CN109627299B - 一种具有广谱抗菌活性的细菌素Gr17及其应用 - Google Patents
一种具有广谱抗菌活性的细菌素Gr17及其应用 Download PDFInfo
- Publication number
- CN109627299B CN109627299B CN201811611740.3A CN201811611740A CN109627299B CN 109627299 B CN109627299 B CN 109627299B CN 201811611740 A CN201811611740 A CN 201811611740A CN 109627299 B CN109627299 B CN 109627299B
- Authority
- CN
- China
- Prior art keywords
- bacteriocin
- enterococcus
- strain
- enterococcus faecium
- product
- 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
- 108010062877 Bacteriocins Proteins 0.000 title claims abstract description 50
- 230000000844 anti-bacterial effect Effects 0.000 title claims abstract description 27
- 241000194032 Enterococcus faecalis Species 0.000 claims abstract description 12
- 241000222122 Candida albicans Species 0.000 claims abstract description 8
- 241000588724 Escherichia coli Species 0.000 claims abstract description 8
- 229940032049 enterococcus faecalis Drugs 0.000 claims abstract description 8
- 241001135265 Cronobacter sakazakii Species 0.000 claims abstract description 6
- 241000589517 Pseudomonas aeruginosa Species 0.000 claims abstract description 6
- 241000191967 Staphylococcus aureus Species 0.000 claims abstract description 6
- 229940095731 candida albicans Drugs 0.000 claims abstract description 6
- 244000063299 Bacillus subtilis Species 0.000 claims abstract description 5
- 235000014469 Bacillus subtilis Nutrition 0.000 claims abstract description 5
- 241000186779 Listeria monocytogenes Species 0.000 claims abstract description 5
- 241000186781 Listeria Species 0.000 claims abstract description 3
- 241000206604 Brochothrix thermosphacta Species 0.000 claims abstract 2
- 125000003275 alpha amino acid group Chemical group 0.000 claims abstract 2
- 241000194033 Enterococcus Species 0.000 claims description 31
- 241000194031 Enterococcus faecium Species 0.000 claims description 16
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 14
- 239000000047 product Substances 0.000 claims description 14
- 238000000034 method Methods 0.000 claims description 13
- 238000000855 fermentation Methods 0.000 claims description 11
- 230000004151 fermentation Effects 0.000 claims description 11
- 238000002360 preparation method Methods 0.000 claims description 6
- 239000011780 sodium chloride Substances 0.000 claims description 6
- 239000000243 solution Substances 0.000 claims description 6
- 239000006228 supernatant Substances 0.000 claims description 6
- 238000002835 absorbance Methods 0.000 claims description 5
- 238000005341 cation exchange Methods 0.000 claims description 5
- 239000001963 growth medium Substances 0.000 claims description 5
- 238000004128 high performance liquid chromatography Methods 0.000 claims description 5
- 238000004519 manufacturing process Methods 0.000 claims description 5
- 239000008055 phosphate buffer solution Substances 0.000 claims description 5
- 229920002684 Sepharose Polymers 0.000 claims description 4
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 claims description 4
- 229910052921 ammonium sulfate Inorganic materials 0.000 claims description 4
- 238000012870 ammonium sulfate precipitation Methods 0.000 claims description 4
- 235000011130 ammonium sulphate Nutrition 0.000 claims description 4
- 230000003385 bacteriostatic effect Effects 0.000 claims description 4
- 238000000502 dialysis Methods 0.000 claims description 4
- 238000004255 ion exchange chromatography Methods 0.000 claims description 4
- 238000009629 microbiological culture Methods 0.000 claims description 4
- 229920005654 Sephadex Polymers 0.000 claims description 3
- 239000012507 Sephadex™ Substances 0.000 claims description 3
- 239000012043 crude product Substances 0.000 claims description 3
- 238000012258 culturing Methods 0.000 claims description 3
- 238000003756 stirring Methods 0.000 claims description 3
- 229910021642 ultra pure water Inorganic materials 0.000 claims description 3
- 239000012498 ultrapure water Substances 0.000 claims description 3
- 238000004321 preservation Methods 0.000 claims description 2
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims 6
- DTQVDTLACAAQTR-UHFFFAOYSA-N Trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 claims 4
- 238000010828 elution Methods 0.000 claims 2
- 241000191940 Staphylococcus Species 0.000 claims 1
- 230000008021 deposition Effects 0.000 claims 1
- 239000007788 liquid Substances 0.000 claims 1
- 238000011068 loading method Methods 0.000 claims 1
- 230000000630 rising effect Effects 0.000 claims 1
- 238000001228 spectrum Methods 0.000 abstract description 3
- 241000193830 Bacillus <bacterium> Species 0.000 abstract 1
- 238000000746 purification Methods 0.000 description 11
- 241000894006 Bacteria Species 0.000 description 6
- 235000013305 food Nutrition 0.000 description 6
- 230000000694 effects Effects 0.000 description 5
- 239000006872 mrs medium Substances 0.000 description 5
- 108090000623 proteins and genes Proteins 0.000 description 5
- 230000000845 anti-microbial effect Effects 0.000 description 4
- 239000008363 phosphate buffer Substances 0.000 description 4
- 102000004169 proteins and genes Human genes 0.000 description 4
- 238000011084 recovery Methods 0.000 description 4
- 108020004465 16S ribosomal RNA Proteins 0.000 description 3
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 3
- 108091005804 Peptidases Proteins 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 230000001580 bacterial effect Effects 0.000 description 3
- 230000015572 biosynthetic process Effects 0.000 description 3
- 210000004027 cell Anatomy 0.000 description 3
- 238000012512 characterization method Methods 0.000 description 3
- 238000011534 incubation Methods 0.000 description 3
- 108090000765 processed proteins & peptides Proteins 0.000 description 3
- 239000004094 surface-active agent Substances 0.000 description 3
- WFIYPADYPQQLNN-UHFFFAOYSA-N 2-[2-(4-bromopyrazol-1-yl)ethyl]isoindole-1,3-dione Chemical compound C1=C(Br)C=NN1CCN1C(=O)C2=CC=CC=C2C1=O WFIYPADYPQQLNN-UHFFFAOYSA-N 0.000 description 2
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 2
- 102000007474 Multiprotein Complexes Human genes 0.000 description 2
- 108010085220 Multiprotein Complexes Proteins 0.000 description 2
- 102000035195 Peptidases Human genes 0.000 description 2
- 239000004365 Protease Substances 0.000 description 2
- 108010076504 Protein Sorting Signals Proteins 0.000 description 2
- DBMJMQXJHONAFJ-UHFFFAOYSA-M Sodium laurylsulphate Chemical compound [Na+].CCCCCCCCCCCCOS([O-])(=O)=O DBMJMQXJHONAFJ-UHFFFAOYSA-M 0.000 description 2
- PBCJIPOGFJYBJE-UHFFFAOYSA-N acetonitrile;hydrate Chemical compound O.CC#N PBCJIPOGFJYBJE-UHFFFAOYSA-N 0.000 description 2
- 150000001413 amino acids Chemical group 0.000 description 2
- AFYNADDZULBEJA-UHFFFAOYSA-N bicinchoninic acid Chemical compound C1=CC=CC2=NC(C=3C=C(C4=CC=CC=C4N=3)C(=O)O)=CC(C(O)=O)=C21 AFYNADDZULBEJA-UHFFFAOYSA-N 0.000 description 2
- 229940088598 enzyme Drugs 0.000 description 2
- 235000013332 fish product Nutrition 0.000 description 2
- 239000005452 food preservative Substances 0.000 description 2
- 235000019249 food preservative Nutrition 0.000 description 2
- 244000078673 foodborn pathogen Species 0.000 description 2
- 210000001035 gastrointestinal tract Anatomy 0.000 description 2
- 230000005764 inhibitory process Effects 0.000 description 2
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 2
- 238000004949 mass spectrometry Methods 0.000 description 2
- 238000001840 matrix-assisted laser desorption--ionisation time-of-flight mass spectrometry Methods 0.000 description 2
- 102000004196 processed proteins & peptides Human genes 0.000 description 2
- 229940024999 proteolytic enzymes for treatment of wounds and ulcers Drugs 0.000 description 2
- 238000004007 reversed phase HPLC Methods 0.000 description 2
- 230000035945 sensitivity Effects 0.000 description 2
- WROMPOXWARCANT-UHFFFAOYSA-N tfa trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F.OC(=O)C(F)(F)F WROMPOXWARCANT-UHFFFAOYSA-N 0.000 description 2
- 241000251468 Actinopterygii Species 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- BECXEHHOZNFFFX-IHRRRGAJSA-N Arg-Ser-Tyr Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O BECXEHHOZNFFFX-IHRRRGAJSA-N 0.000 description 1
- KXFCBAHYSLJCCY-ZLUOBGJFSA-N Asn-Asn-Ser Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(O)=O KXFCBAHYSLJCCY-ZLUOBGJFSA-N 0.000 description 1
- DAYDURRBMDCCFL-AAEUAGOBSA-N Asn-Trp-Gly Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)NCC(=O)O)NC(=O)[C@H](CC(=O)N)N DAYDURRBMDCCFL-AAEUAGOBSA-N 0.000 description 1
- 241000193755 Bacillus cereus Species 0.000 description 1
- 206010012735 Diarrhoea Diseases 0.000 description 1
- BWGNESOTFCXPMA-UHFFFAOYSA-N Dihydrogen disulfide Chemical compound SS BWGNESOTFCXPMA-UHFFFAOYSA-N 0.000 description 1
- 108010067770 Endopeptidase K Proteins 0.000 description 1
- JJKKWYQVHRUSDG-GUBZILKMSA-N Glu-Ala-Lys Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CCCCN)C(O)=O JJKKWYQVHRUSDG-GUBZILKMSA-N 0.000 description 1
- AFODTOLGSZQDSL-PEFMBERDSA-N Glu-Asn-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)N)NC(=O)[C@H](CCC(=O)O)N AFODTOLGSZQDSL-PEFMBERDSA-N 0.000 description 1
- GGEJHJIXRBTJPD-BYPYZUCNSA-N Gly-Asn-Gly Chemical compound NCC(=O)N[C@@H](CC(N)=O)C(=O)NCC(O)=O GGEJHJIXRBTJPD-BYPYZUCNSA-N 0.000 description 1
- PAWIVEIWWYGBAM-YUMQZZPRSA-N Gly-Leu-Ala Chemical compound NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C)C(O)=O PAWIVEIWWYGBAM-YUMQZZPRSA-N 0.000 description 1
- HHRODZSXDXMUHS-LURJTMIESA-N Gly-Met-Gly Chemical compound CSCC[C@H](NC(=O)C[NH3+])C(=O)NCC([O-])=O HHRODZSXDXMUHS-LURJTMIESA-N 0.000 description 1
- MQFGXJNSUJTXDT-QSFUFRPTSA-N Ile-Gly-Ile Chemical compound N[C@@H]([C@@H](C)CC)C(=O)NCC(=O)N[C@@H]([C@@H](C)CC)C(=O)O MQFGXJNSUJTXDT-QSFUFRPTSA-N 0.000 description 1
- VSJXPNCQYGOLFM-XIRDDKMYSA-N Lys-Cys-Trp Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CS)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(O)=O VSJXPNCQYGOLFM-XIRDDKMYSA-N 0.000 description 1
- 244000070406 Malus silvestris Species 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 108090000526 Papain Proteins 0.000 description 1
- 102000057297 Pepsin A Human genes 0.000 description 1
- 108090000284 Pepsin A Proteins 0.000 description 1
- 229920001213 Polysorbate 20 Polymers 0.000 description 1
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 1
- 238000012300 Sequence Analysis Methods 0.000 description 1
- 241000531141 Thermosphaera Species 0.000 description 1
- 241000223230 Trichosporon Species 0.000 description 1
- PEYSVKMXSLPQRU-FJHTZYQYSA-N Trp-Ala-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](C)NC(=O)[C@H](CC1=CNC2=CC=CC=C21)N)O PEYSVKMXSLPQRU-FJHTZYQYSA-N 0.000 description 1
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 1
- OVBMCNDKCWAXMZ-NAKRPEOUSA-N Val-Ile-Ser Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CO)C(=O)O)NC(=O)[C@H](C(C)C)N OVBMCNDKCWAXMZ-NAKRPEOUSA-N 0.000 description 1
- CFIBZQOLUDURST-IHRRRGAJSA-N Val-Tyr-Cys Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)N[C@@H](CS)C(=O)O)N CFIBZQOLUDURST-IHRRRGAJSA-N 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 238000013019 agitation Methods 0.000 description 1
- 108010047495 alanylglycine Proteins 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 229940088710 antibiotic agent Drugs 0.000 description 1
- 230000002368 bacteriocinic effect Effects 0.000 description 1
- 206010006451 bronchitis Diseases 0.000 description 1
- 239000004202 carbamide Substances 0.000 description 1
- 235000013877 carbamide Nutrition 0.000 description 1
- 125000000837 carbohydrate group Chemical group 0.000 description 1
- 125000002091 cationic group Chemical group 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 238000004440 column chromatography Methods 0.000 description 1
- 239000000287 crude extract Substances 0.000 description 1
- 230000016396 cytokine production Effects 0.000 description 1
- 210000004443 dendritic cell Anatomy 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 239000012149 elution buffer Substances 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 235000021107 fermented food Nutrition 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 238000001641 gel filtration chromatography Methods 0.000 description 1
- 238000012268 genome sequencing Methods 0.000 description 1
- 108010019832 glycyl-asparaginyl-glycine Proteins 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 230000000968 intestinal effect Effects 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 108010078274 isoleucylvaline Proteins 0.000 description 1
- 239000004310 lactic acid Substances 0.000 description 1
- 235000014655 lactic acid Nutrition 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 108010009298 lysylglutamic acid Proteins 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 238000000816 matrix-assisted laser desorption--ionisation Methods 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 238000013365 molecular weight analysis method Methods 0.000 description 1
- 230000003472 neutralizing effect Effects 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- 235000005985 organic acids Nutrition 0.000 description 1
- 235000019834 papain Nutrition 0.000 description 1
- 229940055729 papain Drugs 0.000 description 1
- 229940111202 pepsin Drugs 0.000 description 1
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 description 1
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 description 1
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 description 1
- 229920000053 polysorbate 80 Polymers 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 239000003755 preservative agent Substances 0.000 description 1
- 230000002335 preservative effect Effects 0.000 description 1
- 235000019419 proteases Nutrition 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 238000013207 serial dilution Methods 0.000 description 1
- 201000009890 sinusitis Diseases 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 108010061238 threonyl-glycine Proteins 0.000 description 1
- 235000013311 vegetables Nutrition 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
- C07K14/195—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria
- C07K14/315—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria from Streptococcus (G), e.g. Enterococci
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L3/00—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs
- A23L3/34—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals
- A23L3/3454—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals in the form of liquids or solids
- A23L3/3463—Organic compounds; Microorganisms; Enzymes
- A23L3/3544—Organic compounds containing hetero rings
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
- C12P21/02—Preparation of peptides or proteins having a known sequence of two or more amino acids, e.g. glutathione
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Genetics & Genomics (AREA)
- General Health & Medical Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Biochemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Nutrition Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Gastroenterology & Hepatology (AREA)
- Biophysics (AREA)
- Medicinal Chemistry (AREA)
- Peptides Or Proteins (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
本发明涉及一种具有广谱抗菌活性的细菌素Gr17及其应用，其氨基酸序列如SEQ ID NO.3所示。所述的细菌素Gr17的抗菌谱包括但不限于：李斯特菌(Listeria monocytogenes)、金黄色葡萄球菌(Staphylococcus aereu)、枯草芽孢杆菌(Bacillus subtilis)、热死环丝菌(Brochothrix thermosphacta)、大肠杆菌(Escherichia coli)、粪肠球菌(Enterococcus faecalis)、绿脓杆菌(Pseudomonas aeruginosa)、阪崎肠杆菌(Enterobacter sakazakii)、白色念珠菌(Candida albicans)。
Description
技术领域
本发明属于生物技术领域，具体而言，涉及一种具有广谱抗菌活性的细菌素Gr17及其应用。
背景技术
细菌素是核糖体合成的蛋白质和蛋白质复合物，对食源性病原体及相关物种具有抗菌作用(Cleveland等，2001)，但不会损害生产细胞的免疫蛋白(Diep等，2007)。由于无毒，高效和安全性的优点，乳酸菌(LAB)中的细菌素已被广泛用作蔬菜，肉类和其他食品中的食品防腐剂(Gálvez等，2007；Yang等，2014)。还有人提出，它们是未来抗生素的可行替代品(Cotter等，2012)。
属于LAB属的肠球菌通常表征为革兰氏阳性，过氧化氢酶阴性，兼性厌氧和非孢子形成菌(Moraes等，2013)。肠球菌是第一个发现在婴儿胃肠道(GIT)中的LAB(Fanaro等，2010)，在发酵食品和环境中也普遍存在(Foulquie Moreno等，2006)。一些其他的肠球菌属，在商业化和疾病治疗也有广泛应用。例如，屎肠球菌
已用作食品生物防腐剂和腹泻治疗(Kathrani等，2016；Holzapfel等，2018)，粪肠球菌Symbioflor 1具有治疗鼻窦炎或支气管炎的疗效(Habermann等人，2002)，屎肠球菌JWS 833能够增强树突细胞上细胞因子的产生(Choi等人，2012)。因此，肠球菌在人类和动物，食品工业以及环境中发挥着很大的作用。
可以从LAB生产许多抗微生物物质，例如有机酸，过氧化氢和细菌素。细菌素对敏感菌株表现出高度抑制活性。根据它们的结构，细菌素被分为四类(Klaenhammer，1993)。I类由耐热小肽(<5kDa)代表。II类含有热稳定的小的未修饰肽(5-10kDa)。III类包含分子大小超过30kDa的热不稳定蛋白质。IV类被表征为含有脂质或碳水化合物部分的蛋白质复合物。通常，IIa类由保守的YGNGV基序和二硫键连接组成(Perez等，2014)。
肠球菌素是肠球菌属的细菌素。已经报道了许多产生肠球菌素的肠球菌和肠球菌素，例如来自E.faecium CTC492的肠球菌素A(Aymerich等人，1996)，来自E.faecium T136的肠球菌素B(Casaus等人，1997)，来自E.faecium P13的肠球菌素P(Cintas等，1997b)，来自E.faecium L50的肠球菌素Q(Cintas等，2000)。大多数IIa类肠球菌素被合成为具有N-末端信号肽的前体，其被ATP结合盒(ABC)转运蛋白(Havarstein等，1995)或Sec分泌系统(Cintas等，1997a)切割。N-末端信号肽的类型决定了肠球菌素的合成机制。虽然细菌素和生产细胞在食品工业中起着至关重要的作用，但对肠粘素的生物合成机制和实际应用的研究相对较少。
发明内容
在本发明中，我们从中国传统低盐发酵全鱼产品中分离的新菌株屎肠球菌Gr17，并测定了其完整基因组序列。此外，测定了纯化的肠球菌素Gr17的物理化学性质和抗菌活性。因此，屎肠球菌Gr17的基因组信息和肠球菌素Gr17的抗菌性能为食品工业中作为食品防腐剂的潜在用途提供了理论基础。
本发明首先涉及一种细菌素Gr17，所述的细菌素Gr17的氨基酸序列如SEQ IDNO.3所示。
本发明还涉及所述的细菌素Gr17在制备针对如下细菌的抑菌产品和/或杀菌产品中的应用：李斯特菌(Listeria monocytogenes)、金黄色葡萄球菌(Staphylococcusaereu)、枯草芽孢杆菌(Bacillus subtilis)、热死环丝菌(Brochothrixthermosphacta)、大肠杆菌(Escherichia coli)、粪肠球菌(Enterococcus faecalis)、绿脓杆菌(Pseudomonas aeruginosa)、阪崎肠杆菌(Enterobacter sakazakii)、白色念珠菌(Candida albicans)。
本发明还涉及所述的细菌素Gr17的生产方法，所述的方法包括如下步骤：
(1)发酵所述细菌素Gr17的生产菌株屎肠球菌Gr17，分离发酵液；
(2)硫酸铵沉淀发酵液后透析获得细菌素Gr17粗品；
(3)离子交换层析法提纯细菌素Gr17粗品获得细菌素Gr17精制品
(4)HPLC提纯细菌素Gr17精制品获得细菌素Gr17纯品。
发酵生产菌株屎肠球菌Gr17的方法为：
(1)将屎肠球菌Gr17接种在MRS培养基中，在37℃下培养至OD 600＝0.3～0.6，获得种子液，培养过程不搅拌；
(2)按0.5％(v/v)比例接种种子液到MRS培养基中，在37℃下发酵培养24小时，培养过程不搅拌。
所述的硫酸铵沉淀发酵液后透析的方法为：使用4％硫酸铵溶液沉淀上清液，用超纯水透析脱盐。
所述的离子交换层析法提纯粗品的步骤为：
(1)用20mM磷酸盐缓冲液(pH5.5)平衡SP-Sepharose Fast Flow阳离子交换柱，加样，然后用0M NaCl到1M NaCl的线性梯度洗脱，流速为1mL/min，收集约40min处级分；
(2)用20mM磷酸盐缓冲液(pH5.5)平衡Sephadex层析柱，加样，然后用20mM磷酸盐缓冲液以0.5mL/min的流速洗脱，收集约30min处的级分。
所述的HPLC提纯细菌素Gr17精制品的步骤为：
使用C18反相柱，流动相含有0.1％三氟乙酸(TFA)的95％水-乙腈(5％-95％)进行线性梯度洗脱，流速为0.5mL/min，在280nm处监测吸光度，收集约20min处的级分。
本发明还涉及发酵生产肠球菌素Gr17的屎肠球菌Gr17菌株，所述的菌株保藏于中国微生物菌种保藏管理委员会普通微生物中心(CGMCC)，地址，北京市朝阳区北辰西路1号院3号，保藏编号CGMCC No.16677。
本发明还涉及所述的屎肠球菌Gr17菌株在制备抑菌和/或杀菌产品中的应用。
附图说明
图1、肠球菌素Gr17的纯化过程检测谱图。
图2、肠球菌素Gr17的质谱分子量
具体实施方式
样品和细菌培养条件
样品来自中国贵州黎平侗族少数民族地区的中国传统低盐发酵全鱼产品“酸鱼”。所有LAB菌株均在MRS培养基中培养，37℃，无需搅拌。用于指示菌株的培养基列于表1中，在37℃下培养。将所有细菌在-80℃下储存在含有20％甘油(v/v)的培养液中。
实施例1、产生细菌素的LAB的分离
(1)将样品与无菌0.9％NaCl(80mL)混合。用无菌0.9％NaCl进行连续稀释，并将每种稀释液(100μL)涂布在MRS琼脂平板上，在37℃下培养24小时。
(2)将约589个单细菌菌落在到2mL MRS培养基中，37℃下培养24小时。
(3)将上清液在4℃下以8000g离心20分钟，重新调节至pH7.0，并通过0.22μm过滤器过滤获得仅含有分泌的细菌素(不含菌株)的上清液，通过用游标卡尺测量抑制圈的直径来检测细菌素的抗菌活性。
(4)根据对指示菌株(单核细胞增生李斯特菌和大肠杆菌)的测试选出具有抗菌活性的菌株(22株)。
(5)再进一步根据由其他指示菌株(金黄色葡萄球菌，枯草芽孢杆菌和蜡状芽孢杆菌)测试得出的具有广谱抗菌谱和高抗菌活性的菌株命名为Gr17。
实施例2、细菌素产生菌的DNA纯化和鉴定
将菌株Gr17在37℃下(无需搅拌)在MRS培养基中培养。用QIAamp DNA Mini Kit(Qiagen，Germany)进行基因组DNA的纯化。使用NanoDrop 2500分光光度计(ThermoScientific，MA，USA)测试基因组DNA的浓度和纯度。根据16S rRNA基因序列进行菌株基因型鉴定。提取的基因组DNA用作PCR模板，引物如SEQ ID NO.1和SEQ ID NO.2所示：
SEQ ID NO.1：16S rRNA-F：5'-AGAGTTTGATCCTGGCTCAG-3'；
SEQ ID NO.2：16S rRNA-R：5'-GGTTACCTTGTTACGACTT-3'。
扩增的16S rRNA由Sangon Biotech(中国上海)测序，之后对GenBank数据库进行了比对。
基因组测序和组装
使用PacBio平台制备菌株Gr17的完整基因组。完整的基因组序列如Seq ID No.3所示。根据完整基因组和16S rRNA的信息，Gr17菌株被鉴定为屎肠球菌，并命名为屎肠球菌Gr17。在GenBank中的登记号为NO.CP033376和NO.CP033377。
实施例3、细菌素的纯化和鉴定
1、细菌素粗品的制备
(1)将Gr17菌株在100mL MRS培养基中培养至OD 600＝0.4，然后将0.5％(v/v)培养物接种到2L MRS培养基中，在37℃下培养24小时。
(2)在4℃下以8000g离心20分钟除去细菌细胞，用4％硫酸铵沉淀上清液，用超纯水透析脱盐。
(3)测定粗提物的抗菌活性，将细菌素样品保存在-80℃。
2、柱层析制备细菌素纯品
(1)使用
纯化系统(GE，Sweden，USA)将活性提取物进一步纯化。方法如下：用20mM磷酸盐缓冲液(pH5.5)平衡SP-Sepharose Fast Flow阳离子交换柱(16×25mm)后，将通过0.22μm过滤器过滤的样品加载到柱上，并用0M NaCl到1M NaCl的线性梯度洗脱，流速为1mL/min。根据UV吸光度收集级分并测定抗菌活性。
(2)用20mM磷酸盐缓冲液(pH5.5)平衡Sephadex G10柱，用洗脱缓冲液(20mM磷酸盐缓冲液)以0.5mL/min的流速洗脱来自阳离子交换柱的2mL细菌素。根据UV吸光度收集的级分，测定抗菌活性。
(3)使用配备有反相高效液相色谱(RP-HPLC)系统(Agilent，CA，USA)的C18反相柱(5μm，4.6mm×250mm，Agilent，California，USA)进一步进行细菌素的纯化。使用含有0.1％三氟乙酸(TFA)的95％水-乙腈(5％-95％)的线性梯度洗脱，流速为0.5mL/min，在280nm处监测吸光度。收集纯化的细菌素即为肠球菌素Gr17，测定抗菌活性。
(4)根据说明书，通过二辛可宁酸(BCA)试剂盒(Thermo Fisher Scientific，MA，USA)测定肠球菌素Gr17的浓度。
通过硫酸铵沉淀从发酵上清液中提取粗制肠球菌素Gr17。实现了大约2.12倍的纯化效果和85.73％的回收率。在SP-Sepharose Fast Flow阳离子交换柱的纯化过程中(图1A)，活性级分出现在约40分钟处，纯化倍数为17.92倍，回收率为36.08％。Sephadex凝胶过滤色谱的纯化过程具有三种不同的肽级分，活性级分约在30分钟处(图1B)，Sephadex G10凝胶过程增加了65.6倍的抗菌活性，回收率为27％。HPLC过程使抗菌活性增加87.37倍，回收率为5.77％(图1C)。
3、肠球菌Gr17的分子量
纯化的肠球菌素Gr17的分子量通过ABI 4800(MALDI-TOF-MS)质谱(AppliedBiosystems，Foster city，USA)测定。将与基质溶液混合的肠球菌素Gr17点在靶板上并使其干燥。在阳离子模式下进行光谱测定以进行MALDI分析。
结果显示，MALDI-TOF-MS显示肠球菌素Gr17的分子量为4531.01Da(图2)。根据完整的基因组序列和分子量分析，整个肠球菌素Gr17的氨基酸序列如SEQ ID NO.3所示：
SEQ ID NO.3：RSYGNGVYCNNSKCWVNWGEAKENIIGIVISGWATGLAGMGR。
由于形成必需的二硫键(Drider等，2006)，确定的分子量与计算的结果相似。使用针对GenBank的蛋白质BLAST(www.ncbi.nlm.nih.gov/BLAST)，肠球菌素Gr17与已报道的细菌素没有表现出同源性。此外，其他成熟IIa类细菌素的序列比对结果显示肠球菌素Gr17是新的细菌素。因此，来自肠球菌Gr17的肠球菌素Gr17是一种新的IIa类细菌素。
实施例4、肠球菌Gr17的功能鉴定
1、肠球菌素Gr17的抗菌谱
将实施例3纯化的肠球菌Gr17用于测定抗菌谱。测试了针对含有食物腐败细菌和食源性病原体的指示菌株的抗菌谱。结果见表1
表1、肠球菌素Gr17的抗菌谱
a CMCC,National Center for Medical Culture Collections；ATCC,AmericanType Culture Collection；CGMCC,China Center of General Microbial CultureCollection；CVCC,China Center of Veterinary Culture Collection；CICC,ChinaCenter of Industrial Culture Collection.
b抗菌圈直径(mm):+++：>21mm；++：11–20mm；+：1–10mm；-：无抑制
2、肠球菌素Gr17对温度，pH，表面活性剂和蛋白水解酶的敏感性
将实施例3纯化的肠球菌素Gr17进行性质测定。
(1)为了确定温度对抗菌活性的影响，将肠球菌素Gr17在60℃、80℃、100℃温育30分钟，在121℃温育15分钟。测试残留的抗菌活性，37℃的样品作为对照。
(2)通过用1M NaOH或HCl调节pH在2和11之间来测试肠球菌素Gr17的pH稳定性。在pH2或12、37℃下温育3小时后，中和pH至pH6.5，测试残留的抗菌活性。结果显示，肠球菌素Gr17在pH2～10的范围内都有活性，但在pH11时失活。
(3)测试乙二胺四乙酸(EDTA)，十二烷基硫酸钠(SDS)，吐温-20，吐温-80和尿素(1％，v/v，终浓度)对肠球菌素Gr17的影响。在特定的表面活性剂中37℃温育3小时后，测试残留的抗菌活性。结果显示，肠球菌素Gr17的抗菌活性不受表面活性剂的影响。
(4)将80μL的肠球菌素Gr17与20μL酶(1mg/mL，Sigma)(包括胃蛋白酶(pH 3.0)，木瓜蛋白酶(pH 6.5)，蛋白酶K(pH 7.5))混合，测定肠球菌Gr17对各种蛋白水解酶的敏感性。将肠球菌Gr17与选定的酶类在37℃下孵育3小时后检测活性。结果显示，肠球菌素Gr17的抗菌活性不受蛋白酶的影响。
最后需要说明的是，以上实施例仅用作帮助本领域技术人员理解本发明的实质，并不用做对本发明保护范围的限定。
SEQUENCE LISTING
<110> 北京工商大学
<120> 一种具有广谱抗菌活性的细菌素Gr17及其应用
<210> 1
<211> 20
<212> DNA
<213> 人工序列
<400> 1
agagtttgat cctggctcag 20
<210> 2
<211> 19
<212> DNA
<213> 人工序列
<400> 2
ggttaccttg ttacgactt 19
<210> 3
<211> 42
<212> PRT
<213> Enterococcus faecalis
<400> 3
Arg Ser Tyr Gly Asn Gly Val Tyr Cys Asn Asn Ser Lys Cys Trp Val
1 5 10 15
Asn Trp Gly Glu Ala Lys Glu Asn Ile Ile Gly Ile Val Ile Ser Gly
20 25 30
Trp Ala Thr Gly Leu Ala Gly Met Gly Arg
35 40
Claims (9)
1.一种细菌素Gr17，所述的细菌素Gr17的氨基酸序列如SEQ ID NO.3所示。
2.权利要求1所述的细菌素Gr17在制备针对如下细菌的抑菌产品和/或杀菌产品中的应用：李斯特菌（Listeria monocytogenes）、金黄色葡萄球菌（Staphylococcus aereu）、枯草芽孢杆菌（Bacillus subtilis）、热死环丝菌（Brochothrixthermosphacta）、大肠杆菌（Escherichia coli）、粪肠球菌（Enterococcus faecalis）、绿脓杆菌（Pseudomonas aeruginosa）、阪崎肠杆菌（Enterobacter sakazakii）、白色念珠菌（Candida albicans）。
3.权利要求1所述的细菌素Gr17的生产方法，所述的方法包括如下步骤：
（1）发酵所述细菌素Gr17的生产菌株屎肠球菌Gr17，分离发酵液；
（2）硫酸铵沉淀发酵液后透析获得细菌素Gr17粗品；
（3）离子交换层析法提纯细菌素Gr17粗品获得细菌素Gr17精制品
（4）HPLC提纯细菌素Gr17精制品获得细菌素Gr17纯品；
所述屎肠球菌Gr17菌株保藏于中国微生物菌种保藏管理委员会普通微生物中心，地址，北京市朝阳区北辰西路1号院3号，保藏编号 CGMCC No.16677。
4.根据权利要求3所述的方法，其特征在于，发酵生产菌株屎肠球菌Gr17的方法为：
（1）将屎肠球菌Gr17接种在MRS培养基中，在37℃下培养至OD 600 = 0.3～0.6，获得种子液，培养过程不搅拌；
（2）按体积比0.5％比例接种种子液到MRS培养基中，在37℃下发酵培养24小时，培养过程不搅拌。
5.根据权利要求3所述的方法，其特征在于，所述的硫酸铵沉淀发酵液后透析的方法为：使用4％硫酸铵溶液沉淀上清液，用超纯水透析脱盐。
6.根据权利要求3-5任一所述的方法，其特征在于，所述的离子交换层析法提纯粗品的步骤为：
（1）用pH5.5的20mM磷酸盐缓冲液平衡SP-Sepharose Fast Flow阳离子交换柱，加样，然后用0M NaCl到1M NaCl的线性梯度洗脱，流速为1 mL / min，收集40min处级分；
（2）用pH5.5的20mM磷酸盐缓冲液平衡Sephadex 层析柱，加样，然后用20mM磷酸盐缓冲液以0.5mL / min的流速洗脱，收集30min处的级分。
7.根据权利要求6所述的方法，其特征在于，所述的HPLC提纯细菌素Gr17精制品的步骤为：
使用C18反相柱，流动相为含有0.1％三氟乙酸的95％水-5%乙腈至含有0.1％三氟乙酸的5％水-95%乙腈，进行线性梯度洗脱，流速为0.5mL/min，在280nm处监测吸光度，收集20min处的级分。
8.发酵生产肠球菌素Gr17的屎肠球菌Gr17菌株，所述的菌株保藏于中国微生物菌种保藏管理委员会普通微生物中心（CGMCC），地址，北京市朝阳区北辰西路1号院3号，保藏编号CGMCC No.16677。
9.权利要求8所述的屎肠球菌Gr17菌株在制备抑菌和/或杀菌产品中的应用。
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811304246 | 2018-11-03 | ||
| CN2018113042462 | 2018-11-03 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN109627299A CN109627299A (zh) | 2019-04-16 |
| CN109627299B true CN109627299B (zh) | 2022-02-18 |
Family
ID=66078329
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811611740.3A Active CN109627299B (zh) | 2018-11-03 | 2018-12-27 | 一种具有广谱抗菌活性的细菌素Gr17及其应用 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109627299B (zh) |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111972499B (zh) * | 2020-07-20 | 2024-04-05 | 北京工商大学 | 一种花香低盐腐乳膏发酵生产方法 |
| CN111972498B (zh) * | 2020-07-20 | 2024-04-09 | 北京工商大学 | 一种多功能菌协同发酵低盐腐乳的方法与应用 |
| CN114874937B (zh) * | 2022-04-30 | 2024-02-20 | 浙江工商大学 | 清酒乳杆菌产细菌素的分离纯化及抗菌用途和所用乳酸菌 |
| CN115851498B (zh) * | 2022-09-16 | 2023-11-10 | 河南牧业经济学院 | 一种产无前导肽细菌素的枯草芽孢杆菌及制备方法与应用 |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2005007665A2 (en) * | 2003-06-17 | 2005-01-27 | Vicuron Pharmaceuticals Inc. | Lincomycin derivatives possessing antibacterial activity |
| CN103255216A (zh) * | 2013-04-24 | 2013-08-21 | 山东艾克韦生物技术有限公司 | 院内感染多病原体并行检测基因芯片及其制作方法和应用 |
| WO2017084985A1 (en) * | 2015-11-17 | 2017-05-26 | Norwegian University Of Life Sciences | Bacteriocin composition and method |
| CN108503522A (zh) * | 2018-04-24 | 2018-09-07 | 中国科学院华南植物园 | Fasamycins类化合物及其制备方法和在制备抗菌药物中的应用 |
-
2018
- 2018-12-27 CN CN201811611740.3A patent/CN109627299B/zh active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2005007665A2 (en) * | 2003-06-17 | 2005-01-27 | Vicuron Pharmaceuticals Inc. | Lincomycin derivatives possessing antibacterial activity |
| CN103255216A (zh) * | 2013-04-24 | 2013-08-21 | 山东艾克韦生物技术有限公司 | 院内感染多病原体并行检测基因芯片及其制作方法和应用 |
| WO2017084985A1 (en) * | 2015-11-17 | 2017-05-26 | Norwegian University Of Life Sciences | Bacteriocin composition and method |
| CN108503522A (zh) * | 2018-04-24 | 2018-09-07 | 中国科学院华南植物园 | Fasamycins类化合物及其制备方法和在制备抗菌药物中的应用 |
Non-Patent Citations (1)
| Title |
|---|
| 动物双歧杆菌细菌素bifidocin A群体感应合成调控行为分析;刘国荣;《食品科学》;20180620;第39卷(第12期);全文 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN109627299A (zh) | 2019-04-16 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN109627299B (zh) | 一种具有广谱抗菌活性的细菌素Gr17及其应用 | |
| Kalmokoff et al. | Isolation and characterization of a bacteriocin (Butyrivibriocin AR10) from the ruminal anaerobe Butyrivibrio fibrisolvens AR10: evidence in support of the widespread occurrence of bacteriocin-like activity among ruminal isolates of B. fibrisolvens | |
| An et al. | Purification and partial characterization of M1-UVs300, a novel bacteriocin produced by Lactobacillus plantarum isolated from fermented sausage | |
| US6506389B2 (en) | Adherence factors of non pathogenic microorganisms and applications thereof for screening microorganisms for specific probiotic properties; novel pharmaceutical compositions and food additives comprising such microorganisms and adherence factors | |
| Perumal et al. | Purification and characterization of a bacteriocin, BacBS2, produced by Bacillus velezensis BS2 isolated from Meongge Jeotgal | |
| Yeo et al. | Narrow antagonistic activity of antimicrobial peptide from Bacillus subtilis SCK-2 against Bacillus cereus | |
| Fanaei et al. | Microbial assisted (Bacillus mojavensis) production of bio-surfactant lipopeptide with potential pharmaceutical applications and its characterization by MALDI-TOF-MS analysis | |
| Losteinkit et al. | Characterization of bacteriocin N15 produced by Enterococcus faecium N15 and cloning of the related genes | |
| Miteva et al. | Detection and characterization of a novel antibacterial substance produced by a Lactobacillus delbrueckii strain 1043 | |
| AU2006333246B2 (en) | Bacteriocin inducer peptides | |
| CN111394326B (zh) | 呕吐毒素降解酶ddh及其在单端孢霉烯族毒素脱毒中的应用 | |
| US20030165505A1 (en) | Nucleic acid and corresponding protein entitled 161P2F10B useful in treatment and detection of cancer | |
| RU2250267C2 (ru) | Бактериоцин против листерий | |
| Jung et al. | Stability and antibacterial activity of bacteriocins produced by Bacillus thuringiensis and Bacillus thuringiensis ssp. kurstaki | |
| CN110117553B (zh) | 对几种食源性病原细菌有抑菌活性的蜡样芽胞杆菌及其细菌素 | |
| Kindoli et al. | Properties of a bacteriocin from Bacillus subtilis H27 isolated from Cheonggukjang | |
| Sirtori et al. | Purification and partial characterization of an antimicrobial peptide produced by Bacillus sp. strain P45, a bacterium from the Amazon basin fish Piaractus mesopotamicus | |
| CN110746488A (zh) | 一种具有食品防腐保鲜作用的细菌素pe-zyb1及其应用 | |
| CN110885359B (zh) | 一种嗜酸乳杆菌产细菌素及其分离纯化方法 | |
| AU727676B2 (en) | Novel adherence factors of non pathogenic microorganisms and applications thereof for screening microorganisms for specific probiotic properties; novel pharmaceutical compositions and food additives comprising such microorganisms and adherence factors | |
| KR100351622B1 (ko) | 신규 락토바실러스 속 (lactobacillus sp.)mt-1077 및 그로부터 생산되는 신규 박테리오신 | |
| CN115124603A (zh) | 一种细菌素rsq01及其应用 | |
| EP1148064A1 (en) | Lactobacillus johnsonii bacteriocin, active against Helicobacter pylori | |
| CN111269296B (zh) | 一种nLsA蛋白及其结构基因和应用 | |
| CN114106197B (zh) | 一种窄谱抗菌肽及其应用 |
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 |