CN106729610B - Application of N15 polypeptide in inhibiting staphylococcus aureus inhibitor - Google Patents
Application of N15 polypeptide in inhibiting staphylococcus aureus inhibitor Download PDFInfo
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- CN106729610B CN106729610B CN201611169791.6A CN201611169791A CN106729610B CN 106729610 B CN106729610 B CN 106729610B CN 201611169791 A CN201611169791 A CN 201611169791A CN 106729610 B CN106729610 B CN 106729610B
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- polypeptide
- staphylococcus aureus
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- cell proliferation
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- 241000191967 Staphylococcus aureus Species 0.000 title claims abstract description 25
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 25
- 229920001184 polypeptide Polymers 0.000 title claims abstract description 19
- 102000004196 processed proteins & peptides Human genes 0.000 title claims abstract description 19
- 239000003112 inhibitor Substances 0.000 title claims abstract description 6
- 230000002401 inhibitory effect Effects 0.000 title abstract description 12
- 150000001413 amino acids Chemical class 0.000 claims abstract description 3
- 238000002360 preparation method Methods 0.000 claims description 2
- 108090000623 proteins and genes Proteins 0.000 abstract description 6
- 230000035755 proliferation Effects 0.000 abstract description 5
- 108010057466 NF-kappa B Proteins 0.000 abstract description 4
- 102000003945 NF-kappa B Human genes 0.000 abstract description 4
- 230000001105 regulatory effect Effects 0.000 abstract description 4
- 230000008827 biological function Effects 0.000 abstract description 3
- 108050006400 Cyclin Proteins 0.000 abstract description 2
- 230000006820 DNA synthesis Effects 0.000 abstract description 2
- 102000016928 DNA-directed DNA polymerase Human genes 0.000 abstract description 2
- 108010014303 DNA-directed DNA polymerase Proteins 0.000 abstract description 2
- 102000009339 Proliferating Cell Nuclear Antigen Human genes 0.000 abstract description 2
- 230000000903 blocking effect Effects 0.000 abstract description 2
- 230000004663 cell proliferation Effects 0.000 abstract description 2
- 108091092356 cellular DNA Proteins 0.000 abstract description 2
- 230000004069 differentiation Effects 0.000 abstract description 2
- 230000008556 epithelial cell proliferation Effects 0.000 abstract description 2
- 230000003780 keratinization Effects 0.000 abstract description 2
- 238000000034 method Methods 0.000 abstract description 2
- 230000008569 process Effects 0.000 abstract description 2
- 102000004169 proteins and genes Human genes 0.000 abstract description 2
- 238000013518 transcription Methods 0.000 abstract description 2
- 230000035897 transcription Effects 0.000 abstract description 2
- 108020004414 DNA Proteins 0.000 description 10
- 210000004027 cell Anatomy 0.000 description 7
- 230000001580 bacterial effect Effects 0.000 description 6
- 230000000694 effects Effects 0.000 description 6
- 239000007788 liquid Substances 0.000 description 4
- 244000052616 bacterial pathogen Species 0.000 description 3
- 239000001963 growth medium Substances 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 2
- 208000015181 infectious disease Diseases 0.000 description 2
- 230000005764 inhibitory process Effects 0.000 description 2
- 238000009630 liquid culture Methods 0.000 description 2
- 239000008223 sterile water Substances 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 1
- 206010007247 Carbuncle Diseases 0.000 description 1
- 108010065152 Coagulase Proteins 0.000 description 1
- 230000004543 DNA replication Effects 0.000 description 1
- 206010017553 Furuncle Diseases 0.000 description 1
- 108010006464 Hemolysin Proteins Proteins 0.000 description 1
- 108010014603 Leukocidins Proteins 0.000 description 1
- 239000001888 Peptone Substances 0.000 description 1
- 108010080698 Peptones Proteins 0.000 description 1
- 241000607142 Salmonella Species 0.000 description 1
- 206010041925 Staphylococcal infections Diseases 0.000 description 1
- 208000033809 Suppuration Diseases 0.000 description 1
- 241001052560 Thallis Species 0.000 description 1
- 206010048038 Wound infection Diseases 0.000 description 1
- 206010000269 abscess Diseases 0.000 description 1
- 230000000843 anti-fungal effect Effects 0.000 description 1
- 230000001775 anti-pathogenic effect Effects 0.000 description 1
- 229940121375 antifungal agent Drugs 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 235000015278 beef Nutrition 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000000684 flow cytometry Methods 0.000 description 1
- 244000053095 fungal pathogen Species 0.000 description 1
- 239000001046 green dye Substances 0.000 description 1
- 230000009036 growth inhibition Effects 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 239000003228 hemolysin Substances 0.000 description 1
- 238000009776 industrial production Methods 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 230000017074 necrotic cell death Effects 0.000 description 1
- 230000009965 odorless effect Effects 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 235000019319 peptone Nutrition 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000010076 replication Effects 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 230000009885 systemic effect Effects 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 210000005253 yeast cell Anatomy 0.000 description 1
Images
Classifications
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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
- A61K38/04—Peptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof
- A61K38/10—Peptides having 12 to 20 amino acids
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Pharmacology & Pharmacy (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Immunology (AREA)
- Medicinal Chemistry (AREA)
- Gastroenterology & Hepatology (AREA)
- Chemical & Material Sciences (AREA)
- Epidemiology (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Peptides Or Proteins (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses an application of N15 polypeptide in preparing a staphylococcus aureus inhibitor, wherein N15 polypeptide is a short peptide with 15 amino acids, has multiple biological functions, and comprises the capacity of regulating NF-kB transcription factors to combine with target genes and influencing the transcription and expression of NF-kB target gene product protein; blocking PCNA binding to DNA polymerase, thereby preventing cellular DNA synthesis from inhibiting cell proliferation; inhibiting epithelial cell proliferation, regulating epidermal proliferation and differentiation, normalizing skin keratinization process, etc. the present invention has the new biological function and excellent practicability and industrial foreground.
Description
Technical Field
The invention relates to the technical field of biology, in particular to application of an N15 polypeptide in preparation of a staphylococcus aureus inhibitor.
Background
Staphylococcus aureus is most pathogenic, and produces hemolysin, leukocidin, plasma coagulase, etc., which causes many kinds of infection, such as furuncle, carbuncle, abscess and wound infection. Staphylococcus aureus infections are characterized by localized tissue necrosis, thick, yellow, odorless pus. Can also cause systemic infection, influence human health and threaten human life safety. In order to inhibit and destroy pathogenic fungi, efforts have been made to find effective anti-pathogenic bacterial agents.
Disclosure of Invention
The invention aims to provide a polypeptide, which can inhibit the growth of staphylococcus aureus and has application in preparing a pathogenic bacteria inhibitor, and the polypeptide is disclosed by the prior art and is named as N15 polypeptide.
The N15 polypeptide is a short peptide with 15 amino acids, the sequence of the short peptide is methionine-proline-tyrosine-serine-threonine-glutamic acid-leucine-isoleucine-phenylalanine-tyrosine-isoleucine-glutamic acid-methionine-aspartic acid-proline, and the short peptide has multiple biological functions, including regulating the ability of NF-kB transcription factors to combine with target genes and influencing the transcription and expression of NF-kB target gene product proteins; blocking PCNA binding to DNA polymerase, thereby preventing cellular DNA synthesis from inhibiting cell proliferation; inhibiting epithelial cell proliferation, regulating epidermal proliferation and differentiation, and normalizing skin keratinization process.
The inventor finds that the N15 polypeptide can generate inhibition effect on the growth of various pathogenic bacteria in research.
Previous studies have shown that the addition of N15 to Staphylococcus aureus has the effect of inhibiting the proliferation of Staphylococcus aureus. N15 was found to cause the reduction of DNA content in Staphylococcus aureus by detecting the DNA content in Staphylococcus aureus, so that the inhibition of the proliferation of Staphylococcus aureus by N15 could be achieved by inhibiting the replication of genomic DNA. In addition, the experimental results also show that the N15 can inhibit the growth of the salmonella.
The invention also provides an antifungal medicine prepared from the N15 polypeptide, which is an aqueous solution of the N15 polypeptide, and the concentration is more than or equal to 500 mug/mL.
The invention has the beneficial effects that: the N15 polypeptide has clear structure, easy industrial production, capacity of inhibiting the proliferation of pathogenic bacteria, high effect and no toxic side effect.
Drawings
FIG. 1 shows the effect of flow cytometry on the DNA content of Staphylococcus aureus from N15.
Detailed Description
The present invention is further demonstrated and described below with reference to the accompanying drawings and specific experimental examples, which are merely illustrative of the present invention and are not intended to limit the present invention.
Experimental example 1: n15 has effect in inhibiting growth of Staphylococcus aureus
The experimental method comprises the following steps:
the staphylococcus aureus strain is inoculated on the slant of a beef extract peptone culture medium, activated culture is carried out at 37 ℃, an inoculating loop is used for picking bacteria from the slant to prepare a bacterial suspension in sterile water, a liquid extractor is used for absorbing 100 mu L of bacterial liquid into a small sterilized centrifugal tube under the aseptic operation of inoculation culture, the bacterial liquid is quickly injected into a sterilized flat dish after being fully mixed, N15 polypeptide is added into the flat plate, the mixed liquid of the sterile water with the same volume is injected as a blank, the blank is placed in a constant-temperature incubator at 37 ℃ for culture for 48 hours, the growth condition of bacterial colonies is observed, the bacterial colonies are counted, and the result is shown in table 1.
TABLE 1 comparison of the growth inhibition results of N15 on Staphylococcus aureus
The results show that the number of colonies of staphylococcus aureus on the plates to which the N15 polypeptide was added was significantly reduced compared to the control, and that N15 had an inhibitory effect on the growth of staphylococcus aureus.
Experimental example 2: n15 has inhibitory effect on DNA replication of Staphylococcus aureus
The experimental method comprises the following steps:
preparing a YEPD liquid culture medium, culturing staphylococcus aureus, adding N15 into the YEPD liquid culture medium, setting the staphylococcus aureus without N15 polypeptide as a control, adding N15 polypeptide (the final concentration is 500 mu g/mL), sampling 2mL of staphylococcus aureus thalli after 2 hours of treatment, adding Sytox-Green dye to stain the DNA of staphylococcus aureus cells, and then measuring the relative DNA content of the staphylococcus aureus by using a flow cytometer, wherein the result is shown in figure 1.
As shown in the experimental results, the abscissa represents the DNA content in a single S.aureus cell, and the ordinate represents the number of yeast cells. The DNA content in the N15-treated cells was not significantly different from that of the control cells within 2 hours of the N15 treatment, while the DNA content in the N15-treated cells was significantly reduced from that of the control cells after 15 hours of the N15 treatment as time elapsed. This result indicates that N15 causes a decrease in the DNA content of Staphylococcus aureus cells.
Claims (1)
- Use of an N15 polypeptide in the preparation of an inhibitor of staphylococcus aureus; the N15 polypeptide is a short peptide with 15 amino acids, and the sequence of the short peptide is methionine-proline-tyrosine-serine-threonine-glutamic acid-leucine-isoleucine-phenylalanine-tyrosine-isoleucine-glutamic acid-methionine-aspartic acid-proline.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611169791.6A CN106729610B (en) | 2016-12-16 | 2016-12-16 | Application of N15 polypeptide in inhibiting staphylococcus aureus inhibitor |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611169791.6A CN106729610B (en) | 2016-12-16 | 2016-12-16 | Application of N15 polypeptide in inhibiting staphylococcus aureus inhibitor |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106729610A CN106729610A (en) | 2017-05-31 |
| CN106729610B true CN106729610B (en) | 2021-01-29 |
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| Application Number | Title | Priority Date | Filing Date |
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| CN201611169791.6A Active CN106729610B (en) | 2016-12-16 | 2016-12-16 | Application of N15 polypeptide in inhibiting staphylococcus aureus inhibitor |
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Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111803619A (en) * | 2020-07-26 | 2020-10-23 | 武汉益承生物科技有限公司 | Application of polypeptide in preparing medicine for treating wound |
Family Cites Families (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1238052C (en) * | 2003-07-07 | 2006-01-25 | 夏献民 | Obtainment of cell growth inhibiting polypeptide and its usage |
| CN100485038C (en) * | 2005-09-13 | 2009-05-06 | 四川大学华西医院 | Miniaturized anti-drug-resistance staphylococcus aureus polypeptide and application and preparation method thereof |
| CN102321158B (en) * | 2011-08-23 | 2013-03-27 | 常州德健生物科技有限公司 | Polypeptide for preventing cell DNA synthesis and inhibiting cell proliferation and application |
| CN104043102B (en) * | 2014-03-28 | 2017-04-19 | 武汉益承生物科技股份有限公司 | Use of polypeptide in preparation of drug for treating nuclear factor-kB abnormal activation diseases |
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- 2016-12-16 CN CN201611169791.6A patent/CN106729610B/en active Active
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