CN106397568A - 一种抗菌肽Dermaseptin‑M及其应用 - Google Patents
一种抗菌肽Dermaseptin‑M及其应用 Download PDFInfo
- Publication number
- CN106397568A CN106397568A CN201611109715.6A CN201611109715A CN106397568A CN 106397568 A CN106397568 A CN 106397568A CN 201611109715 A CN201611109715 A CN 201611109715A CN 106397568 A CN106397568 A CN 106397568A
- Authority
- CN
- China
- Prior art keywords
- dermaseptin
- cell
- antibacterial peptide
- application
- ermaseptin
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 239000003910 polypeptide antibiotic agent Substances 0.000 title claims abstract description 20
- 241000700605 Viruses Species 0.000 claims abstract description 12
- 241000191967 Staphylococcus aureus Species 0.000 claims abstract description 8
- 241000589517 Pseudomonas aeruginosa Species 0.000 claims abstract description 7
- 241000588724 Escherichia coli Species 0.000 claims abstract description 6
- 125000003275 alpha amino acid group Chemical group 0.000 claims abstract 2
- 241000894006 Bacteria Species 0.000 claims description 20
- 108700032080 Hyalophora cecropia cecropin D Proteins 0.000 claims description 17
- 239000003814 drug Substances 0.000 claims description 11
- 241000192125 Firmicutes Species 0.000 claims description 5
- 208000011580 syndromic disease Diseases 0.000 claims description 5
- 238000009395 breeding Methods 0.000 claims description 4
- 230000001488 breeding effect Effects 0.000 claims description 4
- 238000002360 preparation method Methods 0.000 claims description 4
- 208000023504 respiratory system disease Diseases 0.000 claims description 4
- 108700042778 Antimicrobial Peptides Proteins 0.000 claims description 2
- 102000044503 Antimicrobial Peptides Human genes 0.000 claims description 2
- 230000009385 viral infection Effects 0.000 claims 1
- 230000000694 effects Effects 0.000 abstract description 12
- 230000000844 anti-bacterial effect Effects 0.000 abstract description 8
- 230000003115 biocidal effect Effects 0.000 abstract description 6
- 230000005764 inhibitory process Effects 0.000 abstract description 3
- 238000001228 spectrum Methods 0.000 abstract description 3
- 238000002474 experimental method Methods 0.000 abstract description 2
- 239000003242 anti bacterial agent Substances 0.000 abstract 1
- 230000002779 inactivation Effects 0.000 abstract 1
- 210000004027 cell Anatomy 0.000 description 27
- 241001135989 Porcine reproductive and respiratory syndrome virus Species 0.000 description 19
- 244000063299 Bacillus subtilis Species 0.000 description 13
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 12
- 235000014469 Bacillus subtilis Nutrition 0.000 description 11
- 238000000855 fermentation Methods 0.000 description 11
- 230000004151 fermentation Effects 0.000 description 11
- 239000007788 liquid Substances 0.000 description 11
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 10
- 239000000243 solution Substances 0.000 description 9
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 7
- 239000012531 culture fluid Substances 0.000 description 7
- 108090000765 processed proteins & peptides Proteins 0.000 description 7
- 108090000623 proteins and genes Proteins 0.000 description 7
- 150000001413 amino acids Chemical group 0.000 description 6
- 238000012360 testing method Methods 0.000 description 6
- 241000282898 Sus scrofa Species 0.000 description 5
- 230000002401 inhibitory effect Effects 0.000 description 5
- 239000002609 medium Substances 0.000 description 5
- 239000000843 powder Substances 0.000 description 5
- 239000000047 product Substances 0.000 description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 5
- 238000005119 centrifugation Methods 0.000 description 4
- 239000012530 fluid Substances 0.000 description 4
- 238000000338 in vitro Methods 0.000 description 4
- 208000015181 infectious disease Diseases 0.000 description 4
- 238000011081 inoculation Methods 0.000 description 4
- 230000010354 integration Effects 0.000 description 4
- 102000004196 processed proteins & peptides Human genes 0.000 description 4
- 239000006228 supernatant Substances 0.000 description 4
- 230000001629 suppression Effects 0.000 description 4
- 230000004083 survival effect Effects 0.000 description 4
- 201000003176 Severe Acute Respiratory Syndrome Diseases 0.000 description 3
- 238000002835 absorbance Methods 0.000 description 3
- 235000001014 amino acid Nutrition 0.000 description 3
- 238000004458 analytical method Methods 0.000 description 3
- 239000006285 cell suspension Substances 0.000 description 3
- 238000001816 cooling Methods 0.000 description 3
- 230000009849 deactivation Effects 0.000 description 3
- 108090000454 dermaseptin Proteins 0.000 description 3
- 238000011161 development Methods 0.000 description 3
- 201000010099 disease Diseases 0.000 description 3
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 3
- 229940079593 drug Drugs 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 230000001717 pathogenic effect Effects 0.000 description 3
- 235000018102 proteins Nutrition 0.000 description 3
- 102000004169 proteins and genes Human genes 0.000 description 3
- 210000002966 serum Anatomy 0.000 description 3
- 239000011780 sodium chloride Substances 0.000 description 3
- 239000000725 suspension Substances 0.000 description 3
- JTSDBFGMPLKDCD-XVFHVFLVSA-N tilmicosin Chemical compound O([C@@H]1[C@@H](C)[C@H](O)CC(=O)O[C@@H]([C@H](/C=C(\C)/C=C/C(=O)[C@H](C)C[C@@H]1CCN1C[C@H](C)C[C@H](C)C1)CO[C@H]1[C@@H]([C@H](OC)[C@H](O)[C@@H](C)O1)OC)CC)[C@@H]1O[C@H](C)[C@@H](O)[C@H](N(C)C)[C@H]1O JTSDBFGMPLKDCD-XVFHVFLVSA-N 0.000 description 3
- 229960000223 tilmicosin Drugs 0.000 description 3
- 239000013598 vector Substances 0.000 description 3
- 208000030507 AIDS Diseases 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 208000003322 Coinfection Diseases 0.000 description 2
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 2
- 102000004142 Trypsin Human genes 0.000 description 2
- 108090000631 Trypsin Proteins 0.000 description 2
- 230000000843 anti-fungal effect Effects 0.000 description 2
- 229940121375 antifungal agent Drugs 0.000 description 2
- 238000009360 aquaculture Methods 0.000 description 2
- 244000144974 aquaculture Species 0.000 description 2
- 239000007864 aqueous solution Substances 0.000 description 2
- 230000001580 bacterial effect Effects 0.000 description 2
- 244000309466 calf Species 0.000 description 2
- 229940041514 candida albicans extract Drugs 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- 238000000034 method Methods 0.000 description 2
- 238000005457 optimization Methods 0.000 description 2
- 244000052769 pathogen Species 0.000 description 2
- 239000013600 plasmid vector Substances 0.000 description 2
- 229920001184 polypeptide Polymers 0.000 description 2
- 238000001556 precipitation Methods 0.000 description 2
- 238000004321 preservation Methods 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 230000004044 response Effects 0.000 description 2
- 238000005070 sampling Methods 0.000 description 2
- 210000000582 semen Anatomy 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 230000003595 spectral effect Effects 0.000 description 2
- 230000009466 transformation Effects 0.000 description 2
- 239000012588 trypsin Substances 0.000 description 2
- 239000012137 tryptone Substances 0.000 description 2
- 239000012138 yeast extract Substances 0.000 description 2
- QCVGEOXPDFCNHA-UHFFFAOYSA-N 5,5-dimethyl-2,4-dioxo-1,3-oxazolidine-3-carboxamide Chemical compound CC1(C)OC(=O)N(C(N)=O)C1=O QCVGEOXPDFCNHA-UHFFFAOYSA-N 0.000 description 1
- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- UCDOXFBTMLKASE-HERUPUMHSA-N Ala-Ser-Trp Chemical compound C[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)O)N UCDOXFBTMLKASE-HERUPUMHSA-N 0.000 description 1
- BCADFFUQHIMQAA-KKHAAJSZSA-N Asn-Thr-Val Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(O)=O BCADFFUQHIMQAA-KKHAAJSZSA-N 0.000 description 1
- 241000193755 Bacillus cereus Species 0.000 description 1
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 description 1
- 206010059866 Drug resistance Diseases 0.000 description 1
- 102000002322 Egg Proteins Human genes 0.000 description 1
- 108010000912 Egg Proteins Proteins 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 241000305071 Enterobacterales Species 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- QNAYBMKLOCPYGJ-REOHCLBHSA-N L-alanine Chemical compound C[C@H](N)C(O)=O QNAYBMKLOCPYGJ-REOHCLBHSA-N 0.000 description 1
- ROHFNLRQFUQHCH-YFKPBYRVSA-N L-leucine Chemical compound CC(C)C[C@H](N)C(O)=O ROHFNLRQFUQHCH-YFKPBYRVSA-N 0.000 description 1
- AAKRWBIIGKPOKQ-ONGXEEELSA-N Leu-Val-Gly Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)NCC(O)=O AAKRWBIIGKPOKQ-ONGXEEELSA-N 0.000 description 1
- ROHFNLRQFUQHCH-UHFFFAOYSA-N Leucine Natural products CC(C)CC(N)C(O)=O ROHFNLRQFUQHCH-UHFFFAOYSA-N 0.000 description 1
- GWADARYJIJDYRC-XGEHTFHBSA-N Met-Thr-Ser Chemical compound CSCC[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(O)=O GWADARYJIJDYRC-XGEHTFHBSA-N 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 108010019160 Pancreatin Proteins 0.000 description 1
- 229930182555 Penicillin Natural products 0.000 description 1
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 description 1
- 108091005804 Peptidases Proteins 0.000 description 1
- 239000001888 Peptone Substances 0.000 description 1
- 108010080698 Peptones Proteins 0.000 description 1
- PEFJUUYFEGBXFA-BZSNNMDCSA-N Phe-Lys-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@H](CCCCN)NC(=O)[C@@H](N)CC1=CC=CC=C1 PEFJUUYFEGBXFA-BZSNNMDCSA-N 0.000 description 1
- 241000269431 Phyllomedusa sauvagii Species 0.000 description 1
- 206010067268 Post procedural infection Diseases 0.000 description 1
- 239000004365 Protease Substances 0.000 description 1
- 101710192597 Protein map Proteins 0.000 description 1
- 108010026552 Proteome Proteins 0.000 description 1
- 206010037660 Pyrexia Diseases 0.000 description 1
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- MTCFGRXMJLQNBG-UHFFFAOYSA-N Serine Natural products OCC(N)C(O)=O MTCFGRXMJLQNBG-UHFFFAOYSA-N 0.000 description 1
- UIIMBOGNXHQVGW-DEQYMQKBSA-M Sodium bicarbonate-14C Chemical compound [Na+].O[14C]([O-])=O UIIMBOGNXHQVGW-DEQYMQKBSA-M 0.000 description 1
- SWIKDOUVROTZCW-GCJQMDKQSA-N Thr-Asn-Ala Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](C)C(=O)O)N)O SWIKDOUVROTZCW-GCJQMDKQSA-N 0.000 description 1
- GVMXJJAJLIEASL-ZJDVBMNYSA-N Thr-Pro-Thr Chemical compound C[C@@H](O)[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)O)C(O)=O GVMXJJAJLIEASL-ZJDVBMNYSA-N 0.000 description 1
- BBPCSGKKPJUYRB-UVOCVTCTSA-N Thr-Thr-Leu Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(O)=O BBPCSGKKPJUYRB-UVOCVTCTSA-N 0.000 description 1
- AYFVYJQAPQTCCC-UHFFFAOYSA-N Threonine Natural products CC(O)C(N)C(O)=O AYFVYJQAPQTCCC-UHFFFAOYSA-N 0.000 description 1
- 239000004473 Threonine Substances 0.000 description 1
- AEMPCGRFEZTWIF-IHRRRGAJSA-N Val-Leu-Lys Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(O)=O AEMPCGRFEZTWIF-IHRRRGAJSA-N 0.000 description 1
- 230000001464 adherent effect Effects 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 235000004279 alanine Nutrition 0.000 description 1
- 125000000539 amino acid group Chemical group 0.000 description 1
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 1
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 1
- 235000011130 ammonium sulphate Nutrition 0.000 description 1
- AVKUERGKIZMTKX-NJBDSQKTSA-N ampicillin Chemical compound C1([C@@H](N)C(=O)N[C@H]2[C@H]3SC([C@@H](N3C2=O)C(O)=O)(C)C)=CC=CC=C1 AVKUERGKIZMTKX-NJBDSQKTSA-N 0.000 description 1
- 229960000723 ampicillin Drugs 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 239000003443 antiviral agent Substances 0.000 description 1
- 229940031567 attenuated vaccine Drugs 0.000 description 1
- 235000015278 beef Nutrition 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- NKWPZUCBCARRDP-UHFFFAOYSA-L calcium bicarbonate Chemical compound [Ca+2].OC([O-])=O.OC([O-])=O NKWPZUCBCARRDP-UHFFFAOYSA-L 0.000 description 1
- 229910000020 calcium bicarbonate Inorganic materials 0.000 description 1
- 239000001110 calcium chloride Substances 0.000 description 1
- 229910001628 calcium chloride Inorganic materials 0.000 description 1
- 150000001768 cations Chemical class 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 230000019522 cellular metabolic process Effects 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 230000009194 climbing Effects 0.000 description 1
- 230000009514 concussion Effects 0.000 description 1
- 239000002537 cosmetic Substances 0.000 description 1
- 238000005138 cryopreservation Methods 0.000 description 1
- 230000000120 cytopathologic effect Effects 0.000 description 1
- YFHLIDBAPTWLGU-CTKMSOPVSA-N dermaseptin Chemical group C([C@@H](C(=O)N[C@@H](C)C(=O)NCC(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C)C(=O)N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(=O)NCC(=O)N[C@@H](C)C(=O)N[C@@H](C)C(=O)N[C@@H](C)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCC(N)=O)C(=O)NCC(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCC(N)=O)C(O)=O)[C@@H](C)O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](C)NC(=O)[C@H](CCSC)NC(=O)[C@@H](NC(=O)CNC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCSC)NC(=O)[C@@H](NC(=O)[C@H](CCCCN)NC(=O)[C@H](CC=1C2=CC=CC=C2NC=1)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](C)N)[C@@H](C)O)[C@@H](C)O)C1=CN=CN1 YFHLIDBAPTWLGU-CTKMSOPVSA-N 0.000 description 1
- 229940049701 dermaseptin Drugs 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 230000001627 detrimental effect Effects 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 239000012154 double-distilled water Substances 0.000 description 1
- 238000004043 dyeing Methods 0.000 description 1
- 235000014103 egg white Nutrition 0.000 description 1
- 210000000969 egg white Anatomy 0.000 description 1
- 238000004520 electroporation Methods 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 238000013401 experimental design Methods 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 108010089804 glycyl-threonine Proteins 0.000 description 1
- 125000001165 hydrophobic group Chemical group 0.000 description 1
- 229960003444 immunosuppressant agent Drugs 0.000 description 1
- 230000001861 immunosuppressant effect Effects 0.000 description 1
- 239000003018 immunosuppressive agent Substances 0.000 description 1
- 230000000415 inactivating effect Effects 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- 239000002917 insecticide Substances 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 230000001665 lethal effect Effects 0.000 description 1
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 1
- 235000019341 magnesium sulphate Nutrition 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 238000002844 melting Methods 0.000 description 1
- 230000008018 melting Effects 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 230000035772 mutation Effects 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 231100000028 nontoxic concentration Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 238000011017 operating method Methods 0.000 description 1
- 229940055695 pancreatin Drugs 0.000 description 1
- 231100000915 pathological change Toxicity 0.000 description 1
- 230000036285 pathological change Effects 0.000 description 1
- 229940049954 penicillin Drugs 0.000 description 1
- 235000019319 peptone Nutrition 0.000 description 1
- 239000002574 poison Substances 0.000 description 1
- 231100000614 poison Toxicity 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- 244000144977 poultry Species 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 238000012797 qualification Methods 0.000 description 1
- 230000005855 radiation Effects 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 230000000630 rising effect Effects 0.000 description 1
- 239000011435 rock Substances 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 238000004062 sedimentation Methods 0.000 description 1
- 229920002545 silicone oil Polymers 0.000 description 1
- 239000002356 single layer Substances 0.000 description 1
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 1
- 238000010561 standard procedure Methods 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 239000011550 stock solution Substances 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 239000010902 straw Substances 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 238000004885 tandem mass spectrometry Methods 0.000 description 1
- 238000012549 training Methods 0.000 description 1
- 210000004881 tumor cell Anatomy 0.000 description 1
- 239000005723 virus inoculator Substances 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
- C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
- C07K14/4701—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals not used
- C07K14/4723—Cationic antimicrobial peptides, e.g. defensins
-
- 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
Landscapes
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Molecular Biology (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- Zoology (AREA)
- Genetics & Genomics (AREA)
- Medicinal Chemistry (AREA)
- Gastroenterology & Hepatology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Toxicology (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Peptides Or Proteins (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
本发明属于生物技术领域，具体公开一种抗菌肽Dermaseptin‑M及其应用。其氨基酸序列如SEQ ID No.1所示。通过实验证明本发明抗菌肽不但对大肠杆菌，绿脓杆菌，金黄色葡萄球菌等有明显的抑制作用，而且对病毒也有一定的灭活作用，稳定性好，抗菌活性强，具有高效广谱抑菌作用，可以作为抗生素替代品使用。
Description
技术领域
本发明属于生物技术领域，具体涉及一种抗菌肽Dermaseptin-M及其应用。
背景技术
抗生素在养殖业中的滥用导致药物在动物产品中残留，已经严重影响食品安全，使细菌对抗生素产生耐药性，并通过餐桌潜入人体，成为潜伏在人体内的隐形杀手。其危机是如果发生新的病种将面临无药可用的局面。越来越多的国家开始寻找抗生素的替代品，而抗菌肽以其独特的生物活性和杀菌机制成为最具潜力的抗生素替代品之一。抗菌肽在食品、化妆品领域的应用也不断加深，具有良好的发展前景。
抗菌肽（antibacterial peptides)是一类具有巨大发展潜力的新型抗菌药物。抗菌肽是宿主产生的一类抵抗外界病原体感染的小分子阳离子肽，广泛存在于昆虫、植物、动物及人体内，具有非特异性抗细菌、真菌、病毒等病原体作用，还有抗肿瘤细胞作用，因此又称为肽抗生素（peptide antibitics)。抗菌肽抗菌谱广，对多重耐药菌、肿瘤细胞、艾滋病毒有杀伤作用，甚至还可能有抗重症急性呼吸综合征（severe acute respiratorysyndrome，SARS）病毒的作用。因此有着广阔的开发应用前景，是目前国际学术研究的活跃领域之一。
发明内容
本发明的目的旨在提供一种抗菌肽Dermaseptin-M及其应用。
为实现上述目的，本发明采取的技术方案如下：
一种抗菌肽Dermaseptin-M，其氨基酸序列如SEQ ID No.1所示。
所述抗菌肽Dermaseptin-M在制备抗菌肽制剂中的应用。
所述抗菌肽Dermaseptin-M在制备治疗革兰氏阳性菌、革兰氏阴性菌和/或病毒感染疾病的药物中的应用。
所述革兰氏阴性菌为大肠杆菌或绿脓杆菌，所述革兰氏阳性菌为金黄色葡萄球菌，所述病毒为猪繁殖与呼吸障碍综合征病毒。
有益效果：通过实验证明本发明抗菌肽Dermaseptin-M不但对大肠杆菌，绿脓杆菌，金黄色葡萄球菌等有明显的抑制作用，而且对病毒也有一定的灭活作用，稳定性好，抗菌活性强，具有高效广谱抑菌作用，可以作为抗生素替代品使用。
附图说明
图1：发酵液中Dermaseptin-M蛋白纯度鉴定结果；其中，M为蛋白maker，1，2，3，4均为样品重复；
图2：Dermaseptin-M蛋白质谱图。
具体实施方式
以下结合具体实施例对本发明的技术方案作进一步详细说明，但本发明的保护范围并不局限于此。
实施例1
本发明抗菌肽的氨基酸序列如SEQ ID No.1所示，具体为：
Ala-Ser-Trp-Met-Thr-Ser-Phe-Lys-Lys-Val-Leu-Lys-Thr-Pro-Thr-Lys-Thr-Thr-Leu-Asn-Thr-Val-Leu-Val-Gly-Thr-Asn-Ala。
序列特征：序列为氨基酸序列，含有28个氨基酸残基，链型为α螺旋直链多肽，分子量大小为 3405 .04Da ，等电点是10.25，疏水残基46.42%。
本实施例抗菌肽是发明人在Dermaseptin基因基础上通过氨基酸替代增强其多肽抗微生物能力，将改造后Dermaseptin-M基因转入枯草芽孢杆菌，并通过枯草芽孢杆菌发酵而得；其具体步骤如下：
1. Dermaseptin-M序列合成
Dermaseptin-M是在Dermaseptin序列（如SEQ ID No.2所示）：
GCGTTGTGGATGACCTTGTTAAAAAAGGTTCTTAAAGCTGCCGCAAAGGCTGCGCTGAATGCAGTGTTGGTAGGTGCAAATGCT的基础上，将第37、43、49、52、61、76位碱基G变为A，其编码氨基酸由丙氨酸变为苏氨酸，第5、17位T变成C，其氨基酸由亮氨酸变成丝氨酸，即Dermaseptin-M序列为：
GCGTCGTGGATGACCTCGTTAAAAAAGGTTCTTAAAACTGCCACAAAGACTACGCTGAATACAGTGTTGGTAGGTACAAATGCT（如SEQ ID No.3所示）。替代后，其亲水性增强，更易于表达。序列委托上海生工公司合成。
2. Dermaseptin-M序列整合入枯草芽孢杆菌
2.1 整合载体的构建
将Dermaseptin-M序列按照pUB110载体(购自BioVector质粒载体菌种细胞基因保藏中心）克隆标准操作步骤连接到芽孢杆菌载体pUB110上，得到整合载体pUB110-DM。
2.2 枯草芽孢杆菌基因组的整合
将构建好的整合载体pUB110-DM用电转化法转化至Bacillus subtilis（枯草芽孢杆菌）168 (购自BioVector质粒载体菌种细胞基因保藏中心）感受态，操作步骤如下：
1. 以1：100的比例将10mL Bacillus subtilis（枯草芽孢杆菌）168菌液（市购菌粉用无菌水稀释而得）倒入1000mL LB液体培养基中，37℃、220rpm振摇2-3小时，每半小时测一次OD，当OD值达到0.3-0.4时，停止培养；
2. 将菌液在冰上预冷30分钟，随后将菌液分装到500mL 冰上预冷的离心杯中，4℃、2500rpm离心10分钟；
3. 弃上清，离心杯中加入少量ddH2O，使沉淀悬浮后，再将水注满离心杯，4℃、4000rpm离心10分钟；
4. 弃上清，往离心杯中加入少量10%甘油(灭菌，冰上预冷；甘油与水的体积百分比，下同)，重悬菌体，再加满10%甘油，4℃、4000rpm离心10min；
5. 弃上清，每个离心杯中加入5mL10%的甘油，使沉淀悬浮后，制得感受态；取100 μL感受态加0.01ng pUB110-DM直接电穿孔转化，用0.05%（g/mL，下同）氯霉素抗性的LB固体培养基筛选阳性转化子。
其中，LB液体培养基的配方：胰蛋白胨 10g/L，酵母提取物 5g/L，NaCl 10g/L；
LB固体培养基的配方：胰蛋白胨 10g/L，酵母提取物 5g/L，NaCl 10g/L、琼脂15 g/L。
3 枯草芽孢杆菌优化表达
首先应用 Plackett-Burman 试验设计方法，确定出转速、接种量和总发酵时间为重要影响因素。通过最陡爬坡法接近最大响应面区域，利用 Box-Behnken 试验设计，建立了以Dermaseptin-M蛋白含量为响应值的全变量二次回归方程模型，最终得到枯草芽孢杆菌产蛋白酶的最优发酵条件为：
最佳培养基配方：葡萄糖20g，玉米粉10g，大豆粉10g，酵母粉8g，硅油1g，碳酸氢钙0.5g，氯化钙0.015g，硫酸铵1g，硫酸镁0.15g，加水1000 mL，用碳酸氢钠调节pH至7.0，于121℃条件下灭菌20 min。
最佳发酵条件如下：装液量为500mL的三角瓶中装100mL培养基，初始pH值为7.0，接种菌液量为2%（即每100ml液体培养基中加入2mL 浓度为1×108个/mL的枯草芽孢杆菌悬液，枯草芽孢杆菌悬液是指筛选所得阳性转化子经肉汤培养基培养并调整菌落浓度而得)，培养温度为35℃，摇床转速200rpm，总发酵时间120h。
在摇床条件下，优化后此发酵液的Dermaseptin-M蛋白含量达974.96mg/mL。
鉴定：
（1）、将发酵液取样进行SDS-PAGE鉴别其纯度，结果显示如图1所示，发酵液中几乎没有杂蛋白片段，只有Dermaseptin-M蛋白，灰度扫描计算其纯度为98.9%；
（2）将发酵液取样送于上海中科新生命生物科技有限公司进行质谱鉴定，质谱结果见图2，质谱分析见表1；PMF和MS/MS数据提交到MASCOT数据库进行搜索，分类学检索为Phyllomedusa sauvagii，结果显示，该蛋白与Dermaseptin蛋白相似率为78.57%，即产生了6个突变。结合图谱分析，该蛋白序列为 ：
Ala-Ser-Trp-Met-Thr-Ser-Phe-Lys-Lys-Val-Leu-Lys-Thr-Pro-Thr-Lys-Thr-Thr-Leu-Asn-Thr-Val-Leu-Val-Gly-Thr-Asn-Ala，与发明人预期结果完全一致，说明该蛋白在枯草芽孢杆菌内得到正确表达。
收集发酵液离心，5000rpm，取上清冻干，得到淡黄色粉末状成品即抗菌肽Dermaseptin-M，密封包装，-20°保存可以保存两年，4°可以保存6个月。
4. 发明产品抗菌肽Dermaseptin-M最小抑菌浓度（MIC)的测定
MIC实验选择大肠杆菌、绿脓杆菌和金黄色葡萄球菌等临床常见有害菌进行。分别将大肠杆菌（ATCC25922)、绿脓杆菌（ATCC18642)、金黄色葡萄球菌（ATCC25923）用肉汤培养基37℃培养至对数期，然后用肉汤培养基稀释至2.0×106 cfu/mL。在青霉素瓶中依次加入稀释过的抗菌肽水溶液100μL（浓度为0.5、1、2、4、5、10、15、20mg/mL），向各瓶中加入已经稀释好的菌液100 μL。混匀后置于37℃恒温培养箱静置培养18h，震荡后测定各瓶菌液在600nm处的吸光度值，以100μg/mL氨苄青霉素水溶液作为阳性对照。其中，肉汤培养基的配方为：蛋白胨 10g/L，牛肉浸出粉 3g/L，氯化钠5g/L。
结果判定：以检测不到细菌生长的抗菌肽最小浓度孔作为其最小抑菌浓度，结果如表2所示。
表2可以看出，Dermaseptin-M对大肠杆菌，绿脓杆菌以及金黄色葡萄球菌等临床常见细菌都有很强的抑制效果，其中对金黄色葡萄球菌效果最好，绿脓杆菌次之，具有较好的开发价值。
5. 本发明产品抗菌肽Dermaseptin-M体外抑制猪繁殖与呼吸障碍综合征病毒的活性
猪繁殖与呼吸障碍综合征病毒（PRRSV）是目前困扰养殖业的一大难题，猪群感染后引起猪的免疫抑制，导致免疫失败，还会造成猪瘟、大肠杆菌病等多种疫病混合感染或者继发感染。本发明抗菌肽对PRRSV在体外有一定的抑制作用。
本实验中PRRSV采用HuN4 F112 株弱毒疫苗，购自哈尔滨维科生物技术有限公司，用生理盐水将其病毒滴度稀释至10-7 TCID50/mL，备用。
Marc-145购自中国兽医药品监察所，按如下步骤复苏培养：
1.细胞实验室进行常规消毒，紫外照射40min以上；
2.培养液（DMEM）、0.25％（g/g，下同)胰蛋白酶溶液恒温水浴箱37℃预热20min，备用；
3.从液氮保存罐中取出细胞冻存管，立即放入37℃水浴中，快速摇晃，直至冻存液完全融化；
4.将融化后所得细胞悬液移入15mL离心管，缓慢加入4mL DMEM培养液，离心（1000r/min，5min）；
5.用DMEM培养液混悬沉淀细胞，置于37℃、5% CO2细胞培养箱中培养。每管细胞可以复苏2-3瓶。
48h后，将长满细胞的培养瓶中原来的培养液弃去，加入0.5-1mL 0.25%的胰蛋白酶溶液，使瓶底细胞都浸入溶液中。放在倒置显微镜下观察细胞，在原贴壁细胞逐渐趋于圆形，还未漂起时将胰酶弃去，加入10mL 含1v%小牛血清的DMEM培养液终止消化，用无菌吸管将贴壁细胞吹打成悬液。
5.1 Dermaseptin-M对Marc145细胞最大无毒浓度的测定
将Marc-145细胞悬液接种于96孔板中，每孔0.1mL，每孔接种细胞个数2.5×104。每孔加10v%小牛血清DMEM培养液100μL，置于37℃、5% CO2培养箱中培养，12h后观察细胞贴壁生产情况。将Dermaseptin-M用DMEM培养液稀释，浓度分别为0.5、1、2、4、8、16mg/L，待细胞贴壁后，每孔加DMEM稀释的Dermaseptin-M 100μL。每4个小时观察一次，判定各孔存活细胞的增殖代谢情况及细胞毒性反应，不产生细胞病变的药物最大浓度视为最大无毒剂量。培养96h，测得抗菌肽 Dermaseptin-M对Marc-145细胞的最大安全浓度为8mg/L。即当抗菌肽Dermaseptin-M浓度大于8mg/L时，Marc-145细胞会产生病变。
5.2 体外抗病毒试验
将Marc-145细胞悬液接种于96孔板中，每孔0.1mL，每孔接种细胞个数2.5×104，置于37℃、5% CO2培养箱中培养，48h后细胞形成单层开始进行体外抗病毒试验。
试验分正常细胞对照组（不接种病毒）、PRRSV对照组（只接种PRRSV 0.1mL，不加药物）、药物（替米考星）对照组（20mg/L，用DMEM稀释）和抗菌肽Dermaseptin-M（DM）组（2、4、6mg/L，DMEM液稀释）。试验采用同时感染的方式进行：将PRRSV病毒液用不含血清的DMEM细胞培养基稀释100倍后取100μL，与100μL药物对照组/DM组处理液在37℃、5% CO2培养箱中作用2h后感染Marc-145细胞，检测本发明抗菌肽对PRRSV的直接灭活作用。
试验采用96孔细胞培养板进行，每个处理重复10次，当PRRSV对照组感染阳性率达75%、正常细胞对照组正常时进行MTT染色，用酶标仪测490nm处吸光度值OD490nm。分别计算细胞存活率和DM对PRRSV的抑制率。
细胞存活率=（实验组OD490nm/正常细胞对照组OD490nm）×100%；
抑制率=（实验组OD490nm－PRRSV对照组OD490nm）/PRRSV对照组OD490nm×100%；
抗菌肽Dermaseptin-M对PRRSV抗性结果如表3所示。
表3可以看出，3个不同浓度的DM与PRRSV直接作用后再作用于Marc-145细胞，结果与PRRSV对照组相比，各组的OD490nm显著升高（P＜0.05），各组细胞存活率和对PRRSV的抑制率分别为61.93%、75.29%、85.80%和31.71%、60.12%、82.48%，即三个不同浓度的Dermaseptin-M都有抑制PRRSV的作用，各个浓度间的抑制效应差异显著。Dermaseptin-M在高浓度时的吸光度值、细胞存活率和对PRRSV的抑制率均稍低于替米考星对照组，但差异不显著（P＞0.05）。说明Dermaseptin-M体外可以直接灭活PRRSV，且灭活效果与药物替米考星相当。
综上，本发明抗菌肽Dermaseptin-M不仅对畜禽常见致病菌有显著的抑制作用，对革兰氏阳性菌和阴性菌都有较好的效果，抗菌谱广，而且对PRRSV也有一定的灭活作用，所以本发明抗菌肽具有极大的应用潜力，在制备抗菌抗病毒药物中能够得到广泛的应用。
SEQUENCE LISTING
<110> 河南牧翔动物药业有限公司
河南牧翔科技有限公司
<120> 一种抗菌肽Dermaseptin-M及其应用
<130>
<160> 3
<170> PatentIn version 3.4
<210> 1
<211> 28
<212> PRT
<213> 人工序列
<400> 1
Ala Ser Trp Met Thr Ser Phe Lys Lys Val Leu Lys Thr Pro Thr Lys
1 5 10 15
Thr Thr Leu Asn Thr Val Leu Val Gly Thr Asn Ala
20 25
<210> 2
<211> 84
<212> DNA
<213> 未知
<400> 2
gcgttgtgga tgaccttgtt aaaaaaggtt cttaaagctg ccgcaaaggc tgcgctgaat 60
gcagtgttgg taggtgcaaa tgct 84
<210> 3
<211> 84
<212> DNA
<213> 人工序列
<400> 3
gcgtcgtgga tgacctcgtt aaaaaaggtt cttaaaactg ccacaaagac tacgctgaat 60
acagtgttgg taggtacaaa tgct 84
Claims (4)
1.一种抗菌肽Dermaseptin-M，其特征在于：其氨基酸序列如SEQ ID No.1所示。
2.如权利要求1所述的抗菌肽Dermaseptin-M在制备抗菌肽制剂中的应用。
3.如权利要求1所述的抗菌肽Dermaseptin-M在制备治疗革兰氏阳性菌、革兰氏阴性菌和/或病毒感染疾病的药物中的应用。
4.如权利要求3所述的应用，其特征在于：所述革兰氏阴性菌为大肠杆菌或绿脓杆菌，所述革兰氏阳性菌为金黄色葡萄球菌，所述病毒为猪繁殖与呼吸障碍综合征病毒。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611109715.6A CN106397568B (zh) | 2016-12-02 | 2016-12-02 | 一种抗菌肽Dermaseptin-M及其应用 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611109715.6A CN106397568B (zh) | 2016-12-02 | 2016-12-02 | 一种抗菌肽Dermaseptin-M及其应用 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106397568A true CN106397568A (zh) | 2017-02-15 |
| CN106397568B CN106397568B (zh) | 2019-10-01 |
Family
ID=58084844
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201611109715.6A Active CN106397568B (zh) | 2016-12-02 | 2016-12-02 | 一种抗菌肽Dermaseptin-M及其应用 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106397568B (zh) |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1351667A (zh) * | 1999-03-17 | 2002-05-29 | 维多利亚大学创新和发展公司 | 抗广谱病原体的转基因植物 |
| WO2003010191A1 (en) * | 2001-07-27 | 2003-02-06 | Empresa Brasileira De Pesquisa Agropecuária - Embrapa | New cationic peptides of the dermaseptin family isolated from the skin secretion of phyllomedusa hypochondrialis |
| CN1616483A (zh) * | 2003-10-17 | 2005-05-18 | 上海高科联合生物技术研发有限公司 | 一组人工合成的抗菌肽及其制备方法和应用 |
| WO2015112980A2 (en) * | 2014-01-24 | 2015-07-30 | The Regents Of The University Of Colorado, A Body Corporate | Dermaseptin-type and piscidin-type antimicrobial peptides |
-
2016
- 2016-12-02 CN CN201611109715.6A patent/CN106397568B/zh active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1351667A (zh) * | 1999-03-17 | 2002-05-29 | 维多利亚大学创新和发展公司 | 抗广谱病原体的转基因植物 |
| WO2003010191A1 (en) * | 2001-07-27 | 2003-02-06 | Empresa Brasileira De Pesquisa Agropecuária - Embrapa | New cationic peptides of the dermaseptin family isolated from the skin secretion of phyllomedusa hypochondrialis |
| CN1616483A (zh) * | 2003-10-17 | 2005-05-18 | 上海高科联合生物技术研发有限公司 | 一组人工合成的抗菌肽及其制备方法和应用 |
| WO2015112980A2 (en) * | 2014-01-24 | 2015-07-30 | The Regents Of The University Of Colorado, A Body Corporate | Dermaseptin-type and piscidin-type antimicrobial peptides |
Non-Patent Citations (2)
| Title |
|---|
| 张庆华等: "抗菌肽Dermaseptin S4及其突变体基因在大肠杆菌中的表达", 《中国生物制品学杂志》 * |
| 陈香君等: "新型抗肿瘤杂合抗菌肽的设计、合成及抗肿瘤活性研究", 《宁夏医科大学学报》 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN106397568B (zh) | 2019-10-01 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103834596B (zh) | 一种枯草芽孢杆菌shou003、抗弧菌蛋白及其制备方法与应用 | |
| CN102702333B (zh) | 抗耐药性病原菌感染多肽及其用途 | |
| CN104151415B (zh) | 一种天然抗菌肽Alligatorin4及其应用 | |
| WO2020103511A1 (zh) | 一种拟穴青蟹抗菌肽Scyreprocin及其应用 | |
| CN113583972A (zh) | 一株可降低抗生素耐药性的大肠杆菌噬菌体及其应用 | |
| WO2021203467A1 (zh) | 一类脂肽化合物在抗新型冠状病毒中的应用 | |
| CN103122347B (zh) | 一种杀灭葡萄球菌的裂解酶及其应用 | |
| CN109748972A (zh) | 一种细胞珠蛋白-人源乳铁蛋白肽融合蛋白、基因及应用 | |
| Dantán-González et al. | Actin monoubiquitylation is induced in plants in response to pathogens and symbionts | |
| CN101638661B (zh) | 新城疫病毒hn基因和f基因重组乳酸菌的构建 | |
| KR102247293B1 (ko) | 항균 활성을 가지는 바실러스 서브틸리스 bsc35 균주 및 이의용도 | |
| CN108642029A (zh) | 绿僵菌蛋白酶Pr1C及其基因和应用 | |
| CN104861050A (zh) | 鲍曼不动杆菌锌依赖寡肽a1s_1610蛋白及其制备方法和应用 | |
| CN106397568A (zh) | 一种抗菌肽Dermaseptin‑M及其应用 | |
| CN101381757B (zh) | 抗菌肽的固体发酵制备方法及应用 | |
| CN107502603B (zh) | 一种大肠杆菌裂解酶及其制备方法与应用 | |
| CN105543256A (zh) | 一种噬菌体的裂解酶及杀菌应用 | |
| CN107670023A (zh) | V12cbd蛋白及其编码基因的一种新用途 | |
| CN109200284A (zh) | 一种猪支原体肺炎活疫苗黏膜免疫佐剂、制备方法及其应用 | |
| KR20020036265A (ko) | 길항미생물 버홀더리아 글라디올리 지비-0999 균주, 이를포함하는 미생물제제 및 이를 이용하여 식물의 풋마름병을방제하는 방법 | |
| CN106818885A (zh) | 一种酸性蛋白酶在制备抑制病原菌制剂中的应用 | |
| CN110923289B (zh) | 一种用于治疗柑橘黄龙病的药物的筛选方法 | |
| CN111471670B (zh) | 一种具有体外裂解活性的沙门氏菌宽谱裂解酶及其应用 | |
| CN106937674A (zh) | 用于肉品保鲜的抗菌剂 | |
| KR101971572B1 (ko) | 신규한 살모넬라 엔테리티디스 특이 박테리오파지 및 이를 포함하는 항균 조성물 |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| CB02 | Change of applicant information | ||
| CB02 | Change of applicant information |
Address after: 451162 Airport Road, airport, Henan, Zhengzhou, five Applicant after: HENAN SOAR VETERINARY PHARMACEUTICAL Co.,Ltd. Applicant after: HENAN MUXIANG BIOTECHNOLOGY Co.,Ltd. Address before: 451162 Airport Road, airport, Henan, Zhengzhou, five Applicant before: HENAN SOAR VETERINARY PHARMACEUTICAL Co.,Ltd. Applicant before: HENAN MUXIANG TECHNOLOGY Co.,Ltd. |
|
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| PE01 | Entry into force of the registration of the contract for pledge of patent right | ||
| PE01 | Entry into force of the registration of the contract for pledge of patent right |
Denomination of invention: An antibacterial peptide dermaseptin-m and its application Effective date of registration: 20211224 Granted publication date: 20191001 Pledgee: China Construction Bank Corporation Zhengzhou Railway Sub Branch Pledgor: HENAN SOAR VETERINARY PHARMACEUTICAL Co.,Ltd.|HENAN MUXIANG BIOTECHNOLOGY Co.,Ltd. Registration number: Y2021980016170 |
|
| PC01 | Cancellation of the registration of the contract for pledge of patent right | ||
| PC01 | Cancellation of the registration of the contract for pledge of patent right |
Granted publication date: 20191001 Pledgee: China Construction Bank Corporation Zhengzhou Railway Sub Branch Pledgor: HENAN SOAR VETERINARY PHARMACEUTICAL Co.,Ltd.|HENAN MUXIANG BIOTECHNOLOGY Co.,Ltd. Registration number: Y2021980016170 |
|
| PE01 | Entry into force of the registration of the contract for pledge of patent right | ||
| PE01 | Entry into force of the registration of the contract for pledge of patent right |
Denomination of invention: A antimicrobial peptide Deraseptin-M and its application Granted publication date: 20191001 Pledgee: China Construction Bank Corporation Zhengzhou Railway Sub Branch Pledgor: HENAN SOAR VETERINARY PHARMACEUTICAL Co.,Ltd.|HENAN MUXIANG BIOTECHNOLOGY Co.,Ltd. Registration number: Y2024980004821 |
|
| CP03 | Change of name, title or address | ||
| CP03 | Change of name, title or address |
Address after: No. 9, Yugang Road, Airport Economic Comprehensive Experimental Zone, Zhengzhou City, Henan Province, 451162 Patentee after: Henan Muxiang Biotechnology Co.,Ltd. Country or region after: China Address before: 451162 airport road five, Zhengzhou, Henan Patentee before: HENAN SOAR VETERINARY PHARMACEUTICAL Co.,Ltd. Country or region before: China Patentee before: HENAN MUXIANG BIOTECHNOLOGY Co.,Ltd. |