CN116693607A - Antibacterial polypeptide, application, bacterial growth inhibitor, anti-infective drug or health care product - Google Patents
Antibacterial polypeptide, application, bacterial growth inhibitor, anti-infective drug or health care product Download PDFInfo
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- CN116693607A CN116693607A CN202210173682.0A CN202210173682A CN116693607A CN 116693607 A CN116693607 A CN 116693607A CN 202210173682 A CN202210173682 A CN 202210173682A CN 116693607 A CN116693607 A CN 116693607A
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- staphylococcus aureus
- escherichia coli
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- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 44
- 102000004196 processed proteins & peptides Human genes 0.000 title claims abstract description 41
- 229920001184 polypeptide Polymers 0.000 title claims abstract description 37
- 239000003814 drug Substances 0.000 title claims abstract description 11
- 230000036541 health Effects 0.000 title claims abstract description 8
- 239000003966 growth inhibitor Substances 0.000 title claims description 7
- 229940079593 drug Drugs 0.000 title abstract description 8
- 230000000844 anti-bacterial effect Effects 0.000 title description 14
- 230000001580 bacterial effect Effects 0.000 title description 9
- 230000002924 anti-infective effect Effects 0.000 title description 4
- 241000191967 Staphylococcus aureus Species 0.000 claims abstract description 28
- 241000588724 Escherichia coli Species 0.000 claims abstract description 26
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims abstract description 6
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- 239000004480 active ingredient Substances 0.000 claims description 7
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- 238000002360 preparation method Methods 0.000 claims description 4
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- 239000004599 antimicrobial Substances 0.000 claims 1
- 201000010099 disease Diseases 0.000 claims 1
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- 230000002265 prevention Effects 0.000 claims 1
- 230000002401 inhibitory effect Effects 0.000 abstract description 12
- 241000894006 Bacteria Species 0.000 abstract description 7
- 241000192125 Firmicutes Species 0.000 abstract description 4
- 239000003242 anti bacterial agent Substances 0.000 abstract description 4
- 210000001035 gastrointestinal tract Anatomy 0.000 abstract description 4
- 208000015181 infectious disease Diseases 0.000 abstract description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 9
- 239000007788 liquid Substances 0.000 description 9
- 239000001963 growth medium Substances 0.000 description 8
- 239000003910 polypeptide antibiotic agent Substances 0.000 description 8
- 230000005764 inhibitory process Effects 0.000 description 7
- 102000044503 Antimicrobial Peptides Human genes 0.000 description 6
- 108700042778 Antimicrobial Peptides Proteins 0.000 description 6
- 150000001413 amino acids Chemical class 0.000 description 6
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- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 description 4
- 244000005700 microbiome Species 0.000 description 4
- 239000007787 solid Substances 0.000 description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 4
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 3
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 3
- 238000004458 analytical method Methods 0.000 description 3
- 229940088710 antibiotic agent Drugs 0.000 description 3
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- 239000000758 substrate Substances 0.000 description 3
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 description 2
- 230000003385 bacteriostatic effect Effects 0.000 description 2
- 125000002091 cationic group Chemical group 0.000 description 2
- 238000003776 cleavage reaction Methods 0.000 description 2
- 238000012258 culturing Methods 0.000 description 2
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- WPXFILQZNKUYQO-BZSNNMDCSA-N 2-[[(2s)-2-[[(2s)-1-[(2s)-2-amino-3-(4-hydroxyphenyl)propanoyl]pyrrolidine-2-carbonyl]amino]-4-methylpentanoyl]amino]acetic acid Chemical compound OC(=O)CNC(=O)[C@H](CC(C)C)NC(=O)[C@@H]1CCCN1C(=O)[C@@H](N)CC1=CC=C(O)C=C1 WPXFILQZNKUYQO-BZSNNMDCSA-N 0.000 description 1
- 125000003345 AMP group Chemical group 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- 101710102315 Alpha/beta-gliadin Proteins 0.000 description 1
- 108010077805 Bacterial Proteins Proteins 0.000 description 1
- 206010059866 Drug resistance Diseases 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- SMLDOQHTOAAFJQ-WDSKDSINSA-N Gln-Gly-Ser Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)NCC(=O)N[C@@H](CO)C(O)=O SMLDOQHTOAAFJQ-WDSKDSINSA-N 0.000 description 1
- 108010073032 Grain Proteins Proteins 0.000 description 1
- FFEARJCKVFRZRR-BYPYZUCNSA-N L-methionine Chemical compound CSCC[C@H](N)C(O)=O FFEARJCKVFRZRR-BYPYZUCNSA-N 0.000 description 1
- 206010034133 Pathogen resistance Diseases 0.000 description 1
- 102000007079 Peptide Fragments Human genes 0.000 description 1
- 108010033276 Peptide Fragments Proteins 0.000 description 1
- BRDYYVQTEJVRQT-HRCADAONSA-N Phe-Arg-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CC2=CC=CC=C2)N)C(=O)O BRDYYVQTEJVRQT-HRCADAONSA-N 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
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- 229940124350 antibacterial drug Drugs 0.000 description 1
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- 238000012512 characterization method Methods 0.000 description 1
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- 231100000135 cytotoxicity Toxicity 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 238000011033 desalting Methods 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 239000003937 drug carrier Substances 0.000 description 1
- 230000005686 electrostatic field Effects 0.000 description 1
- 238000010828 elution Methods 0.000 description 1
- IDGUHHHQCWSQLU-UHFFFAOYSA-N ethanol;hydrate Chemical compound O.CCO IDGUHHHQCWSQLU-UHFFFAOYSA-N 0.000 description 1
- 229920006227 ethylene-grafted-maleic anhydride Polymers 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 238000011049 filling Methods 0.000 description 1
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- 239000000796 flavoring agent Substances 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 238000013467 fragmentation Methods 0.000 description 1
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- 230000002452 interceptive effect Effects 0.000 description 1
- 238000011068 loading method Methods 0.000 description 1
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- 230000002503 metabolic effect Effects 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 229930182817 methionine Natural products 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 230000031787 nutrient reservoir activity Effects 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 230000035699 permeability Effects 0.000 description 1
- 150000003904 phospholipids Chemical class 0.000 description 1
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- 239000002994 raw material Substances 0.000 description 1
- 238000010532 solid phase synthesis reaction Methods 0.000 description 1
- 238000005507 spraying Methods 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 108700042752 tyrosyl-prolyl-leucyl-glycine Proteins 0.000 description 1
- 229910021642 ultra pure water Inorganic materials 0.000 description 1
- 239000012498 ultrapure water Substances 0.000 description 1
- 239000003643 water by type Substances 0.000 description 1
Classifications
-
- 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
- C07K7/06—Linear peptides containing only normal peptide links having 5 to 11 amino acids
-
- 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
-
- 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
-
- 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
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- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Molecular Biology (AREA)
- General Health & Medical Sciences (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Medicinal Chemistry (AREA)
- Oncology (AREA)
- Animal Behavior & Ethology (AREA)
- Biophysics (AREA)
- Communicable Diseases (AREA)
- Biochemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Pharmacology & Pharmacy (AREA)
- Genetics & Genomics (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Mycology (AREA)
- Nutrition Science (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Peptides Or Proteins (AREA)
Abstract
The present invention relates to a polypeptide derived from Daqu and having inhibitory activity against gram-negative bacteria and gram-positive bacteria typified by Escherichia coli and Staphylococcus aureus. The amino acid sequence is Tyr-Pro-Leu-Gly-Gln-Gly-Ser-Phe-Arg-Pro. The polypeptide YPLGQGSFRP has good inhibitory activity on gram-negative bacteria and gram-positive bacteria, and has good application prospect in preparing medicines and/or health products for preventing and/or reducing intestinal tract infection diseases caused by escherichia coli and staphylococcus aureus as a natural and safe antibacterial agent.
Description
Technical Field
The invention relates to an antibacterial polypeptide YPLGQGSFRP derived from Daqu and application thereof in preparing a bacterial growth inhibitor, an anti-infective drug and/or a health care product.
Background
Antibiotics are special secondary metabolic organic substances produced by microorganisms, and can inhibit the growth and metabolism of bacteria by inhibiting the formation of nucleic acids of bacteria, inhibiting the synthesis of bacterial proteins, changing the permeability of cell membranes of bacteria, interfering the synthesis of cell walls of bacteria and the like when used at low concentration. Antibiotics are widely used because of their remarkable antibacterial effect, however, overuse also results in the generation of bacterial resistance, making the treatment of bacterial infectious diseases increasingly difficult.
The antibacterial peptide (antimicrobial peptides, AMPs) is used as a novel antibiotic, has very wide sources, and has the characteristics of 10-50 amino acids, strong cationic property, amphipathy, broad-spectrum antibacterial property and the like. Compared with the traditional antibiotics, the antibacterial peptide has the following greatest advantages: the antibacterial peptide is mainly combined to a bacterial cell membrane through electrostatic attraction, and is combined with phospholipid to be inserted into a cytoplasmic membrane through hydrophobic action, so that the structure of the bacterial cell membrane is destroyed. The external force action can not cause the microorganism to generate drug resistance basically. However, the current use of antimicrobial peptides still faces two major challenges: firstly, the antibacterial activity of the natural antibacterial peptide is relatively low, and secondly, the stability and cytotoxicity of the antibacterial peptide are difficult to regulate and control. Thus, the method is applicable to a variety of applications. The development of safe, stable and low-cytotoxicity antibacterial peptide has important significance.
The Daqu is prepared by fermenting grains serving as substrates at high temperature by microorganisms, wherein the antibacterial peptide derived from the grain protein is food-borne, natural and safe, and has good potential as an antibacterial drug. Therefore, the antibacterial peptide derived from the Daqu substrate wheat protein is screened from Daqu, and can be used for preparing and researching medicines and/or health-care products for preventing and/or reducing intestinal tract infection diseases caused by escherichia coli and staphylococcus aureus.
Disclosure of Invention
The invention aims to provide an application of an antibacterial polypeptide YPLGQGSFRP derived from a Daqu substrate wheat protein in preparing medicines and/or health-care products for preventing and/or reducing intestinal tract infection diseases caused by escherichia coli and staphylococcus aureus.
To achieve the above object, the present invention uses the polypeptide YPLGQGSFRP as an active ingredient for inhibiting the growth activity of microorganisms.
It has the sequence table SEQ ID NO:1, the polypeptide YPLGQGSFRP is an active ingredient of the anti-escherichia coli and staphylococcus aureus medicament, and a pharmaceutically acceptable carrier or auxiliary material can be added.
The polypeptide compound YPLGQGSFRP with the inhibitory activity on escherichia coli and staphylococcus aureus has the amino acid sequence of Tyr-Pro-Leu-Gly-Gln-Gly-Ser-Phe-Arg-Pro. The molecular weight is 1121.24Da, white powder is easy to dissolve in water, and has strong inhibition effect on the growth of escherichia coli and staphylococcus aureus.
Compared with the prior art, the invention has the following beneficial effects:
the invention is obtained from Daqu and determines the structure of the polypeptide, verifies that the polypeptide has better antibacterial activity for the first time, and has good application prospect in preparing medicines and/or health care products for preventing and/or reducing intestinal tract infection diseases caused by escherichia coli and staphylococcus aureus.
Drawings
FIG. 1 is a diagram showing the results of inhibition by polypeptide YPLGQGSFRP, wherein FIG. 1 (A) is a diagram showing the results of an experiment for inhibiting E.coli by polypeptide YPLGQGSFRP; FIG. 1 (B) is a graph showing the experimental results of inhibition of Staphylococcus aureus by polypeptide YPLGQGSFRP.
Detailed Description
Example 1 preparation and identification of polypeptide YPLGQGSFRP
A method of combining LC-MS/MS with a bottom-up proteomics technique is adopted. The method is characterized in that commercially available Maotai-flavor high-temperature Daqu (RH-02, luzhou Ruihua biological starter propagation Co., ltd.) is used as a raw material, and peptide fragments with inhibition effect on escherichia coli and staphylococcus aureus are screened by combining structure-activity relationship characteristics through gradient extraction of water and ethanol, centrifugation, ultrafiltration and LC-MS/MS analysis.
The specific method comprises the following steps:
(1) Sample preparation
1g of the Daqu sample was mixed with 20mL of ultrapure water, centrifuged at 200rpm for 15min at 37℃and 4℃and 15000 Xg, the supernatant (designated as supernatant A) was removed, 20mL of 50% ethanol-water solution was added to the precipitate (designated as precipitate A), the mixture was centrifuged at 200rpm for 15min at 37℃and 4℃and 15000 Xg, the supernatant (designated as supernatant B) was removed, the precipitate (designated as precipitate B) was added to 20mL of ethanol, and the mixture was centrifuged at 200rpm for 15min at 37℃and 15000 Xg and 20mL of ethanol was added to the precipitate (designated as supernatant C). The supernatant A, B, C was ultrafiltered with an ultrafiltration membrane having a molecular weight cut-off of 10000Da, and the obtained filtrate A, B, C was desalted with a C18-SPE cartridge (Waters Oasis HLB SPE), respectively. After desalting, the samples were lyophilized and stored at-80 ℃ to obtain the corresponding lyophilized and stored samples A, B, C.
(2) LC-MS/MS analysis
Freeze-dried stored samples A, B, C were reconstituted in 0.1% (v/v) FA-H, respectively 2 In O solution, LTQ-Orbitrap Velos (double partial pressure linear trap and electrostatic field orbit trap combined high-resolution mass spectrum) is used for carrying out mass spectrum analysis on a sample. The loading amounts were 2. Mu.g, respectively, and the mobile phase A of the liquid chromatograph was an aqueous solution (v/v) containing 0.1% formic acid, and the mobile phase B was acetonitrile (v/v) containing 0.1% formic acid. The gradient elution procedure was as follows:
0-80min,7% -27% B (v/v); 80-95min,27% -40% B;95-97min, 40-90% B;97-107min,90% B;107-109min,90% -0% B;109-126min,0% B, flow rate 60. Mu.L/min
The mass spectrum ion source is ESI, the ion transmission temperature is 250 ℃, the spraying voltage is 2.2kV, the normalized collision energy is 35%, the primary spectrum resolution is 60000, the scanning range is 400-2000m/z, and the data acquisition mode is a data dependent mode (DDA). Selecting 20 strongest parent ions in the primary spectrum for secondary fragmentation, wherein dynamic exclusion is set as repeated counting, 2; repeating time, 30s; the exclusion time was 60 seconds.
(3) Data retrieval
RAW file use MaxQuant TM The (v.1.5.3.30) software was retrieved in the wheat (Wheat, 379 proteins) database (http:// www.uniprot.org /). The search parameters are as follows: no enzyme cleavage, maximum number of missed cleavage and fixed modification were set, and the variable modification was set to methionine oxidation (+ 15.9949 Da). The mass tolerance deviation of the parent ion was 20ppm and the fragment ion was 0.5Da. Controlling PSM false positive rate (FDR)<1% of the polypeptides were analyzed as valid data. 475 polypeptides from 39 proteins were identified in the experiment. Since antibacterial peptides generally have the following characteristics, such as generally containing hydrophobic amino acids, basic amino acids; has strong cationic property, amphipathy and the like. Thus, the knotThe structure-activity relationship of the synthetic antimicrobial peptides was used to screen out a portion of potential antimicrobial peptides from the identification results, as shown in table 1:
table 1 results of partial identification of potential antimicrobial peptides
(4) Analysis of polypeptide YPLGQGSFRP Properties
Polypeptide YPLGQGSFRP is derived from main storage protein of wheat seeds, namely alpha/beta gliadin (P02863, f 224-233) peptide isoelectric point is 8.75, and molecular mass is 1121.24Da. Biological information of the polypeptide obtained by an online tool ExpASY shows that the polypeptide has positive charges, an instability coefficient of 40.60, and an aliphatic amino acid index and hydrophilicity of 39.00 and-0.750 respectively.
Information of SEQ ID No.1
(a) Sequence characterization
* Length: 10 amino acids
* Type (2): amino acids
* Chain type: linear single strand
(b) Molecular type: proteins
Sequence description:
SEQ ID No.1
YPLGQGSFRP
EXAMPLE 2 detection of bacteriostatic Activity of polypeptide YPLGQGSFRP
Three potential antimicrobial peptides YPLGQGSFRP, IFWGIPALLK and AAFSPVSLHSALSLLAAGAGS were selected and delegated to the Nanj peptide biotechnology Co.Ltd, with purities of 95.46%, 99.73% and 95.31%, respectively, by solid phase synthesis. The antibacterial activity of three potential antibacterial peptides is examined by taking gram-positive bacteria escherichia coli and gram-negative bacteria staphylococcus aureus as target strains.
(1) Strain and resuscitation
Coli (Escherichia coli K) and staphylococcus aureus (Staphylococcus aureus) were both stored at-80 ℃ by glycerol storage. Before the experiment, one loop of escherichia coli is inoculated into 5mL of liquid LB medium, one loop of staphylococcus aureus is inoculated into 5mL of liquid TSB medium, and the strain is revived by culturing for 12 hours at 37 ℃ and 200 rpm.
After 12h, 100. Mu.L of E.coli bacterial liquid was transferred to 5mL of liquid LB medium, 100. Mu.L of Staphylococcus aureus bacterial liquid was transferred to 5mL of liquid TSB medium, and the mixture was again cultured at 37℃for 12h for resuscitation at 200 rpm. The resuscitating operation was repeated 2-3 times to revive the strain.
(2) Culture medium
The antibacterial experiment used a round petri dish with a diameter of 90 mm. The culture medium is LB solid culture medium and TSB solid culture medium, and the two culture media respectively contain agar with the mass concentration of 1.5%. LB solid medium was mixed to a final concentration of 1X 10 5 CFU/mL of E.coli, TSB solid medium was mixed to a final concentration of 1X 10 5 CFU/mL staphylococcus aureus. The culture medium mixed with the bacterial liquid is respectively poured into six different round culture dishes (the escherichia coli and the staphylococcus aureus are respectively poured into three culture dishes), the filling height of the culture medium in the culture dishes is 4mm, and the culture medium is preserved at 4 ℃ after being sealed, so that six flat plates are obtained.
(3) Antibacterial experiments
3 holes with the diameter of 3mm are respectively manufactured on six flat plates, and the hole spacing is 3cm. Six plates were divided into three groups, one plate containing E.coli and one plate containing Staphylococcus aureus were grouped, and 1.5mg of the same target peptide dissolved in 50. Mu.L of sterile water was added to each of two wells in each group (three target peptides, three groups of experiments), each of which was performed in duplicate. A third well was blank with sterile water and 50. Mu.L was added as well. Standing the plate after sample addition in a refrigerator at 4 ℃ for 4 hours, taking out the plate after the sample liquid in the hole is completely absorbed and diffused, culturing the plate for 12 hours in an inverted mode in a constant-temperature incubator at 37 ℃, and observing and measuring and recording the size of a bacteriostasis zone.
(4) Experimental results
The diameters of the inhibition zones of the three target peptides on escherichia coli and staphylococcus aureus are shown in table 2. The polypeptides IFWGIPALLK and AAFSPVSLHSALSLLAAGAGS have no inhibition effect on escherichia coli and only have weak inhibition effect on staphylococcus aureus; YPLGQGSFRP has inhibiting effect on Escherichia coli and Staphylococcus aureus, and has slightly better inhibiting effect on Escherichia coli than Staphylococcus aureus. The results indicate that polypeptide YPLGQGSFRP has inhibitory activity against both gram-negative and gram-positive bacteria and has higher inhibitory activity against gram-negative bacteria.
Figure 1 shows the inhibitory effect of polypeptide YPLGQGSFRP on escherichia coli and staphylococcus aureus.
Table 2 bacteriostatic effects of polypeptide YPLGQGSFRP
Sequence listing
<110> institute of chemical and physical of Dalian of academy of sciences of China
<120> antibacterial polypeptide, application, bacterial growth inhibitor, anti-infective drug or health care product
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 10
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 1
Tyr Pro Leu Gly Gln Gly Ser Phe Arg Pro
1 5 10
Claims (8)
1. An antimicrobial polypeptide, characterized in that: the polypeptide is YPLGQGSFRP and has a sequence table SEQ ID NO:1, an amino acid sequence in seq id no; the amino acid sequence of the polypeptide is Tyr-Pro-Leu-Gly-Gln-Gly-Ser-Phe-Arg-Pro.
2. Use of the polypeptide of claim 1 for the preparation of a growth inhibitor for escherichia coli and/or staphylococcus aureus.
3. Use of a polypeptide according to claim 1 for the preparation of a medicament and/or health care product for the prevention and/or treatment of a disease of intestinal bacterial infection caused by escherichia coli and/or staphylococcus aureus.
4. A use according to claim 2 or 3, characterized in that: the polypeptide YPLGQGSFRP is taken as an active ingredient, and a pharmaceutically or food acceptable carrier or auxiliary material can be added.
5. An escherichia coli and/or staphylococcus aureus growth inhibitor, characterized by: which comprises the polypeptide of claim 1 as an active ingredient.
6. The growth inhibitor for escherichia coli and/or staphylococcus aureus according to claim 5, wherein: the polypeptide of claim 1 is used as an active ingredient, wherein a pharmaceutically or food acceptable carrier or auxiliary material can be added.
7. A medicine and/or health care product for preventing and/or treating intestinal bacterial infection diseases caused by escherichia coli and/or staphylococcus aureus is characterized in that: which comprises the polypeptide of claim 1 as an active ingredient.
8. The pharmaceutical and/or nutraceutical product according to claim 7, characterized in that: the polypeptide of claim 1 is used as an active ingredient, wherein a pharmaceutically or food acceptable carrier or auxiliary material can be added.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210173682.0A CN116693607A (en) | 2022-02-24 | 2022-02-24 | Antibacterial polypeptide, application, bacterial growth inhibitor, anti-infective drug or health care product |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210173682.0A CN116693607A (en) | 2022-02-24 | 2022-02-24 | Antibacterial polypeptide, application, bacterial growth inhibitor, anti-infective drug or health care product |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN116693607A true CN116693607A (en) | 2023-09-05 |
Family
ID=87841929
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210173682.0A Pending CN116693607A (en) | 2022-02-24 | 2022-02-24 | Antibacterial polypeptide, application, bacterial growth inhibitor, anti-infective drug or health care product |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN116693607A (en) |
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2022
- 2022-02-24 CN CN202210173682.0A patent/CN116693607A/en active Pending
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