CN110156875B - Antibacterial peptide H5-p5, and preparation method and application thereof - Google Patents
Antibacterial peptide H5-p5, and preparation method and application thereof Download PDFInfo
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- CN110156875B CN110156875B CN201910425744.0A CN201910425744A CN110156875B CN 110156875 B CN110156875 B CN 110156875B CN 201910425744 A CN201910425744 A CN 201910425744A CN 110156875 B CN110156875 B CN 110156875B
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Images
Classifications
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/17—Amino acids, peptides or proteins
- A23L33/18—Peptides; Protein hydrolysates
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/64—Proteins; Peptides; Derivatives or degradation products thereof
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q11/00—Preparations for care of the teeth, of the oral cavity or of dentures; Dentifrices, e.g. toothpastes; Mouth rinses
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
- C07K7/04—Linear peptides containing only normal peptide links
- C07K7/08—Linear peptides containing only normal peptide links having 12 to 20 amino acids
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
Abstract
An antibacterial peptide H5-p5, the antibacterial peptide H5-p5 is composed of 18 amino acid residues, the molecular weight is 2423.12 Da, the net charge number is +10, the isoelectric point is 11.78, and the amino acid sequence is Tyr-Ile-Arg-Lys-Ile-Arg-Phe-Phe-Lys-Lys-Leu-Lys-Lys-Ile-Leu-Lys-Lys-NH2. The antibacterial peptide H5-p5 has the advantages of small molecular weight, simple artificial synthesis, remarkable antibacterial effect, strong inhibition effect on staphylococcus aureus and methicillin-resistant staphylococcus aureus, low hemolytic activity and wide application prospect.
Description
Technical Field
The invention belongs to the technical field of biochemistry, and particularly relates to preparation of antibacterial peptide H5-p5 and application of the antibacterial peptide in preparation of antibacterial drugs.
Background
Since the discovery of antibiotics such as penicillin, there has been a fundamental improvement in the treatment of bacterial infectious diseases, and the use of antibiotics has saved the lives of countless patients and has extended the average life span of humans. However, with the use or abuse of antibiotics on a large scale, especially in developing countries, the development and spread of some drug-resistant bacteria, including some very virulent pathogenic bacteria such as methicillin-resistant staphylococcus and streptococcus pneumoniae, etc., have been promoted. Therefore, the search for safe and effective antibacterial drugs that do not easily develop drug resistance properties is a competitive and endeavor among scientists worldwide.
Antimicrobial peptides are highly effective, broad-spectrum immune molecules produced rapidly by the body when organisms are invaded by microorganisms, and generally consist of 12 to 100 amino acids. Antimicrobial peptides are widely present in different types of organisms, and thousands of antimicrobial peptides have been identified in microorganisms, plants, amphibians, marine vertebrates, mammals, and even humans. The antibacterial mechanism of the antibacterial peptide is complex, but most theories believe that the mechanism involves the action of the cationic property and the hydrophobic property of the antibacterial peptide and the negatively charged microbial cell membrane, and after the antibacterial peptide is contacted with the bacterial cell membrane, the membrane permeability is changed, or a transmembrane hole is formed on the bacterial cell membrane, and finally the bacterial content is leaked out and dies. The antibacterial peptide has lethal effect on bacteria, and is not easy to generate drug resistance. Therefore, the efficiency of antibacterial peptides in killing bacteria is much higher than that of conventional antibiotics.
The discovery and rapid development of the antibacterial peptide provide a huge resource library for developing novel antibacterial drugs, also provide great possibility for solving the problem of clinical drug-resistant strains, and have wide application prospects in the fields of medicine and health, agricultural production, food industry and the like. With the further understanding of the antibacterial mechanism of the antibacterial peptide and the discovery of new antibacterial peptide, people can directly separate and purify the antibacterial peptide from organisms to obtain the antibacterial peptide, can also recombine the antibacterial peptide by utilizing a genetic engineering means, and can also directly synthesize a large amount of small-molecule antibacterial peptide in a short time by utilizing a chemical synthesis means.
The natural antibacterial peptide part has immunogenicity due to large molecular weight, has low antibacterial activity, has cytotoxic effect on host cells, or can cause hemolysis and the like, and the popularization and application of the antibacterial peptide as an antibacterial medicament are limited, so that the search for the antibacterial peptide which has smaller molecular weight and stronger antibacterial activity, particularly does not cause hemolysis or cytotoxic effect, is the most key factor for solving the popularization of the antibacterial peptide as the antibacterial medicament. In recent years, scientists have also been dedicated to structural modification or redesign of natural antibacterial peptides, such as replacing some amino acid residues or directly designing the primary structure of the amino acids of the antibacterial peptide as required, while searching for novel antibacterial peptides, in order to obtain antibacterial peptides with higher activity and better pertinence without toxic effect on host cells.
Disclosure of Invention
The invention aims to provide a novel antibacterial peptide H5-p5 with medicinal value and a preparation method thereof.
The invention also aims to provide the application of the antibacterial peptide in the aspect of preparing antibacterial medicaments or health-care products or mouthwash.
An antibacterial peptide H5-p5, wherein the amino acid sequence of the antibacterial peptide H5-p5 is as follows: Tyr-Ile-Arg-Lys-Ile-Arg-Arg-Phe-Phe-Lys-Lys-Leu-Lys-Lys-Ile-Leu-Lys-Lys-NH2
(tyrosine-isoleucine-arginine-lysine-isoleucine-arginine-phenylalanine-lysine-leucine-lysine-isoleucine-leucine-lysine-NH2). The C-terminal amidation of the complete sequence of the antibacterial peptide H5-p 5.
The scheme is characterized in that the scheme comprises 18 amino acid residues, the molecular weight is 2423.12 Da, and the isoelectric point is 11.78.
The preparation method of the antibacterial peptide H5-p5 comprises the steps of synthesizing the whole sequence of the antibacterial peptide by using an automatic polypeptide synthesizer according to the amino acid sequence of the antibacterial peptide, and desalting and purifying by using HPLC reverse phase column chromatography.
An application of the antibacterial peptide H5-p5 in preparing medicines or health products or mouthwash for resisting methicillin-resistant staphylococcus aureus.
The invention has the beneficial effects that the antibacterial peptide H5-p5 is artificially synthesized, has the advantages of small molecular weight, convenient artificial synthesis, strong bactericidal action and the like, and in addition, the antibacterial peptide H5-p5 also has the characteristics of extremely low hemolytic activity and eukaryotic cell toxicity. By aligning with 3072 antimicrobial peptide sequences in the APD database, no complete repeats with the H5-p5 sequence were found. H5-p5 has strong bacteriostatic action on staphylococcus aureus and drug-resistant staphylococcus aureus, but has no strong bacteriostatic action on beneficial bacteria such as streptococcus sanguis, and has weak hemolytic property and low cytotoxicity, which shows that H5-p5 has good antibacterial selectivity and low toxic and side effects. In addition, H5-p5 can inhibit the formation of a staphylococcus aureus biofilm, influence the integrity of a bacterial cell membrane, and can also reduce the expression of genes related to bacterial virulence factors, so that the antibacterial effect is achieved in multiple ways. The antibacterial peptides with the similar sequence to H5-p5 have no report of a multi-path antibacterial mechanism.
Drawings
FIG. 1 is a high performance liquid chromatography HPLC purity chart. FIG. 2 is an electrospray mass spectrum. FIG. 3 shows the hemolytic activity assay (mouse blood) of antimicrobial peptide H5-p5, and the results of the above experiment are the average of three independent experiments. FIG. 4 is a cytotoxicity assay (embryonic kidney cell HEK) for antimicrobial peptide H5-p5, the results of which are the average of three independent experiments. FIG. 5 is an experiment showing the effect of antibacterial peptide H5-p5 on the formation of bacterial biofilms. FIG. 6 is a graph showing the effect of antibacterial peptide H5-p5 on the integrity of bacterial cell membrane. FIG. 7 is an experiment on the effect of antibacterial peptide H5-p5 on the drug resistance related gene spA of bacteria. FIG. 8 is an experiment of the effect of antibacterial peptide H5-p5 on bacterial drug resistance related gene hld.
Detailed Description
The present invention will be better understood from the following examples. However, those skilled in the art will readily appreciate that the description of the embodiments is only for illustrating the present invention and should not be taken as limiting the invention as detailed in the claims.
Example 1: preparation of antibacterial peptide H5-p5
I, chemical synthesis method of antibacterial peptide H5-p 5: the full sequence was synthesized from the corresponding L-amino acid (available from Merck Sigma) and an automated polypeptide synthesizer (433A, applied biosystems, USA) based on the amino acid sequence described in the summary. The extract was purified by desalting by HPLC reverse phase column (WelchXB C184.6X 150 mm) chromatography with pure water (0.1% trifluoroacetic acid) -60% acetonitrile (0.1% trifluoroacetic acid) as mobile phase at a flow rate of 1mL/min and detection at a wavelength of 220nm, and the eluted peak was collected and lyophilized. The L-amino acid does not need to be preserved.
And II, identifying the molecular weight of the synthesized antibacterial peptide H5-p5 by using electrospray mass spectrometry. The sample is injected by a liquid phase system, the mobile phase is 50% H2O/50% CAN, the flow rate is 0.2ml/min, the flow rate of protective gas is 1.5L/min, and the collision energy is 4.5 kV.
III, the purity of the purified antibacterial peptide H5-p5 is identified by a High Performance Liquid Chromatography (HPLC) method (Welch XB C184.6 × 250 mm), the isoelectric point is determined by electrospray mass spectrometry and isoelectric focusing electrophoresis in molecular weight determination, and the amino acid sequence structure is determined by an automatic amino acid sequencer.
The HPLC purity identification result is shown in FIG. 1: antimicrobial peptide H5-p5 showed a single symmetrical peak at 14.277 min.
The electrospray mass spectrometry identification result is shown in figure 2: the molecular weight of the antibacterial peptide H5-p5 is 2406.16 Da.
Through HPLC and mass spectrum identification, the antibacterial peptide H5-p5 comprises 18 amino acid residues, the molecular weight is 2423.12 Da, the isoelectric point is 11.78, and the complete sequence is as follows:
Tyr-Ile-Arg-Lys-Ile-Arg-Arg-Phe-Phe-Lys-Lys-Leu-Lys-Lys-Ile-Leu-Lys-Lys-NH2
(tyrosine-isoleucine-arginine-lysine-isoleucine-arginine-phenylalanine-lysine-leucine-lysine-isoleucine-leucine-lysine-NH 2).
Example 2: antibacterial experiment of antibacterial peptide H5-p 5:
minimum Inhibitory Concentration (MIC): the lowest sample concentration at which no bacterial growth was detected. A double dilution method is adopted, and the specific method is as follows:
the bacteria were inoculated on Luria-Bertani (LB) solid medium and cultured in an inverted state in an incubator at 37 ℃. After the bacterial colony grows out, a single bacterial colony is picked by an inoculating loop and transferred into an LB liquid culture medium, and the single bacterial colony is shake-cultured in a 37 ℃ incubator to logarithmic phase. Detecting bacterial liquid OD on ultraviolet spectrophotometer600According to OD600=1×108CFU/ml bacterial liquid diluted with liquid LB medium to 2 × 105CFU/ml. Adding 100 microliter LB liquid culture medium into each hole of a sterile 96-hole plate in advance, then adding 100 microliter of antibacterial peptide sample which is diluted to a certain concentration and is subjected to filtration sterilization by a 0.22-micrometer microporous membrane into the first hole, uniformly mixing, adding 100 microliter into the 2 nd hole, sequentially diluting by multiple times, sucking 100 microliter from the 12 th hole, and discarding until a sample with double concentration gradient is prepared.
Adding 100 mu L of diluted bacterial liquid into each hole, mixing uniformly, culturing for 16h at 37 ℃ by slow shaking, and measuring the light absorption value at 600 nm. And (4) calculating a result: the average value of the sum of the concentrations of the samples in the wells in which no bacterial growth was detected and the wells adjacent thereto in which bacterial growth was present was taken as the minimum inhibitory concentration of the sample.
In addition, candida albicans is a fungus, the culture medium used for culturing is a PDA culture medium, and other conditions are similar.
The results are shown in Table 1. Table 1 is a table of data for the antimicrobial activity assay for antimicrobial peptide H5-p 5. Wherein H5-p5 MIC: namely the minimum inhibitory concentration of the antibacterial peptide H5-p5, and the result is the average value of three independent repeated experiments.
As can be seen from Table 1, the antimicrobial peptide H5-p5 has very strong killing effect on tested strains, for example, the MIC value of the antimicrobial peptide H5-p5 on staphylococcus aureus and methicillin-resistant staphylococcus aureus can be as low as 8 mu g/mL, which shows that the antimicrobial peptide H5-p5 can inhibit the growth of staphylococcus under extremely low concentration. Meanwhile, the antibacterial peptide H5-p5 has a good bactericidal effect on candida albicans clinically isolated from oral cavities, and the minimum concentration can reach 32 mug/mL; and the minimum inhibitory concentration of the streptococcus sanguis for beneficial oral bacteria is more than 64 mu g/mL, which shows that the antibacterial peptide H5-p5 has obvious effect on gram-positive staphylococcus and good selectivity.
Example 3: hemolytic activity assay of antimicrobial peptide H5-p 5:
blood is collected from the heart of a mouse, collected blood is mixed with Ash Solution (Alsever Solution, 8.0g of sodium citrate, 0.55g of citric acid, 20.5g of glucose and 4.2g of sodium chloride, deionized water is added to 1L, the pH is adjusted to 6.1, the mixture is stored at 4 ℃ after autoclaving) according to the proportion of 1:1 and is placed in a centrifuge tube, the centrifuge tube is centrifuged at 1000rpm for 5min, and normal saline is washed until the supernatant is no longer red. Diluting the washed red blood cells with physiological saline to 108Suspension of concentration. The diluted erythrocyte suspension and samples with different concentrations dissolved in normal saline are insulated at 37 ℃ for 30min, then centrifuged at 1000rpm for 5min, and the supernatant is measured for the absorption value at 540 nm. Physiological saline was used as a negative control, and Triton X-100 was used as a positive control. Hemolytic activity is proportional to the 540nm absorbance.
The results are shown in FIG. 3.
As is clear from FIG. 3, the antimicrobial peptide H5-p5 did not cause hemolysis in the mouse blood even at a high concentration of 320. mu.g/ml.
Example 4: cytotoxicity assay of antimicrobial peptide H5-p 5:
human normal cell embryonic kidney HEK293 cells were cultured in Du's modified (DMEM) medium containing 10% fetal bovine serum and diabodies (100U/mL each of penicillin and streptomycin) to log phase, washed three times with PBS buffer, digested with 0.25% trypsin, suspended in fresh DMEM medium, and cell density adjusted to 1 × 106Spreading 96-well plate at 200 μ L/well, adding samples with different concentrations after cell adherence, co-culturing at 37 deg.C and 5% carbon dioxide for 24 hr, and culturingAfter that, 20. mu.L of 5mg/ml MTT solution (prepared by using cell culture PBS buffer solution) is added into each well of a 96-well cell culture plate, the culture is continued for 4 hours, the liquid in the wells is sucked by a pipette, 100. mu.L of dimethyl sulfoxide (DMSO) is added into each well, the mixture is gently shaken for 10min at room temperature, and the light absorption with the wavelength of 490nm is detected by a microplate reader.
The results are shown in FIG. 4.
As can be seen from FIG. 4, the antimicrobial peptide H5-p5 had a very low cytotoxic effect of about 10% on human normal cell embryonic kidney HEK293 cells even at a concentration of 1000. mu.g/mL.
Example 5: the action mechanism of the antibacterial peptide H5-p5 is explored:
I. staphylococcus aureus and MRSA bacteria were cultured with Tryptic Soy Broth (TSB) and diluted to about 108CFU/mL. 1ml of the bacterial solution was added to a 12-well plate and H5-p5 or physiological saline at MIC concentration was added, after which the plate was incubated at 37 ℃ for 24 hours. The samples were treated overnight by immersion in PBS containing 2.5% glutaraldehyde. After three rinses with PBS, the samples were dehydrated by successive treatments with 30%, 50%, 70%, 80%, 85%, 90%, 95% and 100% ethanol for 5-15 minutes. Finally, the sample was rinsed with 100% hexamethyldisilazane and dried on a glass petri dish overnight before being imaged using field emission scanning electron microscopy.
The results are shown in FIGS. 5 and 6.
As shown in FIG. 5, the antimicrobial peptide H5-p5 can reduce the colony aggregation of Staphylococcus aureus and MRSA bacteria and inhibit the formation of the biofilm of Staphylococcus aureus and MRSA bacteria. As shown in FIG. 6, the antibacterial peptide H5-p5 can affect the integrity of the bacterial cell membrane, so that the bacterial cell membrane is broken, the content flows out, and the bacteria are killed.
II. The overnight grown Staphylococcus aureus and MRSA bacteria were diluted to 108CFU/mL, then incubated for 1 hour in MIC concentration of H5-p5 or physiological saline, extracting bacterial RNA from 5mL bacterial culture-centrifugation at 12,000 × g for 2 minutes at 4 ℃ bacterial culture harvested, washing the pellet with 1mL TRIzol, and lysing with 0.5mL zirconia-silica beads (diameter, 0.1 mm) in a high speed homogenizer, extraction with DNA/RNA extraction reagent (chloroform: isoamyl alcohol = 24: 1), andRNA was isolated sequentially with isopropanol and 70% ethanol and finally dissolved in Diethylpyrocarbonate (DEPC) water.
Genomic DNA contained in RNA was degraded and reverse transcribed to give cDNA using PrimeScriptTM RT kit and gDNA Eraser (Perfect Real Time, Takara). The Staphylococcus aureus virulence factor-associated genes, spA and hld, were then detected by quantitative reverse transcription-PCR (qRT-PCR) using the LightCycler 480II system (Forrens 2,6343 Rotkreuz, Switzerland) and TB GreenTM Premix Ex TaqTMII (Tli RNaseH Plus, Takara) according to the manufacturer's instructions. The expression level of 16S rRNA was used as a control to normalize the expression levels of other genes. All experiments were repeated at least three times.
The results are shown in FIGS. 7 and 8.
As can be seen from FIGS. 7 and 8, the antimicrobial peptide H5-p5 was able to down-regulate the resistance-associated genes spA and hld in Staphylococcus aureus and MRSA bacteria. The spA gene encodes staphylococcus aureus protein A, which is a virulence factor of staphylococcus aureus; the hld gene encodes an exotoxin, a hemolytic toxin. H5-p5 can not only exert an antibacterial effect by inhibiting the formation of bacterial biofilms and influencing the integrity of bacterial cell membranes, but also exert an attenuation antibacterial effect by down-regulating the expression level of genes related to virulence factors.
In conclusion, the antibacterial peptide H5-p5 has the advantages of small molecular weight, simple artificial synthesis, high efficiency in killing methicillin-resistant staphylococcus aureus and low hemolytic activity, and the antibacterial peptide H5-p5 and the composition containing H5-p5 can be used as a methicillin-resistant staphylococcus aureus resistant medicament and can also be applied to the fields of health products, mouthwash and the like.
Sequence listing
<110> institute of biological research of academy of sciences of Shandong province
<120> antibacterial peptide H5-p5, and preparation method and application thereof
<141>2019-05-21
<160>1
<170>SIPOSequenceListing 1.0
<210>1
<211>18
<212>PRT
<213>Artificial Sequence
<220>
<221>AMIDATION
<222>(1)..(18)
<223> C-terminal amidation of the entire sequence of antimicrobial peptide H5-p5
<400>1
Tyr Ile Arg Lys Ile Arg Arg Phe Phe Lys Lys Leu Lys Lys Ile Leu
1 5 10 15
Lys Lys
Sequence listing
<110> institute of biological research of academy of sciences of Shandong province
<120> antibacterial peptide H5-p5, and preparation method and application thereof
<141>2019-05-21
<160>1
<170>SIPOSequenceListing 1.0
<210>1
<211>18
<212>PRT
<213>Artificial Sequence
<220>
<221>AMIDATION
<222>(1)..(18)
<223> C-terminal amidation of the entire sequence of antimicrobial peptide H5-p5
<400>1
Tyr Ile Arg Lys Ile Arg Arg Phe Phe Lys Lys Leu Lys Lys Ile Leu
1 5 10 15
Lys Lys
Claims (4)
1. An antibacterial peptide H5-p5, which is characterized in that the amino acid sequence of the antibacterial peptide H5-p5 is as follows: Tyr-Ile-Arg-Lys-Ile-Arg-Arg-Phe-Phe-Lys-Lys-Leu-Lys-Lys-Ile-Leu-Lys-Lys-NH2。
2. The antibacterial peptide H5-p5 of claim 1, which comprises 18 amino acid residues, has a molecular weight of 2423.12 Da and an isoelectric point of 11.78.
3. The method for preparing the antibacterial peptide H5-p5 as claimed in claim 1, wherein the full sequence of the antibacterial peptide is synthesized by an automatic polypeptide synthesizer according to the amino acid sequence of the antibacterial peptide, and the antibacterial peptide is desalted and purified by HPLC reverse phase column chromatography.
4. The use of the antimicrobial peptide H5-p5 as claimed in claim 1, which is used in the preparation of a methicillin-resistant Staphylococcus aureus (MRSA) resistant medicament or health product or mouthwash.
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| CN104311627A (en) * | 2014-10-29 | 2015-01-28 | 中山大学 | Synthetic method and application of antibacterial peptide |
| CN107312080A (en) * | 2017-07-23 | 2017-11-03 | 复旦大学 | A kind of antibacterial peptide and its application from GSDMD albumen |
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| CN104311627A (en) * | 2014-10-29 | 2015-01-28 | 中山大学 | Synthetic method and application of antibacterial peptide |
| WO2017216373A1 (en) * | 2016-06-16 | 2017-12-21 | Centre National De La Recherche Scientifique | Cxcr4 receptor-binding compounds useful for increasing interferon level |
| CN107312080A (en) * | 2017-07-23 | 2017-11-03 | 复旦大学 | A kind of antibacterial peptide and its application from GSDMD albumen |
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