Antimicrobial peptides derived from lactoferrin and their applications

Abstract

The present invention belongs to the field of biomedicine, and specifically relates to an antimicrobial peptide derived from lactoferrin and its application. The antimicrobial peptide derived from lactoferrin, whose amino acid sequence is shown in any one of SEQ ID NOs: 1 to 98, has a broad-spectrum antimicrobial effect, especially significant antibacterial effect against Pseudomonas putida, Rhodococcus erythropolis, Bacillus subtilis, Malassezia furfur, Cryptococcus neoformans, and Candida albicans, and has great application value.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine, and in particular relates to an antimicrobial peptide derived from lactoferrin and applications thereof. Background Art

[0002] Antibiotic resistance has become a serious problem affecting anti-infection treatment and the development of animal husbandry. The research and development of new antimicrobial substances is the key to solving this problem. To protect the health of humans and animals, antibiotic alternatives can be used. Antimicrobial peptides are considered to be the best alternative to antibiotics because of their broad-spectrum and highly effective antibacterial, antifungal, antiviral, antiparasitic, and antitumor effects, lack of residue, and low resistance to drug resistance. They can also promote wound healing and angiogenesis. (Fischbach MA, Walsh CT. Antibiotics for emerging pathogens [J]. Science, 2009, 325(5944): 1089-1093.) Antimicrobial peptides, also known as host defense peptides or peptide antibiotics, are widely present in nature and are an important component of the innate immune system. In recent years, they have become one of the hot topics in international life science research (Li Jie, Hang Bolin, Qin Aijian, et al. Bioinformatics analysis of bovine hemoglobin-derived antimicrobial peptide P3 and its similar peptides [J]. Chinese Journal of Animal Husbandry and Veterinary Medicine, 2017, 44(1): 59-64.). At present, a large number of antimicrobial peptides of natural origin or artificial synthesis are being extensively and deeply studied. As of May 2022, more than 3,000 antimicrobial peptides have been included in the antimicrobial peptide database (https: / / aps.unmc.edu) (Zhang Shuhui, Luo Lu, Sun Xueyan, et al. Optimization of extraction process and activity study of antimicrobial peptides from Agaricus edulis mycelium [J]. Food and Machinery, 2022, 38(08): 158-165. & Browne K, Chakraborty S, Chen R, et al. A new era of antibiotics: The clinical potential of antimicrobial peptides [J]. International Journal of Molecular Sciences, 2020, 21(19): 7047.).

[0003] Most traditional antimicrobial peptides are isolated from microorganisms, animals, or plants. The isolation process is cumbersome, time-consuming, costly, and risky. Random screening is highly selective and can easily compromise the activity of some antimicrobial peptides. Efficiently discovering more antimicrobial peptides with high activity, stability, and low toxicity is a current research hotspot. However, the protein source of peptides obtained through semi-rational design is unclear, and subsequent applications require addressing safety concerns. With the continued accumulation of research on the activity and related physicochemical properties of antimicrobial peptides and the advancement of bioinformatics, there remains a need for the development of newer, more valuable broad-spectrum antimicrobial peptides.

[0004] Lactoferrin (LF) is a multifunctional iron-binding glycoprotein belonging to the transferrin family. It is abundant in milk and is a major protein in human exocrine secretions, exhibiting diverse biological activities (Fernandes K E, Carter D A. The antifungal activity of lactoferrin and its derived peptides: mechanisms of action and synergy with drugs against fungal pathogens [J]. Frontiers in Microbiology, 2017, 8:2). LF not only possesses broad-spectrum antimicrobial, anti-inflammatory, and immune-regulating properties, but is also natural and non-toxic. Bovine lactoferrin (bLF) is readily available, amenable to large-scale production, and poses no ethical challenges, offering enhanced clinical applications. Most of the lactoferrin currently used in commerce is derived from bovine milk. 0.23 mg of lactoferrin can be extracted from each milliliter of milk, with a purity of 95% (Yang Anshu, Gao Jinyan, Li Xin, et al. Study on the separation and purification of lactoferrin from bovine milk [J]. Food Science and Technology, 2005(3):17-19.). In 2015, the European Food Safety Authority (EFSA) approved bovine lactoferrin as a new food ingredient (Wang Nannan, Cai Tingting, Zhu Wanping. Research progress on the antibacterial effect of lactoferrin [J]. Food Science and Technology, 2020, 45(08):233-237.).In addition to being an active intact protein, LF is also a rich source of antimicrobial peptides cleaved from the polypeptide chain by a variety of proteolytic enzymes (Sinha M, Kaushik S, Kaur P, et al. Antimicrobial lactoferrin peptides: the hidden players in the protective function of amultifunctional protein[J]. International Journal of Peptides, 2013, 2013.), and many have even been shown to have antimicrobial activity exceeding that of the entire LF (Fernandes KE, Payne RJ, Carter DA. Lactoferrin-derived peptide lactofunginis potently synergistic with amphotericin B[J]. Antimicrobial Agents and Chemotherapy, 2020, 64(10): e00842-20.). However, the access to its antimicrobial peptides is limited and the preparation process is complicated. With the development of bioinformatics and big data analysis technologies, the protein sources of antimicrobial peptides obtained through semi-rational design are unknown, and their subsequent applications are subject to safety concerns. Therefore, this invention deeply explores the sequence and function of lactoferrin to obtain new potential antimicrobial peptides. To date, no antimicrobial peptides have been directly analyzed by building an overlapping peptide library based on the full sequence of lactoferrin. Summary of the Invention

[0005] The present invention adopts a bioinformatics method to design a lactoferrin peptide library based on an overlapping peptide library, and uses the Antimicrobial Peptide Scanner vr.2 server to predict and screen antimicrobial peptides with high scores. The antimicrobial peptides can be used as additives and are widely used in the fields of cosmetics or medicine.

[0006] This invention addresses the current challenges of extracting antimicrobial peptides from natural ingredients, including high cost, low efficiency, and low success rate. Based on the existing lactoferrin sequence, this method utilizes bioinformatics methods to screen for antimicrobial peptides, and experimentally validates that it can efficiently and rapidly screen for antimicrobial peptides. The antimicrobial peptides are derived from the lactoferrin sequence, achieving the goals of good biocompatibility, environmental friendliness, and safety.

[0007] In order to achieve the above object, the technical solution provided by the present invention is:

[0008] The present invention provides an antimicrobial peptide derived from lactoferrin, characterized in that its amino acid sequence is shown in any one of SEQ ID NOs: 1 to 98; preferably, its amino acid sequence is shown in any one of SEQ ID NOs: 1 to 8, 31-36, 64-77, and 86-89; more preferably, its amino acid sequence is shown in any one of SEQ ID NO: 2, SEQ ID NOs: 31-32, SEQ ID NO. 36, SEQ ID NO: 64, SEQ ID NOs: 67-73, SEQ ID NO: 77, SEQ ID NO. 86, and SEQ ID NO. 88.

[0009] The present invention also provides a nucleic acid encoding the antimicrobial peptide, and an expression vector or recombinant cell containing the nucleic acid.

[0010] The present invention further provides the use of the antimicrobial peptide in antibacterial or antibacterial activities. The term "antimicrobial or antibacterial activity" refers to an antimicrobial or antibacterial activity against Escherichia coli, Pseudomonas aeruginosa, Pseudomonas putida, Staphylococcus aureus, Propionibacterium acnes, Rhodococcus erythropolis, Bacillus subtilis, Malassezia furfur, Cryptococcus neoformans, or Candida albicans; preferably, the term "antimicrobial or antibacterial activity" refers to an antimicrobial or antibacterial activity against Pseudomonas putida, Rhodococcus erythropolis, Bacillus subtilis, Malassezia furfur, Cryptococcus neoformans, or Candida albicans.

[0011] The present invention also provides the use of the antimicrobial peptide or its encoding nucleic acid in the preparation of an antimicrobial product. Specifically, the product is a medicine, cosmetic, toiletries, or pesticide; preferably, the antimicrobial activity refers to an antimicrobial or antibacterial effect against Escherichia coli, Pseudomonas aeruginosa, Pseudomonas putida, Staphylococcus aureus, Propionibacterium acnes, Rhodococcus erythropolis, Bacillus subtilis, Malassezia furfur, Cryptococcus neoformans, or Candida albicans; more preferably, the antimicrobial activity refers to an antimicrobial or antibacterial effect against Pseudomonas putida, Rhodococcus erythropolis, Bacillus subtilis, Malassezia furfur, Cryptococcus neoformans, or Candida albicans.

[0012] This invention innovatively utilizes overlapping peptide libraries to design a lactoferrin peptide library, enabling the establishment of a complete sequence-based lactoferrin peptide library. This reduces time and costs, improves the accuracy of the hydrolyzate sequence composition, and demonstrates originality in the field of bioinformatics for bioactive peptides. Furthermore, by innovatively utilizing the AMP Scanner vr.2 server to predict and screen for highly active lactoferrin antimicrobial peptides, a high-throughput screening method for lactoferrin antimicrobial peptides is implemented. This method reduces the number of compounds tested in bioactivity assays while saving time and costs associated with peptide sequence purification and identification, thereby increasing the probability of successful screening.

[0013] Specifically, the peptide provided by the present invention has a broad-spectrum antibacterial effect, especially an antibacterial effect on Pseudomonas putida, Rhodococcus erythropolis, Bacillus subtilis, Malassezia furfur, Cryptococcus neoformans and Candida albicans.

[0014] The present invention innovatively implements a method for establishing a lactoferrin peptide library based on bioinformatics. By utilizing overlapping peptide library technology and combining it with food-derived lactoferrin, a full-sequence peptide library can be rapidly established, significantly saving time and cost and improving the accuracy of the results. Furthermore, a method for high-throughput screening of lactoferrin antimicrobial peptides based on bioinformatics is also described. By utilizing the AMPScanner vr.2 server and combining it with the lactoferrin peptide library, highly active antimicrobial peptides can be rapidly screened out, significantly saving manpower and material resources, avoiding the hazards of toxic reagents to experimenters during the screening process, and improving screening efficiency and addressing the limitations of peptide library capacity. The peptides thus obtained by the present invention have a broad-spectrum antimicrobial effect, particularly significant antibacterial activity against Pseudomonas putida, Rhodococcus erythropolis, Bacillus subtilis, Malassezia furfur, Cryptococcus neoformans, and Candida albicans, thereby having a significant application value. DETAILED DESCRIPTION

[0015] Unless otherwise specified, the reagents and instruments used in the following examples are all commercially available products.

[0016] Example 1 Design of lactoferrin peptide library and rapid screening method based on overlapping peptide library

[0017] 1. Establishment of Lactoferrin Peptide Library

[0018] (1) Obtain the amino acid sequence of lactoferrin in the UniProt protein database;

[0019] (2) The above amino acid sequence was designed using the overlapping peptide library method to obtain 1398 overlapping peptide segments with a length of 11, and adjacent peptide segments overlapped by 10 amino acids.

[0020] 2. Bioinformatics prediction of antimicrobial activity sequences

[0021] (1) Submit the above 1398 peptide sequences in a valid FASTA format to the Antimicrobial Peptide (AMP) Scanner vr.2 server to obtain the predicted classification (AMPs or Non-AMPs) and predicted probability of the above predicted peptides;

[0022] (2) Based on the predicted classification in step (1), 268 peptide sequences with AMP activity were screened;

[0023] (3) connecting all x consecutive peptide sequences with overlapping partial amino acid sequences in step (2) into a long peptide chain consisting of y amino acid residues, wherein the C-terminus of the nth amino acid sequence is connected to the non-overlapping sequence of the n+x-1th amino acid sequence, to obtain 72 peptide sequences;

[0024] (4) The peptide sequences obtained in step (3) were submitted to the AMP Scanner vr.2 server according to step (1), and 63 peptide sequences with AMP activity were obtained. The results are shown in Table 1.

[0025] Table 1. 63 peptides with AMP activity

[0026]

[0027]

[0028] (5) Based on the predicted probability of the peptide sequence obtained in step (4) ≥ 0.99, 11 peptide sequences were screened, specifically the reported sequences SEQ ID NO.1, SEQ ID NO.34, and SEQ ID NO.35, and the new sequences SEQ ID NO.2, SEQ ID NO.3, SEQ ID NO.4, SEQ ID NO.5, SEQ ID NO.31, SEQ ID NO.32, SEQ ID NO.33, and SEQ ID NO.36. Sequence ID NO.64 was obtained by removing the N-terminal amino acid residues and adding some C-terminal amino acid residues based on SEQ ID NO.4. The grand average hydropathy (GRAVY) coefficient was predicted using the ProtParam application of the ExPASy database, and the peptide sequences with the smallest GRAVY values ​​were selected. The results are shown in Table 2.

[0029] Table 2. Prediction of the average antibacterial and hydrophilicity coefficients of two peptide sequences

[0030] SEQ ID NO. Peptide sequence Prediction Classification Prediction probability GRAVY 4 MKLFVPALLSLGALGLCLAA AMP 0.9920 1.905 64 KLFVPALLSLGALGLCLAAPRKNVRWC AMP 0.9998 0.889

[0031] (6) The peptide sequences obtained in step (2) were screened based on their predicted probability ≥ 0.99 to obtain 17 peptide sequences;

[0032] (7) The peptide sequences obtained in step (6) were ligated using the peptide ligation method in step (3) to obtain 10 peptide sequences, including the reported sequences SEQ ID NO. 65 and SEQ ID NO. 74. These 10 peptides were submitted to the AMP Scannervr.2 server according to step (1). All sequences had AMP activity, and the predicted probabilities were ≥ 0.99, as shown in Table 3.

[0033] Table 3. 10 peptide sequences with AMP activity (prediction probability ≥ 0.99)

[0034]

[0035]

[0036] (8) The peptide sequences obtained in step (2) were screened based on their predicted probability ≥ 0.95 to obtain 63 peptide sequences;

[0037] (9) The peptide sequences obtained in step (8) were subjected to the peptide linking method of step (3) to obtain 25 peptide sequences; the obtained peptide sequences were submitted to the AMP Scanner vr.2 server according to step (1), and 24 peptide sequences with AMP activity were obtained, as shown in Table 4; based on the predicted probability of the obtained peptide sequences ≥0.99, 7 peptide sequences were obtained, specifically the reported sequences SEQ ID NO.75, SEQ ID NO.87, and SEQ ID NO.89, and the new sequences SEQ ID NO.76, SEQ ID NO.77, SEQ ID NO.86, and SEQ ID NO.88.

[0038] Table 4. 24 peptide sequences with AMP activity

[0039] SEQ ID NO. Peptide sequence length Prediction Classification Prediction probability 75 WFKCRRWQWRMKKLGAP 17 AMP 0.9999 76 GAPSITCVRRAFAL 14 AMP 0.9958 77 RYTRVVWCAVGP 12 AMP 0.9925 78 LAAPRKNVRWC 11 AMP 0.9829 79 DGGYIYTAGKCGL 13 AMP 0.9805 80 PYFGYSGAFKCL 12 AMP 0.9775 81 CLAKLGGRPTY 11 AMP 0.9736 82 GAVAKFFSASCVP 13 AMP 0.9660 83 LLSLGALGLCL 11 AMP 0.9607 84 MKLFVPALLSL 11 AMP 0.9532 85 YYGYTGAFRCL 11 AMP 0.9501 86 TSLTWNSVKGKKSCH 15 AMP 0.9997 87 KVRGPPVSCIKR 12 AMP 0.998 88 VAARRARVVWCAVG 14 AMP 0.9977 89 LAGRRRSVQWCAVS 14 AMP 0.9954 90 PIGTLRPFLNWTG 13 AMP 0.9896 91 VLLFLGALGLCLA 13 AMP 0.9892 92 KLVFLVLLFLGALG 14 AMP 0.9880 93 AGITNLKKCST 11 AMP 0.9570 94 GLKSCHTGLRR 11 AMP 0.9568 95 LRPFLNWTGPP 11 AMP 0.9549 96 ALGLCLAGRRR 11 AMP 0.9537 97 YYGYTGAFRCL 11 AMP 0.9501 98 DGGYVYTAGKCGLV 14 AMP 0.9398

[0040] Example 2 Qualitative Verification of Antibacterial Activity of Polypeptides by Microdilution Method

[0041] 1. Synthetic antimicrobial peptides

[0042] According to the predicted and screened lactoferrin antimicrobial peptide sequence in Example 1, it was synthesized by GenScript Biotech Co., Ltd.

[0043] 2. Antibacterial Testing

[0044] (1) The antimicrobial peptide was dissolved in distilled water to a concentration of 2 mg / ml, and the antimicrobial activity was detected using the microdilution method.

[0045] Preparation of relevant reagents:

[0046] 1) Test bacteria: Escherichia coli (BNCC185254), Pseudomonas aeruginosa (ATCC9027), Pseudomonas putida (BNCC192818), Staphylococcus aureus (BNCC186335), Propionibacterium acnes (BNCC336649), Rhodococcus erythropolis (BNCC337015), Bacillus subtilis subsp. subtilis ... Propionibacterium acnes (BNCC336649), Propionibacterium acnes (BNCC336649), Propionibacterium acnes (BNCC336649), Propionibacterium acnes (BNCC336649), Propionibacterium acnes (BNCC336649), Pro

[0047] 10498), Malassezia furfur ATCC44344, Cryptococcus neoformans ATCC32719, and Candida albicans ATCC10231.

[0048] 2) Preparation of antimicrobial peptide stock solution: Using sterile distilled water as the solvent, prepare an antimicrobial peptide stock solution with a concentration of 2 mg / mL, and then aliquot it into small amounts for storage at -20°C.

[0049] 3) Preparation of bacterial suspension: Activate bacteria or fungi in the corresponding liquid culture medium at 37℃ or 30℃ with shaking for 24 hours, and adjust the bacterial concentration to 2×10 6 CFU / mL (bacteria) or 2×10 5 CFU / mL (fungus) was used as the bacterial stock solution for the experiment.

[0050] Experimental plan:

[0051] Sample#1

[0052] To a sterile 96-well cell culture plate, add 75 μL of bacterial stock solution and 75 μL of 2 mg / mL antimicrobial peptide (final concentration 1 mg / mL) to each well. A negative control, containing distilled water without antimicrobial peptide, was used. The assay was repeated three times. Cultures were shaken at 900 rpm in a microplate at 37°C or 30°C for 24 hours. Three μL of the mixed culture from each well was inoculated onto solid culture medium and cultured for 24 hours. A strong inhibitory effect was considered if the experimental group showed little growth relative to the negative control; a moderate growth rate was considered an inhibitory effect; and no difference in growth between the experimental group and the negative control group was considered a non-inhibitory effect.

[0053] Experimental results:

[0054] The results of the microdilution method for qualitatively determining the antibacterial activity of the antimicrobial peptides are shown in Table 5. The results show that, in addition to SEQ ID NO. 3 and SEQ ID NO. 5, SEQ ID NO. 2, SEQ ID NO. 4, SEQ ID NOs: 6-8, SEQ ID NOs: 31-33, SEQ ID NO. 36, SEQ ID NO. 64, SEQ ID NOs: 67-73, SEQ ID NOs: 76-86, and SEQ ID NO. 88 have antibacterial activities against Gram-negative and Gram-positive bacteria, as well as fungi. Among them, the peptides of SEQ ID NO. 2, SEQ ID NOs: 31-32, SEQ ID NO. 36, SEQ ID NO. 64, SEQ ID NOs: 67-70, SEQ ID NO. 86, and SEQ ID NO. 88 have antibacterial activities against Pseudomonas putida, Rhodococcus erythropolis, Bacillus subtilis, Malassezia furfur, Cryptococcus neoformans, and Candida albicans. SEQ ID NO. 64 also has antibacterial activity against Escherichia coli. In addition, SEQ ID NO. 67 showed significant antibacterial activity against Pseudomonas putida, Bacillus subtilis, and Cryptococcus neoformans. A comprehensive comparison of the overall antibacterial effects of all antimicrobial peptides against Gram-negative bacteria, Gram-positive bacteria, and fungi revealed that 15 antimicrobial peptides (SEQ ID NO. 2, SEQ ID NOs: 31-32, SEQ ID NO. 36, SEQ ID NO. 64, SEQ ID NOs: 67-73, SEQ ID NO. 77, SEQ ID NO. 86, and SEQ ID NO. 88) were more sensitive to Gram-negative bacteria, Gram-positive bacteria, and fungi.

[0055] Table 5. Qualitative inhibition results of microdilution method

[0056]

[0057]

[0058] +: has antibacterial activity; -: has no antibacterial activity.

[0059] Example 3 Quantitative determination of antibacterial activity by microdilution method

[0060] Preparation of relevant reagents:

[0061] 1) Test bacteria: Escherichia coli, Pseudomonas putida, Rhodococcus erythropolis, Bacillus subtilis, Malassezia furfur, Cryptococcus neoformans and Candida albicans.

[0062] 2) Preparation of antimicrobial peptide stock solution: Using sterile distilled water as the solvent, prepare an antimicrobial peptide stock solution with a concentration of 2 mg / mL, and then aliquot it into small amounts for storage at -20°C.

[0063] 3) Preparation of bacterial suspension: Activate bacteria or fungi in the corresponding liquid culture medium at 37℃ or 30℃ with shaking for 24 hours, and adjust the bacterial concentration to 2×10 6 CFU / mL (bacteria) or 2×10 5 CFU / mL (fungus) was used as the bacterial suspension for the experiment.

[0064] Experimental plan:

[0065] Sample#1

[0066] In a sterile 96-well cell culture plate, 75 μL of the experimental bacterial suspension (Escherichia coli, Pseudomonas putida, Rhodococcus erythropolis, Bacillus subtilis, Cryptococcus neoformans, and Candida albicans) and 75 μL of different concentrations of antimicrobial peptides were added to each well of the experimental group. A negative control group containing distilled water without antimicrobial peptides was used. The assay was repeated three times. The cells were incubated in a microplate at 900 rpm and shaking at 37°C or 30°C for 24 hours. After incubation, the OD600 of each well was measured using a multi-functional microplate. The concentration that significantly inhibited microbial growth was defined as the minimum inhibitory concentration (MIC) of the antimicrobial peptide.

[0067] Table 6. Minimum inhibitory concentration of antimicrobial peptides (μg / mL)

[0068]

[0069]

[0070] Sample#2

[0071] In a sterile 96-well cell culture plate, 75 μL of the Malassezia furfur suspension and 75 μL of various concentrations of antimicrobial peptides were added to each well of the experimental group. A negative control group, containing distilled water without antimicrobial peptides, was used. The assay was repeated three times. After shaking in a microplate at 900 rpm at 30°C for 24 hours, the mixed culture was diluted 1000-fold. 100 μL of the diluted solution was evenly spread on MF solid medium and incubated at 30°C for 18–24 hours. The results were observed and the number of colonies on the plate was counted.

[0072] Experimental results:

[0073] Based on the above selection of SEQ ID NO. 2, SEQ ID NO: 31-32, SEQ ID NO: 36, SEQ ID NO: 64, SEQ ID NO: 67-73, SEQ ID NO: 77, SEQ ID NO: 86, and SEQ ID NO: 88, their minimum inhibitory concentrations against the test bacteria were further determined, and the results are shown in Table 6. As can be seen from the table, the 15 antimicrobial peptides all have varying degrees of antibacterial activity against Gram-negative, Gram-positive bacteria, and fungi. Among them, SEQ ID NO. 64 has a minimum inhibitory concentration of 7.3 μg / mL against Gram-positive bacteria Rhodococcus erythropolis and Bacillus subtilis, showing significant antibacterial activity compared to Gram-negative bacteria and fungi. In addition, as can be seen from the table, SEQ ID NO. 2, SEQ ID NO: 31-32, SEQ ID NO: 36, SEQ ID NO: 77, and SEQ ID NO: 86 also have strong antibacterial activity against Pseudomonas putida, Bacillus subtilis, and Cryptococcus neoformans. A comprehensive comparison of the overall antibacterial effects of 15 antimicrobial peptides against Gram-negative bacteria, Gram-positive bacteria and fungi showed that SEQ ID NO.2, SEQ ID NO:31-32, SEQ ID NO.36, SEQ ID NO.64, SEQ ID NO.70, SEQ ID NO.77 and SEQ ID NO.86 had significant antibacterial effects against Gram-negative bacteria, Gram-positive bacteria and fungi.

Claims

1. An antimicrobial peptide derived from lactoferrin, characterized in that Its amino acid sequence is shown in any one of SEQ ID NO: 2, SEQ ID NO: 31-32, SEQ ID NO. 36, SEQ ID NO: 64, SEQ ID NO: 67-73, and SEQ ID NO: 77, SEQ ID NO. 86 and SEQ ID NO.

88.

2. A nucleic acid encoding the antimicrobial peptide according to claim 1.

3. An expression vector or recombinant cell containing the nucleic acid according to claim 2.

4. Use of the antimicrobial peptide according to claim 1 in the preparation of an antimicrobial or antibacterial product, wherein the product is a medicine, cosmetic, toiletries or pesticide; For the antimicrobial peptide with the amino acid sequence shown in SEQ ID NO: 2, the antibacterial or antibacterial effect refers to the antibacterial or antibacterial effect against Pseudomonas putida, Rhodococcus erythropolis, Bacillus subtilis, Malassezia furfur or Cryptococcus neoformans; For the antimicrobial peptides with amino acid sequences shown in any one of SEQ ID NOs: 31-32, 69-70, and 86, the antibacterial or antibacterial effect refers to the antibacterial or antibacterial effect against Pseudomonas putida, Rhodococcus erythropolis, Bacillus subtilis, Malassezia furfur, Cryptococcus neoformans, or Candida albicans; For the antimicrobial peptide with the amino acid sequence shown in SEQ ID NO: 36, the antibacterial or antibacterial effect refers to the antibacterial or antibacterial effect against Pseudomonas putida, Bacillus subtilis, Cryptococcus neoformans or Candida albicans; For the antimicrobial peptide with the amino acid sequence shown in SEQ ID NO: 64, the antibacterial or antibacterial effect refers to the antibacterial or antibacterial effect against Pseudomonas putida, Rhodococcus erythropolis, Bacillus subtilis or Cryptococcus neoformans; For the antimicrobial peptide with the amino acid sequence shown in SEQ ID NO: 67, the antimicrobial or antibacterial effect refers to the antimicrobial or antibacterial effect against Bacillus subtilis or Cryptococcus neoformans; For the antimicrobial peptide with the amino acid sequence shown in SEQ ID NO: 68, the antibacterial or antibacterial effect refers to the antibacterial or antibacterial effect against Rhodococcus erythropolis, Bacillus subtilis, Malassezia furfur, Cryptococcus neoformans or Candida albicans; For the antimicrobial peptide with the amino acid sequence shown in SEQ ID NO: 71, the antimicrobial or antibacterial effect refers to the antimicrobial or antibacterial effect on Bacillus subtilis; For the antimicrobial peptide with the amino acid sequence shown in SEQ ID NO: 72, the antimicrobial or antibacterial effect refers to the antimicrobial or antibacterial effect against Bacillus subtilis, Malassezia furfur or Candida albicans; For the antimicrobial peptide with the amino acid sequence shown in SEQ ID NO: 73, the antimicrobial or antibacterial effect refers to the antimicrobial or antibacterial effect against Malassezia furfur; For the antimicrobial peptide with the amino acid sequence shown in SEQ ID NO: 77, the antibacterial or antibacterial effect refers to the antibacterial or antibacterial effect against Pseudomonas putida, Rhodococcus erythropolis, Bacillus subtilis or Cryptococcus neoformans; For the antimicrobial peptide with the amino acid sequence shown in SEQ ID NO: 88, the antibacterial or bacteriostatic effect refers to the antibacterial or bacteriostatic effect on Bacillus subtilis, Malassezia furfur or Candida albicans.

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