Antibacterial peptide with activity of inhibiting human pathogenic bacteria as well as preparation method and application of antibacterial peptide

Abstract

The invention belongs to the technical field of biology, and discloses an antibacterial peptide with activity of inhibiting human pathogenic bacteria as well as a preparation method and application of the antibacterial peptide. The antibacterial peptide or the salt thereof is disclosed for the first time, and the antibacterial peptide has a good antibacterial effect on common genital tract bacteria (such as staphylococcus aureus, gardnerella and candida albicans), oral pathogenic bacteria (such as fusobacterium nucleatum and streptococcus gordoni) and other pathogenic bacteria (such as escherichia coli, salmonella enteritidis and streptococcus suis); and the compound shows slight cytotoxicity to mammalian cells only at a high concentration.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology, specifically relating to an antimicrobial peptide that inhibits the activity of pathogenic bacteria in humans, its preparation method, and its application. Background Technology

[0002] The female reproductive tract environment is often affected by various factors such as hormonal fluctuations, poor hygiene, seasonal changes, and even emotional instability, leading to flora imbalance. This can result in pathogen invasion and infection by bacteria, fungi, viruses, and other microorganisms, with Gardnerella vaginalis being a common example. Gardnerella vaginalis Candida albicans ( Candida albicans Staphylococcus aureus ( Staphylococcus aureus Reproductive tract infections manifest clinically as vaginal itching and burning, abnormal discharge, frequent and painful urination, and in severe cases, can lead to premature birth and cervical cancer in pregnant women. Statistics show that the infection rate of reproductive tract microorganisms among women of childbearing age in my country is as high as 80%, with the vast majority of patients with bacterial or fungal infections receiving antibiotic treatment. While antibiotics combat pathogens, they also reduce the abundance of beneficial bacteria, which is detrimental to maintaining reproductive tract flora homeostasis and easily leads to drug resistance, causing secondary infections. Therefore, there is an urgent need to develop new drugs to avoid the risk of reproductive tract flora imbalance.

[0003] Oral health is an important aspect of overall human health. The results of the Fourth National Oral Health Epidemiological Survey show that residents' oral health literacy is gradually improving. However, most Chinese people still suffer from a range of oral health problems, such as tooth decay, periodontitis, halitosis, and plaque, which frequently occur and seriously affect their health. Research indicates that oral problems such as tooth decay, periodontitis, halitosis, and plaque are inextricably linked to oral microorganisms. Fusobacterium nucleatum (… Fusobacterium nucleatum *Fusobacterium argyi* is a Gram-negative obligate anaerobic bacterium and the most abundant *Fusobacterium* found in the oral cavity. It can function as a symbiotic bacterium, promoting plaque biofilm formation, or it can induce the production of volatile sulfides, causing halitosis. *Streptococcus Gordonii* (*Fusobacterium argyi*) Streptococcus gordonii It is a common opportunistic pathogen in the oral cavity. It can not only synergistically cause tooth decay and periapical periodontitis with other oral pathogens, but also enter the bloodstream and cause local or systemic diseases, such as skin and soft tissue infections, sepsis, infective endocarditis, and suppurative arthritis.

[0004] The existing treatment drugs for pathogenic bacteria infection are mainly traditional antibiotics, and long-term use can easily cause bacterial drug resistance and bacterial flora imbalance. Therefore, it is urgent to develop new drugs to avoid the risk of bacterial drug resistance. The advantages of antimicrobial peptides (AMPs) compared with traditional antibiotic drugs are numerous, which cannot easily cause pathogenic microorganisms to develop drug resistance, can slow down the problem of antibiotic drug resistance, ensure long-term effectiveness, have high safety, low residual risk, and small impact on the environment and food safety, and can reduce environmental pollution. SUMMARY

[0005] The first aspect of the present application aims to provide an antimicrobial peptide or a salt thereof.

[0006] The second aspect of the present application aims to provide a conjugate.

[0007] The third aspect of the present application aims to provide a fusion protein.

[0008] The fourth aspect of the present application aims to provide a biological material related to the antimicrobial peptide or the salt thereof of the first aspect of the present application, the conjugate of the second aspect of the present application, the fusion protein of the third aspect of the present application.

[0009] The fifth aspect of the present application aims to provide a composition.

[0010] The sixth aspect of the present application aims to provide the use of the antimicrobial peptide or the salt thereof of the first aspect of the present application, the conjugate of the second aspect of the present application, the fusion protein of the third aspect of the present application, the biological material of the fourth aspect of the present application, or the composition of the fifth aspect of the present application.

[0011] The seventh aspect of the present application aims to provide a product.

[0012] The eighth aspect of the present application aims to provide a method for inhibiting the growth of pathogenic microorganisms.

[0013] The ninth aspect of the present application aims to provide a method for producing the antimicrobial peptide or the salt thereof of the first aspect of the present application.

[0014] The tenth aspect of the present application aims to provide a method for preventing and / or treating a disease caused by pathogenic microorganism infection.

[0015] In order to achieve the above-mentioned purposes, the technical solutions adopted by the present application are as follows: The first aspect of the present application provides an antimicrobial peptide or a salt thereof, wherein the amino acid sequence of the antimicrobial peptide is: a1) SEQ ID NO: 1; or a2) an amino acid sequence obtained by substitution and / or deletion and / or addition of one or more amino acids of SEQ ID NO: 1 and having the same function as SEQ ID NO: 1; or a3) An amino acid sequence that shares 99%, 98%, 97%, 96%, 95%, 94%, 93%, 92%, 91%, 90%, 89%, 88%, 87%, 86%, 85%, 84%, 83%, 82%, 81%, and 80% homology with SEQ ID NO:1 and has the same function as SEQ ID NO:1.

[0016] In some embodiments of the present invention, the salt of the antimicrobial peptide includes a metal salt of the antimicrobial peptide, or a salt formed by the antimicrobial peptide and an inorganic or organic acid.

[0017] In some embodiments of the present invention, the metal includes at least one of lithium, sodium, potassium, calcium, magnesium, manganese, copper, zinc or aluminum.

[0018] In some embodiments of the present invention, the organic acid includes at least one selected from acetic acid, citric acid, lactic acid, malonic acid, maleic acid, tartaric acid, fumaric acid, benzoic acid, aspartic acid, glutamic acid, succinic acid, oleic acid, trifluoroacetic acid, or oxalic acid.

[0019] In some embodiments of the present invention, the inorganic acid includes at least one of hydrochloric acid, sulfuric acid, phosphoric acid, diphosphoric acid, hydrobromic acid, boric acid, carbonic acid, or nitric acid.

[0020] A second aspect of the present invention provides a conjugate comprising a modification portion and an antimicrobial peptide of the first aspect of the present invention.

[0021] The modification is an artificially induced drug, performed while preserving the peptide activity. The purposes of the modification include prolonging the half-life, increasing water solubility, and reducing or eliminating toxic side effects.

[0022] In some embodiments of the present invention, the modified portion is located at the N-terminus and / or C-terminus of the antimicrobial peptide.

[0023] In some embodiments of the present invention, the modified portion comprises at least one of chemical modification, targeting portion, fluorescent dye and protein tag.

[0024] In some embodiments of the present invention, the chemical modification includes at least one of amidation, acetylation, aminoation, methylation, phosphorylation, glycosylation, and ubiquitination.

[0025] In some embodiments of the present invention, the targeting portion includes at least one of a ligand, a receptor, and an antibody (such as a full-length antibody, Fab, scFv, F(ab')2, chimeric antibody, etc.).

[0026] In some embodiments of the present invention, the fluorescent dye comprises at least one of the following: Cy series (such as Cy3, Cy5.5, Cy7, etc.), Texas series (such as Texas Red-X succinimide ester, Texas Red Amine, etc.), Alexa series (such as Alexa Fluor 488, Alexa Fluor 555, Alexa Fluor 568, Alexa Fluor 594, Alexa Fluor 633, Alexa Fluor 647, Alexa Fluor 680, Alexa Fluor 750, etc.), Rhodamine, Bodipy, Rox, FAM, FITC, MCA, TAMRA, and Dnp.

[0027] In some embodiments of the present invention, the protein tag includes at least one of His, Flag, GST, MBP, HA, Myc, GFP, and biotin.

[0028] A third aspect of the present invention provides a fusion protein comprising a first domain and a second domain; The first structural domain is the antimicrobial peptide of the first aspect of the present invention; The second domain has the effect of prolonging the in vivo half-life.

[0029] In some embodiments of the present invention, the first domain is located at the N-terminus and / or C-terminus of the antimicrobial peptide.

[0030] In some embodiments of the present invention, the second domain and the antimicrobial peptide are connected to the N-terminus and / or C-terminus of the antimicrobial peptide via a connector.

[0031] In some embodiments of the present invention, the connector includes a flexible linker, including but not limited to GSAS, (GGCAGCGCCAGC, SEQ ID NO:2). n , (GGCGGCGGCAGC, SEQ ID NO:3) n , (GGCGGCGGCGGCAGC, SEQID NO:4) n YAPVDV, (GGGS) n (GGSG) n (GGGGS) n (G) n , where 1≤n≤5, and n is an integer.

[0032] In some embodiments of the present invention, the second structural domain includes (but is not limited to): Immunoglobulin Fc region (such as human immunoglobulin Fc region); and / or Serum albumin (such as human HSA) or fragments thereof, domains that bind to serum albumin (such as anti-serum albumin antibodies, including nanobodies), polyethylene glycol, polyethylene glycol-liposome complexes, or combinations thereof.

[0033] In some embodiments of the present invention, the immunoglobulin is one or more selected from IgG, IgA1, IgA2, IgD, IgE, and IgM.

[0034] In some embodiments of the present invention, the IgG is selected from one or more combinations of IgG1, IgG2, IgG3 or IgG4 subtypes.

[0035] A fourth aspect of the invention provides biomaterials relating to the antimicrobial peptide of the first aspect of the invention, the conjugate of the second aspect of the invention, or the fusion protein of the third aspect of the invention, said biomaterials comprising at least one of b1)-b8): b1) A nucleic acid molecule encoding the antimicrobial peptide of the first aspect of the present invention, the conjugate of the second aspect of the present invention, or the fusion protein of the third aspect of the present invention; b2) An expression cassette containing the nucleic acid molecule described in b1); b3) A carrier containing the nucleic acid molecule described in b1); b4) A carrier containing the expression box described in b2); b5) Transgenic cell lines containing the nucleic acid molecules described in b1); b6) Transgenic cell lines containing the expression cassette described in b2); b7) A transgenic cell line containing the vector described in b3); b8) A transgenic cell line containing the vector described in b4).

[0036] In some embodiments of the present invention, the transgenic cell line does not contain propagation material.

[0037] In some embodiments of the present invention, the vector is independently selected from non-pathogenic viral vectors and viral vectors.

[0038] In some embodiments of the present invention, the viral vector includes at least one of lentiviral vector, adenovirus vector, baculovirus vector, retrovirus vector, poxvirus vector, Sendai virus vector, and herpes simplex virus vector.

[0039] In some embodiments of the present invention, the non-viral vector includes at least one of plasmid vectors, cationic polymer vectors, chitosan, polyethyleneimine, nanoparticle vectors, and liposomes.

[0040] In some embodiments of the present invention, the vector is a plasmid vector, a phage particle, a viral vector, a cell vector, a bacteriophage, a sclerotium, an F sclerotium, or an artificial chromosome.

[0041] In some embodiments of the present invention, the plasmid vector may be an optional plasmid, and the viral vector may be an optional virus.

[0042] In some embodiments of the present invention, the recombinant expression vector uses pET-28a(+) as the original expression vector.

[0043] In some embodiments of the present invention, the cells include prokaryotic cells and eukaryotic cells; the cells are not new plant or animal varieties.

[0044] In some embodiments of the present invention, the prokaryotic cells include bacteria well known in the art, such as Escherichia coli, Streptomyces, and Bacillus subtilis, which are capable of expressing the target protein.

[0045] In some embodiments of the present invention, the eukaryotic cells include at least one of yeast cells, mammalian cells, plant cells, and insect cells.

[0046] In some embodiments of the present invention, the cells include engineered cell lines such as CHO, CHO-K1, and CHO-GS, and lymphocyte lines such as T cells, NK cells, and CIK cells.

[0047] A fifth aspect of the invention provides a composition comprising at least one of c1)-c3): c1) The antimicrobial peptide or its salt according to the first aspect of the present invention; c2) The conjugate of the second aspect of the present invention; c3) The fusion protein of the third aspect of the present invention.

[0048] In some embodiments of the present invention, the composition further comprises other active antibacterial ingredients.

[0049] In some embodiments of the present invention, the other active antibacterial components include at least one of the following: penicillin antibiotics (such as ampicillin, oxacillin, etc.), cephalosporin antibiotics (such as cefadroxil, cefuroxime, cefoperazone, etc.), carbapenem antibiotics (such as imipenem, meropenem, etc.), lipoprotein (glycopeptide) antibiotics (such as polymyxins and vancomycin, etc.), aminoglycoside antibiotics, tetracycline antibiotics (such as doxycycline, minocycline, oxytetracycline, etc.), lincosamide antibiotics (such as lincomycin, clindamycin, etc.), macrolide antibiotics (such as azithromycin, roxithromycin, erythromycin, etc.), fluoroquinolone antibiotics (such as ciprofloxacin, lomefloxacin, etc.), and fusidic acid.

[0050] In some embodiments of the present invention, the other active antibacterial components include at least one of the following natural antibacterial substances: cinnamaldehyde, eugenol, coptis chinensis, berberine, quercetin, baicalin, thymol, andrographolide, curcumin, kaempferol, apigenin, soy isoflavones, gallic acid, protocatechuic acid, chlorogenic acid, emodin, juglone, ellagic acid, condensed tannin, propolis, lysozyme, carvacrol, menthol, eucalyptol, cinnamaldehyde, linalool, artemisinin, berberine, sanguisorbin, quinine, allicin, allyl sulfide, sulforaphane, and lectin.

[0051] In some embodiments of the present invention, the bacteria include at least one of bacteria and fungi.

[0052] In some embodiments of the present invention, the bacteria include at least one of Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus pyogenes, Streptococcus pneumoniae, Enterococcus faecalis / Enterococcus urinaria, Bacillus anthracis, Clostridium tetani, Clostridium perfringens, Clostridium botulinum, Corynebacterium diphtheriae, Listeria monocytogenes, Neisseria meningitidis, Neisseria gonorrhoeae, Escherichia coli, Salmonella typhi, Salmonella non-typhi, Shigella dysenteriae, Vibrio cholerae, Pseudomonas aeruginosa, Bordetella pertussis, Helicobacter pylori, Haemophilus influenzae, Mycobacterium tuberculosis, Streptococcus suis, Salmonella enteritidis, enterotoxic Escherichia coli, Fusobacterium nucleatum, Gardnerella vaginalis, Streptococcus Gordonii, Streptococcus mutans, and Porphyromonas gingivalis.

[0053] In some embodiments of the present invention, the fungus includes at least one of Candida albicans, Trichophyton rubrum, Trichophyton mentagrophytes, Trichophyton tonsurans, Malassezia furfur, Trichophyton nodules, Trichophyton nigricans, and Pneumocystis jirovecii.

[0054] In some embodiments of the present invention, the bacteria are at least one of Escherichia coli, Staphylococcus aureus, Gardnerella vaginalis, Candida albicans, Fusobacterium nucleatum, Streptococcus Gordonii, Salmonella enteritidis, Streptococcus suis, and enterotoxic Escherichia coli.

[0055] A sixth aspect of the invention provides the use of the antimicrobial peptide RK17 of the first aspect of the invention or a salt thereof, the conjugate of the second aspect of the invention, the fusion protein of the third aspect of the invention, the biomaterial of the fourth aspect of the invention, and / or the composition of the fifth aspect of the invention in any one of e1)-e3): e1) Antibacterial; e2) Prepare antibacterial products; e3) Prepare products for preventing and treating diseases caused by pathogenic microorganisms; In some embodiments of the present invention, the bacteria mentioned in e1) includes at least one of bacteria and fungi.

[0056] In some embodiments of the present invention, the bacteria include at least one of Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus pyogenes, Streptococcus pneumoniae, Enterococcus faecalis / Enterococcus urinaria, Bacillus anthracis, Clostridium tetani, Clostridium perfringens, Clostridium botulinum, Corynebacterium diphtheriae, Listeria monocytogenes, Neisseria meningitidis, Neisseria gonorrhoeae, Escherichia coli, Salmonella typhi, Salmonella non-typhi, Shigella dysenteriae, Vibrio cholerae, Pseudomonas aeruginosa, Bordetella pertussis, Helicobacter pylori, Haemophilus influenzae, Mycobacterium tuberculosis, Streptococcus suis, Salmonella enteritidis, enterotoxic Escherichia coli, Fusobacterium nucleatum, Gardnerella vaginalis, Streptococcus Gordonii, Streptococcus mutans, and Porphyromonas gingivalis.

[0057] In some embodiments of the present invention, the fungus includes at least one of Candida albicans, Trichophyton rubrum, Trichophyton mentagrophytes, Trichophyton tonsurans, Malassezia furfur, Trichophyton nodules, Trichophyton nigricans, and Pneumocystis jirovecii.

[0058] In some embodiments of the present invention, the bacteria are at least one of Escherichia coli, Staphylococcus aureus, Gardnerella vaginalis, Candida albicans, Fusobacterium nucleatum, Streptococcus Gordonii, Salmonella enteritidis, Streptococcus suis, and enterotoxic Escherichia coli.

[0059] In some embodiments of the present invention, the product described in e2) comprises at least one of reagents, drugs, feed, feed additives, preservatives, daily chemical products, textiles, paper products, medical materials, and cell culture media.

[0060] In some embodiments of the present invention, the daily chemical products include at least one of hand sanitizer, shower gel, shampoo, mouthwash, toothpaste, soap, cosmetics (including but not limited to facial cleanser, face cream, serum, toner, face cream, etc.), feminine wash, laundry soap, laundry detergent, laundry powder, dish soap, disinfectant, and toilet cleaner.

[0061] In some embodiments of the present invention, the fabric includes at least one of clothing, bedding, disinfectant wipes, dressings, and bandages.

[0062] In some embodiments of the present invention, the paper product includes at least one of sanitary napkins, panty liners, diapers, diaper pads, and wet wipes.

[0063] In some embodiments of the present invention, the medical material comprises at least one of tissue regeneration material, tissue repair material, medical hydrogel material, and medical antibacterial material.

[0064] In some embodiments of the present invention, the dosage form of the drug includes at least one of injection, oral, and topical formulations.

[0065] In some embodiments of the present invention, the product described in e3) includes at least one of pharmaceuticals, daily chemical products, medical devices, and health products.

[0066] In some embodiments of the present invention, the diseases described in e3) include reproductive tract infections (such as urethritis, vaginitis, cervicitis, endometritis, salpingitis, pelvic peritonitis, etc.) and oral diseases (such as dental caries, gingivitis, chronic periodontitis, ulcerative gingivitis, thrush, angular cheilitis, etc.).

[0067] A seventh aspect of the invention provides a product comprising at least one of d1)-d4): d1) The antimicrobial peptide or its salt according to the first aspect of the present invention; d2) The conjugate of the second aspect of the present invention; d3) The fusion protein of the third aspect of the present invention; d4) The composition of the fourth aspect of the present invention; The product is any one of f1)-f2): f1) Antibacterial products; f2) Products for preventing and treating diseases caused by pathogenic microorganisms.

[0068] In some embodiments of the present invention, the bacteria mentioned in f1) includes at least one of bacteria and fungi.

[0069] In some embodiments of the present invention, the bacteria include at least one of Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus pyogenes, Streptococcus pneumoniae, Enterococcus faecalis / Enterococcus urinaria, Bacillus anthracis, Clostridium tetani, Clostridium perfringens, Clostridium botulinum, Corynebacterium diphtheriae, Listeria monocytogenes, Neisseria meningitidis, Neisseria gonorrhoeae, Escherichia coli, Salmonella typhi, Salmonella non-typhi, Shigella dysenteriae, Vibrio cholerae, Pseudomonas aeruginosa, Bordetella pertussis, Helicobacter pylori, Haemophilus influenzae, Mycobacterium tuberculosis, Streptococcus suis, Salmonella enteritidis, enterotoxic Escherichia coli, Fusobacterium nucleatum, Gardnerella vaginalis, Streptococcus Gordonii, Streptococcus mutans, and Porphyromonas gingivalis.

[0070] In some embodiments of the present invention, the fungus includes at least one of Candida albicans, Trichophyton rubrum, Trichophyton mentagrophytes, Trichophyton tonsurans, Malassezia furfur, Trichophyton nodules, Trichophyton nigricans, and Pneumocystis jirovecii.

[0071] In some embodiments of the present invention, the bacteria are at least one of Escherichia coli, Staphylococcus aureus, Gardnerella vaginalis, Candida albicans, Fusobacterium nucleatum, Streptococcus Gordonii, Salmonella enteritidis, Streptococcus suis, and enterotoxic Escherichia coli.

[0072] In some embodiments of the present invention, the product described in f1) comprises at least one of reagents, drugs, feed, feed additives, preservatives, daily chemical products, textiles, paper products, medical materials, and cell culture media.

[0073] In some embodiments of the present invention, the daily chemical products include at least one of hand sanitizer, shower gel, shampoo, mouthwash, toothpaste, soap, cosmetics (including but not limited to facial cleanser, face cream, serum, toner, face cream, etc.), feminine wash, laundry soap, laundry detergent, laundry powder, dish soap, disinfectant, and toilet cleaner.

[0074] In some embodiments of the present invention, the fabric includes at least one of clothing, bedding, disinfectant wipes, dressings, and bandages.

[0075] In some embodiments of the present invention, the paper product includes at least one of sanitary napkins, panty liners, diapers, diaper pads, and wet wipes.

[0076] In some embodiments of the present invention, the medical material comprises at least one of tissue regeneration material, tissue repair material, medical hydrogel material, and medical antibacterial material.

[0077] In some embodiments of the present invention, the dosage form of the drug includes at least one of injection, oral, and topical formulations.

[0078] In some embodiments of the present invention, the product described in f2) includes at least one of pharmaceuticals, daily chemical products, medical devices, and health products.

[0079] In some embodiments of the present invention, the diseases described in f2) include reproductive tract infections (such as urethritis, vaginitis, cervicitis, endometritis, salpingitis, pelvic peritonitis, etc.) and oral diseases (such as dental caries, gingivitis, chronic periodontitis, ulcerative gingivitis, thrush, angular cheilitis, etc.).

[0080] An eighth aspect of the present invention provides a method for inhibiting the growth of pathogenic microorganisms, comprising treating a sample to be treated with an antimicrobial peptide or a salt thereof of the first aspect of the present invention, a conjugate of the second aspect of the present invention, a fusion protein of the third aspect of the present invention, a biomaterial of the fourth aspect of the present invention, a composition of the fifth aspect of the present invention, or a product of the seventh aspect of the present invention; wherein the sample to be treated comprises pathogenic microorganisms or a sample containing pathogenic microorganisms.

[0081] In some embodiments of the present invention, the pathogenic microorganism comprises at least one of bacteria and fungi.

[0082] In some embodiments of the present invention, the bacteria include at least one of Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus pyogenes, Streptococcus pneumoniae, Enterococcus faecalis / Enterococcus urinaria, Bacillus anthracis, Clostridium tetani, Clostridium perfringens, Clostridium botulinum, Corynebacterium diphtheriae, Listeria monocytogenes, Neisseria meningitidis, Neisseria gonorrhoeae, Escherichia coli, Salmonella typhi, Salmonella non-typhi, Shigella dysenteriae, Vibrio cholerae, Pseudomonas aeruginosa, Bordetella pertussis, Helicobacter pylori, Haemophilus influenzae, Mycobacterium tuberculosis, Streptococcus suis, Salmonella enteritidis, enterotoxic Escherichia coli, Fusobacterium nucleatum, Gardnerella vaginalis, Streptococcus Gordonii, Streptococcus mutans, and Porphyromonas gingivalis.

[0083] In some embodiments of the present invention, the fungus includes at least one of Candida albicans, Trichophyton rubrum, Trichophyton mentagrophytes, Trichophyton tonsurans, Malassezia furfur, Trichophyton nodules, Trichophyton nigricans, and Pneumocystis jirovecii.

[0084] In some embodiments of the present invention, the pathogenic microorganism includes at least one of Gardnerella vaginalis, Staphylococcus aureus, Streptococcus, Candida albicans, Escherichia coli, Salmonella, and Fusobacterium nucleatum.

[0085] In some embodiments of the present invention, the pathogenic microorganism is at least one of Escherichia coli, Staphylococcus aureus, Gardnerella vaginalis, Candida albicans, Fusobacterium nucleatum, Streptococcus Gordonii, Salmonella enteritidis, Streptococcus suis, and enterotoxic Escherichia coli.

[0086] A ninth aspect of the present invention provides a method for producing the antimicrobial peptide or a salt thereof of the first aspect of the present invention, the method comprising chemical synthesis or by culturing the transgenic cell line of the fourth aspect of the present invention.

[0087] In some embodiments of the present invention, the chemical synthesis includes polypeptide solid-phase synthesis.

[0088] A tenth aspect of the present invention provides a method for preventing and / or treating diseases caused by pathogenic microorganism infection, comprising the steps of administering to a subject an effective dose of an antimicrobial peptide of the first aspect of the present invention or a salt thereof, a conjugate of the second aspect of the present invention, a fusion protein of the third aspect of the present invention, a biomaterial of the fourth aspect of the present invention, a composition of the fifth aspect of the present invention, or a product of the seventh aspect of the present invention.

[0089] In some embodiments of the present invention, the antimicrobial peptide or its salts, conjugates, fusion proteins, compositions and / or products are administered at least once daily.

[0090] In some embodiments of the present invention, when the number of administrations is more than twice a day, the same dose is administered each time.

[0091] In some embodiments of the present invention, when the administration frequency is more than twice a day, a different dose is administered each time.

[0092] The term "effective dose" refers to the amount that is effective for treatment or prevention in the subject being treated. The precise amount of a therapeutic composition also depends on the practitioner's judgment and may be unique to each individual. Appropriate regimens for initial dosing and synergists are also variable, but are represented by initial and subsequent dosing. Factors affecting dosage include the patient's physical and clinical condition, route of administration, intended therapeutic goal (symptom relief or cure), and the efficacy, stability, and toxicity of the specific therapeutic substance or the efficacy of other treatments the subject may be receiving.

[0093] The term "subject" refers to any mammal. A subject described as "in need" refers to a mammal requiring treatment (or prevention) for a disease. This includes, for example, any one or more animals within the class Mammalia, such as Monotremes, Marsupials, Insectivora, Flying Lepidoptera, Climbing Therapsids, Dermoptera, Chiroptera, Primates, Phenotypica, Pholidota, Lagomorpha, Rodentia, Carnivora, Sirenia, Hyracodontia, Tubularodontia, Perissodactylus, Artiodactylus, Cetacea, etc. Common mammals include Rodentia (e.g., mice, rats, hamsters, guinea pigs), Lagomorpha (e.g., rabbits), Perissodactylus (e.g., horses, donkeys), Artiodactylus (e.g., sheep, goats, camels, cattle, pigs), Primates (e.g., monkeys, gorillas, chimpanzees, humans), and Carnivora (e.g., dogs, cats). It is understood that a subject may be healthy or may have a disease at any stage of development.

[0094] The beneficial effects of this invention are: This invention discloses for the first time an antimicrobial peptide (GFGRGLWKGVKRVLSRAGVFSAGGRNL, SEQ ID NO:1) or its salt, which has good antibacterial effects against common genital bacteria (such as Staphylococcus aureus, Gardnerella vaginalis, and Candida albicans), oral pathogens (such as Fusobacterium nucleatum and Streptococcus Gordonii), and other pathogens (such as Escherichia coli, Salmonella enteritidis, and Streptococcus suis), and only exhibits slight cytotoxicity to mammalian cells at high concentrations.

[0095] Specifically, the antimicrobial peptide provided by this invention has the following characteristics compared to existing antimicrobial drugs: 1. Reduced Drug Resistance: Antimicrobial peptides interfere with the survival environment of pathogens through multiple mechanisms of action, reducing the development of drug resistance. Compared to traditional antibiotics, antimicrobial peptides effectively avoid drug resistance problems and ensure efficacy during long-term use.

[0096] 2. Regulating the balance of the microecology: Antimicrobial peptides can selectively inhibit the growth of harmful bacteria, protect and promote the proliferation of probiotics, thereby helping to maintain the stability of the microecology and reduce the risk of recurrence and reinfection.

[0097] 3. High safety and no significant side effects: Antimicrobial peptides are widely available and naturally exist in living organisms. They have no obvious toxic side effects on the human body and do not damage liver and kidney function during use, thus avoiding the safety risks associated with antibiotics.

[0098] 4. Precision treatment: Antimicrobial peptides have strong selectivity and can act on specific pathogens, thereby achieving the purpose of precision treatment without widely affecting the growth of other beneficial microorganisms.

[0099] 5. Potential to replace antibiotics: With the increasing severity of antibiotic abuse, antimicrobial peptides, as a natural and green alternative, meet the needs of modern health, safety and environmental protection, and have broad application prospects. Attached Figure Description

[0100] The present invention will be further described below with reference to the accompanying drawings and embodiments, wherein: Figure 1 The structure shown is the tertiary structure of the antimicrobial peptide (GFGRGLWKGVKRVLSRAGVFSAGGRNL, SEQ ID NO:1) of this invention, which is the result predicted by the AlphaFold3 protein prediction tool.

[0101] Figure 2 The antimicrobial peptides of this invention exhibit antibacterial effects at a level of 10 μmol / L against genital tract bacteria (Staphylococcus aureus ATCC6538, Gardnerella vaginalis ATCC14018, Candida albicans ATCC10231), oral pathogens (Fusobacterium nucleatum ATCC25586, Streptococcus Gordonii ATCC10558), and at a level of 8 μmol / L against other pathogens (Escherichia coli ATCC25922, Salmonella enteritidis CICC 20082847, Streptococcus suis CVCC4268, and enterotoxic Escherichia coli K88).

[0102] Figure 3 The results show the minimum inhibitory concentration (MIC) test results of the antimicrobial peptides of this invention against Escherichia coli ATCC25922 and Staphylococcus aureus ATCC 6538.

[0103] Figure 4 The results show the minimum inhibitory concentration (MIC) test results of the antimicrobial peptides of this invention against enterotoxic Escherichia coli K88, Salmonella enteritidis CICC 20082847, and Streptococcus suis CVCC 4268.

[0104] Figure 5 The scanning electron microscopy characterization of the antimicrobial peptides of this invention disrupting the structural integrity of bacteria (enterotoxic Escherichia coli K88) is shown with a scale bar of 200 μm.

[0105] Figure 6The results show the hemolytic effect of the antimicrobial peptides of this invention (using porcine erythrocytes as test cells).

[0106] Figure 7 This is the result of the cytotoxicity test (using HT-29 cells as test cells) of the antimicrobial peptide of the present invention. Detailed Implementation

[0107] The following will describe the concept and technical effects of the present invention clearly and completely with reference to embodiments, so as to fully understand the purpose, features and effects of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, not all embodiments. Other embodiments obtained by those skilled in the art based on the embodiments of the present invention without creative effort are all within the scope of protection of the present invention.

[0108] Unless otherwise specified in the examples, the procedures should be performed under standard conditions or conditions recommended by the manufacturer. Reagents or instruments whose manufacturers are not specified are all commercially available products.

[0109] The features and performance of the present invention will be further described in detail below with reference to embodiments.

[0110] Example 1: Discovery and Screening of Antimicrobial Peptides By mining and screening potential antimicrobial peptide genes in the pig's gut microbiome, an antimicrobial peptide was obtained. The amino acid sequence of this antimicrobial peptide is GFGRGLWKGVKRVLSRAGVFSAGGRNL (SEQ ID NO:1).

[0111] Example 2 Physicochemical Properties and Structural Biological Analysis of Antimicrobial Peptides Based on the amino acid sequence of the antimicrobial peptide in Example 1, the physicochemical properties of the antimicrobial peptide were analyzed and statistically analyzed in this example. At the same time, the three-dimensional structure of this antimicrobial peptide was predicted using the Alphafold protein three-dimensional structure prediction tool.

[0112] The physicochemical properties of the antimicrobial peptides are shown in Table 1, and their three-dimensional structures are shown in Table 1. Figure 1 As shown, the antimicrobial peptide exhibits a distinct α-helix structure.

[0113] Table 1 Physicochemical properties of antimicrobial peptides

[0114] The above-mentioned antimicrobial peptides were further synthesized using solid-phase peptide synthesis (SPPS), and the general process is as follows: (1) Calculate the weight of each raw material (with protection) based on the weight of the target polypeptide; (2) Place the resin in a 150 ml reactor and add 50 ml of dichloromethane to soak for 2 hours; (3) Wash the resin with dichloromethane and then dry it. Repeat this process four times to dry the resin. (4) Weigh the first amino acid (with protection) at the C-terminus of the target polypeptide into the reactor, add dichloromethane and N-diisopropylethylamine, and then place the reactor in a shaker at 30°C for 2 hours. (5) Seal with methanol solution (methanol: N-diisopropylethylamine: dichloromethane = 1:1:2) for half an hour, then wash with dichloromethane four times and dry; (6) Add a 20% piperidine solution to the reactor to remove the 9-fluorenylmethoxycarbonyl protecting group. After removing the protecting group, wash four times with dichloromethane and then dry. (7) Take a small amount of resin and test it using the ninhydrin method. If the resin is colored, it indicates that the protecting group has been successfully removed. (8) Weigh the second amino acid (with protection) at the C-terminus and add it to the reactor. Then add 1-hydroxybenzotriazole and 1,3-diisopropylcarbodiimide. Then place the reactor in a shaker at 30°C and react for 1 hour. (9) Take a small amount of resin for testing. Use the ninhydrin method to test. If the resin is colored, it indicates that the condensation is incomplete and the reaction should continue. If the resin is colorless, it indicates that the reaction is complete. After the reaction is complete, wash the resin four times with dichloromethane and then dry it. (10) Add a certain amount of 20% piperidine (piperidine / dichloromethane = 1:4) to the reactor, and shake it on a decolorizing shaker for 20 minutes to remove the 9-fluorenylmethoxycarbonyl protecting group on the resin. After removing the protecting group, wash it four times with dichloromethane, and then dry it to check whether the protection has been removed; (11) Take a small amount of resin and test it using the ninhydrin method. If the resin is colored, it indicates that the protecting group has been successfully removed. (12) Connect the amino acids sequentially according to steps (8)-(11); (13) Add Boc anhydride (Di-tert-butyl dicarbonate) to protect the N-terminal amino group; (14) Take a small amount of resin for testing. Use the ninhydrin method to test. If the resin is colored, it indicates that the condensation is incomplete and the reaction should continue. If the resin is colorless, it indicates that the reaction is complete. After the reaction is complete, wash the resin four times with dichloromethane and then dry it. (15) Remove the special protecting group from the lysine side chain and connect the small molecules with the side chain sequentially according to steps (8)-(11); (16) Use a cleavage reagent to remove all the protecting groups of the polypeptide and cut the polypeptide off the resin for further purification.

[0115] Example 3: Verification of the antibacterial effect of antimicrobial peptides This embodiment is used to verify the antibacterial effect of the antimicrobial peptide in Example 1, as detailed below: 1. Preliminary confirmation of the antibacterial activity of antimicrobial peptides Nine common pathogens causing genital tract, oral cavity and other infections were used in this test (Table 2).

[0116] Table 2 Pathogenic bacteria used in antibacterial tests

[0117] The antimicrobial effect of the antimicrobial peptide from Example 1 at a fixed concentration (10 μmol / L) was preliminarily confirmed as follows: For bacteria, the pathogens were inoculated into Mueller Hinton broth (MHB) medium and cultured overnight at 37°C with shaking at 220 rpm. For fungi, the pathogens were inoculated into YPD liquid medium and cultured overnight at 30°C with shaking at 220 rpm. For anaerobic bacteria, the pathogens were inoculated into BHIS liquid medium and cultured overnight in an anaerobic incubator at 37°C. The culture was then diluted to the initial concentration (OD). 600 =0.1, approximately equivalent to 1×10 8 CFU / mL), then diluted 100-fold to obtain the bacterial suspension. Antimicrobial peptides (final concentration 8 μmol / L or 10 μmol / L) were added to the bacterial suspension. 200 μL of the bacterial suspension containing the antimicrobial peptides (final concentration 8 μmol / L or 10 μmol / L) was added to a 96-well plate. A blank control group (200 μL MHB solution) and a negative control group (100 μL of peptide-free bacterial suspension + 100 μL MHB / YPD solution; the culture medium was selected according to the pathogens mentioned above) were also set up. After incubation at 37℃ for 24 hours, the OD was measured. 600 The OD value of Candida albicans was determined after incubation at 30°C for 14 hours. 600 Value. The effective value of antimicrobial peptides is defined as OD. 600 The mean value was less than 70% of that of the negative control group, and the difference between the two groups was statistically significant. P Value < 0.05). All experiments were performed in triplicate.

[0118] The results are as follows Figure 2 As shown, at the selected concentrations (8 μmol / L or 10 μmol / L), the antimicrobial peptides exhibited significant antimicrobial effects against all tested pathogens.

[0119] 2. Minimum inhibitory concentration (MIC) test of antimicrobial peptides Five pathogenic bacteria were used in this test, namely Escherichia coli (E. coli) Escherichia coli ATCC 25922), Staphylococcus aureus ( Staphylococcus aureusATCC 6538), enterotoxic Escherichia coli (ETEC K88), Salmonella enteritidis (ATCC 6538), enterotoxic Escherichia coli (ETEC K88), enterotoxic Salmonella ... Salmonella enteritidis CICC 20082847), Streptococcus suis ( Streptococcus suits (CVCC4268) The inhibitory effects of a series of antimicrobial peptide solutions at various concentrations on these bacteria were tested. The specific procedure was as follows: The pathogen was inoculated into Mueller Hinton broth (MHB) medium and cultured overnight at 37°C with shaking at 220 rpm. The culture was then diluted to the initial concentration (OD). 600 =0.1, approximately equivalent to 1×10 8 CFU / mL), then diluted 100-fold. Antimicrobial peptides (Escherichia coli ATCC 25922, Staphylococcus aureus ATCC 6538, final concentrations of 0.1, 0.3, 1, 3, 10 μmol / L; enterotoxic Escherichia coli K88, Salmonella enteritidis CICC 20082847, Streptococcus suis CVCC 4268, final concentrations of 0.125, 0.25, 0.5, 1, 2, 4, 8 μmol / L) were added to the bacterial suspension. 200 μL of bacterial suspension containing different concentrations of antimicrobial peptides was added to a 96-well plate. A blank control group (200 μL MHB solution) and a negative control group (100 μL of sterile peptide-free bacterial suspension + 100 μL MHB solution) were also established. OD was measured after incubation at 37℃ for 24 hours. 600 The MIC was defined as the lowest concentration of antimicrobial peptide that differed from the mean of the negative control group by three standard deviations. All experiments were performed in triplicate.

[0120] The results are as follows Figures 3-4 As can be seen, the antibacterial ability of the antimicrobial peptide solution gradually increases with increasing concentration, and the OD... 600 The values ​​gradually decreased. The MIC values ​​of the antimicrobial peptides against Escherichia coli ATCC25922 and Staphylococcus aureus ATCC6538 were both 3 μmol / L, the MIC value against enterotoxic Escherichia coli K88 was 1 μmol / L, and the MIC value against Salmonella enteritidis CICC20082847 and Streptococcus suis CVCC 4268 was 4 μmol / L (Table 3).

[0121] Table 3. Minimum Inhibitory Concentration (μmol / L)

[0122] 3. Scanning electron microscopy characterization of antimicrobial peptides disrupting bacterial structural integrity Enterotoxic Escherichia coli K88 was selected as the test strain to investigate the mechanism of action of antimicrobial peptides on its bactericidal activity. Specifically, following standard scanning electron microscopy sample preparation procedures, the enterotoxic Escherichia coli K88 strain was washed with PBS, resuspended, and adjusted to OD250. 600The concentration of antimicrobial peptide was 0.3, and 4× MIC was added. The mixture was incubated at 37°C for 1 to 3 hours. Bacterial cells were collected by centrifugation at 8000 rpm for 3 minutes, and 0.5 mL of 2.5% (v / v) glutaraldehyde was added. The mixture was fixed at 4°C for 24 hours. The fixed samples were washed three times with PBS (15 minutes each time), dehydrated with a gradient of ethanol (30%-100% concentrations / 15 minutes each), and dried. The dried samples were fixed with conductive carbon gel, then ion-sputtered with platinum for 2 minutes. Finally, the samples were observed using a Hitachi Reguilus 8100 field emission scanning electron microscope.

[0123] The results are as follows Figure 5 As shown, with the extension of the incubation time between the antimicrobial peptide and bacterial cells, the bacterial cell structure is gradually destroyed. The electron micrographs after an incubation time of 3 hours show obvious cell structure destruction, indicating that the antimicrobial peptide achieves its bactericidal purpose by destroying the integrity of the bacterial cell structure.

[0124] Example 4 Biosafety Testing of Antimicrobial Peptides 1. Hemolysis test Using porcine erythrocytes as test cells, the hemolytic effect of the antimicrobial peptides of Example 1 at different concentration gradients (128, 64, 32, 16, 8, 4, 2, 1, 0.5, 0.25 μmol / L) was detected to investigate the toxic effects of the antimicrobial peptides on erythrocytes. The specific experimental procedures were as follows: A 4% porcine erythrocyte suspension (Solarbio) was centrifuged at 2000 rpm for 5 minutes, and the supernatant was discarded. The erythrocytes were then resuspended in PBS buffer containing gradient concentrations (128, 64, 32, 16, 8, 4, 2, 1, 0.5, 0.25 µmol / L) of antimicrobial peptides and incubated at 37°C for 1 hour. Triton X-100 was used as a positive control (highest concentration 1%, subsequent concentrations 2-fold dilutions). After the reaction, the cells were centrifuged at 3000 rpm for 10 minutes, and the absorbance of the supernatant was measured at 570 nm. All experiments were independently repeated three times.

[0125] The results are as follows Figure 6 As shown, at the highest concentration of 128 μmol / L, the antimicrobial peptide only exhibited a slight hemolytic reaction, indicating that the antimicrobial peptide of Example 1 has low toxicity to mammalian erythrocytes.

[0126] 2. Toxicity test on mammalian cells The toxic effects of the antimicrobial peptide of Example 1 on HT-29 cells were tested using different concentration gradients (128, 64, 32, 16, 8, 4, 2, 1, 0.5, 0.25 μmol / L) of the HT-29 cell line (human colorectal cell line). The specific experimental procedures were as follows: HT-29 cells in logarithmic growth phase were seeded in 96-well plates and cultured in DMEM medium (Gibco) containing 10% (v / v) fetal bovine serum (FBS, Gibco) and 1% (v / v) penicillin-streptomycin antibiotic (Solarbio); then incubated at 37°C in a 5% CO2 incubator for 12 hours. After discarding the original medium, the medium was replaced with DMEM medium containing 10% (v / v) FBS and gradient concentrations (128, 64, 32, 16, 8, 4, 2, 1, 0.5, 0.25 µmol / L) of antimicrobial peptides and cultured for another 24 hours; Triton X-100 was used as a positive control (highest concentration 1%, followed by 2-fold dilutions); finally, cell viability was assessed using the MTT assay kit (Beyotime), with absorbance measured at 570 nm. Each antimicrobial peptide concentration was tested in four independent replicates.

[0127] The results are as follows Figure 7 As shown, the antimicrobial peptide exhibited certain toxicity to HT-29 cells with increasing concentration. At a concentration 4 times the MIC (16 μmol / L), the survival rate of mammalian cells was higher than 70%, indicating that the antimicrobial peptide had low cytotoxicity to mammalian cells at its effective concentration.

[0128] The embodiments of the present invention have been described in detail above with reference to the accompanying drawings. However, the present invention is not limited to the above embodiments, and various changes can be made within the scope of knowledge possessed by those skilled in the art without departing from the spirit of the present invention. Furthermore, the embodiments of the present invention and the features thereof can be combined with each other unless otherwise specified.

Claims

1. An antimicrobial peptide or a salt thereof, wherein the amino acid sequence of the antimicrobial peptide is as follows: a1) SEQ ID NO:1; or a2) An amino acid sequence of SEQ ID NO:1 that has undergone substitution and / or deletion and / or addition of one or more amino acids and has the same function as SEQ ID NO:1; or a3) An amino acid sequence that has at least 80% homology with SEQ ID NO:1 and has the same function as SEQ ID NO:

1.

2. A conjugate comprising a modification portion and the antimicrobial peptide of claim 1; Preferably, the modified portion is located at the N-terminus and / or C-terminus of the antimicrobial peptide; Preferably, the modified portion comprises at least one of chemical modification, targeting portion, fluorescent dye, and protein tag; Preferably, the chemical modification includes at least one of amidation, acetylation, amination, methylation, phosphorylation, glycosylation, and ubiquitination; Preferably, the targeting portion comprises at least one of a ligand, a receptor, and an antibody; Preferably, the protein tag includes at least one of His, Flag, GST, MBP, HA, Myc, GFP, and biotin.

3. A fusion protein comprising a first domain and a second domain; The first structural domain is the antimicrobial peptide according to claim 1; The second domain has the effect of prolonging the in vivo half-life; Preferably, the first domain is located at the N-terminus and / or C-terminus of the antimicrobial peptide; Preferably, the second domain and the antimicrobial peptide are connected to the N-terminus and / or C-terminus of the antimicrobial peptide via a connector.

4. A biomaterial relating to the antimicrobial peptide of claim 1, the conjugate of claim 2, or the fusion protein of claim 3, wherein the biomaterial comprises any one of b1)-b8): b1) A nucleic acid molecule encoding the antimicrobial peptide of claim 1, the conjugate of claim 2, or the fusion protein of claim 3; b2) An expression cassette containing the nucleic acid molecule described in b1); b3) A carrier containing the nucleic acid molecule described in b1); b4) A carrier containing the expression box described in b2); b5) Transgenic cell lines containing the nucleic acid molecules described in b1); b6) Transgenic cell lines containing the expression cassette described in b2); b7) A transgenic cell line containing the vector described in b3); b8) A transgenic cell line containing the vector described in b4).

5. A composition comprising at least one of c1)-c3): c1) The antimicrobial peptide or a salt thereof as described in claim 1; c2) The conjugate according to claim 2; c3) The fusion protein according to claim 3; Preferably, the composition further comprises other active antibacterial ingredients; Preferably, the other active antibacterial components include at least one of penicillin antibiotics, cephalosporin antibiotics, carbapenem antibiotics, lipopeptide antibiotics, aminoglycoside antibiotics, tetracycline antibiotics, lincosamide antibiotics, macrolide antibiotics, fluoroquinolone antibiotics, and fusidic acid. Preferably, the other active antibacterial components include at least one of cinnamaldehyde, eugenol, coptis chinensis, berberine, quercetin, baicalin, thymol, andrographolide, curcumin, kaempferol, apigenin, soy isoflavones, gallic acid, protocatechuic acid, chlorogenic acid, emodin, juglone, ellagitannin, condensed tannin, propolis, lysozyme, carvacrol, menthol, eucalyptol, cinnamaldehyde, linalool, artemisinin, berberine, sanguisorbin, quinine, allicin, allyl sulfide, sulforaphane, and lectin.

6. The use of the antimicrobial peptide of claim 1 or its salt, the conjugate of claim 2, the fusion protein of claim 3, the biomaterial of claim 4, and / or the composition of claim 5 in any one of e1)-e3): e1) Antibacterial; e2) Prepare antibacterial products; e3) Prepare products for preventing and treating diseases caused by pathogenic microorganisms; Preferably, the product described in e2) comprises at least one of the following: reagents, drugs, feed, feed additives, preservatives, daily chemical products, cleaning and disinfection products, textiles, paper products, medical materials, and cell culture media; Preferably, the daily chemical products include at least one of hand sanitizer, shower gel, shampoo, mouthwash, toothpaste, soap, cosmetics, feminine wash, laundry soap, laundry detergent, laundry powder, dish soap, disinfectant, and toilet cleaner; Preferably, the fabric comprises at least one of clothing, bedding, disinfectant wipes, dressings, and bandages; Preferably, the paper product includes at least one of sanitary napkins, panty liners, diapers, and diaper pads; Preferably, the medical material comprises at least one of tissue regeneration materials, tissue repair materials, medical hydrogel materials, and medical antibacterial materials; Preferably, the product described in e3) includes at least one of the following: pharmaceuticals, daily chemical products, medical devices, and health products.

7. A product comprising at least one of d1)-d4): d1) The antimicrobial peptide or a salt thereof as described in claim 1; d2) The conjugate according to claim 2; d3) The fusion protein according to claim 3; d4) The composition according to claim 5; The product is any one of f1)-f2): f1) Antibacterial products; f2) Products for preventing and treating diseases caused by pathogenic microorganisms.

8. The product according to claim 7, characterized in that: The products described in f1) include at least one of the following: reagents, drugs, feed, feed additives, preservatives, daily chemical products, cleaning and disinfection products, textiles, paper products, medical materials, and cell culture media; Preferably, the daily chemical products include at least one of hand sanitizer, shower gel, shampoo, mouthwash, toothpaste, soap, cosmetics, feminine wash, laundry soap, laundry detergent, laundry powder, dish soap, disinfectant, and toilet cleaner; Preferably, the fabric comprises at least one of clothing, bedding, disinfectant wipes, dressings, and bandages; Preferably, the paper product includes at least one of sanitary napkins, panty liners, diapers, and diaper pads; Preferably, the medical material comprises at least one of tissue regeneration materials, tissue repair materials, medical hydrogel materials, and medical antibacterial materials; Preferably, the product described in f2) includes at least one of the following: pharmaceuticals, daily chemical products, medical devices, and health products.

9. A method for inhibiting the growth of pathogenic microorganisms, comprising treating a sample to be treated with the antimicrobial peptide of claim 1 or a salt thereof, the conjugate of claim 2, the fusion protein of claim 3, the biomaterial of claim 4, the composition of claim 5, or the product of claim 7; wherein the sample to be treated comprises pathogenic microorganisms or a sample containing pathogenic microorganisms; Preferably, the pathogenic microorganism includes at least one of bacteria and fungi; Preferably, the pathogenic microorganism includes at least one of Gardnerella vaginalis, Staphylococcus aureus, Streptococcus, Candida albicans, Escherichia coli, Salmonella, and Fusobacterium nucleatum.

10. A method for producing the antimicrobial peptide of claim 1 or a salt thereof, the method comprising chemical synthesis or by culturing the transgenic cell line of claim 4; Preferably, the chemical synthesis includes polypeptide solid-phase synthesis.

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