A new antimicrobial peptide and its application

Abstract

The present invention discloses a novel antimicrobial peptide and its application. The present invention optimizes the structure of Epi-1, cuts off the non-α regions at both ends, and obtains a target antimicrobial peptide with a 16-amino acid sequence by replacing individual amino acids, thereby obtaining a novel antimicrobial polypeptide F-3 with a shorter sequence, higher antimicrobial activity, and better biocompatibility. A nucleic acid molecule encoding the antimicrobial peptide and an expression vector or recombinant cell containing the nucleic acid molecule are also provided. In vitro verification found that compared with the parent peptide, the derived peptide F-3 has higher antimicrobial activity or a higher bactericidal rate, and the minimum inhibitory concentration of F-3 is less than that of the parent peptide Epi-1; F-3 can also effectively inhibit or eliminate the formation of Enterococcus faecalis biofilm, and has low toxicity and good biocompatibility, and can be used to prepare drugs for preventing and treating diseases caused by Enterococcus faecalis infection; and the shorter amino acid sequence reduces the synthesis cost, and has good application prospects.

Description

Technical Field

[0001] The present invention belongs to the field of biotechnology, and in particular relates to a novel antimicrobial peptide and application thereof. Background Art

[0002] Endodontic disease and periapical disease are common oral infections, primarily caused by microbial infections such as bacteria within the root canal system. Currently, the most effective and commonly used treatment for these conditions is root canal therapy. Root canal therapy involves mechanical cleaning and disinfection to remove infected material from the root canal, followed by a thorough root canal filling. This isolates the root canal system from the oral and periapical environments, controlling infection, promoting healing of periapical lesions, and preventing their occurrence, ultimately saving the affected tooth. However, residual microbial infection within the root canal system can still lead to treatment failure. Studies have shown that the success rate of root canal therapy for teeth without periapical lesions is 92%, while the success rate drops to 74% for teeth with periapical disease. One of the main reasons for root canal treatment failure in most cases is the presence of bacteria in the root canal after treatment.

[0003] Enterococcus faecalis (E. faecalis) is a Gram-positive, facultative anaerobic bacterium commonly found in the human mouth, gastrointestinal tract, and vagina. E. faecalis is the most commonly detected species in teeth following failed root canal treatment, with multiple studies demonstrating a detection rate as high as 90%. Residual E. faecalis in the root canal can cause persistent infection of the periapical tissues and is closely associated with clinical symptoms such as pain. In this persistent infection state, the detection rate of E. faecalis in the root canal is nine times higher than in primary infected root canals, demonstrating the importance of E. faecalis in root canal infection.

[0004] Antimicrobial peptides (AMPs) are a class of small polypeptides with antibacterial and antiviral properties. AMPs are widely present in the epithelial barrier and immune defense systems of multicellular eukaryotic organisms and are sometimes referred to as host-defense peptides (HDPs). They possess diverse biological activities, such as immunomodulation, cancer cell inhibition, and wound healing promotion, and they also exhibit potent inhibitory effects against bacteria resistant to traditional antibiotics. AMPs have garnered extensive attention and research as a potential alternative to antibiotics for the next generation of antimicrobial agents. AMPs' antimicrobial mechanisms primarily include membrane-targeted and non-membrane-targeted approaches. After adsorbing and accumulating on bacterial surfaces, AMPs can penetrate the cell membrane, disrupting its integrity and directly killing the bacteria. AMPs can also bind to lipid II, a precursor of the cell wall, disrupting its integrity and rapidly killing the bacteria. Furthermore, AMPs can directly cross the cell membrane and accumulate within the cell, where they bind to negatively charged macromolecules, inhibiting key bacterial physiological processes, such as protein and nucleic acid synthesis or directly inhibiting enzyme activity, thereby achieving non-membrane-targeted bacterial killing.

[0005] Currently, a variety of AMPs with antibacterial effects against E. faecalis have been discovered or synthesized. LL-37 exhibits antimicrobial effects against planktonic E. faecalis in vitro; Nisin, when combined with calcium hydroxide as an intracanal sealant, significantly reduces E. faecalis in an in vitro root canal model; HBD3 gel inhibits the proliferation of E. faecalis on a dentin model in vitro; DJK-5, when combined with NaClO and EDTA, effectively removes E. faecalis infection and smear layers in an in vitro root canal model; in vitro studies have shown that GH12 can inhibit the activity of E. faecalis by downregulating the gene expression of multiple virulence factors. Although these antimicrobial peptides exhibit antimicrobial effects against E. faecalis, they are often used in combination. Epinecidin-1 is a natural antimicrobial peptide derived from orange grouper and belongs to the piscidin family. The pro-peptide of epinecidin-1 is 67 amino acids long and consists of three domains: the first 22 amino acids constitute a hydrophobic signal peptide, the middle 25 amino acids form a cationic mature peptide, and the remaining 20 amino acids form an anionic C-terminal prodomain. To investigate the pharmacological activity of epinecidin-1, researchers successfully synthesized and purified amino acids 22 to 42 (a total of 21 amino acids) of the epinecidin-1 sequence and named it Epi-1. Epi-1 exhibits strong antimicrobial activity both in vitro and in vivo, with some inhibitory effects against anaerobic bacteria. Epi-1 also exhibits significant antimicrobial activity against multidrug-resistant strains. When used in combination with amoxicillin, metronidazole, and clarithromycin, Epi-1 exhibits synergistic inhibitory effects against multidrug-resistant Helicobacter pylori. Epi-1 is more effective than vancomycin against methicillin-resistant Staphylococcus aureus. Animal studies have shown that Epi-1 can protect mice from septicemia caused by Pseudomonas aeruginosa and reduce mortality in pigs with septicemia caused by methicillin-resistant Staphylococcus aureus. In terms of immunomodulation, Epi-1 has anti-inflammatory effects. Epi-1 can reduce MyD88 expression and inhibit Toll-like receptor activation, thereby suppressing inflammatory responses. Epi-1 can also alleviate macrophage inflammatory responses induced by Escherichia coli lipopolysaccharide. Epi-1 can also inhibit activation of the Akt / p38 signaling pathway in macrophages induced by lipoteichoic acid. In a pig model of thermal skin injury, Epi-1 promotes wound healing by promoting vascularization, epithelial cell proliferation, and collagen synthesis in the wound area. These findings suggest that Epi-1 has multiple biological activities and suggest that Epi-1 may have excellent antibacterial activity against E. faecalis, potentially aiding in the control of root canal infections.

[0006] Both natural antimicrobial peptides and their derivative peptides have long peptide chains, which not only causes the problems of high synthesis consumption and difficulty in clinical transformation, but also due to the existence of factors such as steric hindrance, the antimicrobial effect itself may not be fully reflected; most antimicrobial peptides have poor inhibitory effects on biofilms and limited control over infections.

[0007] In summary, this project intends to select Epi-1 as the mother peptide template and attempt to optimize it, aiming to obtain new antibacterial peptides with shorter sequences, higher antibacterial activity and better biocompatibility. Through in vitro studies and infected root canal models, the antibacterial effects of Epi-1 and its derivative peptides on E. faecalis will be explored, providing a theoretical and application basis for it as a new root canal irrigant. Summary of the Invention

[0008] The purpose of the present invention is to provide a novel antibacterial polypeptide with a shorter sequence, higher antibacterial activity and better biocompatibility.

[0009] The technical solution adopted by the present invention is:

[0010] In a first aspect, the present invention provides a polypeptide, wherein the amino acid sequence of the polypeptide is shown in any one of SEQ ID NOs: 1 to 2.

[0011] The second aspect of the present invention provides a nucleic acid molecule, which encodes the polypeptide described in the first aspect of the present invention.

[0012] In some embodiments of the present invention, the nucleic acid molecule may be the nucleic acid molecule shown in 1) or 2) or 3) or 4) below:

[0013] 1) a DNA molecule encoding the polypeptide;

[0014] 2) cDNA molecules encoding the polypeptide

[0015] 3) a cDNA molecule or genomic DNA molecule that has 75% or more identity with the nucleotide sequence defined in 1) or 2) and encodes the polypeptide;

[0016] 4) A cDNA molecule or genomic DNA molecule that hybridizes under stringent conditions with the nucleotide sequence defined in 1) or 2) and encodes the polypeptide.

[0017] The third aspect of the present invention provides a vector, wherein the vector comprises the nucleic acid molecule described in the second aspect of the present invention.

[0018] In some embodiments of the present invention, the vector comprises a plasmid, a cosmid, a phage or a viral vector.

[0019] The fourth aspect of the present invention provides a cell, wherein the cell comprises the vector described in the third aspect of the present invention.

[0020] In some embodiments of the invention, the cell comprises yeast, bacteria, algae, or fungi.

[0021] In a fifth aspect, the present invention provides use of the polypeptide according to the first aspect of the present invention, the nucleic acid molecule according to the second aspect of the present invention, the vector according to the third aspect of the present invention, or the cell according to the fourth aspect of the present invention in at least one of the following:

[0022] a) Preparation of bacteriostatic, bactericidal or antibacterial products;

[0023] b) bacteriostatic or bactericidal or antimicrobial;

[0024] c) inhibit or eliminate bacterial biofilm formation;

[0025] d) preparing products that inhibit or eliminate bacterial biofilm formation;

[0026] e) Preparation of products for the prevention and / or treatment of diseases caused by bacterial infections;

[0027] f) preparing additives;

[0028] g) preparing oral products.

[0029] In some embodiments of the present invention, the bacteria include at least one of Enterococcus faecalis, Streptococcus mutans, and Porphyromonas gingivalis.

[0030] In some embodiments of the present invention, the disease comprises an oral infectious disease.

[0031] In some embodiments of the present invention, the disease comprises dental caries, periapical disease, and endodontic disease.

[0032] In some embodiments of the present invention, the additive is a food additive, a feed additive, a cosmetic additive or a sanitary product additive.

[0033] The sixth aspect of the present invention provides a product, which comprises the polypeptide described in the first aspect of the present invention, the nucleic acid molecule described in the second aspect of the present invention, the vector described in the third aspect of the present invention, or the cell described in the fourth aspect of the present invention.

[0034] In some embodiments of the present invention, the product includes a medicine, a reagent, an oral product, or an additive.

[0035] In some embodiments of the present invention, the oral products include toothpaste, mouthwash, tooth strips, oral dressings, and oral sprays.

[0036] The beneficial effects of the present invention are:

[0037] The present invention optimizes the structure of Epi-1, truncates the non-α regions at both ends, and replaces individual amino acids to obtain a target antimicrobial peptide with a 16-amino acid sequence. This results in a novel antimicrobial peptide F-3 with a shorter sequence, higher antimicrobial activity, and improved biocompatibility. The invention also provides a nucleic acid molecule encoding the antimicrobial peptide, as well as an expression vector or recombinant cell containing the nucleic acid molecule. In vitro validation has shown that the derivative peptide F-3 exhibits higher antimicrobial activity or a higher bactericidal rate than the parent peptide, with a minimum inhibitory concentration (MIC) lower than that of the parent peptide Epi-1. F-3 can also effectively inhibit or eliminate the formation of Enterococcus faecalis biofilms, has low toxicity, and good biocompatibility, making it suitable for the preparation of drugs to prevent and control diseases caused by Enterococcus faecalis infections. Furthermore, the shorter sequence reduces synthesis costs, suggesting promising application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0038] Figure 1 The structural domains of Epi-1. h: α-helical region; c: disordered region; e: extended region.

[0039] Figure 2 The α-helical diagram of Epi-1 and F-3. Yellow represents hydrophobic amino acids; blue represents hydrophilic amino acids; and gray represents hydrophobic non-polar amino acids.

[0040] Figure 3 Inhibitory test of Epi-1, F-3, CHX and NaClO on E. faecalis ATCC 29212 biofilm (compared with the control group: a P < 0.05; compared with Epi-1: #P < 0.05, ##P < 0.01; compared with the F-3 group: *P < 0.05, **P < 0.01).

[0041] Figure 4 24-h E. faecalis ATCC 29212 biofilm clearance experiment by Epi-1, F-3, CHX, and NaClO (compared with the control group: a P < 0.05; compared with Epi-1: # P < 0.05; compared with F-3 group: * P < 0.05).

[0042] Figure 5 This is the result of the hemolytic activity of EPI-1 and F-3 on rabbit erythrocytes. DETAILED DESCRIPTION

[0043] The following will clearly and completely describe the concept and technical effects of the present invention in conjunction with the embodiments to fully understand the purpose, features and effects of the present invention. Obviously, the embodiments described are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, other embodiments obtained by those skilled in the art without creative work are all within the scope of protection of the present invention.

[0044] Design of antimicrobial peptides: Based on the design concept of amphipathic positively charged α-helical antimicrobial peptides, namely the design characteristics of positive charge, simultaneous hydrophobicity, amphipathicity and imperfect amphipathicity, the antimicrobial peptide F-3 derived from the mother peptide Epi-1 was designed. At the same time, the anti-Enterococcus faecalis activity of F-3 was verified, and its hemolytic activity against rabbit erythrocytes was evaluated.

[0045] Statistical analysis: Each experiment was repeated three times, and the results of each group are expressed as mean ± standard deviation (mean ± SD). Student's t-test or one-way analysis of variance was performed using SPSS 26 software. The test level was α = 0.05, and P < 0.05 indicated a statistically significant result.

[0046] Molecular design and sequence analysis of Example 1F-3

[0047] Epinecidin-1 is a natural antimicrobial peptide derived from orange grouper. It is 67 amino acids long and consists of three domains: the first 22 amino acids constitute the hydrophobic signal peptide, the last 25 amino acids constitute the cationic mature peptide, and the remaining 20 amino acids constitute the anionic C-terminal prodomain. Scholars successfully synthesized and purified the amino acid sequence of Epinecidin-1 from positions 22 to 42 (21 amino acids) and named it Epi-1. Its sequence is: GFIFHIIKGLFHAGKMIHGLV (SEQ ID NO.4).

[0048] The secondary structure of Epi-1 was obtained from the Prabi website and the α-helical region was obtained. Figure 2 (Left), the coiled structures at both ends were truncated, retaining only the central active region, resulting in a new sequence of 16 amino acids, designated FI-0. The amino acid distribution characteristics of FI-0 (FIFHIIKGLFHAGKMI (SEQ ID NO. 5)) were analyzed using the Heliquest website, and further adjustments to the hydrophilicity and hydrophobicity, polarity, and non-polarity were made on this basis. To enhance the antimicrobial activity of the target antimicrobial peptide and reduce its hemolytic activity, lysine and leucine residues were substituted for the non-hydrophobic residues on the non-polar face to obtain the perfect amphipathic F-1 (FIFRIIKRLFRLLKRI (SEQ ID NO. 3)). By replacing single tryptophan residues at positions 7 and 11, the perfect amphipathic nature of F-1 was disrupted, shortening the hydrophobic moment of the target antimicrobial peptide, resulting in F-2 (FIFRIIWRLFWLLKRI (SEQ ID NO. 2)) and F-3 (FIFRIIKWRFRLRKRI (SEQ ID NO. 1)).

[0049] Specifically, the modification was continued in the following three ways: (1) using arginine to replace the non-positively charged amino acids on the polar face to increase the positive charge of the polar face and generate electrostatic interaction with the negatively charged lipid molecules on the surface of the bacterial cell membrane; (2) using tryptophan to replace the non-polar hydrophobic amino acids on the non-polar face to increase the hydrophobicity of the non-polar face, making it easier to destroy the bacterial cell membrane; (3) inserting hydrophilic amino acids into the continuous hydrophobic amino acids to shorten the hydrophobic distance and reduce the toxicity of the peptide. In this way, an α-helical antimicrobial peptide F-3 with high activity and low toxicity can be obtained. The helical diagram ( Figure 2 ), the amino acid sequence of F-3 is FIFRIIKWRFRLRKRI.

[0050] Example 2 Bacterial sensitivity test

[0051] To evaluate the inhibitory effects of Epi-1, F-1, F-2, and F-3 against E. faecalis ATCC 29212 planktonic bacteria, their minimum inhibitory concentrations (MICs) and minimum bactericidal concentrations (MBCs) were tested according to the experimental methods recommended by the Clinical and Laboratory Standards Institute (CLSI) to assess the bacterial sensitivity of the antimicrobial peptides.

[0052] The specific experimental steps are as follows: the drug diluted in RPMI 1640 medium was placed in a 96-well plate, and then E. faecalis ATCC 29212 in the logarithmic phase, which had been diluted in RPMI 1640 medium, was added to make the final concentration of the drug 1.6-400 μg / mL and the final concentration of the bacterial solution 1×10 6 CFU / mL, with a final solution volume of 200 μL / well. Positive controls consisted of NaClO and CHX groups, negative controls contained only bacterial solution and culture medium, and blank controls contained only culture medium. Three replicates were set up for each concentration in each group. Finally, the 96-well plate was placed in a 37°C incubator under anaerobically incubated conditions (80% N2, 10% CO2, and 10% H2). After 24 hours, the turbidity of the mixed liquid in the plate was directly observed. The lowest concentration well with no significant difference in turbidity from the blank control was considered the drug concentration corresponding to its MIC.

[0053] 150 μL of the mixed solution from all wells that showed no visible turbidity was evenly spread onto BHI agar plates. The plates were then incubated in an anaerobic environment (80% N₂, 10% CO₂, and 10% H₂) at 37°C for 48 hours. The plate surface was then observed, and the concentration of the antimicrobial peptide corresponding to the first BHI agar plate without colonies was considered its MBC. The results are shown in Table 1. The results show that the minimum inhibitory concentrations of F-2 and F-3 were lower than that of the parent peptide Epi-1.

[0054] Table 1 Minimum inhibitory concentration and minimum bactericidal concentration of Epi-1 and F-3

[0055]

[0056] Example 3 Experiment on Inhibition of Enterococcus faecalis Biofilm Formation

[0057] The inhibitory effects of Epi-1 and F-3 on biofilm formation of E. faecalis ATCC 29212 were determined by semi-quantitative detection method using crystal violet staining.

[0058] The specific experimental steps are as follows: the peptide diluted in RPMI 1640 medium was placed in a 96-well plate, and then E. faecalis ATCC 29212 in the logarithmic phase after adjusting the growth in RPMI 1640 medium was added to make the final peptide concentrations 0.6×, 0.8×, 1×, and 2× MICs, respectively, and the bacterial concentration was 1×10 6 CFU / mL, with a final volume of 200 μL / well. Positive controls consisted of NaClO and CHX, negative controls contained culture medium and bacterial suspension, and blank controls contained culture medium alone. Three replicates were set up for each concentration. A 96-well plate was incubated in an anaerobic environment (80% N2, 10% CO2, and 10% H2) at 37°C for 24 hours. The supernatant was carefully removed, and free E. faecalis ATCC 29212 was gently washed with PBS. The plates were fixed with methanol for 5 minutes and stained with 0.1% (w / v) crystal violet solution for 15 minutes. Excess stain was then washed with PBS until no obvious staining was observed in the blank control wells. After air-drying, 95% ethanol was added, 200 μL per well, and gently shaken for 30 minutes in the dark to facilitate complete dissolution of the biofilm. The OD value was measured at a wavelength of 595 nm using a microplate reader.

[0059] The results were expressed as biofilm formation rate, and the formula was: biofilm formation rate = (OD value of experimental group - OD value of blank control group) / (OD value of negative control group - OD value of blank control group) × 100%.

[0060] The results are as follows Figure 3As shown in the figure, F-3 exhibited a concentration-dependent inhibitory effect on E. faecalis ATCC 29212 biofilm formation, with inhibition rates higher than those of Epi-1 (0.6×MIC: 8.55% vs. 5.54%; 0.8×MIC: 41.07% vs. 33.39%; 1×MIC: 97.20% vs. 89.41%). At a concentration of 2×MIC, F-3 and Epi-1 achieved biofilm inhibition rates of 99.48% and 99.16%, respectively.

[0061] Example 4 Experiment on Removal of Mature Enterococcus Faecalis Biofilm

[0062] To investigate the effects of Epi-1 and F-3 on the clearing of established E. faecalis ATCC 29212 biofilms.

[0063] The specific experimental steps are as follows: Use RPMI 1640 medium to adjust the logarithmic phase E. faecalis ATCC 29212 to a final concentration of 1×10 6 CFU / mL were plated in a 96-well plate in a final volume of 200 μL / well. The 96-well plate was incubated at 37°C under an anaerobic environment (80% N2, 10% CO2, and 10% H2) for 24 hours. The supernatant was removed and non-adherent E. faecalis ATCC29212 was gently washed with PBS. Each drug group was diluted to 1×, 2×, 4×, 8×, and 10× the corresponding MIC and added to the 96-well plate in a final volume of 200 μL / well. Positive controls consisted of NaClO and CHX, while negative controls contained culture medium and bacterial suspension. Three replicates were set up for each group at different concentrations. The 96-well plate was incubated at 37°C under an anaerobic environment. After 24 hours, the supernatant was removed and non-adherent bacteria were gently washed with PBS. The remaining biofilm on the bottom was scraped with a sterile pipette tip, rinsed with PBS, and collected in an EP tube. The biofilm was then broken up by ultrasonic vibration to release the bacteria inside the biofilm. After gradient dilution with PBS, the samples were spread on BHI agar plates and incubated at 37°C under anaerobic conditions for 48 h before colony counts. The results were expressed as the number of colonies in the biofilm (log).

[0064] The results are as follows Figure 4 As shown. At the MIC concentration, there were statistical differences in the number of viable bacteria remaining after the treatment of F-3, Epi-1, CHX, and NaClO compared with the negative control group. As the concentration increased, the clearance effect of F-3, CHX, and NaClO on mature biofilms gradually increased, while Epi-1 did not show a similar trend. When different concentrations of F-3 were used to treat the formed E. faecalis ATCC 29212 biofilm for 24 hours, the number of viable bacteria decreased by 10%, ... 0.67 , 10 0.79 , 100.48 , 10 1.83 and 10 1.97 At concentrations ≥4×MIC, the number of surviving biofilm bacteria in the F-3 group was significantly lower than that in the Epi-1 group. Only at 4×MIC was the number of surviving biofilm bacteria in the F-3 group slightly higher than that in the CHX group (P < 0.05). At other concentrations, there was no statistical difference between the two groups. Compared with the NaClO group, the effect of F-3 was weaker than that of NaClO, with a statistically significant difference starting at 2×MIC.

[0065] Example 5 Hemolysis Experiment

[0066] The cytotoxicity of antimicrobial peptides is one of the criteria for evaluating their biosafety and clinical applicability. Their hemolytic activity can be used to indicate cytotoxicity. Therefore, this experiment investigated the hemolytic effect of Epi-1 and its derivative peptide F-3 on rabbit erythrocytes.

[0067] The specific procedure was as follows: Fresh rabbit erythrocytes were carefully washed three times with PBS and resuspended (centrifugation conditions: 1000 × g, 4°C, 10 min). In a 96-well plate, the washed and resuspended rabbit erythrocytes were added to serially diluted drug solutions in PBS at concentrations of 0, 1 / 4, 1 / 2, 1, 2, 4, and 8 × the MIC, respectively. The final volume of the mixture was 200 μL / well. A positive control group consisted of 1% Triton-X 100 solution, a negative control group contained only rabbit erythrocytes and PBS, and a blank control group contained only PBS. Three replicates were set up for each concentration. The 96-well plate was incubated in a 37°C incubator for 1 h. After centrifugation (under the same conditions as above), 180 μL of the supernatant was transferred to another 96-well plate. The OD value at a wavelength of 405 nm was measured using a microplate reader. This reading represents the amount of hemoglobin released after rupture of the rabbit erythrocytes after drug treatment.

[0068] The results were expressed as hemolysis rate, and the formula was: rabbit erythrocyte hemolysis rate = (OD value of experimental group - OD value of blank control group) / (OD value of positive control group - OD value of blank control group) × 100%.

[0069] Epi-1 and its derivative peptide F-3 exhibited concentration-dependent hemolytic activity against rabbit erythrocytes. Epi-1 exhibited significant hemolytic activity (P < 0.05) at an MIC of 25 μg / mL, resulting in a hemolytic rate of 14.1%. At the MBC of 100 μg / mL, the hemolytic rate reached 68.2%. F-3, on the other hand, showed virtually no hemolytic activity at an MIC of 6.25 μg / mL, and even at 64 times the MIC (400 μg / mL), the hemolytic rate was only 5.07%.

[0070] Epi-1 and F-3 were compared with the commonly used root canal disinfectants NaClO and CHX. Figure 5 As shown in the data, even at 1 / 4×MIC (31.25μg / mL), the hemolytic activity of NaClO reached 100%, and CHX also showed significant hemolytic activity at 2×MIC (6.25μg / mL) (P<0.05), with a hemolytic rate of 5.06%.

[0071] The above specific embodiments provide a detailed description of the present invention. However, the present invention is not limited to the above embodiments. Various modifications may be made within the scope of knowledge possessed by a person skilled in the art without departing from the spirit of the present invention. Furthermore, the embodiments of the present invention and the features thereof may be combined with each other unless there is a conflict.

Claims

1. A polypeptide, characterized in that The amino acid sequence of the polypeptide is shown in any one of SEQ ID NOs: 1 to 2.

2. A nucleic acid molecule, characterized in that The nucleic acid molecule encodes the polypeptide according to claim 1.

3. A carrier, characterized in that The vector comprises the nucleic acid molecule according to claim 2.

4. A cell, characterized in that The cell comprises the vector of claim 3.

5. Use of the polypeptide according to claim 1, the nucleic acid molecule according to claim 2, the vector according to claim 3, or the cell according to claim 4 in at least one of the following: a) Preparation of bacteriostatic, bactericidal or antimicrobial products; b) bacteriostatic or bactericidal or antimicrobial; c) inhibit or eliminate bacterial biofilm formation; d) preparing products for inhibiting or eliminating bacterial biofilm formation; e) Preparation of products for the prevention and / or treatment of diseases caused by bacterial infections; f) preparing additives; g) preparing oral products; The application is not an application for disease diagnosis and treatment methods; The bacteria is at least one of Enterococcus faecalis, Streptococcus mutans, and Porphyromonas gingivalis; The diseases include oral infectious diseases; The additive is a feed additive, a cosmetic additive or a sanitary product additive; The oral products include toothpaste, mouthwash, tooth strips, oral dressings, and oral sprays.

6. The use according to claim 5, characterized in that The diseases include: dental caries, periapical disease, and pulp disease.

7. A product, characterized in that The product comprises the polypeptide according to claim 1.

8. The product according to claim 7, characterized in that The products include reagents, oral products, and additives.

9. The product according to claim 8, characterized in that The oral products include toothpaste, mouthwash, tooth strips, oral dressings, and oral sprays.

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