Multi-mechanism synergistic efficient broad-spectrum antibacterial peptide and application thereof

Abstract

The invention discloses a multi-mechanism synergistic high-efficiency broad-spectrum antibacterial peptide and application thereof, belongs to the technical field of biological medicines, and provides the multi-mechanism synergistic high-efficiency broad-spectrum antibacterial peptide, and the amino acid sequence of the antibacterial peptide is one of SEQ ID NO.1, SEQ ID NO.2, SEQ ID NO.3, SEQ ID NO.4, SEQ ID NO.5 or SEQ ID NO.6. The invention further discloses a preparation method of the multi-mechanism synergistic high-efficiency broad-spectrum antibacterial peptide. Phenylboronic acid molecules are introduced into the C end of the antibacterial peptide, arginine and lysine exist in the antibacterial peptide, high-density positive charges can be combined with negatively charged bacteria, hydrophobic chains contained in the antibacterial peptide can be inserted into a phospholipid bilayer to break cell membranes, a phenylboronic acid structure interacts with polysaccharide on the surface of the bacteria, and the antibacterial peptide has a good antibacterial effect. The permeability of bacterial cell membranes is changed, so that substances in cells are leaked, and the growth and reproduction of bacteria are inhibited.

Description

Technical Field

[0001] The present invention belongs to the technical field of biomedicine, and particularly relates to a multi-mechanism synergistic highly efficient broad-spectrum antimicrobial peptide and its application. Background Art

[0002] The extensive use of antibiotics has led to the emergence of bacterial drug resistance and even the generation of superbugs. However, the development cost of antibiotics is high, and the development of new antibiotics lags far behind the emergence of new drug-resistant bacteria. In addition, antibiotics may cause adverse reactions in the host. Peptides play a key role in regulating the biological processes of living organisms and are of great significance in aspects such as biology, medicine, and industry. Antimicrobial peptides (AMPs) are an important part of the natural immune defense mechanism of most organisms against pathogens. Compared with other antimicrobial drugs, they have the advantages of a broad antibacterial spectrum, low drug resistance, low toxicity, strong rapid bactericidal ability, and good biocompatibility. Compared with traditional antibiotics, the antibacterial mechanism of antimicrobial peptides is unique. They usually kill bacteria by destroying the integrity of the bacterial cell membrane rather than acting on the metabolic pathways of bacteria. This mechanism makes it difficult for bacteria to develop drug resistance through simple gene mutations. Therefore, antimicrobial peptides have significant advantages in dealing with drug-resistant bacterial infections. In summary, antimicrobial peptides have significant advantages in antibacterial spectrum, drug resistance, toxicity, thermal stability, and biocompatibility, making them ideal alternatives to traditional antibacterial drugs, especially in dealing with drug-resistant bacterial infections, with broad application prospects. Summary of the Invention

[0003] To solve the above technical problems, the present invention proposes a multi-mechanism synergistic highly efficient broad-spectrum antimicrobial peptide and its application. A phenylboronic acid molecule is introduced at the C-terminus of the antimicrobial peptide of the present invention. There are arginine and lysine in the antimicrobial peptide, and there is a high density of positive charges that can bind to negatively charged bacteria. And the antimicrobial peptide contains a hydrophobic chain that can insert into the phospholipid bilayer to disrupt the cell membrane. The interaction between the phenylboronic acid structure and the polysaccharides on the bacterial surface changes the permeability of the bacterial cell membrane, resulting in the leakage of intracellular substances, thereby inhibiting the growth and reproduction of bacteria. It can well inhibit Gram-negative bacteria and Gram-positive bacteria, playing a highly efficient synergistic antibacterial role.

[0004] To achieve the above object, the present invention provides a multi-mechanism synergistic highly efficient broad-spectrum antimicrobial peptide, and the amino acid sequence of the multi-mechanism synergistic highly efficient broad-spectrum antimicrobial peptide is one of those shown in SEQ ID NO.1, SEQ ID NO.2, SEQ ID NO.3, SEQ ID NO.4, SEQ ID NO.5 or SEQ ID NO.6.

[0005] Preferably, phenylboronic acid is introduced at the C-terminus of the multi-mechanism synergistic highly efficient broad-spectrum antimicrobial peptide.

[0006] The present invention also provides an application of the multi - mechanism synergistic highly efficient broad - spectrum antibacterial peptide in the preparation of antibacterial products.

[0007] Preferably, the antibacterial product is a product against Gram - negative bacteria and / or Gram - positive bacteria.

[0008] Preferably, the antibacterial product is a product against Escherichia coli and / or Staphylococcus aureus.

[0009] The present invention also provides a multi - mechanism synergistic highly efficient broad - spectrum antibacterial peptide solution, which is prepared by dissolving the multi - mechanism synergistic highly efficient broad - spectrum antibacterial peptide in ultrapure water; the amino acid sequence of the multi - mechanism synergistic highly efficient broad - spectrum antibacterial peptide is one of those shown in SEQ ID NO.1, SEQ ID NO.2, SEQ ID NO.3, SEQ ID NO.4, SEQ ID NO.5 or SEQ ID NO.6.

[0010] Preferably, the concentration of the multi - mechanism synergistic highly efficient broad - spectrum antibacterial peptide solution is 0.96 - 7.8 μM.

[0011] The present invention also provides an application of the multi - mechanism synergistic highly efficient broad - spectrum antibacterial peptide solution in the preparation of antibacterial products.

[0012] Preferably, the antibacterial product is a product against Gram - negative bacteria and / or Gram - positive bacteria.

[0013] Preferably, the antibacterial product is a product against Escherichia coli and / or Staphylococcus aureus.

[0014] Compared with the prior art, the present invention has the following advantages and technical effects: The multi - mechanism synergistic highly efficient broad - spectrum antibacterial peptide of the present invention is composed of 13 amino acids, including phenylalanine, glycine, proline, leucine, alanine, lysine and arginine. The positions and types of amino acids in each antibacterial peptide are different, but a phenylboronic acid molecule is introduced at the C - terminus. The antibacterial peptide of the present invention has good water solubility and excellent antibacterial properties. The antibacterial peptide of the present invention is rich in positively charged arginine and lysine, with a high density of positive charges that can bind to negatively charged bacteria. The positive charges can bind to the negatively charged bacterial cell membrane through electrostatic adsorption and destroy the bacterial cell membrane. It also contains hydrophobic amino acids and their boron ions. The hydrophobic chain can insert into the phospholipid bilayer to disrupt the cell membrane. The interaction between the phenylboronic acid structure and the polysaccharides on the bacterial surface changes the permeability of the bacterial cell membrane, resulting in the leakage of intracellular substances, thereby inhibiting the growth and reproduction of bacteria and playing a synergistic antibacterial role.

[0015] In the embodiments of the present invention, the minimum inhibitory concentration of the antibacterial peptide against Gram-positive bacteria and Gram-negative bacteria was verified, demonstrating that the antibacterial peptide has very obvious antibacterial activity against both Gram-negative bacteria and Gram-positive bacteria. It can be used as a highly effective broad-spectrum antibacterial peptide. Moreover, the polypeptide sequence of the antibacterial peptide described in the present invention is relatively short, only containing 13 amino acids, which is convenient for production and has a low production cost, facilitating application and promotion. It is also expected to become a new antibacterial candidate drug. The present invention studied the antibacterial activity of the antibacterial peptide against Escherichia coli and Staphylococcus aureus. Through the synergistic antibacterial effect of multiple mechanisms, in the plate count experiment, when the concentration of six antibacterial peptides was 7.8 μM, the percentage reduction in CFU of Gram-negative Escherichia coli and Gram-positive Staphylococcus aureus both reached 100%. Among them, the MIC of one antibacterial peptide against Escherichia coli and Staphylococcus aureus was only 0.98 μM. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] In order to more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the following will briefly introduce the drawings required for use in the embodiments. Obviously, the drawings described below are only some embodiments of the present invention. For those of ordinary skill in the art, without creative efforts, other drawings can also be obtained based on these drawings.

[0017] Figure 1 It is the structural formula of the multi-mechanism synergistic highly effective broad-spectrum antibacterial peptide of the present invention. Among them, a is the antibacterial peptide with the amino acid sequence as shown in SEQ ID NO.1, b is the antibacterial peptide with the amino acid sequence as shown in SEQ ID NO.2, c is the antibacterial peptide with the amino acid sequence as shown in SEQ ID NO.3, d is the antibacterial peptide with the amino acid sequence as shown in SEQ ID NO.4, e is the antibacterial peptide with the amino acid sequence as shown in SEQ ID NO.5, and f is the antibacterial peptide with the amino acid sequence as shown in SEQ ID NO.6; Figure 2 It is the percentage reduction in CFU of Escherichia coli and Staphylococcus aureus measured by the plate count method for the antibacterial peptide with the amino acid sequence as shown in SEQ ID NO.1 at different concentrations. Among them, a is the percentage reduction in CFU of Escherichia coli, and b is the percentage reduction in CFU of Staphylococcus aureus; Figure 3 It is the percentage reduction in CFU of Escherichia coli and Staphylococcus aureus measured by the plate count method for the antibacterial peptide with the amino acid sequence as shown in SEQ ID NO.2 at different concentrations. Among them, a is the percentage reduction in CFU of Escherichia coli, and b is the percentage reduction in CFU of Staphylococcus aureus; Figure 4To measure the percentage reduction of CFU of Escherichia coli and Staphylococcus aureus by antimicrobial peptides with different concentrations of amino acid sequences shown in SEQ ID NO.3 by the plate counting method, where a is the percentage reduction of CFU of Escherichia coli and b is the percentage reduction of CFU of Staphylococcus aureus; Figure 5 To measure the percentage reduction of CFU of Escherichia coli and Staphylococcus aureus by antimicrobial peptides with different concentrations of amino acid sequences shown in SEQ ID NO.4 by the plate counting method, where a is the percentage reduction of CFU of Escherichia coli and b is the percentage reduction of CFU of Staphylococcus aureus; Figure 6 To measure the percentage reduction of CFU of Escherichia coli and Staphylococcus aureus by antimicrobial peptides with different concentrations of amino acid sequences shown in SEQ ID NO.5 by the plate counting method, where a is the percentage reduction of CFU of Escherichia coli and b is the percentage reduction of CFU of Staphylococcus aureus; Figure 7 To measure the percentage reduction of CFU of Escherichia coli and Staphylococcus aureus by antimicrobial peptides with different concentrations of amino acid sequences shown in SEQ ID NO.6 by the plate counting method, where a is the percentage reduction of CFU of Escherichia coli and b is the percentage reduction of CFU of Staphylococcus aureus; Figure 8 The MIC of the multi-mechanism synergistic highly efficient broad-spectrum antimicrobial peptides of the present invention against Escherichia coli and Staphylococcus aureus, where BFFK4 represents the antimicrobial peptide with the amino acid sequence shown in SEQ ID NO.1, BK4FF represents the antimicrobial peptide with the amino acid sequence shown in SEQ ID NO.2, BFFR4 represents the antimicrobial peptide with the amino acid sequence shown in SEQ ID NO.3, BR4FF represents the antimicrobial peptide with the amino acid sequence shown in SEQ ID NO.4, BFFRKRK represents the antimicrobial peptide with the amino acid sequence shown in SEQ ID NO.5, and BRKRKFF represents the antimicrobial peptide with the amino acid sequence shown in SEQ ID NO.6; Figure 9 The ZETA potential of different concentrations of the multi-mechanism synergistic highly efficient broad-spectrum antimicrobial peptides of the present invention, where a is B-FF-K4 representing the antimicrobial peptide with the amino acid sequence shown in SEQ ID NO.1 and B-K4-FF representing the antimicrobial peptide with the amino acid sequence shown in SEQ ID NO.2, b is B-FF-R4 representing the antimicrobial peptide with the amino acid sequence shown in SEQ ID NO.3 and B-R4-FF representing the antimicrobial peptide with the amino acid sequence shown in SEQ ID NO.4, and c is B-FF-RKRK representing the antimicrobial peptide with the amino acid sequence shown in SEQ ID NO.5 and B-RKRK-FF representing the antimicrobial peptide with the amino acid sequence shown in SEQ ID NO.6. Detailed implementation methods

[0018] The various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as a limitation of the present invention, but rather as a more detailed description of certain aspects, features, and implementation manners of the present invention.

[0019] It should be understood that the terms described in the present invention are only for describing specific embodiments and are not used to limit the present invention. Additionally, for the numerical ranges in the present invention, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Each intermediate value within any stated value or stated range, as well as each smaller range between any other stated value or intermediate value within the stated range, is also included in the present invention. The upper and lower limits of these smaller ranges may be independently included or excluded from the range.

[0020] Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by those of ordinary skill in the art to which the present invention pertains. Although the present invention only describes preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials related to the said documents. In case of conflict with any incorporated document, the content of this specification shall prevail.

[0021] Without departing from the scope or spirit of the present invention, various improvements and changes can be made to the specific embodiments of the present invention specification, which are obvious to those skilled in the art. Other embodiments obtained from the specification of the present invention are obvious to those skilled in the art. The specification and examples of the present invention are merely exemplary.

[0022] Regarding the use of "comprising", "including", "having", "containing", etc. in this article, they are all open-ended terms, meaning including but not limited to.

[0023] Sources of materials used in the present invention: Escherichia coli ( E.coli ) and Staphylococcus aureus ( S.aureus ) used in the present invention were purchased from Beijing BioWin Biotechnology Co., Ltd. Escherichia coli ( E.coli ) is Escherichia coli bio-67405, and Staphylococcus aureus ( S.aureus ) is Staphylococcus aureus bio-52471.

[0024] Formulation of the liquid broth medium used in the present invention: 10 g of LB broth powder, add 400 mL of distilled water, stir to dissolve, adjust its pH to 7, sterilize at 121 °C for 20 min, and store at room temperature for later use after sterilization.

[0025] The formula of the solid broth medium used in the present invention: 10 g of LB broth powder, 7 g of agar powder, add 400 mL of distilled water, stir until completely dissolved, adjust the pH to 7, sterilize at 121 °C for 20 min, and store at room temperature for later use after sterilization.

[0026] Example 1 The six antimicrobial peptides described in the present invention were all synthesized by Shanghai Jiepeptide Biotechnology Co., Ltd. using the solid-phase method, and all the chemicals used were of analytical grade.

[0027] The amino acid sequences and structural formulas of the 6 antimicrobial peptides provided by the present invention are as Figure 1 shown. Among them, the red structural part is rich in amino acids with high-density positive charges: arginine (R) and lysine (K). The positively charged amino acids can bind to the negative charges on the phospholipid molecules of the cell membrane through electrostatic adsorption, thereby achieving the effect of disrupting the cell membrane. And all 6 antimicrobial peptides contain hydrophobic amino acids, which can insert into the bacterial cell membrane, further disrupting the cell membrane. Combining with the bactericidal effect of boron ions, it plays a synergistic and highly efficient bactericidal effect. The abbreviations and amino acid sequences of the six antimicrobial peptides are: BFFK4: B(OH)2-FFGPLGLAGKKKK-NH2 (SEQ ID NO.1), and the specific structural formula is as Figure 1 shown in a; BK4FF: B(OH)2-KKKKGPLGLAGFF-NH2 (SEQ ID NO.2), and the specific structural formula is as Figure 1 shown in b; BFFR4: B(OH)2-FFGPLGLAGRRRR-NH2 (SEQ ID NO.3), and the specific structural formula is as Figure 1 shown in c; BR4FF: B(OH)2-RRRRGPLGLAGFF-NH2 (SEQ ID NO.4), and the specific structural formula is as Figure 1 shown in d; BFFRKRK: B(OH)2-FFGPLGLAGRKRK-NH2 (SEQ ID NO.5), and the specific structural formula is as Figure 1 shown in e; BRKRKFF: B(OH)2-RKRKGPLGLAGFF-NH2 (SEQ ID NO.6), and the specific structural formula is as Figure 1 shown in f.

[0028] Example 2 Preparation of the antimicrobial peptide solution: Weigh 5.74 mg of BFFK4 or 5.74 mg of BK4FF separately, dissolve them in 2 mL of ultrapure water respectively, and adjust the pH of the two antimicrobial peptides to 7 with 0.1 M NaOH to prepare a 2 mM BFFK4 polypeptide solution or a 2 mM BK4FF polypeptide solution, and place them in a refrigerator at 4 °C for refrigerated storage; weigh 6.19 mg of BFFR4 or 6.19 mg of BR4FF separately, dissolve them in 2 mL of ultrapure water respectively, and adjust the pH of the two antimicrobial peptides to 7 with 0.1 M NaOH to prepare a 2 mM BFFR4 polypeptide solution or a 2 mM BR4FF polypeptide solution, and place them in a refrigerator at 4 °C for refrigerated storage; weigh 5.96 mg of BFFRKRK and 5.96 mg of BRKRKFF separately, dissolve them in 2 mL of ultrapure water respectively, and adjust the pH of the two antimicrobial peptides to 7 with 0.1 M NaOH to prepare a 2 mM BFFRKRK polypeptide solution or a 2 mM BRKRKFF polypeptide solution, and place them in a refrigerator at 4 °C for refrigerated storage.

[0029] Preparation of Escherichia coli suspension and Staphylococcus aureus suspension: Take out Escherichia coli and Staphylococcus aureus, dilute them to an appropriate multiple and then plate them. After culturing on the plate for 18 h, pick out the appropriate single colonies growing on the agar plate. Pick the Escherichia coli single colonies and Staphylococcus aureus separately into 20 mL of liquid broth medium, and culture them in a shaker at 37 °C and 170 rpm for 8 h. After culturing, store them at 4 °C; take 1 mL of the cultured Escherichia coli liquid and Staphylococcus aureus liquid respectively, dilute them by different multiples and plate them. After plating, check the number of colonies grown from the liquid with different dilution multiples, and the number of colonies contained in the original liquid can be deduced by reverse calculation.

[0030] Measurement of the percentage reduction of CFU of antimicrobial peptides against Gram-negative bacterium Escherichia coli by the plate counting method: Use the dilution method for testing. When testing the MBC, dilute the prepared 6 kinds of 2 mM antimicrobial peptide solutions in centrifuge tubes to 31.25 µM, 7.81 µM, 1.95 µM, 0.98 µM, 0.49 µM and 0.25 µM. Dilute the Escherichia coli grown to the logarithmic phase with sterile water to 1×10 6 CFU / mL. Add an equal volume of Escherichia coli suspension to the centrifuge tubes with antimicrobial peptide solutions of different concentrations, incubate them in a shaker at 37 °C and 170 rpm for 2 h, then take out 100 µL of the sample, spread it on the culture plate, and culture it at 37 °C for 18 h. Set an Escherichia coli suspension without adding antimicrobial peptide as a negative control. The percentage reduction of CFU of different concentrations of polypeptides against Escherichia coli can be known by checking the number of colonies on the agar plate.

[0031] The reduction percentage of CFU of antimicrobial peptides against Gram-positive bacterium Staphylococcus aureus was measured by plate counting method at different concentrations of antimicrobial peptides: Staphylococcus aureus grown to the logarithmic phase was diluted to 1×10 6 CFU / mL. During the test, six kinds of 2 mM antimicrobial peptide solutions were diluted in centrifuge tubes to 7.81 µM, 3.9 µM, 1.95 µM, 0.98 µM, 0.49 µM and 0.25 µM. Equal volumes of Staphylococcus aureus suspension were added to the centrifuge tubes with antimicrobial peptide solutions at different concentrations. After co-incubation at 37 °C and 170 rpm in a shaker for 2 h, 100 µL of the sample was taken out and spread on the culture plate, and cultured at 37 °C for 18 h. A Staphylococcus aureus suspension without antimicrobial peptide was set as a negative control. The reduction percentage of CFU of antimicrobial peptides against Staphylococcus aureus at different concentrations can be obtained by counting the number of colonies on the agar plate.

[0032] The test results are as Figure 2 shown in a, Figure 3 shown in a, Figure 4 shown in a, Figure 5 shown in a, Figure 6 shown in a and Figure 7 shown in a. When the concentration of the six antimicrobial peptides is 7.8 µM, the reduction percentage of CFU against Escherichia coli can reach 100%. Among them, when the concentration of BFFR4 is reduced to 0.98 µM, the reduction percentage of CFU against Escherichia coli can also reach 100%. The test results are as Figure 2 shown in b, Figure 3 shown in b, Figure 4 shown in b, Figure 5 shown in b, Figure 6 shown in b and Figure 7 shown in b. When the concentration of the six antimicrobial peptides is 7.8 µM, the reduction percentage of CFU against Staphylococcus aureus can reach 100%. Among them, when the concentrations of BFFR4 and BR4FF are reduced to 0.98 µM, the reduction percentages of CFU against Staphylococcus aureus can both reach 100%.

[0033] As Figure 8As shown, among the six antimicrobial peptides in the present invention, the minimum inhibitory concentration of BFFR4 against Escherichia coli is 0.96 μM, the minimum inhibitory concentration of BR4FF against Escherichia coli is 1.95 μM, the minimum inhibitory concentration of BFFK4 against Escherichia coli is 1.95 μM, the minimum inhibitory concentration of BK4FF against Escherichia coli is 7.8 μM, the minimum inhibitory concentration of BFFRKRK against Escherichia coli is 1.95 μM, and the minimum inhibitory concentration of BRKRKFF against Escherichia coli is 1.95 μM; among the six antimicrobial peptides in the present invention, the minimum inhibitory concentration of BFFR4 against Staphylococcus aureus is 0.96 μM, the minimum inhibitory concentration of BR4FF against Staphylococcus aureus is 0.96 μM, the minimum inhibitory concentration of BFFK4 against Staphylococcus aureus is 3.9 μM, the minimum inhibitory concentration of BK4FF against Staphylococcus aureus is 7.8 μM, the minimum inhibitory concentration of BFFRKRK against Staphylococcus aureus is 1.95 μM, and the minimum inhibitory concentration of BRKRKFF against Staphylococcus aureus is 1.95 μM.

[0034] Example 3 Determination of ZETA potential of polypeptide solutions with different concentrations: The six prepared 2 mM antimicrobial peptide solutions were respectively diluted to 1 mM, 0.5 mM, 0.25 mM, 0.1 mM, and 0.05 mM for testing. A DTS1060C type transparent sample cell was used, the excitation wavelength was set at 633 nm, the dispersion medium was protein, and the refractive index was 1.450. The experiment was carried out at 25 °C, the minimum and maximum number of runs were 10 and 100, each antimicrobial peptide sample was measured three times and the average value was taken, and the antimicrobial peptide sample was equilibrated for 120 s before each measurement.

[0035] The results are as Figure 9 shown in a of Figure 9 shown in b of Figure 9 shown in c of, indicating that the potentials of the six antimicrobial peptides are all positive values, indicating that the molecular surfaces of the six antimicrobial peptides are all positively charged; secondly, with the increase in concentration, the potential on the surface of the antimicrobial peptide also gradually increases; the positive charges on the surface of the antimicrobial peptide can electrostatically bind to the negative charges on the surface of bacteria, achieving the effect of disrupting the bacterial cell membrane and killing bacteria.

[0036] The above-described embodiments are only descriptions of the preferred embodiments of the present invention and do not limit the scope of the present invention. Without departing from the design spirit of the present invention, various deformations and improvements made by those of ordinary skill in the art to the technical solutions of the present invention shall fall within the protection scope determined by the claims of the present invention.

Claims

1. A multi-mechanism collaborative highly efficient broad-spectrum antimicrobial peptide, characterized in that, The amino acid sequence of the multi-mechanism collaborative highly efficient broad-spectrum antibacterial peptide is one of those shown in SEQ ID NO.1, SEQ ID NO.2, SEQ ID NO.3, SEQ ID NO.4, SEQ ID NO.5 or SEQ ID NO.

6.

2. The multi-mechanism collaborative highly efficient broad-spectrum antimicrobial peptide according to claim 1, characterized in that, Phenylboronic acid is introduced at the C-terminus of the multi-mechanism collaborative highly efficient broad-spectrum antibacterial peptide.

3. The application of the multi-mechanism collaborative highly efficient broad-spectrum antimicrobial peptide according to claim 1 or claim 2 in the preparation of antimicrobial products.

4. The application according to claim 3, characterized in that, The antibacterial product is a product against Gram-negative bacteria and / or Gram-positive bacteria.

5. The application according to claim 3, characterized in that, The antibacterial product is a product against Escherichia coli and / or Staphylococcus aureus.

6. A multi-mechanism collaborative highly efficient broad-spectrum antimicrobial peptide solution, characterized in that, It is prepared by dissolving the multi-mechanism collaborative highly efficient broad-spectrum antibacterial peptide in ultrapure water; the amino acid sequence of the multi-mechanism collaborative highly efficient broad-spectrum antibacterial peptide is one of those shown in SEQ ID NO.1, SEQ ID NO.2, SEQ ID NO.3, SEQ ID NO.4, SEQ ID NO.5 or SEQ ID NO.

6.

7. The multi-mechanism collaborative highly efficient broad-spectrum antimicrobial peptide solution according to claim 6, characterized in that, The concentration of the multi-mechanism collaborative highly efficient broad-spectrum antibacterial peptide solution is 0.96 - 7.8 μM.

8. The application of the multi-mechanism collaborative highly efficient broad-spectrum antimicrobial peptide solution according to claim 6 or claim 7 in the preparation of antimicrobial products.

9. The application according to claim 8, characterized in that, The antibacterial product is a product against Gram-negative bacteria and / or Gram-positive bacteria.

10. The application according to claim 8, characterized in that, The antibacterial product is a product against Escherichia coli and / or Staphylococcus aureus.

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