600 value, OD representing the control group 600 value, and These represent the initial OD values ​​of the sample treatment group and the control group, respectively. 600 value.

[0084] like Figure 12 As shown, both ZIF-8 and ZIF-8@ALP in Example 1 exhibited antibacterial effects against Bacillus vivax. Furthermore, the bacterial survival rate decreased with increasing concentration, but ZIF-8@ALP showed superior antibacterial activity compared to ZIF-8. At a concentration of 150 mg / L, the bacterial survival rate of ZIF-8@ALP decreased to 7.57%, while that of ZIF-8 was 11.3%. Figure 13 As shown, both ZIF-8 and ZIF-8@ALP in Example 1 can exert antibacterial effects against Pseudomonas aeruginosa. As the concentration increases, the bacterial survival rate decreases, but the antibacterial effect of ZIF-8@ALP is better than that of ZIF-8. When the concentration is 150 mg / L, the bacterial survival rate of ZIF-8@ALP decreases to 8.95%, while the bacterial survival rate of ZIF-8 is 11.79%.

[0085] Example 2

[0086] A long-lasting metal-organic framework antifouling agent mediated by amyloid protein, wherein amyloid protein self-assembles on the surface of MOF-5 to form a dense protective shell.

[0087] A method for preparing a long-acting metal-organic framework antifouling agent mediated by amyloid protein specifically includes the following steps:

[0088] Preparation of MOFs:

[0089] Zn 2+Zn and terephthalic acid were added separately to N,N-dimethylformamide and stirred to form a homogeneous solution. 2+ The molar ratio of Zn to terephthalic acid is 1:10, then Zn 2+ The solution was added to a 2-methylimidazole solution and magnetically stirred until homogeneous. The mixture was reacted at 120°C for 24 hours. After the reaction was completed, the mixture was centrifuged at 8000 rpm for 10 minutes. The precipitate was collected and washed three times with N,N-dimethylformamide. The precipitate was then dried at 80°C to obtain MOF-5 particles.

[0090] Preparation of amyloid proteins:

[0091] 25 mg / mL bovine serum albumin was dissolved in 30 mM Tris buffer and magnetically stirred to form a homogeneous solution. 50 mM of the reducing agent L-cysteine ​​was added to completely break the disulfide bonds of the protein. The two were then mixed to obtain ALP.

[0092] Surface modification of MOF:

[0093] 10 mg of MOF-5 particles were placed in 4 mL of amyloid protein solution and incubated in a simple one-step method. The amyloid protein binds to the surface of MOF-5 particles through strong chemical bonds. After reacting for 60 min, the mixture was centrifuged at 8000 rpm for 15 min, the precipitate was collected and washed three times with deionized water, and then dried at 80 °C to obtain MOF-5@ALP.

[0094] Characterization tests:

[0095] Using the same characterization and testing methods as in Example 1, Example 2 successfully synthesized MOF-5 with clear morphology and uniform particle size. MOF-5@ALP was completely encapsulated by nanoscale particles, exhibiting strong binding, and amyloid-like proteins spontaneously assembled on the MOF-5 surface. The synthesized MOF-5 had an octahedral structure with clear boundaries, and a stable shell of amyloid-like proteins could be observed forming on the MOF-5 surface. The elemental distribution of MOF-5 and MOF-5@ALP was uniform, and the amyloid-like proteins were evenly distributed on the MOF-5 surface. Comparison with the diffraction peaks on the standard MOF-5 XRD diffraction card showed characteristic diffraction peaks at 6.8°, 9.7°, and 3.7°, indicating that the amyloid-like protein modification did not alter the crystal form of MOF-5. MOF-5 and MOF-5@ALP showed characteristic diffraction peaks at 1580 cm⁻¹. -1 and 1390cm -1 The absorption peaks at 750 cm⁻¹ are attributed to the asymmetric and symmetric stretching vibrations of the coordination between the terephthalic acid ligand and Zn, respectively. -1 The peak at 1640 cm⁻¹ belongs to the out-of-plane bending vibration of CH at ortho-substituted benzene ring. ALP and MOF-5@ALP peaks at 1640 cm⁻¹ are associated with this vibration. -1The absorption peak at that location is a characteristic protein peak of the β-sheet of amyloid protein.

[0096] Performance testing:

[0097] Using the same performance testing methods as in Example 1, the sustained-release effect of MOF-5@ALP in Example 2 was significantly improved compared to MOF-5. Under harsh conditions, MOF-5@ALP maintained its morphological integrity and stability compared to MOF-5, exhibiting superior stability. Both MOF-5 and MOF-5@ALP demonstrated antibacterial effects against Bacillus vivax, and while bacterial survival decreased with increasing concentration, MOF-5@ALP showed superior antibacterial efficacy at a concentration of 150 mg. At a concentration of 150 mg / L, the bacterial survival rate of MOF-5@ALP decreased to 6.98%, while that of MOF-5 was 11.03%. Both MOF-5 and MOF-5@ALP exhibited antibacterial effects against Pseudomonas aeruginosa, and the bacterial survival rate decreased with increasing concentration. However, MOF-5@ALP showed better antibacterial effects than MOF-5. At a concentration of 150 mg / L, the bacterial survival rate of MOF-5@ALP decreased to 7.07%, while that of MOF-5 was 11.37%.

[0098] Example 3

[0099] A long-lasting metal-organic framework antifouling agent mediated by amyloid protein, wherein amyloid protein self-assembles on the surface of HKUST-1 to form a dense protective shell.

[0100] A method for preparing a long-acting metal-organic framework antifouling agent mediated by amyloid protein specifically includes the following steps:

[0101] Preparation of MOFs:

[0102] Cu 2+ 1,3,5-Benzotricarboxylic acid and ethanol were added separately and stirred to form a homogeneous solution, wherein Cu 2+ The molar ratio of Cu to 1,3,5-benzenetricarboxylic acid is 1:12, then Cu 2+ The solution was added to a 1,3,5-benzenetricarboxylic acid solution and magnetically stirred until homogeneous. The mixture was then reacted at 100°C for 12 hours. After the reaction was completed, the mixture was centrifuged at 10,000 rpm for 10 minutes. The precipitate was collected, washed three times with ethanol, and dried at 60°C to obtain HKUST-1 particles.

[0103] Preparation of amyloid proteins:

[0104] Dissolve 15 mg / mL of lysozyme in 20 mM Hepes buffer, stir magnetically to form a homogeneous solution, add 50 mM of the reducing agent L-cysteine ​​to completely open the disulfide bonds of the protein, and mix the two to obtain ALP.

[0105] Surface modification of MOF:

[0106] 10 mg of HKUST-1 particles were placed in 6 mL of amyloid protein solution and incubated in a simple one-step method. The amyloid protein binds to the surface of HKUST-1 particles through strong chemical bonds. After reacting for 40 min, the mixture was centrifuged at 12000 rpm for 15 min, the precipitate was collected and washed three times with deionized water, and then dried at 60 °C to obtain HKUST-1@ALP.

[0107] Characterization tests:

[0108] Using the same characterization and testing methods as in Example 1, Example 3 successfully synthesized HKUST-1 with clear morphology and uniform particle size. HKUST-1@ALP was completely encapsulated by nanoscale particles, exhibiting strong binding, and amyloid proteins spontaneously assembled on the HKUST-1 surface. The synthesized HKUST-1 had an octahedral structure with clear boundaries, and a stable shell of amyloid proteins could be observed forming on the HKUST-1 surface. HKUST-1 and HKUST-1@ALP showed uniform elemental distribution, and amyloid proteins were uniformly distributed on the HKUST-1 surface. Comparison with the diffraction peaks on the standard XRD diffraction card for HKUST-1 showed characteristic diffraction peaks at 6.7°, 9.5°, and 11.6°, indicating that amyloid protein modification did not alter the crystal form of HKUST-1. HKUST-1 and HKUST-1@ALPP showed characteristic diffraction peaks at 1580 cm⁻¹. -1 and 1390cm -1 The absorption peaks at 750 cm⁻¹ are attributed to the asymmetric and symmetric stretching vibrations of the 1,3,5-benzenetricarboxylic acid ligand coordinated with Cu, respectively. -1 The peak at 1640 cm⁻¹ belongs to the out-of-plane bending vibration of CH4 associated with ortho-substituted benzene ring in 1,3,5-benzenetricarboxylic acid. ALP and HKUST-1@ALP peaks at 1640 cm⁻¹ are also observed. -1 The absorption peak at that location is a characteristic protein peak of the β-sheet of amyloid protein.

[0109] Performance testing:

[0110] Using the same performance testing method as in Example 1, the sustained-release effect of HKUST-1@ALP in Example 3 was significantly improved compared to HKUST-1. Under harsh conditions, MOF-5@ALP maintained its morphological integrity and stability compared to HKUST-1, exhibiting superior stability. Both HKUST-1 and HKUST-1@ALP demonstrated antibacterial effects against Bacillus vivax, and the bacterial survival rate decreased with increasing concentration. However, the antibacterial effect of HKUST-1@ALP was superior to that of HKUST-1 at a concentration of 150 mg / L. At concentration L, the bacterial survival rate of HKUST-1@ALP decreased to 7.83%, while that of HKUST-1 was 12.05%. Both HKUST-1 and HKUST-1@ALP were effective against Pseudomonas aeruginosa, and the bacterial survival rate decreased with increasing concentration. However, the antibacterial effect of HKUST-1@ALP was better than that of HKUST-1. At a concentration of 150 mg / L, the bacterial survival rate of HKUST-1@ALP decreased to 6.91%, while that of HKUST-1 was 11.78%.

[0111] Example 4

[0112] A long-lasting metal-organic framework antifouling agent mediated by amyloid protein, wherein amyloid protein self-assembles on the surface of Ag-MOF to form a dense protective shell.

[0113] A method for preparing a long-acting metal-organic framework antifouling agent mediated by amyloid protein specifically includes the following steps:

[0114] Preparation of MOFs:

[0115] Ag + 1,3,5-Benzotricarboxylic acid and N,N-dimethylformamide were added separately and stirred to form a homogeneous solution, wherein Ag + The molar ratio of Ag to 1,3,5-benzenetricarboxylic acid is 1:6. + The solution was added to a 1,3,5-benzenetricarboxylic acid solution and magnetically stirred until homogeneous. The mixture was then reacted at 120°C for 24 hours. After the reaction was completed, the mixture was centrifuged at 8000 rpm for 20 minutes. The precipitate was collected and washed three times with N,N-dimethylformamide. The precipitate was then dried at 60°C to obtain Ag-MOF particles.

[0116] Preparation of amyloid proteins:

[0117] Dissolve 35 mg / mL of oat protein in 40 mM Hepes buffer, stir magnetically to form a homogeneous solution, add 50 mM of the reducing agent L-cysteine ​​to completely open the disulfide bonds of the protein, and mix the two to obtain ALP.

[0118] Surface modification of MOF:

[0119] 10 mg of Ag-MOF particles were placed in 2 mL of amyloid protein solution and incubated in a simple one-step method. The amyloid protein was strongly bonded to the surface of Ag-MOF particles through various chemical bonds. After reacting for 20 min, the mixture was centrifuged at 8000 rpm for 5 min, the precipitate was collected and washed three times with deionized water, and then dried at 60 °C to obtain Ag-MOF@ALP.

[0120] Characterization tests:

[0121] Using the same characterization and testing methods as in Example 1, Example 4 successfully synthesized Ag-MOF with clear morphology and uniform particle size. Ag-MOF@ALP was completely encapsulated by nanoscale particles, exhibiting strong binding, and amyloid-like proteins spontaneously assembled on the Ag-MOF surface. The synthesized Ag-MOF had a cubic structure with clear boundaries, and a stable shell of amyloid-like proteins could be observed forming on the Ag-MOF surface. Ag-MOF and Ag-MOF@ALP showed uniform elemental distribution, and amyloid-like proteins were uniformly distributed on the Ag-MOF surface. Comparison with the diffraction peaks on the standard Ag-MOF XRD diffraction card showed characteristic diffraction peaks at 5.9°, 10.2°, and 13.8° for both Ag-MOF and Ag-MOF@ALP, indicating that amyloid-like protein modification did not alter the crystal form of Ag-MOF. Ag-MOF and Ag-MOF@ALP showed characteristic diffraction peaks at 1570 cm⁻¹. -1 and 1370cm -1 The absorption peaks at 720 cm⁻¹ are attributed to the asymmetric and symmetric stretching vibrations of the coordination between the 1,3,5-benzenetricarboxylic acid ligand and Ag, respectively. -1 The peak at 1640 cm⁻¹ belongs to the out-of-plane bending vibration of CH4 associated with ortho-substituted benzene ring in 1,3,5-benzenetricarboxylic acid. ALP and Ag-MOF@ALP peaks at 1640 cm⁻¹... -1 The absorption peak at that location is a characteristic protein peak of the β-sheet of amyloid protein.

[0122] Performance testing:

[0123] Using the same performance testing method as in Example 1, the Ag-MOF@ALP in Example 4 showed a significantly improved sustained-release effect compared to Ag-MOF. Under harsh conditions, Ag-MOF@ALP maintained its morphological integrity and stability compared to Ag-MOF, exhibiting superior stability. Both Ag-MOF and Ag-MOF@ALP demonstrated antibacterial effects against Bacillus vivax, and bacterial survival decreased with increasing concentration. However, Ag-MOF@ALP showed superior antibacterial efficacy compared to Ag-MOF at a concentration of 150 mg. At a concentration of 150 mg / L, the bacterial survival rate of Ag-MOF@ALP decreased to 6.58%, while that of Ag-MOF was 10.98%. Both Ag-MOF and Ag-MOF@ALP exhibited antibacterial effects against Pseudomonas aeruginosa, and the bacterial survival rate decreased with increasing concentration. However, Ag-MOF@ALP showed better antibacterial effects than Ag-MOF. At a concentration of 150 mg / L, the bacterial survival rate of Ag-MOF@ALP decreased to 6.63%, while that of Ag-MOF was 11.35%.

[0124] Example 5

[0125] A long-lasting metal-organic framework antifouling agent mediated by amyloid protein, wherein amyloid protein self-assembles on the surface of UiO-66 to form a dense protective shell.

[0126] A method for preparing a long-acting metal-organic framework antifouling agent mediated by amyloid protein specifically includes the following steps:

[0127] Preparation of MOFs:

[0128] Zr 4+ Zr and terephthalic acid were added separately to N,N-dimethylformamide and stirred to form a homogeneous solution. 4+ The molar ratio of Zr to terephthalic acid is 1:4, then Zr 4+ The solution was added to the terephthalic acid solution and magnetically stirred until homogeneous. The mixture was reacted at 120°C for 36 hours. After the reaction was completed, the mixture was centrifuged at 12000 rpm for 10 minutes. The precipitate was collected and washed three times with N,N-dimethylformamide. The precipitate was then dried at 60°C to obtain UiO-66 particles.

[0129] Preparation of amyloid proteins:

[0130] 20 mg / mL of α-lactalbumin was dissolved in 20 mM Hepes buffer and magnetically stirred to form a homogeneous solution. 50 mM of the reducing agent L-cysteine ​​was added to completely break the disulfide bonds of the protein. The two were then mixed to obtain ALP.

[0131] Surface modification of MOF:

[0132] 10 mg of UiO-66 particles were placed in 5 mL of amyloid protein solution and incubated in a simple one-step method. The amyloid protein was strongly bonded to the surface of UiO-66 particles through various chemical bonds. After reacting for 80 min, the mixture was centrifuged at 8000 rpm for 20 min, the precipitate was collected and washed three times with deionized water, and then dried at 60 °C to obtain UiO-66@ALP.

[0133] Characterization tests:

[0134] Using the same characterization and testing methods as in Example 1, Example 5 successfully synthesized UiO-66 with clear morphology and uniform particle size. UiO-66@ALP was completely encapsulated by nanoscale particles, exhibiting strong binding, and amyloid protein spontaneously assembled on the UiO-66 surface. The synthesized UiO-66 had an octahedral structure with clear boundaries, and a stable shell of amyloid protein was observed forming on the UiO-66 surface. The elemental distribution of UiO-66 and UiO-66@ALP was uniform, and the amyloid protein was evenly distributed on the UiO-66 surface. Comparison with the diffraction peaks on the standard XRD diffraction card of UiO-66 showed characteristic diffraction peaks at 7.3°, 8.4°, and 25.6°, indicating that the amyloid protein modification did not change the crystal form of UiO-66. UiO-66 and UiO-66@ALP showed characteristic diffraction peaks at 1580 cm⁻¹. -1 and 1390cm -1 The absorption peaks at 665 cm⁻¹ are attributed to the asymmetric and symmetric stretching vibrations of the coordination between the terephthalic acid ligand and Zr, respectively. -1 The peak at 1640 cm⁻¹ belongs to the stretching vibration of the Zr-O-Zr bridge bond. ALP and UiO-66@ALP are at 1640 cm⁻¹. -1 The absorption peak at that location is a characteristic protein peak of the β-sheet of amyloid protein.

[0135] Performance testing:

[0136] Using the same performance testing method as in Example 1, the sustained-release effect of UiO-66@ALP in Example 5 was significantly improved compared to UiO-66. Under harsh conditions, UiO-66@ALP maintained its morphological integrity and stability compared to UiO-66, exhibiting superior stability. Both UiO-66 and UiO-66@ALP demonstrated antibacterial effects against Bacillus vinifera, and while bacterial survival decreased with increasing concentration, UiO-66@ALP showed superior antibacterial effect at a concentration of 150 mg. At a concentration of 150 mg / L, the bacterial survival rate of UiO-66@ALP decreased to 7.32%, while that of UiO-66 was 11.55%. Both UiO-66 and UiO-66@ALP exhibited antibacterial effects against Pseudomonas aeruginosa, and the bacterial survival rate decreased with increasing concentration. However, the antibacterial effect of UiO-66@ALP was superior to that of UiO-66. At a concentration of 150 mg / L, the bacterial survival rate of UiO-66@ALP decreased to 7.07%, while that of UiO-66 was 11.39%.

[0137] Example 6

[0138] A long-lasting metal-organic framework antifouling agent mediated by amyloid protein, wherein amyloid protein self-assembles on the surface of MOF-74 to form a dense protective shell.

[0139] A method for preparing a long-acting metal-organic framework antifouling agent mediated by amyloid protein specifically includes the following steps:

[0140] Preparation of MOFs:

[0141] Zn 2+ Zn and 2,5-dihydroxyterephthalic acid were added separately to N,N-dimethylformamide and stirred to form a homogeneous solution. 2+ The molar ratio of Zn to 2,5-dihydroxyterephthalic acid is 1:8. 2+ The solution was added to a 2,5-dihydroxyterephthalic acid solution and magnetically stirred until homogeneous. The mixture was reacted at 100°C for 24 hours. After the reaction was completed, the mixture was centrifuged at 8000 rpm for 10 minutes. The precipitate was collected and washed three times with N,N-dimethylformamide. The precipitate was then dried at 60°C to obtain MOF-74 particles.

[0142] Preparation of amyloid proteins:

[0143] 40 mg / mL of β-lactoglobulin was dissolved in 50 mM Tris buffer and magnetically stirred to form a homogeneous solution. 50 mM of the reducing agent L-cysteine ​​was added to completely break the disulfide bonds of the protein. The two were then mixed to obtain ALP.

[0144] Surface modification of MOF:

[0145] 10 mg of MOF-74 particles were placed in 1 mL of amyloid protein solution and incubated in a simple one-step method. The amyloid protein was strongly bonded to the surface of MOF-74 particles through various chemical bonds. After reacting for 10 min, the mixture was centrifuged at 9000 rpm for 15 min, the precipitate was collected and washed three times with deionized water, and dried at 60 °C to obtain MOF-74@ALP.

[0146] Characterization tests:

[0147] Using the same characterization and testing methods as in Example 1, Example 6 successfully synthesized MOF-74 with clear morphology and uniform particle size. MOF-74@ALP was completely encapsulated by nanoscale particles, exhibiting strong binding, and amyloid proteins spontaneously assembled on the MOF-74 surface. The synthesized MOF-74 exhibited a hexagonal prism structure with clear boundaries, and a stable shell of amyloid proteins could be observed forming on the MOF-74 surface. The elemental distribution of MOF-74 and MOF-74@ALP was uniform, and the amyloid proteins were evenly distributed on the MOF-74 surface. Comparison with the diffraction peaks on the standard XRD diffraction card for MOF-74 showed characteristic diffraction peaks at 6.5°, 11.5°, and 12.8°, indicating that the amyloid protein modification did not alter the crystal form of MOF-74. MOF-74 and MOF-74@ALP showed characteristic diffraction peaks at 1560 cm⁻¹. -1 and 1390cm -1 The absorption peaks at 3450 cm⁻¹ are attributed to the asymmetric and symmetric stretching vibrations of the 2,5-dihydroxyterephthalic acid ligand coordinated with Zn, respectively. -1 The broad peak at 1640 cm⁻¹ belongs to the stretching vibration of the ligand hydroxyl group; ALP and MOF-74@ALP at 1640 cm⁻¹... -1 The absorption peak at that location is a characteristic protein peak of the β-sheet of amyloid protein.

[0148] Performance testing:

[0149] Using the same performance testing method as in Example 1, the sustained-release effect of MOF-74@ALP in Example 6 was significantly improved compared to MOF-74. Under harsh conditions, MOF-74@ALP maintained its morphological integrity and stability compared to MOF-74, exhibiting superior stability. Both MOF-74 and MOF-74@ALP demonstrated antibacterial effects against Bacillus vinifera, and bacterial survival decreased with increasing concentration. However, MOF-74@ALP showed superior antibacterial effect compared to MOF-74 at a concentration of 150 mg. At a concentration of 150 mg / L, the bacterial survival rate of MOF-74@ALP decreased to 6.28%, while that of MOF-74 was 10.87%. Both MOF-74 and MOF-74@ALP exhibited antibacterial effects against Pseudomonas aeruginosa, and the bacterial survival rate decreased with increasing concentration. However, MOF-74@ALP showed better antibacterial effects than MOF-74. At a concentration of 150 mg / L, the bacterial survival rate of MOF-74@ALP decreased to 6.47%, while that of MOF-74 was 11.01%.

[0150] Example 7

[0151] A long-lasting metal-organic framework antifouling agent mediated by amyloid protein, wherein amyloid protein self-assembles on the surface of PCN-222 to form a dense protective shell.

[0152] A method for preparing a long-acting metal-organic framework antifouling agent mediated by amyloid protein specifically includes the following steps:

[0153] Preparation of MOFs:

[0154] Zr 4+ Zr and porphyrin tetracarboxylic acid were added separately to N,N-dimethylformamide and stirred to form a homogeneous solution, wherein Zr 4+ The molar ratio of Zr to porphyrin tetracarboxylic acid is 1:10, then Zr 4+ The solution was added to a porphyrin tetracarboxylic acid solution and magnetically stirred until homogeneous. The mixture was reacted at 120°C for 24 hours. After the reaction was completed, the mixture was centrifuged at 8000 rpm for 10 minutes. The precipitate was collected and washed three times with N,N-dimethylformamide. The precipitate was then dried at 60°C to obtain PCN-22 particles.

[0155] Preparation of amyloid proteins:

[0156] Dissolve 25 mg / mL insulin in 20 mM Hepes buffer, stir magnetically to form a homogeneous solution, add 50 mM of the reducing agent L-cysteine ​​to completely break the disulfide bonds of the protein, and mix the two to obtain ALP.

[0157] Surface modification of MOF:

[0158] 10 mg of PCN-222 particles were placed in 4 mL of amyloid protein solution and incubated in a simple one-step method. The amyloid protein binds to the surface of PCN-222 particles through strong chemical bonds. After reacting for 80 min, the mixture was centrifuged at 9000 rpm for 15 min, the precipitate was collected and washed three times with deionized water, and then dried at 60 °C to obtain PCN-222@ALP.

[0159] Characterization tests:

[0160] Using the same characterization and testing methods as in Example 1, Example 7 successfully synthesized PCN-222 with clear morphology and uniform particle size. PCN-222@ALP was completely encapsulated by nanoscale particles, exhibiting strong binding, and amyloid-like proteins spontaneously assembled on the PCN-222 surface. The synthesized PCN-222 exhibited a rod-like structure with clear boundaries, and a stable shell of amyloid-like proteins was observed forming on the PCN-222 surface. The elemental distribution of PCN-222 and PCN-222@ALP was uniform, and the amyloid-like proteins were evenly distributed on the PCN-222 surface. Comparison with the diffraction peaks on the standard XRD diffraction card for PCN-222 showed characteristic diffraction peaks at 4.2°, 6.9°, and 9.1°, indicating that the amyloid-like protein modification did not alter the crystal form of PCN-222. PCN-222 and PCN-222@ALP showed characteristic diffraction peaks at 1600 cm⁻¹. -1 The absorption peak at 1400 cm⁻¹ is attributed to the stretching vibration of the C=N bond in the porphyrin tetracarboxylic acid ligand. -1 The peak at 1640 cm⁻¹ belongs to the symmetric stretching vibration of the coordination between the porphyrin tetracarboxylic acid ligand and Zr. ALP and PCN-222@ALP peaks at 1640 cm⁻¹ are also observed. -1 The absorption peak at that location is a characteristic protein peak of the β-sheet of amyloid protein.

[0161] Performance testing:

[0162] Using the same performance testing method as in Example 1, PCN-222@ALP in Example 7 showed a significantly improved sustained-release effect compared to PCN-222. Under harsh conditions, PCN-222@ALP maintained its morphological integrity and stability compared to PCN-222, exhibiting superior stability. Both PCN-222 and PCN-222@ALP demonstrated antibacterial effects against Bacillus vivax, and bacterial survival decreased with increasing concentration. However, PCN-222@ALP showed superior antibacterial effect compared to PCN-222 at a concentration of 150 mg. At a concentration of 150 mg / L, the bacterial survival rate of PCN-222@ALP decreased to 7.38%, while that of PCN-222 was 12.04%. Both PCN-222 and PCN-222@ALP exhibited antibacterial effects against Pseudomonas aeruginosa, and the bacterial survival rate decreased with increasing concentration. However, the antibacterial effect of PCN-222@ALP was superior to that of PCN-222. At a concentration of 150 mg / L, the bacterial survival rate of PCN-222@ALP decreased to 7.45%, while that of PCN-222 was 11.59%.

[0163] Example 8

[0164] A long-lasting metal-organic framework antifouling agent mediated by amyloid protein, wherein amyloid protein self-assembles on the surface of MIL-100 to form a dense protective shell.

[0165] A method for preparing a long-acting metal-organic framework antifouling agent mediated by amyloid protein specifically includes the following steps:

[0166] Preparation of MOFs:

[0167] Fe 3+ 1,3,5-Benzoic acid and 1,3,5-benzenetricarboxylic acid were added separately to deionized water and stirred to form a homogeneous solution, wherein Fe 3+ The molar ratio of Fe to 1,3,5-benzenetricarboxylic acid is 1:6, then Fe 3+ The solution was added to a 1,3,5-benzenetricarboxylic acid solution and magnetically stirred until homogeneous. The mixture was then reacted at 60°C for 24 hours. After the reaction was completed, the mixture was centrifuged at 12,000 rpm for 10 minutes. The precipitate was collected, washed three times with deionized water, and dried at 120°C to obtain MIL-100 particles.

[0168] Preparation of amyloid proteins:

[0169] 5 mg / mL of soy protein was dissolved in 10 mM Hepes buffer and magnetically stirred to form a homogeneous solution. 50 mM of the reducing agent L-cysteine ​​was added to completely break the disulfide bonds of the protein. The two were then mixed to obtain ALP.

[0170] Surface modification of MOF:

[0171] 10 mg of MIL-100 particles were placed in 8 mL of amyloid protein solution and incubated in a simple one-step method. The amyloid protein binds to the surface of MIL-100 particles through strong chemical bonds. After reacting for 80 min, the mixture was centrifuged at 9000 rpm for 10 min, the precipitate was collected and washed three times with deionized water, and dried at 120 °C to obtain MIL-100@ALP.

[0172] Characterization tests:

[0173] Using the same characterization and testing methods as in Example 1, Example 8 successfully synthesized MIL-100 with clear morphology and uniform particle size. MIL-100@ALP was completely encapsulated by nanoscale particles, exhibiting strong binding, and amyloid proteins spontaneously assembled on the surface of MIL-100. The synthesized MIL-100 had an octahedral structure with clear boundaries, and a stable shell of amyloid proteins could be observed forming on the surface of MIL-100. The elemental distribution of MIL-100 and MIL-100@ALP was uniform, and amyloid proteins were evenly distributed on the surface of MIL-100. Comparison with the diffraction peaks on the standard MIL-100 XRD diffraction card showed characteristic diffraction peaks at 5.2°, 5.9°, and 10.5° for both MIL-100 and MIL-100@ALP, indicating that the amyloid protein modification did not alter the crystal form of MIL-100. MIL-100 and MIL-100@ALP showed characteristic diffraction peaks at 1610 cm⁻¹. -1 and 1370cm -1 The absorption peaks at 580 cm⁻¹ are attributed to the asymmetric and symmetric stretching vibrations of the coordination of the 1,3,5-benzenetricarboxylic acid ligand with Fe, respectively. -1 The peak at 1640 cm⁻¹ belongs to the stretching vibration of the Fe-O-Fe bridging bond. ALP and MIL-100@ALP peaks at 1640 cm⁻¹ are also observed. -1 The absorption peak at that location is a characteristic protein peak of the β-sheet of amyloid protein.

[0174] Performance testing:

[0175] Using the same performance testing method as in Example 1, the sustained-release effect of MIL-100@ALP in Example 8 was significantly improved compared to MIL-100. Under harsh conditions, MIL-100@ALP maintained its morphological integrity and stability compared to MIL-100, exhibiting superior stability. Both MIL-100 and MIL-100@ALP demonstrated antibacterial effects against Bacillus vivax, and while bacterial survival decreased with increasing concentration, the antibacterial effect of MIL-100@ALP was superior to that of MIL-100 at a concentration of 150 mg. At a concentration of 150 mg / L, the bacterial survival rate of MIL-100@ALP decreased to 7.71%, while that of MIL-100 was 12.12%. Both MIL-100 and MIL-100@ALP were effective against Pseudomonas aeruginosa, and the bacterial survival rate decreased with increasing concentration. However, the antibacterial effect of MIL-100@ALP was better than that of MIL-100. At a concentration of 150 mg / L, the bacterial survival rate of MIL-100@ALP decreased to 8.03%, while that of MIL-100 was 11.84%.

Claims

1. A method for preparing a long-lasting metal-organic framework antifouling agent mediated by amyloid-like proteins, characterized in that, The method comprises the following steps: Preparation of MOF: Metal ions and organic ligands are respectively added into a polar solvent to form a uniform solution by stirring, and then the metal ion solution is added into the organic ligand solution to form a MOF structure by coordination bonding between the metal ions and the organic ligands, and the MOF particles are obtained by centrifugation, collection, precipitation, washing and drying after the reaction. Preparation of amyloid-like proteins: The protein is dissolved in a buffer to form a uniform solution by stirring, and then an excess of water-soluble organic reducing substances are added, and the natural protein is completely opened by the reducing agent to form an unfolded protein chain, expose a large number of chemical groups, and self-assemble into an interface-active nanoscale oligomer, and then the amyloid-like protein solution is obtained by self-assembly. Surface modification of MOF: The MOF particles are incubated in the amyloid-like protein solution, and the unfolded nanoscale protein oligomers are combined with the surface of the MOF particles by strong chemical bond interaction, and the long-acting metal organic framework antifouling agent mediated by amyloid-like proteins is obtained by centrifugation, collection, precipitation, washing and drying after the reaction.

2. The method for preparing a long-lasting metal-organic framework antifouling agent mediated by amyloid protein as described in claim 1, characterized in that, The metal ion in the preparation of the MOF is Zn 2+ , Cu 2+ , Ag + , Fe 3+ or Zr 4+ ; The organic ligand is 2-methylimidazole, terephthalic acid, porphyrin tetracarboxylic acid, 2,5-dihydroxyterephthalic acid or 1,3,5-benzene tricarboxylic acid. The solvent is H2O, ethanol or N,N-dimethylformamide.

3. The method for preparing a long-lasting metal-organic framework antifouling agent mediated by amyloid protein as described in claim 1, characterized in that, In the preparation of MOF, the molar ratio of metal ions to organic ligands is 1:(4-12), the reaction time is 4h-36h, the reaction temperature is 20℃-120℃, the centrifugation condition is 8000rpm-12000rpm for 5min-20min, and the drying temperature is 60℃-120℃.

4. The method for preparing a long-lasting metal-organic framework antifouling agent mediated by amyloid protein as described in claim 1, characterized in that, In the preparation of amyloid-like proteins, the protein is serum albumin, lysozyme, α-lactalbumin, insulin, β-lactoglobulin, oat protein or soybean protein. The buffer is Hepes buffer or Tris buffer. The water-soluble organic reducing substance is tris(2-carboxyethyl)phosphonium hydrochloride or L-cysteine.

5. The method for preparing a long-lasting metal-organic framework antifouling agent mediated by amyloid protein as described in claim 1, characterized in that, In the preparation of amyloid-like proteins, the protein concentration is 0.1mg / mL-40mg / mL. The buffer concentration is 10mM-50mM.

6. The method for preparing a long-lasting metal-organic framework antifouling agent mediated by amyloid protein as described in claim 1, characterized in that, In the surface modification of MOF, a simple one-step method is used for incubation. The mass-volume ratio of MOF particles to amyloid-like proteins is 10:(1-10). In the surface modification of MOF, the incubation reaction time is 10min-120min. The centrifugation condition is 8000rpm-12000rpm for 5min-20min. The drying temperature is 60℃-120℃.

7. A long-acting amyloid-like protein-mediated metal-organic framework antifouling agent, prepared by the method of claim 1, characterized in that, Based on the MOF with antifouling function, the amyloid-like proteins self-assemble on the surface of the MOF to form a dense protective shell.

8. The long-acting metal organic framework antifouling agent mediated by amyloid-like proteins according to claim 7 is applied in the field of marine antifouling.

9. The use of the amyloid-like protein-mediated long-acting metal-organic framework antifouling agent according to claim 8 in the field of marine antifouling, when the addition amount of the amyloid-like protein-mediated long-acting metal-organic framework antifouling agent is ≥ 150 mg / L, the bacterial survival rate of anti-Bacillus vietnamiensis and anti-Pseudomonas aeruginosa is < 9%.

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