WO2022021497A1

WO2022021497A1 - Self-assembled polypeptide molecule having bacterial flocculation and antibacterial properties and use thereof

Info

Publication number: WO2022021497A1
Application number: PCT/CN2020/110115
Authority: WO
Inventors: 李新明; 张纪坤
Original assignee: 苏州大学
Priority date: 2020-07-29
Filing date: 2020-08-20
Publication date: 2022-02-03
Also published as: CN111909239A; CN111909239B

Abstract

Disclosed are a self-assembled polypeptide molecule having bacterial flocculation and antibacterial properties and use thereof. On the basis of abundant non-covalent action capability of tryptophan molecule and good interaction capability with a lipid membrane, a self-assembled polypeptide molecule capable of flocculating both gram positive bacteria and gram negative bacteria and having antibacterial activity is designed and synthesized. The series of molecules of the present invention can be subjected to self-assembly gelation in an aqueous solution having a pH of 7.4 under the action of an alkaline phosphatase to form a stable hydrogel material, and the dephosphorylation and self-assembly of the molecules can enhance the flocculation activity thereof.

Description

Self-assembled polypeptide molecules with bacterial flocculation and antibacterial properties and their applications

technical field

The invention relates to a self-assembled polypeptide molecule with bacterial flocculation and antibacterial properties and its application, belonging to the technical field of flocculants.

Background technique

Flocculation is the process of bringing together colloids or other particles suspended in a liquid to form larger particles (or flocs) to facilitate the settling of these particles from a stable suspension. The mechanism of bacterial flocculation is closely related to the mechanism of bacterial adhesion to the surface. For example, the flocculation of bacteria by polymers can be regarded as a process in which bacteria adhere to each other through flocculants adsorbed on their surfaces. The current research explains the phenomenon of bacterial flocculation. Mainly for the charge attraction neutralization and bridging between the flocculant and bacteria. Suspended colloidal particles and pathogenic microorganisms are usually the main pollutants in raw water. Therefore, effective turbidity removal and effective sterilization are the main tasks of drinking water treatment. Traditional processes for producing drinking water include coagulation/flocculation, sedimentation, sand filtration and disinfection. By adding a flocculant, the turbidity can be effectively removed, thereby initiating the coagulation/flocculation process, thereby clarifying the water. Disinfectants such as chlorine commonly used in water treatment plants can effectively control and kill most microorganisms. However, with the rapid development of global industry, the quality of raw water has seriously deteriorated. In order to meet the national drinking water hygiene standards, flocculants and disinfectants must be used in large quantities. Not only does this mean higher disposal costs, but it also means a greater risk of secondary contamination with adverse consequences for human health, such as disinfection by-products produced during chlorine disinfection. In addition, different types of chemicals are required in different water treatment procedures. For example, during coagulation/flocculation and disinfection, the basic steps of drinking water treatment, coagulants/flocculants and disinfectants should be used accordingly. However, traditional medicaments with a single function are often difficult to handle and require high doses, thereby increasing the final cost. Moreover, there may be mutual inhibition between the different chemicals used, reducing the actual treatment efficiency. Therefore, the application of water treatment agents with two or more functions such as flocculation and antibacterial has scientific and practical significance. Flocculation is usually the first step in water purification, and it has become one of the most commonly used and important technologies in water treatment technology due to its simplicity, economy, and high efficiency. The flocculant is one of the key factors that determines the flocculation effect and even the whole water treatment effect. Therefore, it is of great significance to develop new flocculants that are efficient, low-cost, environmentally friendly, and multifunctional.

At present, flocculants used to treat microorganisms such as bacteria in water can generally be divided into three categories: i) inorganic flocculants, such as alum and polyaluminum chloride; ii) synthetic organic flocculants, such as polyacrylamide (PAM) and polymer Ethyleneimine (PEI); iii) Natural polymer flocculants, such as chitosan, starch, sodium alginate, cellulose, lignin, tannin, etc.

Among the commonly used flocculants in water treatment, traditional flocculants, such as inorganic metal flocculants and synthetic organic polymer flocculants, have no obvious bactericidal effect. In addition, residual metal ions or harmful polymerized monomers are released into the target water during use, which inherently carry certain health risks. For example, acrylamide monomer is carcinogenic and neurotoxic to humans, and aluminum salt flocculants can induce Alzheimer's disease. Most of the natural polymer flocculants have poor water solubility, such as cellulose is insoluble in water, chitosan is only soluble in acidic water, starch is insoluble in cold water, etc.; starch), its flocculation and antibacterial properties are often achieved or enhanced by complex chemical grafting modifications using cationic groups such as quaternary ammonium salt derivatives.

In addition, although there have been a lot of reports on natural or synthetic polypeptides with antibacterial activity, there are still very few reports on the flocculation or aggregation of bacteria caused by polypeptides, especially self-assembling short peptides.

SUMMARY OF THE INVENTION

In view of the current common bacterial flocculation in water treatment, inorganic metal flocculants and synthetic organic polymer flocculants have no antibacterial activity and are potentially toxic, and natural polymer flocculants generally have poor water solubility, often requiring additional chemical modification to enhance bacterial flocculation and The problem of antibacterial activity, and there are still few reports on polypeptides with both bacterial flocculation and antibacterial activities, especially self-assembled short peptides. Based on the interaction ability, a self-assembling polypeptide molecule that can flocculate Staphylococcus aureus (Gram-positive bacteria) and Escherichia coli (Gram-negative bacteria) and has antibacterial activity was designed and synthesized.

The first object of the present invention is to provide a self-assembled polypeptide molecule with bacterial flocculation and antibacterial properties, which has the following general formula:

Wherein, R ₁ and R ₂ are respectively selected from

one of the.

The second object of the present invention is to provide a method for preparing the self-assembled polypeptide molecule. The method is to use a solid-phase synthesis method to sequentially connect phosphorylated tyrosine, lysine, amino acid to be synthesized and 2-naphthalene. Acetic acid is used to synthesize the self-assembling polypeptide molecule, and the amino acid to be synthesized includes at least one tryptophan.

The third object of the present invention is to provide the application of the self-assembled polypeptide molecule in an antibacterial flocculant.

Further, the application is to add the self-assembled polypeptide molecule solution to the solution to be treated, and stir to perform flocculation.

Further, the added concentration of the self-assembling polypeptide molecule is not less than 25 μg/mL.

Further, the application further includes adding alkaline phosphatase to the self-assembling polypeptide molecule solution for incubation before adding the self-assembling polypeptide molecule solution to the solution to be treated.

In the bacterial flocculation application of self-assembled polypeptide molecules, the self-assembled polypeptide molecules are generally directly used to interact with Staphylococcus aureus and Escherichia coli to achieve significant bacterial flocculation; alkaline phosphatase can also be added. When the concentration is low, the molecules are not enough to form a gel. After dephosphorylation by alkaline phosphatase, self-assembly occurs to form a nanofibrous structure. There is no need to form a gel during the process.

The fourth object of the present invention is to provide the application of the self-assembled polypeptide molecule in the preparation of hydrogel materials.

Further, the hydrogel material is prepared by adding alkaline phosphatase to the self-assembling polypeptide molecule solution.

Further, the pH of the self-assembled polypeptide molecule solution is 6-8.

The fifth object of the present invention is to provide a hydrogel material prepared from the self-assembled polypeptide molecule.

Beneficial effects of the present invention:

The series of molecules of the present invention can undergo self-assembly and gelation in an aqueous solution of pH 7.4 under the action of alkaline phosphatase to form a stable hydrogel material, and the dephosphorylation and self-assembly of the molecules can enhance their flocculation activity. The self-assembled polypeptide molecule of the present invention has good bacterial flocculation and antibacterial activities, has a single component, and does not require complicated additional chemical modification. Due to its biological origin and short sequence, this series of molecules has the advantages of good biocompatibility, biodegradability, and easy synthesis and production.

Description of drawings

Fig. 1 is the solid phase synthesis step of polypeptide molecule;

Figure 2 shows the gelation process of polypeptide molecules, (A, C, E) gel factor precursors FWYp, WWYp, WFYp at the lowest gel concentration (pH=7.4) and alkaline phosphatase (10units/mL) Triggered formation of FWY, WWY, and WFY supramolecular hydrogels (B, D, F);

Fig. 3 is the dynamic rheological test chart of polypeptide hydrogel (1.0wt%); (A) rheological mechanics test under stress sweep; (B) rheological mechanics test under rate sweep;

Fig. 4 is the bacterial flocculation effect of different concentrations of polypeptide molecule solutions on (A) Staphylococcus aureus and (B) Escherichia coli;

Fig. 5 is the antibacterial activity of different concentrations of polypeptide molecules on Staphylococcus aureus (A, B) and Escherichia coli (C, D) dilution coating plate colony count statistics and pictures of bacterial colonies formed on agar plates;

Fig. 6 is (A) the cytotoxicity of different concentrations of polypeptide molecules and (B) 1wt% polypeptide self-assembled hydrogel;

Fig. 7 shows the change of bacterial flocculation rate with time for each sample at the same concentration (200 μg/mL) for (A) Staphylococcus aureus and (B) Escherichia coli.

detailed description

The present invention will be further described below with reference to the accompanying drawings and specific embodiments, so that those skilled in the art can better understand the present invention and implement it, but the embodiments are not intended to limit the present invention.

Example 1: Solid Phase Synthesis of Polypeptide Molecules

The synthesis process of the polypeptide molecule is shown in Figure 1. In the process of molecular synthesis, according to the sequence of the designed molecule, phosphorylated tyrosine [Fmoc-Tyr(H ₂ PO ₃ )-OH)], lysine [Fmoc-Lys(Boc )-OH], phenylalanine (Fmoc-Phe-OH) or tryptophan [Fmoc-Trp(Boc)-OH], tryptophan [Fmoc-Trp(Boc)-OH] or phenylalanine ( Fmoc-Phe-OH) and 2-naphthaleneacetic acid molecular structural unit, the specific preparation process is as follows:

i) Quickly weigh 1000mg of 2-chlorotrityl chloride resin (mesh 100～200 mesh, degree of substitution 0.8-1.5mmol/g) in a dry solid-phase synthesis reactor, add an appropriate amount of anhydrous dichloride Methane (obtained from analytically pure dichloromethane by atmospheric distillation), swelled the resin under the action of nitrogen for 30 minutes, then, removed anhydrous dichloromethane and added anhydrous DMF (HPLC grade) to wash the resin three times;

ii) Weigh 1691.9 mg (3.5 mmol) of Fmoc-Tyr(H ₂ PO ₃ )-OH and dissolve it in anhydrous DMF, then add 1.52 mL (8.75 mmol) of DIEA, dissolve it by ultrasonic and mix it well and add it to the reactor , reacted for 2 hours under nitrogen agitation, removed the reaction solution and washed the resin three times with anhydrous DMF;

iii) Add the Block solution prepared by DCM:MeOH:DIEA=16:3:1 (v/v) to the resin, and remove the Block solution after 10 minutes of nitrogen shock reaction. This step is performed twice. Afterwards, the Block solution was removed and the resin was washed three times with anhydrous DMF;

iv) prepare a 20% piperidine DMF solution, add an appropriate volume to the resin, pass nitrogen to shake for 30 minutes, and sequentially wash the resin three times with 20% piperidine DMF solution and anhydrous DMF;

v) Weigh 1639.9 mg (3.5 mmol) of Fmoc-Lys(Boc)-OH and 1327.3 mg (3.5 mmol) of HBTU, dissolve them in an appropriate amount of anhydrous DMF by ultrasonic, add 1.52 mL of DIEA, and quickly add them to the reactor after mixing evenly. After 2 hours of nitrogen shaking, the reaction solution was removed and the resin was washed three times with anhydrous DMF;

vi) adding an appropriate amount of 20% piperidine DMF solution to the resin, and after 30 minutes of nitrogen oscillation, wash the resin three times with 20% piperidine DMF solution and anhydrous DMF successively;

vii) Weigh Fmoc-Phe-OH 1356.0 mg or Fmoc-Trp(Boc)-OH 1843.0 mg (3.5 mmol) and HBTU 1327.3 mg (3.5 mmol), dissolve them in an appropriate amount of anhydrous DMF, add DIEA 1.52 mL, and mix well Then, it was quickly added to the reactor, and after 2 hours of nitrogen shock, the reaction solution was removed and the resin was washed three times with anhydrous DMF;

viii) adding an appropriate amount of 20% piperidine DMF solution to the resin, and after 30 minutes of nitrogen oscillation, wash the resin three times with 20% piperidine DMF solution and anhydrous DMF in turn;

ix) Weigh Fmoc-Trp(Boc)-OH 1843.0mg or Fmoc-Phe-OH 1356.0mg (3.5mmol) and HBTU 1327.3mg (3.5mmol) and dissolve them in an appropriate amount of anhydrous DMF by ultrasonic, add DIEA 1.52mL, mix well Then, it was quickly added to the reactor, and after 2 hours of nitrogen shock, the reaction solution was removed and the resin was washed three times with anhydrous DMF;

x) Add an appropriate amount of 20% piperidine DMF solution to the resin, and after 30 minutes of nitrogen shock, wash the resin three times with 20% piperidine DMF solution and anhydrous DMF successively;

xi) Weigh 651.7 mg (3.5 mmol) of 2-naphthalene acetic acid and 1327.3 mg (3.5 mmol) of HBTU and dissolve them in an appropriate amount of anhydrous DMF by ultrasonic, add 1.52 mL of DIEA, and add them to the reactor quickly after mixing, and oscillate with nitrogen for 2 hours , remove the reaction solution;

xii) Wash the resin five times with anhydrous DMF, anhydrous dichloromethane, anhydrous methanol and n-hexane, respectively. After washing, blow dry the resin with nitrogen;

xiii) Add 95% TFA aqueous solution (TFA:water=95:5) to the blown-dried resin, after 2 hours of nitrogen shaking, collect the above-reacted liquid in a beaker, wash the resin twice with 95% TFA aqueous solution and collect liquid;

xiv) Blow dry with an air pump to remove the solvent in the beaker, add an appropriate amount of ice anhydrous ether and settle overnight at -20°C, filter with suction, collect the powder-like solid, and use an analytical and semi-preparative high performance liquid chromatograph (HPLC) After separation and purification (water:acetonitrile=80:20～0:100), after concentration and drying in a freeze dryer, a white solid powder was obtained, and the total yield was about 40%.

According to the above steps, 2-naphthaleneacetic acid-phenylalanine-tryptophan-lysine-phosphotyrosine (NapFWKYp, represented by FWYp), 2-naphthaleneacetic acid-tryptophan-tryptophan were synthesized respectively. -Lysine-phosphotyrosine (NapWWKYp, represented by WWYp) and 2-naphthaleneacetic acid-tryptophan-phenylalanine-lysine-phosphotyrosine (NapWFKYp, represented by WFYp) three polypeptide molecules.

Example 2: Gel testing of polypeptide molecules

Weigh a certain amount of purified polypeptide sample powder into a small sample bottle, add an appropriate amount of ultrapure water, sonicate and adjust the pH to 7.4 with NaOH (1M) and HCl (1M) to fully dissolve the sample, and then add 2 μL of alkaline Phosphatase (ALP), the final volume is 200 μL to obtain a polypeptide sample solution with a certain concentration, and it is allowed to stand at room temperature.

The results are shown in Figure 2. The designed three molecules, FWYp, WWYp, and WFYp, were triggered by alkaline phosphatase (10 units/mL) in an aqueous solution of pH 7.4, and the lowest concentrations of self-assembled to form stable supramolecular hydrogels were 0.3, respectively. wt%, 0.6 wt%, 0.8 wt%.

Example 3: Mechanical Properties Test of Polypeptide Self-Assembled Hydrogel

Pipette 200 μL of the peptide self-assembled hydrogel (with a concentration of 1 wt%) on the sample stage, and use a Thermo Scientific HAAKE RheoStress 6000 rheometer for rheological experiments. During the test, the model selected is PP20H rotor, and the distance between the plates is 0.2mm. Under the condition of 25°C, the frequency is set to 6.282rad/s and the dynamic strain sweep test is carried out in the range of 0.1-10% strain sweep. The dynamic frequency sweep test was performed at a fixed stress of 1.0% in the frequency sweep range of 20 to 0.1 rad/s to ensure the linearity of dynamic viscoelasticity.

The results are shown in Figure 3. In both the stress sweep (Figure 3A) and frequency sweep (Figure 3B) dynamic rheological tests, the storage modulus (G') of the sample is much higher than its loss modulus (G"), That is to say, the three molecules formed stable hydrogels through self-assembly and exhibited remarkable viscoelastic properties.

Example 4: Bacterial flocculation activity test of polypeptide molecules

A stock solution (pH=7) of each polypeptide molecule sample with a concentration of 5000 μg/mL was prepared. The overnight cultured Escherichia coli (ATCC 25922) and Staphylococcus aureus (ATCC 12600) were removed from the medium by centrifugation (5000 rpm, 5 min), washed twice with physiological saline (0.9% NaCl solution), and then washed with physiological Resuspend in saline and dilute until the optical density (OD) value (wavelength is 600nm) measured by a microplate reader is about 1, respectively add it to each 2mL centrifuge tube, and add a certain amount of each sample mother liquor and physiological saline to each tube to prepare A bacterial suspension containing specific concentrations of sample molecules was prepared (test concentrations: 0, 25, 50, 100, 200, 300, 400, 500 μg/mL). Stir with a vortex stirrer (2000 rpm, 5 seconds) to ensure uniform mixing, collect samples at a depth of 1.0 cm below the liquid surface, and measure the OD value (OD ₀ ) with a microplate reader. Afterwards, shake quickly for 5 minutes at 200 rpm on a shaker, and then slowly shake at 50 rpm for 15 minutes. Finally, the mixture was left to stand for 120 minutes. The samples were collected at a depth of 1.0 cm in the supernatant, and the OD value (OD ₁ ) was determined with a microplate reader. The bacterial flocculation rate is defined as the relative reduction percentage of the optical density of the bacterial solution, that is, the flocculation rate (%)=[(OD ₀ -OD ₁ )/OD ₀ ]*100. In this experiment, the commercial flocculant polyaluminum chloride (PAC) was selected as the experimental control.

The results are shown in Figure 4. FWYp, WWYp and WFYp showed a certain flocculation effect at the lowest concentration of 25 μg/mL. At this time, their flocculation rates against Staphylococcus aureus were 17.2%, 9.0% and 12.5%, respectively. ; while the flocculation rates for Escherichia coli were 40.4%, 37.2% and 23.8%, respectively. Further increasing the concentration of these polypeptide samples can improve bacterial flocculation efficiency. For example, when the molecular concentration was increased to 500 μg/mL, the flocculation rates of FWYp, WWYp, and WFYp against Staphylococcus aureus were 83.5, 83.7, and 80.6%, respectively, and the flocculation rates against Escherichia coli were 78.8, 76.4, and 75.9%. The results showed that these polypeptides could interact with Staphylococcus aureus and Escherichia coli and produce significant bacterial flocculation with an efficiency comparable to the commercial flocculant polyaluminum chloride (PAC).

Example 5: Antibacterial activity test of polypeptide molecules

Escherichia coli (ATCC 25922) and Staphylococcus aureus (ATCC 12600) were selected as experimental bacteria, and the antibacterial effect of each polypeptide molecule was determined by dilution coating plate method. A stock solution of each polypeptide molecule sample with a concentration of 5000 μg/mL was prepared. The bacterial suspension cultured to the exponential growth phase was centrifuged (5000 rpm, 5 min) to remove the medium, washed twice with normal saline, resuspended and diluted in normal saline. A certain amount of sample stock solution and physiological saline were added to an equal amount of bacterial suspension (5×10 ⁵ CFU/mL) to a specific concentration, and incubated with shaking in a 37° C. incubator for 2 hours. After 2 hours, the co-incubated bacterial solution was serially diluted 200-fold, and 100 μL was pipetted and spread evenly on the agar plate. Subsequently, the formed colonies were counted and photographed, and only the bacteria incubated with physiological saline were used as a blank control group.

The results are shown in Figure 5. Each polypeptide molecule showed a certain antibacterial activity against Staphylococcus aureus (Figure 5A, 5B) and Escherichia coli (Figure 5C, 5D), and its antibacterial activity gradually increased with the increase of the concentration. . In addition, the antibacterial activity of each polypeptide molecule against Staphylococcus aureus was stronger than that of Escherichia coli.

Example 6: CCK-8 cytotoxicity experiment of polypeptide molecules and self-assembled hydrogels

Human umbilical vein endothelial cells (HUVEC) were selected as the cell model for cytotoxicity experiments. HUVEC cells (2×10 ⁴ CFU/mL) were planted in a 96-well plate, and a certain amount of pre-prepared polypeptide molecule sample solution (test sample concentration: 250, 500, 750 μg/mL) was added to the culture medium, and placed in a 37° C. Incubate in a constant temperature incubator for 24 hours. For gel samples, each polypeptide hydrogel with a concentration of 1 wt% was first prepared in each well of a 96-well plate (75 μL per well), and allowed to stand for 24 hours to stabilize the gel. Cells were seeded into each well containing the gel and cultured in a constant temperature incubator at 37°C for 24 hours, 48 hours and 72 hours, respectively. During the incubation period, the medium was changed once a day. After incubation, CCK-8 stain was added to each well and incubated in a 37°C incubator for 2 hours. After the cells were stained, the optical density was measured by using a microplate reader at 450 nm. The viability of the cells measured in the experiment was expressed as the percentage of cell viability between the experimental sample group and the untreated control group. The cell activity of the untreated control group was set to 100%. All samples had at least 5 parallel experimental groups and the experiment was repeated at least three times.

The results are shown in Fig. 6. HUVEC cells were co-cultured with different concentrations of FWYp, WWYp and WFYp for 24 h, and the cell viability was higher than 90% (Fig. 6A). And cells were implanted on the surface of FWY, WWY or WFY (1 wt%) self-assembled hydrogels, and the cells still had high viability after incubation at 37°C for 24, 48 and 72 h (Fig. 6B). The results of this study show that each polypeptide molecule and its self-assembled hydrogel have good cellular biocompatibility.

Example 7:

A sample stock solution (pH=7) of each polypeptide molecule and its self-assembly at a concentration of 2000 μg/mL was prepared. The overnight cultured Escherichia coli (ATCC 25922) and Staphylococcus aureus (ATCC 12600) were removed from the medium by centrifugation (5000 rpm, 5 min), washed twice with physiological saline (0.9% NaCl solution), and then washed with physiological The saline solution was resuspended and diluted until the optical density (OD) value (wavelength was 600nm) measured by the microplate reader was about 1, respectively added to each 10mL centrifuge tube, and a certain amount of each sample mother liquor and physiological saline was added to each tube to prepare Bacterial suspensions containing samples with a molecular concentration of 200 μg/mL. Stir with a vortex stirrer (2000 rpm, 5 seconds) to ensure uniform mixing, collect samples at a depth of 2.0 cm below the liquid surface, and measure the OD value (OD ₀ ) with a microplate reader. Afterwards, shake quickly for 5 minutes at 200 rpm on a shaker, and then slowly shake at 50 rpm for 15 minutes. Finally, the mixture was allowed to stand and samples were collected at a depth of 2.0 cm below the liquid surface every 20 minutes, and the OD value (OD ₁ ) was measured with a microplate reader (sampling time points: 20, 40, 60, 80, 100, 120, 140, 160, 180min). The bacterial flocculation rate is defined as the relative reduction percentage of the optical density of the bacterial solution, that is, the flocculation rate (%)=[(OD ₀ -OD ₁ )/OD ₀ ]*100. In this experiment, the commercial flocculant polyaluminum chloride (PAC) was selected as the experimental control.

The results are shown in Figure 7. In the absence of polypeptide samples, bacteria in both S. aureus and E. coli suspensions were well dispersed. However, addition of FWYp, WWYp or WFYp to the suspensions of S. aureus and E. coli resulted in bacterial flocculation, and their flocculation rates gradually increased with time within 180 min. Furthermore, after FWYp, WWYp or WFYp and ALP were added together to bacterial suspensions of S. aureus and E. coli, the bacteria in the S. aureus and E. coli suspensions were more abundant than those treated with FWYp, WWYp or WFYp Coagulation occurs rapidly and bacterial flocculation occurs. This suggests that self-assembly of the polypeptide enhances its flocculation activity due to the increased cationic charge density of the molecule and the formation of self-assembled nanostructures after enzyme-triggered dephosphorylation.

The above-mentioned embodiments are only preferred embodiments for fully illustrating the present invention, and the protection scope of the present invention is not limited thereto. Equivalent substitutions or transformations made by those skilled in the art on the basis of the present invention are all within the protection scope of the present invention. The protection scope of the present invention is subject to the claims.

Claims

A self-assembled polypeptide molecule with bacterial flocculation and antibacterial properties, characterized in that it has the following general formula:

Wherein, R 1 and R 2 are respectively selected from
one of the.
A method for preparing a self-assembling polypeptide molecule according to claim 1, wherein the method is to use a solid-phase synthesis method to sequentially connect phosphorylated tyrosine, lysine, amino acid to be synthesized and 2-naphthalene Acetic acid is used to synthesize the self-assembling polypeptide molecule, and the amino acid to be synthesized includes at least one tryptophan.
Application of the self-assembled polypeptide molecule of claim 1 in an antibacterial flocculant.
The application according to claim 3, characterized in that, the application is to add the self-assembled polypeptide molecule solution to the solution to be treated, and stir to perform flocculation.
The application according to claim 4, wherein the added concentration of the self-assembling polypeptide molecule is not less than 25 μg/mL.
The application according to claim 4, wherein the application further comprises adding alkaline phosphatase to the self-assembling polypeptide molecule solution for incubation before adding the self-assembling polypeptide molecule solution to the solution to be treated.
The application of the self-assembled polypeptide molecule of claim 1 in the preparation of hydrogel materials.
The application according to claim 7, wherein the hydrogel material is prepared by adding alkaline phosphatase to the self-assembled polypeptide molecule solution.
The application according to claim 8, wherein the pH of the self-assembled polypeptide molecule solution is 6-8.
A hydrogel material prepared by the self-assembling polypeptide molecule of claim 1.