CN108148123B - 烧土火丝菌天然抗菌肽及其应用 - Google Patents
烧土火丝菌天然抗菌肽及其应用 Download PDFInfo
- Publication number
- CN108148123B CN108148123B CN201711248627.9A CN201711248627A CN108148123B CN 108148123 B CN108148123 B CN 108148123B CN 201711248627 A CN201711248627 A CN 201711248627A CN 108148123 B CN108148123 B CN 108148123B
- Authority
- CN
- China
- Prior art keywords
- antibacterial
- gly
- ala
- antibacterial peptide
- peptide
- 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.)
- Active
Links
- 239000003910 polypeptide antibiotic agent Substances 0.000 title claims abstract description 35
- 241000233866 Fungi Species 0.000 title description 5
- 239000002689 soil Substances 0.000 title description 2
- 230000000844 anti-bacterial effect Effects 0.000 claims abstract description 18
- 108090000765 processed proteins & peptides Proteins 0.000 claims abstract description 17
- 102000004196 processed proteins & peptides Human genes 0.000 claims abstract description 16
- 241000191967 Staphylococcus aureus Species 0.000 claims abstract description 10
- 241000192125 Firmicutes Species 0.000 claims abstract description 8
- 241000194021 Streptococcus suis Species 0.000 claims abstract description 4
- 125000003275 alpha amino acid group Chemical group 0.000 claims abstract 3
- 241000894006 Bacteria Species 0.000 claims description 9
- 229920001184 polypeptide Polymers 0.000 claims description 8
- 239000003814 drug Substances 0.000 claims description 5
- 239000000203 mixture Substances 0.000 claims description 4
- 238000002360 preparation method Methods 0.000 claims description 4
- 241000223198 Humicola Species 0.000 claims 1
- 241000235058 Komagataella pastoris Species 0.000 abstract description 11
- 238000003259 recombinant expression Methods 0.000 abstract description 7
- 229940124350 antibacterial drug Drugs 0.000 abstract description 4
- 235000006662 Lansium Nutrition 0.000 abstract description 3
- 241001156382 Lansium Species 0.000 abstract description 3
- 239000003674 animal food additive Substances 0.000 abstract description 3
- 239000002537 cosmetic Substances 0.000 abstract description 3
- 235000013373 food additive Nutrition 0.000 abstract description 3
- 239000002778 food additive Substances 0.000 abstract description 3
- 108020004705 Codon Proteins 0.000 abstract description 2
- 239000003755 preservative agent Substances 0.000 abstract description 2
- 238000012216 screening Methods 0.000 abstract description 2
- 108010058643 Fungal Proteins Proteins 0.000 abstract 1
- 238000010835 comparative analysis Methods 0.000 abstract 1
- 238000005457 optimization Methods 0.000 abstract 1
- 108090000623 proteins and genes Proteins 0.000 description 23
- 241000215602 Pyronema Species 0.000 description 12
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 12
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 12
- 108700042778 Antimicrobial Peptides Proteins 0.000 description 11
- 102000044503 Antimicrobial Peptides Human genes 0.000 description 11
- 239000001963 growth medium Substances 0.000 description 11
- 238000000034 method Methods 0.000 description 11
- 238000001962 electrophoresis Methods 0.000 description 9
- 239000013604 expression vector Substances 0.000 description 9
- 239000007788 liquid Substances 0.000 description 9
- 239000013598 vector Substances 0.000 description 9
- 108010077435 glycyl-phenylalanyl-glycine Proteins 0.000 description 8
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 8
- 108020004414 DNA Proteins 0.000 description 7
- SMLCYZYQFRTLCO-UWJYBYFXSA-N Tyr-Cys-Ala Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CS)C(=O)N[C@@H](C)C(O)=O SMLCYZYQFRTLCO-UWJYBYFXSA-N 0.000 description 7
- 230000014509 gene expression Effects 0.000 description 7
- 239000002609 medium Substances 0.000 description 7
- JPHYJQHPILOKHC-ACZMJKKPSA-N Glu-Asp-Asp Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O JPHYJQHPILOKHC-ACZMJKKPSA-N 0.000 description 6
- 241000282414 Homo sapiens Species 0.000 description 6
- ISHNZELVUVPCHY-ZETCQYMHSA-N Lys-Gly-Gly Chemical compound NCCCC[C@H](N)C(=O)NCC(=O)NCC(O)=O ISHNZELVUVPCHY-ZETCQYMHSA-N 0.000 description 6
- RATXDYWHIYNZLE-DCAQKATOSA-N Met-Lys-Cys Chemical compound CSCC[C@@H](C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CS)C(=O)O)N RATXDYWHIYNZLE-DCAQKATOSA-N 0.000 description 6
- 239000012634 fragment Substances 0.000 description 6
- 239000011780 sodium chloride Substances 0.000 description 6
- 239000007787 solid Substances 0.000 description 6
- QHASENCZLDHBGX-ONGXEEELSA-N Ala-Gly-Phe Chemical compound C[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 QHASENCZLDHBGX-ONGXEEELSA-N 0.000 description 5
- OFQPMRDJVWLMNJ-CIUDSAMLSA-N Asn-His-Cys Chemical compound C1=C(NC=N1)C[C@@H](C(=O)N[C@@H](CS)C(=O)O)NC(=O)[C@H](CC(=O)N)N OFQPMRDJVWLMNJ-CIUDSAMLSA-N 0.000 description 5
- NNKLKUUGESXCBS-KBPBESRZSA-N Lys-Gly-Tyr Chemical compound [H]N[C@@H](CCCCN)C(=O)NCC(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O NNKLKUUGESXCBS-KBPBESRZSA-N 0.000 description 5
- XIFAHCUNWWKUDE-DCAQKATOSA-N Val-Cys-Lys Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CS)C(=O)N[C@@H](CCCCN)C(=O)O)N XIFAHCUNWWKUDE-DCAQKATOSA-N 0.000 description 5
- 108010084455 Zeocin Proteins 0.000 description 5
- 238000000246 agarose gel electrophoresis Methods 0.000 description 5
- 210000004027 cell Anatomy 0.000 description 5
- 238000012258 culturing Methods 0.000 description 5
- 108010069495 cysteinyltyrosine Proteins 0.000 description 5
- 238000001976 enzyme digestion Methods 0.000 description 5
- 230000002538 fungal effect Effects 0.000 description 5
- XBGGUPMXALFZOT-UHFFFAOYSA-N glycyl-L-tyrosine hemihydrate Natural products NCC(=O)NC(C(O)=O)CC1=CC=C(O)C=C1 XBGGUPMXALFZOT-UHFFFAOYSA-N 0.000 description 5
- 108010010096 glycyl-glycyl-tyrosine Proteins 0.000 description 5
- 238000009630 liquid culture Methods 0.000 description 5
- CWCMIVBLVUHDHK-ZSNHEYEWSA-N phleomycin D1 Chemical compound N([C@H](C(=O)N[C@H](C)[C@@H](O)[C@H](C)C(=O)N[C@@H]([C@H](O)C)C(=O)NCCC=1SC[C@@H](N=1)C=1SC=C(N=1)C(=O)NCCCCNC(N)=N)[C@@H](O[C@H]1[C@H]([C@@H](O)[C@H](O)[C@H](CO)O1)O[C@@H]1[C@H]([C@@H](OC(N)=O)[C@H](O)[C@@H](CO)O1)O)C=1N=CNC=1)C(=O)C1=NC([C@H](CC(N)=O)NC[C@H](N)C(N)=O)=NC(N)=C1C CWCMIVBLVUHDHK-ZSNHEYEWSA-N 0.000 description 5
- 239000000843 powder Substances 0.000 description 5
- 239000000047 product Substances 0.000 description 5
- 235000018102 proteins Nutrition 0.000 description 5
- 102000004169 proteins and genes Human genes 0.000 description 5
- 239000000243 solution Substances 0.000 description 5
- 239000006228 supernatant Substances 0.000 description 5
- 108010051110 tyrosyl-lysine Proteins 0.000 description 5
- 229920001817 Agar Polymers 0.000 description 4
- CXRCVCURMBFFOL-FXQIFTODSA-N Ala-Ala-Pro Chemical compound C[C@H](N)C(=O)N[C@@H](C)C(=O)N1CCC[C@H]1C(O)=O CXRCVCURMBFFOL-FXQIFTODSA-N 0.000 description 4
- FBPFZTCFMRRESA-FSIIMWSLSA-N D-Glucitol Natural products OC[C@H](O)[C@H](O)[C@@H](O)[C@H](O)CO FBPFZTCFMRRESA-FSIIMWSLSA-N 0.000 description 4
- 102000000541 Defensins Human genes 0.000 description 4
- 108010002069 Defensins Proteins 0.000 description 4
- 108090000790 Enzymes Proteins 0.000 description 4
- 102000004190 Enzymes Human genes 0.000 description 4
- IGOYNRWLWHWAQO-JTQLQIEISA-N Gly-Phe-Gly Chemical compound OC(=O)CNC(=O)[C@@H](NC(=O)CN)CC1=CC=CC=C1 IGOYNRWLWHWAQO-JTQLQIEISA-N 0.000 description 4
- JOSAKOKSPXROGQ-BJDJZHNGSA-N Lys-Ser-Ile Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O JOSAKOKSPXROGQ-BJDJZHNGSA-N 0.000 description 4
- GPVLSVCBKUCEBI-KKUMJFAQSA-N Met-Gln-Phe Chemical compound [H]N[C@@H](CCSC)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O GPVLSVCBKUCEBI-KKUMJFAQSA-N 0.000 description 4
- KZNQNBZMBZJQJO-UHFFFAOYSA-N N-glycyl-L-proline Natural products NCC(=O)N1CCCC1C(O)=O KZNQNBZMBZJQJO-UHFFFAOYSA-N 0.000 description 4
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 4
- 239000008272 agar Substances 0.000 description 4
- 229940041514 candida albicans extract Drugs 0.000 description 4
- 238000005520 cutting process Methods 0.000 description 4
- 238000001514 detection method Methods 0.000 description 4
- 238000010828 elution Methods 0.000 description 4
- 108010077515 glycylproline Proteins 0.000 description 4
- 239000006994 mh medium Substances 0.000 description 4
- 239000013612 plasmid Substances 0.000 description 4
- 239000000600 sorbitol Substances 0.000 description 4
- 239000012138 yeast extract Substances 0.000 description 4
- HAVKMRGWNXMCDR-STQMWFEESA-N Arg-Gly-Phe Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)NCC(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O HAVKMRGWNXMCDR-STQMWFEESA-N 0.000 description 3
- OIMUAKUQOUEPCZ-WHFBIAKZSA-N Cys-Asn-Gly Chemical compound SC[C@H](N)C(=O)N[C@@H](CC(N)=O)C(=O)NCC(O)=O OIMUAKUQOUEPCZ-WHFBIAKZSA-N 0.000 description 3
- 241000588724 Escherichia coli Species 0.000 description 3
- YDWZGVCXMVLDQH-WHFBIAKZSA-N Gly-Cys-Asn Chemical compound NCC(=O)N[C@@H](CS)C(=O)N[C@H](C(O)=O)CC(N)=O YDWZGVCXMVLDQH-WHFBIAKZSA-N 0.000 description 3
- ISSDODCYBOWWIP-GJZGRUSLSA-N Gly-Pro-Trp Chemical compound [H]NCC(=O)N1CCC[C@H]1C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(O)=O ISSDODCYBOWWIP-GJZGRUSLSA-N 0.000 description 3
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 3
- JBCLFWXMTIKCCB-UHFFFAOYSA-N H-Gly-Phe-OH Natural products NCC(=O)NC(C(O)=O)CC1=CC=CC=C1 JBCLFWXMTIKCCB-UHFFFAOYSA-N 0.000 description 3
- WMKXFMUJRCEGRP-SRVKXCTJSA-N His-Asn-His Chemical compound C1=C(NC=N1)C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CC2=CN=CN2)C(=O)O)N WMKXFMUJRCEGRP-SRVKXCTJSA-N 0.000 description 3
- PARSHQDZROHERM-NHCYSSNCSA-N Ile-Lys-Gly Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)NCC(=O)O)N PARSHQDZROHERM-NHCYSSNCSA-N 0.000 description 3
- DZQYZKPINJLLEN-KKUMJFAQSA-N Lys-Cys-Tyr Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)O)NC(=O)[C@H](CS)NC(=O)[C@H](CCCCN)N)O DZQYZKPINJLLEN-KKUMJFAQSA-N 0.000 description 3
- DXWNFNOPBYAFRM-IHRRRGAJSA-N Phe-Val-Cys Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CS)C(=O)O)NC(=O)[C@H](CC1=CC=CC=C1)N DXWNFNOPBYAFRM-IHRRRGAJSA-N 0.000 description 3
- BNUKRHFCHHLIGR-JYJNAYRXSA-N Pro-Trp-Asp Chemical compound C1C[C@H](NC1)C(=O)N[C@@H](CC2=CNC3=CC=CC=C32)C(=O)N[C@@H](CC(=O)O)C(=O)O BNUKRHFCHHLIGR-JYJNAYRXSA-N 0.000 description 3
- 206010041925 Staphylococcal infections Diseases 0.000 description 3
- 239000003242 anti bacterial agent Substances 0.000 description 3
- 229940088710 antibiotic agent Drugs 0.000 description 3
- 108010038633 aspartylglutamate Proteins 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 238000003776 cleavage reaction Methods 0.000 description 3
- 238000010276 construction Methods 0.000 description 3
- 239000008367 deionised water Substances 0.000 description 3
- 229910021641 deionized water Inorganic materials 0.000 description 3
- 229940079593 drug Drugs 0.000 description 3
- 238000000855 fermentation Methods 0.000 description 3
- 230000004151 fermentation Effects 0.000 description 3
- 230000002401 inhibitory effect Effects 0.000 description 3
- 208000015688 methicillin-resistant staphylococcus aureus infectious disease Diseases 0.000 description 3
- 238000012986 modification Methods 0.000 description 3
- 230000004048 modification Effects 0.000 description 3
- 238000000746 purification Methods 0.000 description 3
- 108091008146 restriction endonucleases Proteins 0.000 description 3
- 230000007017 scission Effects 0.000 description 3
- CWEAKSWWKHGTRJ-BQBZGAKWSA-N Ala-Gly-Met Chemical compound [H]N[C@@H](C)C(=O)NCC(=O)N[C@@H](CCSC)C(O)=O CWEAKSWWKHGTRJ-BQBZGAKWSA-N 0.000 description 2
- AHWRSSLYSGLBGD-CIUDSAMLSA-N Asp-Pro-Glu Chemical compound OC(=O)C[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCC(O)=O)C(O)=O AHWRSSLYSGLBGD-CIUDSAMLSA-N 0.000 description 2
- 206010011409 Cross infection Diseases 0.000 description 2
- NIXHTNJAGGFBAW-CIUDSAMLSA-N Cys-Lys-Ser Chemical compound C(CCN)C[C@@H](C(=O)N[C@@H](CO)C(=O)O)NC(=O)[C@H](CS)N NIXHTNJAGGFBAW-CIUDSAMLSA-N 0.000 description 2
- 102000012410 DNA Ligases Human genes 0.000 description 2
- 108010061982 DNA Ligases Proteins 0.000 description 2
- 108700007698 Genetic Terminator Regions Proteins 0.000 description 2
- XQDGOJPVMSWZSO-SRVKXCTJSA-N Gln-Pro-Leu Chemical compound CC(C)C[C@@H](C(=O)O)NC(=O)[C@@H]1CCCN1C(=O)[C@H](CCC(=O)N)N XQDGOJPVMSWZSO-SRVKXCTJSA-N 0.000 description 2
- 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 2
- SCWYHUQOOFRVHP-MBLNEYKQSA-N Gly-Ile-Thr Chemical compound NCC(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)O)C(O)=O SCWYHUQOOFRVHP-MBLNEYKQSA-N 0.000 description 2
- AIQWYVFNBNNOLU-RHYQMDGZSA-N Leu-Thr-Val Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(O)=O AIQWYVFNBNNOLU-RHYQMDGZSA-N 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- 206010029803 Nosocomial infection Diseases 0.000 description 2
- 229910019142 PO4 Inorganic materials 0.000 description 2
- 239000001888 Peptone Substances 0.000 description 2
- 108010080698 Peptones Proteins 0.000 description 2
- LHALYDBUDCWMDY-CIUDSAMLSA-N Pro-Glu-Ala Chemical compound C[C@H](NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H]1CCCN1)C(O)=O LHALYDBUDCWMDY-CIUDSAMLSA-N 0.000 description 2
- BTKUIVBNGBFTTP-WHFBIAKZSA-N Ser-Ala-Gly Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C)C(=O)NCC(O)=O BTKUIVBNGBFTTP-WHFBIAKZSA-N 0.000 description 2
- 241001052560 Thallis Species 0.000 description 2
- NHQVWACSJZJCGJ-FLBSBUHZSA-N Thr-Thr-Ile Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O NHQVWACSJZJCGJ-FLBSBUHZSA-N 0.000 description 2
- CKTMJBPRVQWPHU-JSGCOSHPSA-N Val-Phe-Gly Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)NCC(=O)O)N CKTMJBPRVQWPHU-JSGCOSHPSA-N 0.000 description 2
- 108010070944 alanylhistidine Proteins 0.000 description 2
- 150000001413 amino acids Chemical group 0.000 description 2
- 230000001580 bacterial effect Effects 0.000 description 2
- 244000052616 bacterial pathogen Species 0.000 description 2
- 230000003385 bacteriostatic effect Effects 0.000 description 2
- 238000009835 boiling Methods 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 238000000502 dialysis Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 239000012149 elution buffer Substances 0.000 description 2
- 239000008103 glucose Substances 0.000 description 2
- 108010049041 glutamylalanine Proteins 0.000 description 2
- 108010050848 glycylleucine Proteins 0.000 description 2
- 108010087823 glycyltyrosine Proteins 0.000 description 2
- 230000036541 health Effects 0.000 description 2
- 230000001939 inductive effect Effects 0.000 description 2
- 208000015181 infectious disease Diseases 0.000 description 2
- 239000002054 inoculum Substances 0.000 description 2
- 244000144972 livestock Species 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 238000010369 molecular cloning Methods 0.000 description 2
- 239000013642 negative control Substances 0.000 description 2
- 235000019319 peptone Nutrition 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 2
- 239000010452 phosphate Substances 0.000 description 2
- 239000008363 phosphate buffer Substances 0.000 description 2
- 244000144977 poultry Species 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 238000011084 recovery Methods 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 238000002864 sequence alignment Methods 0.000 description 2
- 229910000162 sodium phosphate Inorganic materials 0.000 description 2
- 241000894007 species Species 0.000 description 2
- 239000008223 sterile water Substances 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 230000009466 transformation Effects 0.000 description 2
- 230000001131 transforming effect Effects 0.000 description 2
- 239000012137 tryptone Substances 0.000 description 2
- 239000007222 ypd medium Substances 0.000 description 2
- 229920000936 Agarose Polymers 0.000 description 1
- BUANFPRKJKJSRR-ACZMJKKPSA-N Ala-Ala-Gln Chemical compound C[C@H]([NH3+])C(=O)N[C@@H](C)C(=O)N[C@H](C([O-])=O)CCC(N)=O BUANFPRKJKJSRR-ACZMJKKPSA-N 0.000 description 1
- YAXNATKKPOWVCP-ZLUOBGJFSA-N Ala-Asn-Ala Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](C)C(O)=O YAXNATKKPOWVCP-ZLUOBGJFSA-N 0.000 description 1
- BTYTYHBSJKQBQA-GCJQMDKQSA-N Ala-Asp-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CC(=O)O)NC(=O)[C@H](C)N)O BTYTYHBSJKQBQA-GCJQMDKQSA-N 0.000 description 1
- HUUOZYZWNCXTFK-INTQDDNPSA-N Ala-His-Pro Chemical compound C[C@@H](C(=O)N[C@@H](CC1=CN=CN1)C(=O)N2CCC[C@@H]2C(=O)O)N HUUOZYZWNCXTFK-INTQDDNPSA-N 0.000 description 1
- QCTFKEJEIMPOLW-JURCDPSOSA-N Ala-Ile-Phe Chemical compound C[C@H](N)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 QCTFKEJEIMPOLW-JURCDPSOSA-N 0.000 description 1
- MEFILNJXAVSUTO-JXUBOQSCSA-N Ala-Leu-Thr Chemical compound C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O MEFILNJXAVSUTO-JXUBOQSCSA-N 0.000 description 1
- 108010011667 Ala-Phe-Ala Proteins 0.000 description 1
- REWSWYIDQIELBE-FXQIFTODSA-N Ala-Val-Ser Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CO)C(O)=O REWSWYIDQIELBE-FXQIFTODSA-N 0.000 description 1
- RZVVKNIACROXRM-ZLUOBGJFSA-N Asn-Ala-Asp Chemical compound C[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)O)NC(=O)[C@H](CC(=O)N)N RZVVKNIACROXRM-ZLUOBGJFSA-N 0.000 description 1
- YRTOMUMWSTUQAX-FXQIFTODSA-N Asn-Pro-Asp Chemical compound NC(=O)C[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CC(O)=O)C(O)=O YRTOMUMWSTUQAX-FXQIFTODSA-N 0.000 description 1
- NJIKKGUVGUBICV-ZLUOBGJFSA-N Asp-Ala-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CC(O)=O NJIKKGUVGUBICV-ZLUOBGJFSA-N 0.000 description 1
- IXIWEFWRKIUMQX-DCAQKATOSA-N Asp-Arg-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@@H](N)CC(O)=O IXIWEFWRKIUMQX-DCAQKATOSA-N 0.000 description 1
- VHWNKSJHQFZJTH-FXQIFTODSA-N Asp-Asp-Met Chemical compound CSCC[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)O)NC(=O)[C@H](CC(=O)O)N VHWNKSJHQFZJTH-FXQIFTODSA-N 0.000 description 1
- XAJRHVUUVUPFQL-ACZMJKKPSA-N Asp-Glu-Asp Chemical compound OC(=O)C[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O XAJRHVUUVUPFQL-ACZMJKKPSA-N 0.000 description 1
- RWHHSFSWKFBTCF-KKUMJFAQSA-N Asp-His-Phe Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)O)NC(=O)[C@H](CC2=CN=CN2)NC(=O)[C@H](CC(=O)O)N RWHHSFSWKFBTCF-KKUMJFAQSA-N 0.000 description 1
- 241000283690 Bos taurus Species 0.000 description 1
- 238000009010 Bradford assay Methods 0.000 description 1
- 229920000298 Cellophane Polymers 0.000 description 1
- 108700010070 Codon Usage Proteins 0.000 description 1
- 208000035473 Communicable disease Diseases 0.000 description 1
- PRXCTTWKGJAPMT-ZLUOBGJFSA-N Cys-Ala-Ser Chemical compound [H]N[C@@H](CS)C(=O)N[C@@H](C)C(=O)N[C@@H](CO)C(O)=O PRXCTTWKGJAPMT-ZLUOBGJFSA-N 0.000 description 1
- DZLQXIFVQFTFJY-BYPYZUCNSA-N Cys-Gly-Gly Chemical compound SC[C@H](N)C(=O)NCC(=O)NCC(O)=O DZLQXIFVQFTFJY-BYPYZUCNSA-N 0.000 description 1
- MXZYQNJCBVJHSR-KATARQTJSA-N Cys-Lys-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CS)N)O MXZYQNJCBVJHSR-KATARQTJSA-N 0.000 description 1
- 230000004544 DNA amplification Effects 0.000 description 1
- 238000001712 DNA sequencing Methods 0.000 description 1
- 206010059866 Drug resistance Diseases 0.000 description 1
- 101710146739 Enterotoxin Proteins 0.000 description 1
- 206010016952 Food poisoning Diseases 0.000 description 1
- 208000019331 Foodborne disease Diseases 0.000 description 1
- ZRXBYKAOFHLTDN-GUBZILKMSA-N Gln-Cys-His Chemical compound C1=C(NC=N1)C[C@@H](C(=O)O)NC(=O)[C@H](CS)NC(=O)[C@H](CCC(=O)N)N ZRXBYKAOFHLTDN-GUBZILKMSA-N 0.000 description 1
- QBLMTCRYYTVUQY-GUBZILKMSA-N Gln-Leu-Asp Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(O)=O)C(O)=O QBLMTCRYYTVUQY-GUBZILKMSA-N 0.000 description 1
- FNAJNWPDTIXYJN-CIUDSAMLSA-N Gln-Pro-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@@H]1CCCN1C(=O)[C@@H](N)CCC(N)=O FNAJNWPDTIXYJN-CIUDSAMLSA-N 0.000 description 1
- YPFFHGRJCUBXPX-NHCYSSNCSA-N Gln-Pro-Val Chemical compound CC(C)[C@H](NC(=O)[C@@H]1CCCN1C(=O)[C@@H](N)CCC(N)=O)C(O)=O YPFFHGRJCUBXPX-NHCYSSNCSA-N 0.000 description 1
- CGOHAEBMDSEKFB-FXQIFTODSA-N Glu-Glu-Ala Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(O)=O CGOHAEBMDSEKFB-FXQIFTODSA-N 0.000 description 1
- MZZSCEANQDPJER-ONGXEEELSA-N Gly-Ala-Phe Chemical compound NCC(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 MZZSCEANQDPJER-ONGXEEELSA-N 0.000 description 1
- BUEFQXUHTUZXHR-LURJTMIESA-N Gly-Gly-Pro zwitterion Chemical compound NCC(=O)NCC(=O)N1CCC[C@H]1C(O)=O BUEFQXUHTUZXHR-LURJTMIESA-N 0.000 description 1
- INLIXXRWNUKVCF-JTQLQIEISA-N Gly-Gly-Tyr Chemical compound NCC(=O)NCC(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 INLIXXRWNUKVCF-JTQLQIEISA-N 0.000 description 1
- UHPAZODVFFYEEL-QWRGUYRKSA-N Gly-Leu-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)CN UHPAZODVFFYEEL-QWRGUYRKSA-N 0.000 description 1
- DNVDEMWIYLVIQU-RCOVLWMOSA-N Gly-Val-Asp Chemical compound NCC(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(O)=O)C(O)=O DNVDEMWIYLVIQU-RCOVLWMOSA-N 0.000 description 1
- HXKZJLWGSWQKEA-LSJOCFKGSA-N His-Ala-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CC1=CN=CN1 HXKZJLWGSWQKEA-LSJOCFKGSA-N 0.000 description 1
- HAPWZEVRQYGLSG-IUCAKERBSA-N His-Gly-Glu Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)NCC(=O)N[C@@H](CCC(O)=O)C(O)=O HAPWZEVRQYGLSG-IUCAKERBSA-N 0.000 description 1
- DLTCGJZBNFOWFL-LKTVYLICSA-N His-Tyr-Ala Chemical compound C[C@@H](C(=O)O)NC(=O)[C@H](CC1=CC=C(C=C1)O)NC(=O)[C@H](CC2=CN=CN2)N DLTCGJZBNFOWFL-LKTVYLICSA-N 0.000 description 1
- OTSVBELRDMSPKY-PCBIJLKTSA-N Ile-Phe-Asn Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC(=O)N)C(=O)O)N OTSVBELRDMSPKY-PCBIJLKTSA-N 0.000 description 1
- YKZAMJXNJUWFIK-JBDRJPRFSA-N Ile-Ser-Ala Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](C)C(=O)O)N YKZAMJXNJUWFIK-JBDRJPRFSA-N 0.000 description 1
- KWTVLKBOQATPHJ-SRVKXCTJSA-N Leu-Ala-Lys Chemical compound C[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CC(C)C)N KWTVLKBOQATPHJ-SRVKXCTJSA-N 0.000 description 1
- OIARJGNVARWKFP-YUMQZZPRSA-N Leu-Asn-Gly Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(=O)NCC(O)=O OIARJGNVARWKFP-YUMQZZPRSA-N 0.000 description 1
- FIJMQLGQLBLBOL-HJGDQZAQSA-N Leu-Asn-Thr Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O FIJMQLGQLBLBOL-HJGDQZAQSA-N 0.000 description 1
- FQZPTCNSNPWHLJ-AVGNSLFASA-N Leu-Gln-Lys Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCCCN)C(O)=O FQZPTCNSNPWHLJ-AVGNSLFASA-N 0.000 description 1
- HNDWYLYAYNBWMP-AJNGGQMLSA-N Leu-Ile-Lys Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CC(C)C)N HNDWYLYAYNBWMP-AJNGGQMLSA-N 0.000 description 1
- KPYAOIVPJKPIOU-KKUMJFAQSA-N Leu-Lys-Lys Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(O)=O KPYAOIVPJKPIOU-KKUMJFAQSA-N 0.000 description 1
- SVBJIZVVYJYGLA-DCAQKATOSA-N Leu-Ser-Val Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(O)=O SVBJIZVVYJYGLA-DCAQKATOSA-N 0.000 description 1
- XFIHDSBIPWEYJJ-YUMQZZPRSA-N Lys-Ala-Gly Chemical compound OC(=O)CNC(=O)[C@H](C)NC(=O)[C@@H](N)CCCCN XFIHDSBIPWEYJJ-YUMQZZPRSA-N 0.000 description 1
- GAOJCVKPIGHTGO-UWVGGRQHSA-N Lys-Arg-Gly Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CCCNC(N)=N)C(=O)NCC(O)=O GAOJCVKPIGHTGO-UWVGGRQHSA-N 0.000 description 1
- DLCAXBGXGOVUCD-PPCPHDFISA-N Lys-Thr-Ile Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O DLCAXBGXGOVUCD-PPCPHDFISA-N 0.000 description 1
- RCMDUFDXDYTXOK-CIUDSAMLSA-N Met-Gln-Cys Chemical compound CSCC[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CS)C(O)=O RCMDUFDXDYTXOK-CIUDSAMLSA-N 0.000 description 1
- 206010067482 No adverse event Diseases 0.000 description 1
- 108091028043 Nucleic acid sequence Proteins 0.000 description 1
- 206010034133 Pathogen resistance Diseases 0.000 description 1
- 241001494479 Pecora Species 0.000 description 1
- MMYUOSCXBJFUNV-QWRGUYRKSA-N Phe-Gly-Cys Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)NCC(=O)N[C@@H](CS)C(=O)O)N MMYUOSCXBJFUNV-QWRGUYRKSA-N 0.000 description 1
- APJPXSFJBMMOLW-KBPBESRZSA-N Phe-Gly-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H](N)CC1=CC=CC=C1 APJPXSFJBMMOLW-KBPBESRZSA-N 0.000 description 1
- MSSXKZBDKZAHCX-UNQGMJICSA-N Phe-Thr-Val Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(O)=O MSSXKZBDKZAHCX-UNQGMJICSA-N 0.000 description 1
- 241000235648 Pichia Species 0.000 description 1
- UTAUEDINXUMHLG-FXQIFTODSA-N Pro-Asp-Ala Chemical compound C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)O)NC(=O)[C@@H]1CCCN1 UTAUEDINXUMHLG-FXQIFTODSA-N 0.000 description 1
- XUSDDSLCRPUKLP-QXEWZRGKSA-N Pro-Asp-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)[C@@H]1CCCN1 XUSDDSLCRPUKLP-QXEWZRGKSA-N 0.000 description 1
- ZPPVJIJMIKTERM-YUMQZZPRSA-N Pro-Gln-Gly Chemical compound OC(=O)CNC(=O)[C@H](CCC(=O)N)NC(=O)[C@@H]1CCCN1 ZPPVJIJMIKTERM-YUMQZZPRSA-N 0.000 description 1
- CHYAYDLYYIJCKY-OSUNSFLBSA-N Pro-Thr-Ile Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O CHYAYDLYYIJCKY-OSUNSFLBSA-N 0.000 description 1
- RSTWKJFWBKFOFC-JYJNAYRXSA-N Pro-Trp-Asn Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](CC(N)=O)C(O)=O RSTWKJFWBKFOFC-JYJNAYRXSA-N 0.000 description 1
- 108010026552 Proteome Proteins 0.000 description 1
- 241000736246 Pyrola Species 0.000 description 1
- 108091058545 Secretory proteins Proteins 0.000 description 1
- 102000040739 Secretory proteins Human genes 0.000 description 1
- NVNPWELENFJOHH-CIUDSAMLSA-N Ser-Ser-His Chemical compound C1=C(NC=N1)C[C@@H](C(=O)O)NC(=O)[C@H](CO)NC(=O)[C@H](CO)N NVNPWELENFJOHH-CIUDSAMLSA-N 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- 108010006785 Taq Polymerase Proteins 0.000 description 1
- NDZYTIMDOZMECO-SHGPDSBTSA-N Thr-Thr-Ala Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C)C(O)=O NDZYTIMDOZMECO-SHGPDSBTSA-N 0.000 description 1
- IXEGQBJZDIRRIV-QEJZJMRPSA-N Trp-Asn-Glu Chemical compound [H]N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O IXEGQBJZDIRRIV-QEJZJMRPSA-N 0.000 description 1
- VTCKHZJKWQENKX-KBPBESRZSA-N Tyr-Lys-Gly Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CCCCN)C(=O)NCC(O)=O VTCKHZJKWQENKX-KBPBESRZSA-N 0.000 description 1
- SYSWVVCYSXBVJG-RHYQMDGZSA-N Val-Leu-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](C(C)C)N)O SYSWVVCYSXBVJG-RHYQMDGZSA-N 0.000 description 1
- LTTQCQRTSHJPPL-ZKWXMUAHSA-N Val-Ser-Asp Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(=O)O)C(=O)O)N LTTQCQRTSHJPPL-ZKWXMUAHSA-N 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 206010048038 Wound infection Diseases 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 239000000853 adhesive Substances 0.000 description 1
- 230000001070 adhesive effect Effects 0.000 description 1
- 108010087924 alanylproline Proteins 0.000 description 1
- 230000003698 anagen phase Effects 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 238000003975 animal breeding Methods 0.000 description 1
- 230000000845 anti-microbial effect Effects 0.000 description 1
- 108010077245 asparaginyl-proline Proteins 0.000 description 1
- 108010040443 aspartyl-aspartic acid Proteins 0.000 description 1
- 108010093581 aspartyl-proline Proteins 0.000 description 1
- 235000015278 beef Nutrition 0.000 description 1
- 230000003115 biocidal effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 238000002815 broth microdilution Methods 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 108010079058 casein hydrolysate Proteins 0.000 description 1
- 238000005341 cation exchange Methods 0.000 description 1
- 238000005277 cation exchange chromatography Methods 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 239000013599 cloning vector Substances 0.000 description 1
- NKLPQNGYXWVELD-UHFFFAOYSA-M coomassie brilliant blue Chemical compound [Na+].C1=CC(OCC)=CC=C1NC1=CC=C(C(=C2C=CC(C=C2)=[N+](CC)CC=2C=C(C=CC=2)S([O-])(=O)=O)C=2C=CC(=CC=2)N(CC)CC=2C=C(C=CC=2)S([O-])(=O)=O)C=C1 NKLPQNGYXWVELD-UHFFFAOYSA-M 0.000 description 1
- 108010016616 cysteinylglycine Proteins 0.000 description 1
- 235000013365 dairy product Nutrition 0.000 description 1
- 230000007123 defense Effects 0.000 description 1
- 238000010612 desalination reaction Methods 0.000 description 1
- 231100000249 enterotoxic Toxicity 0.000 description 1
- 230000002242 enterotoxic effect Effects 0.000 description 1
- 239000000147 enterotoxin Substances 0.000 description 1
- 231100000655 enterotoxin Toxicity 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 238000001502 gel electrophoresis Methods 0.000 description 1
- 108010037850 glycylvaline Proteins 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 108010040030 histidinoalanine Proteins 0.000 description 1
- 210000005260 human cell Anatomy 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 238000004255 ion exchange chromatography Methods 0.000 description 1
- 230000009191 jumping Effects 0.000 description 1
- 230000002147 killing effect Effects 0.000 description 1
- 230000031700 light absorption Effects 0.000 description 1
- 108010054155 lysyllysine Proteins 0.000 description 1
- 208000004396 mastitis Diseases 0.000 description 1
- 238000001840 matrix-assisted laser desorption--ionisation time-of-flight mass spectrometry Methods 0.000 description 1
- 238000000691 measurement method Methods 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 235000013336 milk Nutrition 0.000 description 1
- 239000008267 milk Substances 0.000 description 1
- 210000004080 milk Anatomy 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 230000008520 organization Effects 0.000 description 1
- 230000035515 penetration Effects 0.000 description 1
- 108010078656 plectasin Proteins 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 238000002331 protein detection Methods 0.000 description 1
- 239000012474 protein marker Substances 0.000 description 1
- 238000001742 protein purification Methods 0.000 description 1
- 235000020185 raw untreated milk Nutrition 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 238000007711 solidification Methods 0.000 description 1
- 230000008023 solidification Effects 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 108010061238 threonyl-glycine Proteins 0.000 description 1
- 230000009261 transgenic effect Effects 0.000 description 1
- 108010005834 tyrosyl-alanyl-glycine Proteins 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/37—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from fungi
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/142—Amino acids; Derivatives thereof
- A23K20/147—Polymeric derivatives, e.g. peptides or proteins
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
- 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
- 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/10—General cosmetic use
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Polymers & Plastics (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- General Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Veterinary Medicine (AREA)
- Food Science & Technology (AREA)
- Mycology (AREA)
- Engineering & Computer Science (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- Molecular Biology (AREA)
- Medicinal Chemistry (AREA)
- Genetics & Genomics (AREA)
- Biophysics (AREA)
- Biochemistry (AREA)
- Epidemiology (AREA)
- Birds (AREA)
- Animal Husbandry (AREA)
- Zoology (AREA)
- Gastroenterology & Hepatology (AREA)
- Dermatology (AREA)
- Nutrition Science (AREA)
- Peptides Or Proteins (AREA)
Abstract
本发明公开了一组烧土火丝菌天然抗菌肽，其氨基酸序列分别如SEQ ID NO:5‑8所示。通过检索真菌蛋白数据库，筛选分析获得抗菌肽成熟肽序列，通过对比分析，该组天然抗菌肽与目前已知所有抗菌肽的氨基酸序列存在明显差异，属于新型抗菌肽。该组抗菌肽经密码子优化后实现其在毕赤酵母中的重组表达。抗菌实验结果表明，本发明的抗菌肽对革兰氏阳性菌，尤其是金黄色葡萄球菌ATCC43300，猪链球菌CVCC 3928，CVCC 606具有显著抑菌活性，可应用于抗菌药物、食品添加剂、化妆品、饲料添加剂、防腐剂等领域，具有广阔的应用价值和市场前景。
Description
技术领域
本发明涉及生物医药领域，，具体地说，涉及烧土火丝菌天然抗菌肽及其应用。
背景技术
抗生素对感染性疾病的治疗发挥了重要作用，但由于抗生素滥用，耐药菌株不断增加，现已出现多种超级细菌，病原菌耐药性问题已被世界卫生组织(WHO)列为未来人类健康面临的主要威胁和挑战之一。金黄色葡萄球菌是人类重要病原菌，常引起食物中毒和伤口感染，在动物养殖业中主要引起牛、羊的乳房炎。原料奶中金黄色葡萄球菌易产生金黄色葡萄球菌肠毒素等有害物质而污染乳制品(
等，EnterotoxigenicStaphylococcus aureus in bulk milk in Norway.Journal of AppliedMicrobiology.2005,99:158-166.)，影响乳制品的质量安全，同时对消费者带来安全隐患。耐药性金黄色葡萄球菌(MRSA、VRSA等)能在畜禽和人之间实现交叉感染，很多引起人类感染的MRSA菌株最终发现均来源于动物体内。另外，有研究表明，耐药性金黄色葡萄球菌能在畜禽和人之间实现交叉感染，很多引起人类感染的MRSA菌株最终发现均来源于动物体内。因而寻找新型抗菌物质取代或辅助传统抗生素以应对细菌耐药性问题至关重要。
抗菌肽(antimicrobial peptide)是广泛存在于生物体内具有抗菌活性的多肽，形成生物体第一道先天防御系统，对细菌、真菌、病毒和原虫等均具有杀灭作用。在诸多的抗菌肽中，真菌防御素的研究正逐渐受到关注。真菌防御素对革兰氏阳性细菌具有独特高效的杀菌作用，与传统抗生素之间不存在交叉耐受性，且对人的细胞没有毒害作用。真菌防御素易于通过基因表达进行大量生产，解决了抗菌肽生产成本高的问题。然而，现存的抗菌肽中，仅有0.46％来自真菌。因此，寻找真菌中天然抗菌肽并进行制备是本发明研究的重要内容。
发明内容
本发明的目的是提供烧土火丝菌天然抗菌肽及其应用。
为了实现本发明目的，本发明提供的烧土火丝菌天然抗菌肽，其特征在于，所述抗菌肽选自氨基酸序列分别如SEQ ID NO:5-8所示的多肽P1-P4中的任一个。
本发明还提供上述抗菌肽的编码基因，多肽P1-P4编码基因的核苷酸序列分别如SEQ ID NO:9-12所示。
本发明还提供表达盒、表达载体、克隆载体、转基因细胞系或工程菌，其包括包含编码所述抗菌肽基因序列的核酸。
本发明还提供一种重组毕赤酵母，其是用携带有所述抗菌肽编码基因的表达载体(例如pPICZαA)转化毕赤酵母工程菌，筛选阳性克隆得到的。
在本发明的一个具体实施方式中，所述重组毕赤酵母的构建如下：所述抗菌肽的编码基因经密码子优化后，在优化后的基因序列(分别如SEQ ID NO:9-12所示)的5’端添加XhoI酶切位点和Kex2切割位点，在3’端添加TAA和TAG终止子序列以及XbaI酶切位点，得到基因构建物，再将所述基因构建物插入载体pPICZαA的XhoI和XbaI酶切位点之间构建得到重组表达载体，最后用所述重组表达载体转化毕赤酵母菌X-33，筛选阳性克隆。
本发明还提供利用所述重组毕赤酵母发酵生产天然抗菌肽的方法，其是将上述的重组毕赤酵母发酵培养，分泌产生抗菌肽。
本发明还提供所述抗菌肽在制备广谱抗菌药物或组合物中的应用。
本发明还提供由所述抗菌肽制备的广谱抗菌药物或组合物。其中，所述菌为革兰氏阳性菌，包括但不限于金黄色葡萄球菌、猪链球菌。
本发明进一步提供所述抗菌肽在食品添加剂、化妆品、饲料添加剂领域中的应用。
本发明首次提供了一组来源于烧土火丝菌的天然抗菌肽及其在抗菌方面的应用，尤其针对革兰氏阳性菌具有很好的杀菌效果，尤其是金黄色葡萄球菌ATCC43300，猪链球菌CVCC 3928，CVCC 606具有显著抑菌活性，可应用于抗菌药物、食品添加剂、化妆品、饲料添加剂、防腐剂等领域，具有广阔的应用价值和市场前景。
附图说明
图1为本发明实施例5中P1-P4摇瓶表达120h Tricine-SDS-PAGE电泳图，M：超低分子量蛋白Marker；A,B,C,D中1-6：分别为1-4号阳性转化子在摇瓶水平发酵0,24,48,72,96和120h上清(20μL)。
具体实施方式
以下实施例用于说明本发明，但不用来限制本发明的范围。若未特别指明，实施例均按照常规实验条件，如Sambrook等分子克隆实验手册(Sambrook J&Russell DW,Molecular Cloning:a Laboratory Manual,2001)，或按照制造厂商说明书建议的条件。
以下实施例中使用的酶和试剂：限制性内切酶、TaqDNA聚合酶、T4DNA连接酶等分别购自Biolabs、Invitrogen和Promega公司。四种dNTP购自Promega公司。DNA和蛋白质分子量标准为Biolabs产品。其它常规试剂采用进口分装或国产分析纯。
以下实施例中涉及的培养基和缓冲液配方：
LB培养基：胰蛋白胨10g/L，酵母浸提取物5g/L，NaCl 10g/L；固体LB培养基则加入2％的琼脂糖。
低盐LB培养基：胰蛋白胨10g/L，酵母浸提取物5g/L，NaCl 5g/L；固体低盐LB培养基则加入2％的琼脂粉。
MH培养基：酪蛋白水解物17.5g/L，牛肉浸粉5g/L，淀粉1.5g/L。
MHA培养基：固体MH培养基则加入2％的琼脂粉。
YPD培养基：蛋白胨20g/L，酵母浸提取物10g/L，葡萄糖20g/L；固体YPD培养基则加入2％琼脂粉。
YPDS培养基：蛋白胨20g/L，酵母浸提取物10g/L，山梨醇182.2g/L，葡萄糖20g/L，琼脂粉20g/L。
有关LB培养基、低盐LB、MH、YPD、YPDS等培养基的使用参照Invitrogen公司提供的毕赤酵母操作手册。
20mM磷酸盐缓冲液(A液)：0.4654g Na2HPO4，2.9172g NaH2PO4，加去离子水至950mL，置于磁力搅拌器至完全溶解后调pH5.7，定容至1000mL。
1M NaCl 20mM磷酸盐缓冲液(B液)：0.4654g Na2HPO4，2.9172g NaH2PO4，58.44gNaCl，加去离子水至950mL，置于磁力搅拌器至完全溶解后调pH 5.7，定容至1000mL。
以下实施例中涉及的基因扩增及转化子鉴定方法为PCR法及DNA测序法。
以下实施例中涉及的蛋白检测方法为Tricine-SDS-PAGE，参照(
H.Tricine-SDS-PAGE.Nat protoc,2006,1(1):16-22)。
以下实施例中涉及的蛋白质浓度测定方法为考马斯亮蓝法。
以下实施例中涉及的蛋白分子量的确定方法为MALDI-TOF MS法。
以下实施例中涉及的蛋白纯化的方法为基于离子层析法。
以下实施例中涉及的菌种和质粒见表1。这些菌种是公众可以得到的，无需进行保藏。
表1供试菌种和质粒
实施例1天然抗菌肽与Plectasin类似真菌防御素的发现
通过检索，从UniProt数据库(http://www.uniprot.org/)真菌蛋白组数据中发现来源于烧土火丝菌(Pyronema omphalodes)的四条多肽序列(U4LUG1,U4LWE8,U4LNY7,U4LND7)。该组多肽序列及其功能区域分别如SEQ ID NO:1-4和SEQ ID NO:5-8所示。
将该组多肽全长基因序列用NCBI网站的blastX软件(https://blast.ncbi.nlm.nih.gov/Blast.cgi？PROGRAM＝blastx&PAGE_TY PE＝BlastSearch&LINK_LOC＝blasthome)进行序列比对分析结果表明，该基因的编码产物可能为真菌防御素家族抗菌肽前体。进一步将该组基因编码的多肽前体序列用NCBI网站的protein blast软件(https://blast.ncbi.nlm.nih.gov/Blast.cgi？PROGRAM＝blastp&PAGE_TY PE＝BlastSearch&LINK_LOC＝blasthome)进行序列比对，并与之前发现的其它真菌防御素家族抗菌肽序列进行比较分析，确定该组抗菌肽成熟过程中酶切位点为-R-G-，从而获得该组抗菌肽成熟肽序列SEQ ID NO:5-8，命名为P1-P4。P1-P4与目前已知所有抗菌肽的氨基酸序列存在差异，属于新型抗菌肽。
实施例2抗菌肽P1-P4基因合成
依据毕赤酵母密码子偏爱性设计抗菌肽P1-P4基因序列(SEQ ID NO:9-12)。为保证表达过程中序列的完整性，在突变体基因序列5’端添加XhoI酶切位点和Kex2切割位点，在3’端添加TAA，TAG终止子序列和XbaI酶切位点。以上序列由上海生工生物工程股份有限公司完成。
实施例3毕赤酵母诱导型表达载体pPICP1～pPICP4的构建
用限制性内切酶XhoI和XbaI分别对合成的基因片段和载体pPICZαA进行双酶切，回收pPICZαA载体片段和抗菌肽基因片段，连接，得到载体pPICP1～pPICP4；
载体pPICP1～pPICP4详细构建过程：利用限制性内切酶XhoI和XbaI分别对合成的基因片段和pPICZαA进行双酶切，酶切体系和条件如下：
以上酶切体系加样完毕后于37℃反应3个小时，2％琼脂糖凝胶电泳检测，电泳条件：130V，30min。电泳完毕在紫外灯下利用手术刀分别切割载体片段与基因片段对应电泳条带并利用天根生物技术有限公司胶回收试剂盒进行DNA片段回收，按试剂盒提供说明书的相关细节进行操作。
利用琼脂糖凝胶电泳检测回收片段并使用定量软件(GeneTools)进行初步定量，按照片段/载体(5:1)的摩尔比例使用T4DNA连接酶进行连接，体系和条件如下：
以上连接体系加样完毕后于16℃过夜连接，转化E.coli DH5α。转化操作细节如下：
1)连接产物加入100μL E.coli DH5α感受态细胞，冰浴30min；
2)42℃热激90s，立即冰浴2～3min；
3)加入900μL 37℃预热的低盐LB培养基，37℃，80-100rpm恢复培养1h；
4)4000rpm离心5min，吸去700μL上清；
5)重悬菌体后取100-200μL涂布含有25μg/mL Zeocin的LB低盐固体培养基；
6)37℃倒置培养12-16h。
挑取阳性转化子，根据基因序列设计引物，进行菌液PCR验证转化子正确性，PCR体系、条件如下：
PCR体系：
PCR条件：
2％琼脂糖凝胶电泳检测阳性转化子菌液PCR产物，电泳条件：130V，30min。15％甘油管保存含有重组表达载体的E.coli并提取质粒，为线性化电转P.pastoris准备，相关实验细节按照质粒提取试剂盒(天根生物科技有限公司)说明书操作。
实施例4含pPICP1～pPICP4重组酵母菌株的构建
(1)重组载体pPICP1～pPICP4的线性化
利用PmeI对诱导型重组表达载体pPICP1～pPICP4进行线性化，线性化体系和反应条件如下：
以上酶切体系加样完毕后于37℃反应3个小时，2％琼脂糖凝胶电泳检测，电泳条件：130V，30min。电泳完毕检测重组表达载体正确线性化后利用DNA回收试剂盒回收线性化的重组表达载体。
(2)线性化载体的毕赤酵母电转化与鉴定
1)挑取YPD平板上的X-33单菌落，接种至10mL YPD液体培养基，30℃，250rpm，过夜培养；
2)取50μL过夜培养液接种至100mL YPD液体培养基，30℃，250rpm，培养至OD600吸光值为1.2；
3)50mL培养物，4℃，4000rpm，5min离心后加入50mL无菌水重悬；
4)4℃，4000rpm，5min离心后加入25mL无菌水重悬；
5)4℃，4000rpm，5min离心后加入2mL 1M山梨醇重悬；
6)4℃，4000rpm，5min离心后加入100μL 1M山梨醇重悬后即为X-33感受态细胞(以上6步操作须在冰上操作，动作轻柔)；
7)预混80μL X-33感受态细胞和5-10μg线性化载体，转移至冰上预冷的0.2cm电转杯中，冰上放置5min后电转仪操作(1200V，25μF，400Ω)；
8)立即加入1mL 1M山梨醇，混匀；
9)30℃温育1-2h；
10)取100μL温育后的X-33细胞涂布含有100μg/mL Zeocin的YPDS平板，30℃倒置培养2-4天。
挑取YPDS平板上的单菌落接种至100μg/mL Zeocin的YPD液体培养基中，30℃，250rpm，过夜培养。取1mL过夜培养物4℃，12000rpm，5min离心后PBS重悬，沸水浴10min，快速放置于-70℃30min，立即再次沸水浴10min，4℃，12000rpm，5min离心后取上清作为模板进行PCR验证阳性转化子，PCR体系与条件如下：
PCR体系：
PCR条件：
2％琼脂糖凝胶电泳检测阳性转化子菌液PCR产物，电泳条件：120V，30min。将大小正确的阳性转化子一一对应转移至含有100μg/mL Zeocin的YPD平板上以备进一步表达。
实施例5天然抗菌肽P1-P4的表达
(1)转化子的表达
挑取阳性转化子，接种至含有100μg/mL Zeocin的YPD液体培养基，30℃，250rpm振荡培养18-20h；0.5-1％接种量转接至50mL YPD液体培养基，30℃，250rpm振荡培养1天后以4层灭菌纱布替换玻璃纸封口膜包裹摇瓶口，30℃，250rpm振荡培养3天至发酵结束。
(2)重组酵母发酵液抗菌活性检测
抑菌圈实验分析：挑取S.aureusATCC 25923单菌落接种于10mL MH培养基中，37℃，250rpm培养至OD600nm≈0.4，1％接种量转移至50mL MH固体培养基中，混匀，迅速倒19cm×19cm的方形培养皿中，待凝固后，在培养基表面小心放置牛津杯，加入50μL发酵液上清。
(3)重组酵母分泌蛋白水平Tricine-SDS-PAGE检测
对所获得的高活性重组酵母菌株进一步采用Tricine-SDS-PAGE分析重组突变体表达水平，电泳方法参照(
2006)。结果见图1。
实施例6天然抗菌肽P1-P4的纯化
(1)阳离子交换层析纯化
Bradford法测定蛋白浓度，4℃，12000rpm离心10min后取上清。将HiPrep SP FF阳离子交换柱(长度16mm，内径10mm，GE Healthcare)利用A液平衡5-10个柱体积后上样。进样完毕后，先用含有20mM，pH6.7的磷酸盐洗脱缓冲液(A液)进行洗脱，待穿透峰洗脱完后，用含有1M NaCl的20mM，pH6.7的磷酸盐洗脱缓冲液(B液)进行洗脱，收集洗脱峰。洗脱步骤为：100％A液，洗脱5个柱体积，为穿透峰；20％A，60％B液，洗脱5个柱体积，为洗脱峰。利用UV215nm监测洗脱情况，Tricine-SDS-PAGE和检测目标产物纯化情况。
(2)1kDa透析袋除盐
收集到的洗脱峰，经1kDa截留分子量透析袋4℃透析，每2h换水一次，换水6次。收集透析后的透析液，低温真空冷冻干燥机(-54℃，0.016MPa)冻干，通过Tricine-SDS-PAGE凝胶电泳检测。
实施例7抗革兰氏阳性菌活性检测
天然抗菌肽P1-P4最小抑菌浓度(MIC，minimum inhibitory concentration)的测定参照临床和实验室标准协会(CLSI，Clinical andLaboratory Standards Institute)制定的方法(WIEGAND等，Agar and broth dilution methods to determine the minimalinhibitory concentration(MIC)of antimicrobial substances.Nature protocols,2008,3(2):163-175)，根据具体情况略有改动，操作细节如下：
受试菌株的单菌落挑至MH液体培养基中，37℃ 250rpm振荡过夜培养活化后，转接至MH液体培养基中培养至对数生长期(OD600nm＝0.4～0.6)，然后制备成105CFU/mL的菌液，加入96孔无菌细胞培养板内，每孔90μL。
用PBS通过2倍倍比稀释法对抗菌肽P1-P4进行稀释，每孔10μL抗菌肽，使其终浓度分别为128、64、32、16、8、4、2、1、0.5、0.25、0.125和0.0625μg/mL，阴性对照组为PBS代替抗菌肽的受试菌液，空白对照组为无菌MH培养基。每个处理三个平行样。
将培养板置于37℃恒温培养箱孵育16～18h，直至阴性对照孔出现肉眼可见的明显浑浊菌液，能够完全抑制细菌生长的最低浓度即为抗菌肽对受试菌株的MIC值。如出现跳孔或平行样间结果不一致情况，则重新测试。
结果如表2所示，抗菌肽P1-P4对各革兰氏阳性菌均体现出不同程度的较好的抑菌效果。
表2天然抗菌肽P1-P4对革兰氏阳性菌的抗菌活性
虽然，上文中已经用一般性说明及具体实施方案对本发明作了详尽的描述，但在本发明基础上，可以对之做一些修改或改进，这对本领域技术人员而言是显而易见的。因此，在不偏离本发明精神的基础上所做的这些修改或改进，均属于本发明要求保护的范围。
序列表
<110> 北京生泰云科技有限公司
<120> 烧土火丝菌天然抗菌肽及其应用
<130> KHP171113995.7
<160> 12
<170> SIPOSequenceListing 1.0
<210> 1
<211> 95
<212> PRT
<213> 烧土火丝菌(Pyronema omphalodes)
<400> 1
Met Gln Phe Pro Thr Ile Leu Thr Val Ala Ile Phe Gly Leu Leu Asn
1 5 10 15
Ile Ser Ala Leu Ala Ala Pro Gln Pro Val Glu Glu Ala His Gly Glu
20 25 30
Ile Phe Asn Pro Asp Val Gln Leu Asp Ala Asp Thr Asn Ala Asp Asp
35 40 45
Ser Ser His Leu Ala Lys Arg Gly Phe Gly Cys Asn Gly Pro Trp Asp
50 55 60
Glu Asp Asp Met Lys Cys His Asn His Cys Lys Ser Ile Lys Gly Tyr
65 70 75 80
Lys Gly Gly Tyr Cys Ala Lys Ala Gly Phe Val Cys Lys Cys Tyr
85 90 95
<210> 2
<211> 92
<212> PRT
<213> 烧土火丝菌(Pyronema omphalodes)
<400> 2
Met Gln Phe Thr Thr Ile Leu Ala Leu Thr Leu Thr Val Phe Gly Leu
1 5 10 15
Gly Ile Thr Ala Ala Pro Gln Pro Ala Pro Glu Ala His Ala Val Ser
20 25 30
Asp Pro Glu Ala His Pro Asp His Phe Ala Gly Met Asp Ala Ser Gln
35 40 45
Leu Lys Lys Arg Gly Phe Gly Cys Asn Gly Pro Trp Asp Glu Asp Asp
50 55 60
Met Lys Cys His Asn His Cys Lys Ser Ile Lys Gly Tyr Lys Gly Gly
65 70 75 80
Tyr Cys Ala Ser Ala Gly Phe Val Cys Lys Cys Tyr
85 90
<210> 3
<211> 94
<212> PRT
<213> 烧土火丝菌(Pyronema omphalodes)
<400> 3
Met Gln Phe Thr Thr Ala Phe Thr Val Val Leu Thr Val Phe Gly Leu
1 5 10 15
Leu Asn Gly Val Leu Ala Ala Pro Gln Pro Leu Pro Gln Gly His Ala
20 25 30
Val Ser Asp Pro Asp Ala Asn Pro Asp His Tyr Ala Gly Val Asp Asn
35 40 45
Ala Leu Ile Lys Lys Arg Gly Phe Gly Cys Gly Gly Pro Trp Asn Glu
50 55 60
Asp Asp Met Gln Cys His Asn His Cys Lys Ser Ile Lys Gly Tyr Lys
65 70 75 80
Gly Gly Tyr Cys Ala Ser Ala Gly Phe Val Cys Lys Cys Tyr
85 90
<210> 4
<211> 95
<212> PRT
<213> 烧土火丝菌(Pyronema omphalodes)
<400> 4
Met Gln Phe Thr Thr Ile Leu Ser Val Gly Ile Thr Val Phe Gly Leu
1 5 10 15
Leu Asn Thr Gly Ala Phe Ala Ala Pro Gln Pro Leu Pro Glu Ala His
20 25 30
Ala Val Ser Asp Pro Glu Ala Asn Ala Asp Arg Leu Ala Gly Met Asp
35 40 45
Ala Ala Gln Leu Gln Lys Arg Gly Phe Gly Cys Asn Gly Pro Trp Asp
50 55 60
Glu Asp Asp Met Lys Cys His Asn His Cys Lys Thr Ile Lys Gly Tyr
65 70 75 80
Lys Gly Gly Tyr Cys Ala Ser Ala Gly Phe Val Cys Lys Cys Tyr
85 90 95
<210> 5
<211> 40
<212> PRT
<213> 烧土火丝菌(Pyronema omphalodes)
<400> 5
Gly Phe Gly Cys Asn Gly Pro Trp Asp Glu Asp Asp Met Lys Cys His
1 5 10 15
Asn His Cys Lys Ser Ile Lys Gly Tyr Lys Gly Gly Tyr Cys Ala Lys
20 25 30
Ala Gly Phe Val Cys Lys Cys Tyr
35 40
<210> 6
<211> 40
<212> PRT
<213> 烧土火丝菌(Pyronema omphalodes)
<400> 6
Gly Phe Gly Cys Asn Gly Pro Trp Asp Glu Asp Asp Met Lys Cys His
1 5 10 15
Asn His Cys Lys Ser Ile Lys Gly Tyr Lys Gly Gly Tyr Cys Ala Ser
20 25 30
Ala Gly Phe Val Cys Lys Cys Tyr
35 40
<210> 7
<211> 40
<212> PRT
<213> 烧土火丝菌(Pyronema omphalodes)
<400> 7
Gly Phe Gly Cys Gly Gly Pro Trp Asn Glu Asp Asp Met Gln Cys His
1 5 10 15
Asn His Cys Lys Ser Ile Lys Gly Tyr Lys Gly Gly Tyr Cys Ala Ser
20 25 30
Ala Gly Phe Val Cys Lys Cys Tyr
35 40
<210> 8
<211> 40
<212> PRT
<213> 烧土火丝菌(Pyronema omphalodes)
<400> 8
Gly Phe Gly Cys Asn Gly Pro Trp Asp Glu Asp Asp Met Lys Cys His
1 5 10 15
Asn His Cys Lys Thr Ile Lys Gly Tyr Lys Gly Gly Tyr Cys Ala Ser
20 25 30
Ala Gly Phe Val Cys Lys Cys Tyr
35 40
<210> 9
<211> 120
<212> DNA
<213> 烧土火丝菌(Pyronema omphalodes)
<400> 9
ggttttggtt gtaacggtcc atgggatgaa gatgatatga agtgtcataa ccattgtaag 60
tctattaagg gttacaaggg tggttactgt gctaaggctg gttttgtttg taagtgttac 120
<210> 10
<211> 120
<212> DNA
<213> 烧土火丝菌(Pyronema omphalodes)
<400> 10
ggttttggtt gtaacggtcc atgggatgaa gatgatatga agtgtcataa ccattgtaag 60
tctattaagg gttacaaggg tggttactgt gcttctgctg gttttgtttg taagtgttac 120
<210> 11
<211> 120
<212> DNA
<213> 烧土火丝菌(Pyronema omphalodes)
<400> 11
ggttttggtt gtggtggtcc atggaacgaa gatgatatgc aatgtcataa ccattgtaag 60
tctattaagg gttacaaggg tggttactgt gcttctgctg gttttgtttg taagtgttac 120
<210> 12
<211> 120
<212> DNA
<213> 烧土火丝菌(Pyronema omphalodes)
<400> 12
ggttttggtt gtaacggtcc atgggatgaa gatgatatga agtgtcataa ccattgtaag 60
actattaagg gttacaaggg tggttactgt gcttctgctg gttttgtttg taagtgttac 120
Claims (1)
1.烧土火丝菌天然抗菌肽在制备广谱抗菌药物或组合物中的应用，其中，所述菌为革兰氏阳性菌，选自金黄色葡萄球菌、猪链球菌；
烧土火丝菌天然抗菌肽选自氨基酸序列分别如SEQ ID NO:5-8所示的多肽P1-P4中的任一个。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711248627.9A CN108148123B (zh) | 2017-12-01 | 2017-12-01 | 烧土火丝菌天然抗菌肽及其应用 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711248627.9A CN108148123B (zh) | 2017-12-01 | 2017-12-01 | 烧土火丝菌天然抗菌肽及其应用 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN108148123A CN108148123A (zh) | 2018-06-12 |
| CN108148123B true CN108148123B (zh) | 2020-11-03 |
Family
ID=62465966
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201711248627.9A Active CN108148123B (zh) | 2017-12-01 | 2017-12-01 | 烧土火丝菌天然抗菌肽及其应用 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108148123B (zh) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111320678B (zh) * | 2020-03-09 | 2023-06-09 | 安亭生物有限责任公司 | 抗菌肽突变体及其应用 |
| CN112569344A (zh) * | 2020-12-31 | 2021-03-30 | 中国农业科学院饲料研究所 | 抗菌肽p2在制备停乳链球菌抗菌药物中的应用 |
| CN113880931B (zh) * | 2021-10-20 | 2022-06-03 | 中国科学院深海科学与工程研究所 | 抗菌肽及其制备方法和应用 |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1606567A (zh) * | 2001-11-20 | 2005-04-13 | 诺和酶股份有限公司 | 来自假黑盘菌的抗微生物多肽 |
| CN101189256A (zh) * | 2005-06-06 | 2008-05-28 | 诺维信公司 | 具有抗微生物活性的多肽和编码该多肽的多核苷酸 |
-
2017
- 2017-12-01 CN CN201711248627.9A patent/CN108148123B/zh active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1606567A (zh) * | 2001-11-20 | 2005-04-13 | 诺和酶股份有限公司 | 来自假黑盘菌的抗微生物多肽 |
| CN101189256A (zh) * | 2005-06-06 | 2008-05-28 | 诺维信公司 | 具有抗微生物活性的多肽和编码该多肽的多核苷酸 |
Non-Patent Citations (2)
| Title |
|---|
| Jiajia Wu等.The Fungal Defensin Family Enlarged.《Pharmaceuticals (Basel)》.2014,第7卷(第8期),866-880. * |
| The Fungal Defensin Family Enlarged;Jiajia Wu等;《Pharmaceuticals (Basel)》;20140831;第7卷(第8期);第866-867、872页、图2、图5和表1 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN108148123A (zh) | 2018-06-12 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN107857803A (zh) | 天然抗菌肽及其应用 | |
| Chen et al. | Cloning and heterologous expression of a bacteriocin sakacin P from Lactobacillus sakei in Escherichia coli | |
| CN105238773B (zh) | 一种抗葡萄球菌的宽谱噬菌体嵌合裂解酶及其制备方法与应用 | |
| CN108148123B (zh) | 烧土火丝菌天然抗菌肽及其应用 | |
| CN104250293A (zh) | 抗菌肽mp1106、其制备方法及应用 | |
| US20210171585A1 (en) | Antimicrobial peptide id13, preparation method and use thereof | |
| Neshani et al. | Preparation and evaluation of a new biopesticide solution candidate for plant disease control using pexiganan gene and Pichia pastoris expression system | |
| CN111420037A (zh) | 噬菌体裂解酶Lysep3在制备广谱抗菌药物中的应用 | |
| CN103319586B (zh) | 抗耐药性金黄色葡萄球菌的抗菌肽mp1102、其制备方法及应用 | |
| CN115747075A (zh) | 一种能胞外分泌抗菌肽的三角褐指藻构建方法 | |
| KR101650869B1 (ko) | 엔지니어링 박테리아 재조합 단백질의 고효율 발현방법 및 응용 | |
| CN103320461A (zh) | 在Pichia pastoris中组成型表达靶向抗菌肽AgPlectasin的方法 | |
| CN105238809A (zh) | 抗菌肽cc31在枯草芽孢杆菌中的表达方法 | |
| CN101812459A (zh) | 编码抗菌肽Lactoferricin B的核酸及其原核表达方法 | |
| CN110468143B (zh) | 抗菌肽nzx的制备方法及应用 | |
| CN103333912B (zh) | 在Pichia pastoris中组成型表达菌丝霉素衍生物MP1102的方法 | |
| CN112094833A (zh) | 一种抑菌蛋白及其编码基因、用途和菌株法 | |
| CN112662698A (zh) | 新型抗菌肽dlp4的制备方法 | |
| CN108314718A (zh) | 一种家蝇抗菌肽maf-1a肽聚体、其编码基因及其表达和应用 | |
| CN103724412A (zh) | 中国明对虾抗脂多糖因子及其制备和应用 | |
| CN108752431A (zh) | 杂合抗菌肽Mel-MytB及其应用 | |
| Huang et al. | Efficient production of human goose-type lysozyme 2 in the methylotrophic yeast Pichia pastoris | |
| CN106146629B (zh) | 一组胰蛋白酶抗性抗菌肽及其制备方法 | |
| CN102433341B (zh) | 一种斜带石斑鱼抗菌肽的原核表达产物及其制备方法 | |
| Zhao et al. | Effect of tandem repeats of antimicrobial peptide CC34 on production of target proteins and activity of Pichia pastoris |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |