CN117143782A - 一种唾液链球菌嗜热亚种vb331及其应用 - Google Patents
一种唾液链球菌嗜热亚种vb331及其应用 Download PDFInfo
- Publication number
- CN117143782A CN117143782A CN202311419206.3A CN202311419206A CN117143782A CN 117143782 A CN117143782 A CN 117143782A CN 202311419206 A CN202311419206 A CN 202311419206A CN 117143782 A CN117143782 A CN 117143782A
- Authority
- CN
- China
- Prior art keywords
- thermophilus
- streptococcus salivarius
- subspecies
- powder
- streptococcus
- 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
- 241000194020 Streptococcus thermophilus Species 0.000 title claims abstract description 30
- 241000193468 Clostridium perfringens Species 0.000 claims abstract description 17
- 210000001035 gastrointestinal tract Anatomy 0.000 claims abstract description 14
- 239000006041 probiotic Substances 0.000 claims abstract description 10
- 235000018291 probiotics Nutrition 0.000 claims abstract description 10
- 241000607142 Salmonella Species 0.000 claims abstract description 9
- 238000004321 preservation Methods 0.000 claims abstract description 9
- 241000588724 Escherichia coli Species 0.000 claims abstract description 8
- 241000607768 Shigella Species 0.000 claims abstract description 8
- 241000191967 Staphylococcus aureus Species 0.000 claims abstract description 8
- 241000186779 Listeria monocytogenes Species 0.000 claims abstract description 7
- 208000035143 Bacterial infection Diseases 0.000 claims abstract description 6
- 241000194032 Enterococcus faecalis Species 0.000 claims abstract description 6
- 208000022362 bacterial infectious disease Diseases 0.000 claims abstract description 6
- 229940032049 enterococcus faecalis Drugs 0.000 claims abstract description 6
- 238000009629 microbiological culture Methods 0.000 claims abstract description 5
- 230000000529 probiotic effect Effects 0.000 claims abstract description 5
- 239000000843 powder Substances 0.000 claims description 18
- 241000894006 Bacteria Species 0.000 claims description 15
- 238000000855 fermentation Methods 0.000 claims description 13
- 230000004151 fermentation Effects 0.000 claims description 12
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims description 9
- 241000194024 Streptococcus salivarius Species 0.000 claims description 9
- 239000000203 mixture Substances 0.000 claims description 9
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 8
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 8
- 235000013305 food Nutrition 0.000 claims description 8
- 238000002360 preparation method Methods 0.000 claims description 8
- 230000036541 health Effects 0.000 claims description 7
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 6
- 230000001580 bacterial effect Effects 0.000 claims description 6
- 229940041514 candida albicans extract Drugs 0.000 claims description 6
- 239000000463 material Substances 0.000 claims description 6
- 239000012138 yeast extract Substances 0.000 claims description 6
- 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 claims description 5
- 235000013361 beverage Nutrition 0.000 claims description 5
- 239000008103 glucose Substances 0.000 claims description 5
- 235000013336 milk Nutrition 0.000 claims description 5
- 239000008267 milk Substances 0.000 claims description 5
- 210000004080 milk Anatomy 0.000 claims description 5
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 claims description 5
- 235000013618 yogurt Nutrition 0.000 claims description 5
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 claims description 4
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 4
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 claims description 4
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 claims description 4
- 229910052799 carbon Inorganic materials 0.000 claims description 4
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims description 4
- 239000003814 drug Substances 0.000 claims description 4
- 229910017053 inorganic salt Inorganic materials 0.000 claims description 4
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 4
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 4
- 230000000813 microbial effect Effects 0.000 claims description 4
- 229910052757 nitrogen Inorganic materials 0.000 claims description 4
- 239000001888 Peptone Substances 0.000 claims description 3
- 108010080698 Peptones Proteins 0.000 claims description 3
- 239000000654 additive Substances 0.000 claims description 3
- 229910000396 dipotassium phosphate Inorganic materials 0.000 claims description 3
- 235000019797 dipotassium phosphate Nutrition 0.000 claims description 3
- 229940079593 drug Drugs 0.000 claims description 3
- 229940099596 manganese sulfate Drugs 0.000 claims description 3
- 239000011702 manganese sulphate Substances 0.000 claims description 3
- 235000007079 manganese sulphate Nutrition 0.000 claims description 3
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 claims description 3
- 235000019319 peptone Nutrition 0.000 claims description 3
- 239000011780 sodium chloride Substances 0.000 claims description 3
- 235000002639 sodium chloride Nutrition 0.000 claims description 3
- JLPULHDHAOZNQI-ZTIMHPMXSA-N 1-hexadecanoyl-2-(9Z,12Z-octadecadienoyl)-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCCCCCC\C=C/C\C=C/CCCCC JLPULHDHAOZNQI-ZTIMHPMXSA-N 0.000 claims description 2
- OWEGMIWEEQEYGQ-UHFFFAOYSA-N 100676-05-9 Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC2C(OC(O)C(O)C2O)CO)O1 OWEGMIWEEQEYGQ-UHFFFAOYSA-N 0.000 claims description 2
- GUBGYTABKSRVRQ-XLOQQCSPSA-N Alpha-Lactose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@H](O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-XLOQQCSPSA-N 0.000 claims description 2
- 235000017060 Arachis glabrata Nutrition 0.000 claims description 2
- 244000105624 Arachis hypogaea Species 0.000 claims description 2
- 235000010777 Arachis hypogaea Nutrition 0.000 claims description 2
- 235000018262 Arachis monticola Nutrition 0.000 claims description 2
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 claims description 2
- 229920000742 Cotton Polymers 0.000 claims description 2
- 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 claims description 2
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 claims description 2
- FBPFZTCFMRRESA-JGWLITMVSA-N D-glucitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-JGWLITMVSA-N 0.000 claims description 2
- 108010068370 Glutens Proteins 0.000 claims description 2
- 244000068988 Glycine max Species 0.000 claims description 2
- 235000010469 Glycine max Nutrition 0.000 claims description 2
- 229910021578 Iron(III) chloride Inorganic materials 0.000 claims description 2
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 claims description 2
- 239000005913 Maltodextrin Substances 0.000 claims description 2
- 229920002774 Maltodextrin Polymers 0.000 claims description 2
- GUBGYTABKSRVRQ-PICCSMPSSA-N Maltose Natural products O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-PICCSMPSSA-N 0.000 claims description 2
- 229930195725 Mannitol Natural products 0.000 claims description 2
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims description 2
- 235000019764 Soybean Meal Nutrition 0.000 claims description 2
- 229920002472 Starch Polymers 0.000 claims description 2
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 2
- 229930006000 Sucrose Natural products 0.000 claims description 2
- 240000008042 Zea mays Species 0.000 claims description 2
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 claims description 2
- 235000002017 Zea mays subsp mays Nutrition 0.000 claims description 2
- 235000020167 acidified milk Nutrition 0.000 claims description 2
- 150000003863 ammonium salts Chemical class 0.000 claims description 2
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 claims description 2
- 229910052921 ammonium sulfate Inorganic materials 0.000 claims description 2
- 235000011130 ammonium sulphate Nutrition 0.000 claims description 2
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 claims description 2
- GUBGYTABKSRVRQ-QUYVBRFLSA-N beta-maltose Chemical compound OC[C@H]1O[C@H](O[C@H]2[C@H](O)[C@@H](O)[C@H](O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@@H]1O GUBGYTABKSRVRQ-QUYVBRFLSA-N 0.000 claims description 2
- 229910000019 calcium carbonate Inorganic materials 0.000 claims description 2
- 235000010216 calcium carbonate Nutrition 0.000 claims description 2
- 239000001110 calcium chloride Substances 0.000 claims description 2
- 229910001628 calcium chloride Inorganic materials 0.000 claims description 2
- 235000011148 calcium chloride Nutrition 0.000 claims description 2
- 239000002775 capsule Substances 0.000 claims description 2
- 239000004202 carbamide Substances 0.000 claims description 2
- 235000013877 carbamide Nutrition 0.000 claims description 2
- 235000013351 cheese Nutrition 0.000 claims description 2
- 235000013409 condiments Nutrition 0.000 claims description 2
- 229910000365 copper sulfate Inorganic materials 0.000 claims description 2
- ARUVKPQLZAKDPS-UHFFFAOYSA-L copper(II) sulfate Chemical compound [Cu+2].[O-][S+2]([O-])([O-])[O-] ARUVKPQLZAKDPS-UHFFFAOYSA-L 0.000 claims description 2
- 235000005822 corn Nutrition 0.000 claims description 2
- 239000003937 drug carrier Substances 0.000 claims description 2
- 239000011790 ferrous sulphate Substances 0.000 claims description 2
- 235000003891 ferrous sulphate Nutrition 0.000 claims description 2
- 235000021312 gluten Nutrition 0.000 claims description 2
- 235000011187 glycerol Nutrition 0.000 claims description 2
- 230000002401 inhibitory effect Effects 0.000 claims description 2
- RBTARNINKXHZNM-UHFFFAOYSA-K iron trichloride Chemical compound Cl[Fe](Cl)Cl RBTARNINKXHZNM-UHFFFAOYSA-K 0.000 claims description 2
- BAUYGSIQEAFULO-UHFFFAOYSA-L iron(2+) sulfate (anhydrous) Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 claims description 2
- 229910000359 iron(II) sulfate Inorganic materials 0.000 claims description 2
- 239000008101 lactose Substances 0.000 claims description 2
- 229940035034 maltodextrin Drugs 0.000 claims description 2
- 239000000594 mannitol Substances 0.000 claims description 2
- 235000010355 mannitol Nutrition 0.000 claims description 2
- 235000013379 molasses Nutrition 0.000 claims description 2
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 2
- 235000019796 monopotassium phosphate Nutrition 0.000 claims description 2
- 235000015097 nutrients Nutrition 0.000 claims description 2
- 235000020232 peanut Nutrition 0.000 claims description 2
- 239000001103 potassium chloride Substances 0.000 claims description 2
- 235000011164 potassium chloride Nutrition 0.000 claims description 2
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 claims description 2
- 235000013406 prebiotics Nutrition 0.000 claims description 2
- 239000001509 sodium citrate Substances 0.000 claims description 2
- 239000000600 sorbitol Substances 0.000 claims description 2
- 235000010356 sorbitol Nutrition 0.000 claims description 2
- 229940083466 soybean lecithin Drugs 0.000 claims description 2
- 239000004455 soybean meal Substances 0.000 claims description 2
- 239000003549 soybean oil Substances 0.000 claims description 2
- 235000012424 soybean oil Nutrition 0.000 claims description 2
- 239000008107 starch Substances 0.000 claims description 2
- 235000019698 starch Nutrition 0.000 claims description 2
- 239000005720 sucrose Substances 0.000 claims description 2
- 239000003826 tablet Substances 0.000 claims description 2
- HRXKRNGNAMMEHJ-UHFFFAOYSA-K trisodium citrate Chemical compound [Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O HRXKRNGNAMMEHJ-UHFFFAOYSA-K 0.000 claims description 2
- 229940038773 trisodium citrate Drugs 0.000 claims description 2
- 235000019263 trisodium citrate Nutrition 0.000 claims description 2
- NWONKYPBYAMBJT-UHFFFAOYSA-L zinc sulfate Chemical compound [Zn+2].[O-]S([O-])(=O)=O NWONKYPBYAMBJT-UHFFFAOYSA-L 0.000 claims description 2
- 229910000368 zinc sulfate Inorganic materials 0.000 claims description 2
- 229960001763 zinc sulfate Drugs 0.000 claims description 2
- 239000008176 lyophilized powder Substances 0.000 claims 1
- 230000003449 preventive effect Effects 0.000 claims 1
- 230000035755 proliferation Effects 0.000 claims 1
- 230000001225 therapeutic effect Effects 0.000 claims 1
- 230000005764 inhibitory process Effects 0.000 abstract description 10
- 230000000694 effects Effects 0.000 abstract description 7
- 244000052616 bacterial pathogen Species 0.000 abstract description 5
- 230000003115 biocidal effect Effects 0.000 abstract 1
- 239000001963 growth medium Substances 0.000 description 10
- 239000002609 medium Substances 0.000 description 8
- 241000282414 Homo sapiens Species 0.000 description 6
- 239000003242 anti bacterial agent Substances 0.000 description 6
- 230000000844 anti-bacterial effect Effects 0.000 description 6
- 229940088710 antibiotic agent Drugs 0.000 description 6
- 238000000034 method Methods 0.000 description 6
- 239000000047 product Substances 0.000 description 6
- 238000012258 culturing Methods 0.000 description 5
- 239000007788 liquid Substances 0.000 description 5
- 238000009630 liquid culture Methods 0.000 description 5
- 210000003296 saliva Anatomy 0.000 description 5
- 108020004465 16S ribosomal RNA Proteins 0.000 description 4
- 238000004519 manufacturing process Methods 0.000 description 4
- 244000005700 microbiome Species 0.000 description 4
- 238000003752 polymerase chain reaction Methods 0.000 description 4
- 241000894007 species Species 0.000 description 4
- 230000001954 sterilising effect Effects 0.000 description 4
- 241000194017 Streptococcus Species 0.000 description 3
- 230000003385 bacteriostatic effect Effects 0.000 description 3
- 230000009286 beneficial effect Effects 0.000 description 3
- 239000011248 coating agent Substances 0.000 description 3
- 238000000576 coating method Methods 0.000 description 3
- 208000015181 infectious disease Diseases 0.000 description 3
- 239000007787 solid Substances 0.000 description 3
- 229920001817 Agar Polymers 0.000 description 2
- 238000007400 DNA extraction Methods 0.000 description 2
- 102000016928 DNA-directed DNA polymerase Human genes 0.000 description 2
- 108010014303 DNA-directed DNA polymerase Proteins 0.000 description 2
- 206010016952 Food poisoning Diseases 0.000 description 2
- 208000019331 Foodborne disease Diseases 0.000 description 2
- 239000008272 agar Substances 0.000 description 2
- 235000015278 beef Nutrition 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 210000004027 cell Anatomy 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 239000000284 extract Substances 0.000 description 2
- 235000021472 generally recognized as safe Nutrition 0.000 description 2
- 230000000968 intestinal effect Effects 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 238000012163 sequencing technique Methods 0.000 description 2
- 238000007711 solidification Methods 0.000 description 2
- 230000008023 solidification Effects 0.000 description 2
- 238000010257 thawing Methods 0.000 description 2
- 241000193403 Clostridium Species 0.000 description 1
- 208000037384 Clostridium Infections Diseases 0.000 description 1
- 206010009657 Clostridium difficile colitis Diseases 0.000 description 1
- 206010054236 Clostridium difficile infection Diseases 0.000 description 1
- 206010012735 Diarrhoea Diseases 0.000 description 1
- 208000004232 Enteritis Diseases 0.000 description 1
- 241000194033 Enterococcus Species 0.000 description 1
- 241000287828 Gallus gallus Species 0.000 description 1
- 201000000628 Gas Gangrene Diseases 0.000 description 1
- 208000018522 Gastrointestinal disease Diseases 0.000 description 1
- 108010033040 Histones Proteins 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 206010034133 Pathogen resistance Diseases 0.000 description 1
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 238000000137 annealing Methods 0.000 description 1
- 230000003110 anti-inflammatory effect Effects 0.000 description 1
- 230000002790 anti-mutagenic effect Effects 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- KLOIYEQEVSIOOO-UHFFFAOYSA-N carbocromen Chemical compound CC1=C(CCN(CC)CC)C(=O)OC2=CC(OCC(=O)OCC)=CC=C21 KLOIYEQEVSIOOO-UHFFFAOYSA-N 0.000 description 1
- 210000003850 cellular structure Anatomy 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 239000012531 culture fluid Substances 0.000 description 1
- 238000012136 culture method Methods 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000029182 enterotoxemia Diseases 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 230000007407 health benefit Effects 0.000 description 1
- 230000007366 host health Effects 0.000 description 1
- 230000028993 immune response Effects 0.000 description 1
- 210000000987 immune system Anatomy 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 238000010369 molecular cloning Methods 0.000 description 1
- 210000000214 mouth Anatomy 0.000 description 1
- 230000001338 necrotic effect Effects 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 230000004792 oxidative damage Effects 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 230000001766 physiological effect Effects 0.000 description 1
- 229920000136 polysorbate Polymers 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 230000005180 public health Effects 0.000 description 1
- 238000007790 scraping Methods 0.000 description 1
- 239000001632 sodium acetate Substances 0.000 description 1
- 235000017281 sodium acetate Nutrition 0.000 description 1
- 230000000638 stimulation Effects 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 230000002195 synergetic effect Effects 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 239000003053 toxin Substances 0.000 description 1
- 231100000765 toxin Toxicity 0.000 description 1
- 108700012359 toxins Proteins 0.000 description 1
- 239000012137 tryptone Substances 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
- A23C9/00—Milk preparations; Milk powder or milk powder preparations
- A23C9/12—Fermented milk preparations; Treatment using microorganisms or enzymes
- A23C9/123—Fermented milk preparations; Treatment using microorganisms or enzymes using only microorganisms of the genus lactobacteriaceae; Yoghurt
- A23C9/1236—Fermented milk preparations; Treatment using microorganisms or enzymes using only microorganisms of the genus lactobacteriaceae; Yoghurt using Leuconostoc, Pediococcus or Streptococcus sp. other than Streptococcus Thermophilus; Artificial sour buttermilk in general
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23G—COCOA; COCOA PRODUCTS, e.g. CHOCOLATE; SUBSTITUTES FOR COCOA OR COCOA PRODUCTS; CONFECTIONERY; CHEWING GUM; ICE-CREAM; PREPARATION THEREOF
- A23G9/00—Frozen sweets, e.g. ice confectionery, ice-cream; Mixtures therefor
- A23G9/32—Frozen sweets, e.g. ice confectionery, ice-cream; Mixtures therefor characterised by the composition containing organic or inorganic compounds
- A23G9/36—Frozen sweets, e.g. ice confectionery, ice-cream; Mixtures therefor characterised by the composition containing organic or inorganic compounds containing microorganisms or enzymes; containing paramedical or dietetical agents, e.g. vitamins
- A23G9/363—Frozen sweets, e.g. ice confectionery, ice-cream; Mixtures therefor characterised by the composition containing organic or inorganic compounds containing microorganisms or enzymes; containing paramedical or dietetical agents, e.g. vitamins containing microorganisms, enzymes
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/16—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
- A23K10/18—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions of live microorganisms
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
- A61K35/741—Probiotics
- A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/46—Streptococcus ; Enterococcus; Lactococcus
-
- 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
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biotechnology (AREA)
- General Health & Medical Sciences (AREA)
- Polymers & Plastics (AREA)
- Medicinal Chemistry (AREA)
- Wood Science & Technology (AREA)
- Food Science & Technology (AREA)
- Genetics & Genomics (AREA)
- Animal Behavior & Ethology (AREA)
- Veterinary Medicine (AREA)
- Pharmacology & Pharmacy (AREA)
- Mycology (AREA)
- Public Health (AREA)
- Biomedical Technology (AREA)
- Biochemistry (AREA)
- Tropical Medicine & Parasitology (AREA)
- Nutrition Science (AREA)
- Virology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- General Engineering & Computer Science (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Molecular Biology (AREA)
- Animal Husbandry (AREA)
- Communicable Diseases (AREA)
- Oncology (AREA)
- Physiology (AREA)
- Epidemiology (AREA)
- Inorganic Chemistry (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
本发明提供了一种唾液链球菌嗜热亚种(Streptococcus salivarius subsp.thermophilus)VB331及其应用，该菌种保藏于中国微生物菌种保藏管理委员会普通微生物中心，保藏编号为CGMCC NO.28407。该唾液链球菌嗜热亚种能有效抑制多种消化道病原菌的定植、繁殖与生长，包括大肠杆菌、金黄色葡萄球菌、沙门氏菌、粪肠球菌、单核细胞增生李斯特氏菌、志贺氏菌，尤其是对产气荚膜梭菌具有强大的抑制作用。该菌种为天然菌种，安全性强，能够为对抗多重有害细菌感染的抗菌、抑菌等功能产品开发提供新的益生菌源。
Description
技术领域
本发明属于益生菌及其应用技术领域，具体涉及一株唾液链球菌嗜热亚种VB331及其在抗菌抑菌产品中的应用，尤其是针对产气荚膜梭菌。
背景技术
细菌感染会造成重大的健康风险并导致沉重的经济负担。传统治疗有害细菌感染疾病的方法是使用抗生素，随着抗生素的过度使用，细菌的耐药性已成为一个日益严重的公共卫生问题，近年来越来越多的国家限制或禁止使用抗生素。且抗生素可能同时杀死有益和有害的细菌，破坏人体整体的微生物平衡，发生严重的副作用。为了解决这些限制，研究人员一直在探索控制有害微生物的替代方法。
益生菌在世界范围内被普遍接受的定义是“当施用足量时对宿主健康有益的活的微生物”。人体胃肠道是益生菌定植并发挥作用的主要场所，益生菌在肠道微环境中进行代谢活动，影响人体的食物药物成分代谢、细胞更新、免疫反应等诸多生理活动。研究表明，益生菌和益生菌制剂在多种胃肠道疾病的防治中发挥作用，可用于治疗腹泻、防治艰难梭菌感染及减少氧化损伤等。
唾液链球菌嗜热亚种(Streptococcus salivarius subsp. thermophilus)为一种革兰氏阳性菌，属于口腔正常菌群，同时是用于酸奶发酵的两种重要菌株之一。唾液链球菌嗜热亚种在其进化过程中失去了所有致病基因，并适应了牛奶这样明确、狭窄的生态位，被视为适应食物环境而进化了逆行演化。目前唾液链球菌嗜热亚种已获得美国GRAS(Generally Recognized as Safe)和欧洲QPS(Qualified Presumption of Safety)的安全认证，也是我国卫生部2010年65号公告批准使用的食品生产加工的菌种之一。以往研究中唾液链球菌嗜热亚种的健康益处包括产生抗氧化化合物、降低某些类型癌症的风险、抗炎作用、抗诱变作用和刺激肠道免疫系统等。
发明内容
本发明提供了一种从成人唾液中筛选得到的唾液链球菌嗜热亚种(Streptococcus salivarius subsp. thermophilus) VB331，于2023年9月8日保藏于中国微生物菌种保藏管理委员会普通微生物中心，保藏编号为CGMCC NO.28407。以成人唾液为分离源，获得的菌种具有更高的安全性。该菌株已被证实具有显著的抗菌、抑菌活性，能有效抑制多种消化道病原菌的定植、繁殖与生长，包括大肠杆菌、金黄色葡萄球菌、沙门氏菌、粪肠球菌、单核细胞增生李斯特氏菌、志贺氏菌，尤其是对产气荚膜梭菌具有强大的抑制作用。
本发明提供的唾液链球菌嗜热亚种，其为活细胞形式，例如菌株为细菌培养液的形式。
本发明提供的唾液链球菌嗜热亚种，其为非活细胞形式，例如菌株为冻干粉末形式。
本发明提供的唾液链球菌嗜热亚种可用于制备用于制备预防与治疗消化道有害细菌感染的微生物制剂、药品、保健品、食品、饮料、添加剂、饲料中的一个或多个，且优选通过抑制消化道有害细菌的定植、繁殖、生长发挥预防与治疗作用。
在针对消化道有害细菌引起的感染时，有害菌为大肠杆菌、金黄色葡萄球菌、沙门氏菌、粪肠球菌、单核细胞增生李斯特氏菌、志贺氏菌和产气荚膜梭菌中的一种或多种，优选产气荚膜梭菌。
本发明提供一种包含上述唾液链球菌嗜热亚种的组合物，该制剂可以包含另外的益生菌材料，还可以包含益生元材料。
本发明提供的组合物，其还包含可摄入的载体，可摄入的载体可以是药学上可接受的载体，如胶囊、片剂或粉末；也可以是食品，例如酸化乳、酸奶、冷冻酸奶、乳粉、浓缩乳、奶酪酱、调味品或饮料。
本发明提供的唾液链球菌嗜热亚种可以在含有可同化的碳源和/或氮源的营养培养基里，进行厌氧发酵。
优选技术方案中可同化的碳源选自淀粉、麦芽糊精、葡萄糖、蔗糖、乳糖、麦芽糖、工业糖蜜、甘油、豆油、山梨醇、甘露醇之一或者任意几种的组合；优选为葡萄糖。
优选技术方案中可同化的氮源选自酵母提取物、酵母粉、酵母膏、大豆卵磷脂、黄豆饼粉、棉籽饼粉、花生饼粉、麸质粉、玉米浆干粉、豆粕、蛋白胨、尿素、铵盐之一或者任意几种的组合；优选为酵母提取物。
优选技术方案中发酵培养基还包括无机盐，所述无机盐选自柠檬酸三钠、磷酸二氢钾、磷酸氢二钾、硫酸铵、碳酸钙、硫酸亚铁、硫酸锌、硫酸铜、氯化钠、氯化钾、氯化钙、硫酸镁、氯化铁、硫酸锰之一或任意几种的组合；优选为磷酸氢二钾、硫酸镁。
有益效果：
产气荚膜梭菌(Clostridium perfringens)是一种兼性厌氧的革兰氏阳性菌，根据分泌毒的
不同分为A~G等7种毒素型，广泛分布于外界环境和大多数动物肠道中，同时也是一种条件性致病菌，对人和动物都具有极强的致病性，A型产气荚膜梭菌引起人类气性坏疽和食物中毒，C型产气荚膜梭菌导致人食物中毒事件也有报告，G型产气荚膜梭菌引起肉鸡坏死性肠炎(又称肠毒血症)，由此病原菌引起的疾病对动物生产及人类健康有着极大危害。
本发明提供的唾液链球菌嗜热亚种VB331，该菌株已被证实具有显著的抗菌、抑菌活性，能有效抑制多种消化道病原菌的定植、繁殖与生长。该菌种的广谱高效的抑菌特性，尤其是对产气荚膜梭菌强大的抑制作用，可作为益生菌用于制备预防与治疗胃肠道有害细菌感染的微生物制剂、药品、保健品、食品、饮料、添加剂、饲料。
唾液链球菌嗜热亚种VB331作为天然存在益生菌，同时具有广谱高效的抑菌特性，在消化道细菌感染的预防和治疗中能够发挥协同效应，尤其是产气荚膜梭菌的感染，能够安全快速恢复胃肠道健康，减少抗生素的使用，同时避免抗生素使用的副作用。
本发明的附加方面和优点将在下面的描述中部分给出，部分将从下面的描述中变得明显，或通过本发明的实践了解到。
保藏信息：
菌株名称：VB331
保藏日期：2023年09月08日
保藏单位：中国普通微生物菌种保藏管理中心（CGMCC）
保藏编号：CGMCC No.28407
附图说明
本发明的上述和/或附加的方面和优点从结合下面附图对实施例的描述中将变得明显和容易理解，其中：
图1 VB331对不同致病菌的抑菌圈大小比较；
图2 基于16S rDNA序列构建的VB331种群邻接法系统进化树，其中，ALIF_s K12、GL698454_s C150、JN682090_s Brum 177均为Streptococcus，翻译为链球菌。
具体实施方式
下述实施例中所用的实验方法如无特殊说明，均为常规方法。
下述实施例中采用的材料、试剂等如无特殊说明，皆为普通市售品，皆可于市场购得。
其中所用MAGEN细菌基因组DNA提取试剂盒购自上海迈跟生物科技有限公司。Phusion高保真DNA聚合酶购自Thermo Scientific。
下面将通过实施例对本发明作进一步的描述，这些描述并不是对本发明内容作进一步的限定。本领域的技术人员应理解，对本发明内容所作的等同替换，或相应的改进，仍属于本发明的保护范围之内。
以下实施例所使用的培养基中，物料配比所用的百分数（%）计量单位，除特殊说明外均为质量百分比。
实施例1 菌种来源
唾液链球菌嗜热亚种(Streptococcus salivarius subsp. thermophilus)VB331是从成人唾液中分离获得的。取成人新鲜唾液在无菌容器中加入灭菌PBS缓冲液，充分震荡后稀释涂布在MRS固体培养基平板上，培养48h后挑取大小形态不一的单菌落在新鲜的平板上划线，然后挑取单菌落到斜面，斜面培养好后镜检菌落形态，镜检无杂菌后进行鉴定。以成人唾液为分离源，获得的菌种具有更高的安全性，更适合用于保健品的生产。
实施例2 菌种鉴定
参照《分子克隆实验指南》书中的有关内容进行实验。纯化后的菌种在MRS培养基中培养1天后取1 mL菌液10000 rpm离心1 min，弃上清，收集菌体，按照MAGEN细菌基因组DNA提取试剂盒操作步骤提取DNA。以提取的DNA为模板，采用通用引物27F(5′AGAGTTTGATCCTGGCTCAG 3′)和1492R (5′ TACGGYTACCTTGTTAYGACTT 3′)通过PCR(聚合酶链式反应) 进行16S rDNA扩增。PCR的反应体系为50 μL，按照Phusion高保真DNA聚合酶说明书配置反应体系和设置程序，退火温度55℃。PCR产物送去上海生工测序。
菌株所测的16S rDNA的序列经校队后，与ECBioCloud数据库中相关种、属的序列进行同源序列BLAST比较，以确定该菌株的分类地位。经过比对发现，其与Streptococcus thermophilusATCC 19258(T)同源性高达99.1%，与Streptococcus salivariusNCTC 8618(T)同源性高达98.83%，故将该菌株鉴定为唾液链球菌嗜热亚种(Streptococcus salivarius subsp. thermophilus) 菌株。该菌种内部编号为VB331，已于2022年9月8日保藏于中国微生物菌种保藏管理委员会普通微生物中心，保藏地址为北京市朝阳区北辰西路1号院3号，保藏编号为CGMCC NO.28407。
采用基于k-mer频率指纹的方法。从16S rDNA序列中提取长度为16的k-mer并统计其出现次数，进而生成k-mer频率指纹。利用余弦距离，计算了序列间的相似性并构建了系统发育树，见附图2。VB331与Streptococcus-thermophilus-ATCC19258、Streptococcus- salivarius-NCTC8618、Streptococcus-vestibularis-ATCC49124和 ALIF_s-K12之间存在着非常近的亲缘关系。它与Streptococcus-porcorum-682-03和其他几个菌种进一步结合，表明VB331与这些菌之间有相对较近的亲缘关系。
实施例3 有组蛋白包裹的游离DNA文库的构建
1.菌种活化
取VB331原始菌种甘油管，在室温下解冻，吸取0.1ml菌悬液接种到MRS固体平板，涂布均匀，平皿放入厌氧袋，置于37℃培养箱内培养48h，得到活化好的单菌落。2.种子制备
取活化好的单菌落，用接种环刮取一环菌落，然后接种到盛有100mlMRS液体培养基的500ml三角瓶中，包扎好放入厌氧袋，37℃、70rpm，摇床上振荡培养48h，控制种子液OD值≥2.0。MRS液体培养基消毒前调pH至7.0±0.2，消毒条件为121~123℃、30min。3.发酵制备
培养好的种子液，以3-5%(V/W)的比例接种到盛有100 mlMRS液体培养基的500ml三角瓶中，包扎好放入厌氧袋，37℃、70rpm，摇床上振荡培养24-48h，控制发酵液OD值≥3.0。MRS液体培养基消毒前调pH至7.0±0.2，消毒条件为121~123℃、30min。
MRS液体培养基：酵母提取物0. 5 %、蛋白胨1. 0%、牛肉膏1. 0%、葡萄糖2. 0%、磷酸氢二钾0. 2%、柠檬酸氢二铵0. 2%、乙酸钠0.5%、硫酸镁0. 02%、硫酸锰0. 005 %（以上为质量百分比）、吐温80 0. 1%（体积/质量比）。
MRS固体培养基：在MRS液体培养基的基础上，添加1. 5%琼脂粉。
实施例4：抑菌活性
采用优化后的双层板培养法对唾液链球菌嗜热亚种VB331菌株进行多种微生物抑制的研究，包括以下步骤：
下层唾液链球菌嗜热亚种VB331的制备：将活化好的单菌冷冻管解冻后，接种于pH为6.9的MRS平板中，37℃厌氧培养3天。菌种接种量：2ul/点。
上层致病菌层的制备：灭菌的产气荚膜培养基冷却至40-50℃，接入产气荚膜梭菌，使培养基中最终浓度约106/ml，混匀。将该培养基按7ml/皿加入已长好唾液链球菌嗜热亚种VB331的平板中，待凝固后，37℃厌氧培养1天。
灭菌的LB培养基冷却至40-50℃，接入大肠杆菌、金黄色葡萄球菌、沙门氏菌、粪肠球菌、单核细胞增生李斯特氏菌、志贺氏菌使培养基中，最终浓度约106/ml，混匀。将该培养基按7ml/皿加入已长好唾液链球菌嗜热亚种VB331的平板中，待凝固后，37℃有氧培养1天。
待上层板长好后观察唾液链球菌嗜热亚种VB331周围是否有抑菌圈，判断是否有抑菌能力。
产气荚膜梭菌培养基：胰蛋白胨 1.5%，牛肉浸膏0.3%，氯化钠0.5%，葡萄糖1%，琼脂0.5%，PH7.0。实验结果见表1：
表1唾液链球菌嗜热亚种VB331抑菌圈试验结果
其中：大肠杆菌(E. coli)的抑菌圈大小为26mm；金黄色葡萄球菌(Staphylococcus aureus) 的抑菌圈大小为23mm；沙门氏菌(Salmonella)的抑菌圈大小为22mm；粪肠球菌(Enterococcus) 的抑菌圈大小为17mm；单核细胞增生李斯特氏菌(Listeria monocytogenes)的抑菌圈大小为16mm；志贺氏菌(Shigella) 的抑菌圈大小为14mm；产气荚膜梭菌(Clostridium perfringens)的抑菌圈大小为40mm，是所有菌中最大的。
可以明显看出，VB331针对消化道有害细菌大肠杆菌、金黄色葡萄球菌、沙门氏菌、粪肠球菌、单核细胞增生李斯特氏菌、志贺氏菌和产气荚膜梭菌产气荚膜梭菌均有显著的抑制作用，尤其是对产气荚膜梭菌的抑制作用尤其突出。
VB331的16S测序序列:
AGGTGGGGGCGTGCCCTATACTGCAGTAGAACGCTGAAGAGAGGAGCTTGCTCTTCTTGGATGAGTTGCGAACGGGTGAGTAACGCGTAGGTAACCTGCCTTGTAGCGGGGGATAACTATTGGAAACGATAGCTAATACCGCATAACAATGGATGACACATGTCATTTATTTGAAAGGGGCAATTGTTCCACTACAAGATGGACCTGCGTTGTATTAGCTAGTAGGTGAGGTAATGGCTCACCTAGGCGACGATACATAGCCGACCTGAGAGGGTGATCGGCCACACTGGGACTGAGACACGGCCCAGACTCCTACGGGAGGCAGCAGTAGGGAATCTTCGGCAATGGGGGCAACCCTGACCGAGCAACGCCGCGTGAGTGAAGAAGGTTTTCGGATCGTAAAGCTCTGTTGTAAGTCAAGAACGGGTGTGAGAGTGGAAAGTTCACACTGTGACGGTAGCTTACCAGAAAGGGACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTCCCGAGCGTTGTCCGGATTTATTGGGCGTAAAGCGAGCGCAGGCGGTTTGATAAGTCTGAAGTTAAAGGCTGTGGCTCAACCATAGTTCGCTTTGGAAACTGTCAAACTTGAGTGCAGAAGGGGAGAGTGGAATTCCATGTGTAGCGGTGAAATGCGTAGATATATGGAGGAACACCGGTGGCGAAAGCGGCTCTCTGGTCTGTAACTGACGCTGAGGCTCGAAAGCGTGGGGAGCGAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGAGTGCTAGGTGTTGGATCCTTTCCGGGATTCAGTGCCGCAGCTAACGCATTAAGCACTCCGCCTGGGGAGTACGACCGCAAGGTTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAACGCGAAGAACCTTACCAGGTCTTGACATCCCGATGCTATTTCTAGAGATAGAAAGTTACTTCGGTACATCGGTGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCCTATTGTTAGTTGCCATCATTCAGTTGGGCACTCTAGCGAGACTGCCGGTAATAAACCGGAGGAAGGTGGGGATGACGTCAAATCATCATGCCCCTTATGACCTGGGCTACACACGTGCTACAATGGTTGGTACAACGAGTTGCGAGTCGGTGACGGCGAGCTAATCTCTTAAAGCCAATCTCAGTTCGGATTGTAGGCTGCAACTCGCCTACATGAAGTCGGAATCGCTAGTAATCGCGGATCAGCACGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCACGAGAGTTTGTAACACCCGAAGTCGGTGAGGTAACCTTTGGAGCCAGCCGCCTAAGGTGAACCC（SEQ ID NO:1）。
Claims (19)
1.一种唾液链球菌嗜热亚种(Streptococcus salivarius subsp. thermophilus)VB331，保藏于中国微生物菌种保藏管理委员会普通微生物中心，保藏编号为CGMCCNO.28407，保藏日期为2023年9月8日。
2.如权利要求1所述的唾液链球菌嗜热亚种，其为活细胞形式。
3.如权利要求2所述的唾液链球菌嗜热亚种，其中所述唾液链球菌嗜热亚种为细菌培养液的形式。
4.如权利要求1所述的唾液链球菌嗜热亚种，其为非活细胞形式。
5.如权利要求4所述的唾液链球菌嗜热亚种，其中所述唾液链球菌嗜热亚种为冻干粉末形式。
6.一种发酵液，其特征在于：通过权利要求1所述的唾液链球菌嗜热亚种发酵而来。
7.一种权利要求1-5中任一项所述的唾液链球菌嗜热亚种或权利要求6所述的发酵液的用途，其特征在于：用于制备预防与治疗消化道有害细菌感染的微生物制剂、药品、保健品、食品、饮料、添加剂、饲料中的一个或多个。
8.如权利要求7所述的用途，其特征在于所述唾液链球菌嗜热亚种或所述发酵液通过抑制消化道有害细菌的定植、繁殖、生长发挥预防与治疗作用。
9.如权利要求8所述的用途，其特征在于所述消化道有害细菌为大肠杆菌、金黄色葡萄球菌、沙门氏菌、粪肠球菌、单核细胞增生李斯特氏菌、志贺氏菌和产气荚膜梭菌中的至少之一。
10.一种组合物，其包含权利要求1-5中任一项所述的唾液链球菌嗜热亚种。
11.如权利要求10所述的组合物，其还包含另外的益生菌材料。
12.如权利要求11所述的组合物，其还包含益生元材料。
13.如权利要求12所述的组合物，其还包含可摄入的载体。
14.如权利要求13所述的组合物，其中所述可摄入的载体是药学上可接受的载体，如胶囊、片剂或粉末。
15.如权利要求13所述的组合物，其中所述可摄入的载体是食品，如酸化乳、酸奶、冷冻酸奶、乳粉、浓缩乳、奶酪酱、调味品或饮料。
16.一种权利要求1-5中任一项所述的唾液链球菌嗜热亚种的发酵方法，其特征在于，所述的发酵在含有可同化的碳源和/或氮源的营养培养基里，进行无氧发酵。
17.如权利要求16所述的发酵方法，其特征在于：
所述可同化的碳源选自淀粉、麦芽糊精、葡萄糖、蔗糖、乳糖、麦芽糖、工业糖蜜、甘油、豆油、山梨醇、甘露醇之一或者任意几种的组合。
18.如权利要求16所述的发酵方法，其特征在于：
所述可同化的氮源选自酵母提取物、酵母粉、酵母膏、大豆卵磷脂、黄豆饼粉、棉籽饼粉、花生饼粉、麸质粉、玉米浆干粉、豆粕、蛋白胨、尿素、铵盐之一或者任意几种的组合。
19.如权利要求16所述的发酵方法，其特征在于：
所述发酵培养基还包括无机盐，所述无机盐选自柠檬酸三钠、磷酸二氢钾、磷酸氢二钾、硫酸铵、碳酸钙、硫酸亚铁、硫酸锌、硫酸铜、氯化钠、氯化钾、氯化钙、硫酸镁、氯化铁、硫酸锰之一或任意几种的组合。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311419206.3A CN117143782B (zh) | 2023-10-30 | 2023-10-30 | 一种唾液链球菌嗜热亚种vb331及其应用 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311419206.3A CN117143782B (zh) | 2023-10-30 | 2023-10-30 | 一种唾液链球菌嗜热亚种vb331及其应用 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN117143782A true CN117143782A (zh) | 2023-12-01 |
| CN117143782B CN117143782B (zh) | 2024-03-22 |
Family
ID=88884808
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202311419206.3A Active CN117143782B (zh) | 2023-10-30 | 2023-10-30 | 一种唾液链球菌嗜热亚种vb331及其应用 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN117143782B (zh) |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH04271777A (ja) * | 1991-02-28 | 1992-09-28 | Snow Brand Milk Prod Co Ltd | 低生酸性乳酸菌、スターター、これを用いた発酵乳及びその製造法 |
| KR20040084167A (ko) * | 2003-03-26 | 2004-10-06 | 주식회사 프로바이오닉 | 유해 병원성 미생물의 생육 억제능을 가지며, 식품보존력이 뛰어난 식품보존용 조성물 |
| CN1710088A (zh) * | 2005-06-24 | 2005-12-21 | 南京农业大学 | 利用唾液链球菌嗜热亚种生产γ－氨基丁酸的方法 |
| CN104666366A (zh) * | 2013-12-03 | 2015-06-03 | 青岛碧水蓝天生物技术有限公司 | 抑制感染组合物及含有该组合物的饮食品 |
| US20200368218A1 (en) * | 2018-02-13 | 2020-11-26 | European Molecular Biology Laboratory | Repurposing compounds for the treatment of infections and for modulating the composition of the gut microbiome |
| CN116656578A (zh) * | 2023-07-26 | 2023-08-29 | 杭州微致生物科技有限公司 | 一种发酵粘液乳杆菌vb216及其应用 |
-
2023
- 2023-10-30 CN CN202311419206.3A patent/CN117143782B/zh active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH04271777A (ja) * | 1991-02-28 | 1992-09-28 | Snow Brand Milk Prod Co Ltd | 低生酸性乳酸菌、スターター、これを用いた発酵乳及びその製造法 |
| KR20040084167A (ko) * | 2003-03-26 | 2004-10-06 | 주식회사 프로바이오닉 | 유해 병원성 미생물의 생육 억제능을 가지며, 식품보존력이 뛰어난 식품보존용 조성물 |
| CN1710088A (zh) * | 2005-06-24 | 2005-12-21 | 南京农业大学 | 利用唾液链球菌嗜热亚种生产γ－氨基丁酸的方法 |
| CN104666366A (zh) * | 2013-12-03 | 2015-06-03 | 青岛碧水蓝天生物技术有限公司 | 抑制感染组合物及含有该组合物的饮食品 |
| US20200368218A1 (en) * | 2018-02-13 | 2020-11-26 | European Molecular Biology Laboratory | Repurposing compounds for the treatment of infections and for modulating the composition of the gut microbiome |
| CN116656578A (zh) * | 2023-07-26 | 2023-08-29 | 杭州微致生物科技有限公司 | 一种发酵粘液乳杆菌vb216及其应用 |
Non-Patent Citations (2)
| Title |
|---|
| NATAŠA GOLIĆ ET AL.: "In vitro and in vivo antagonistic activity of new probiotic culture against Clostridium difficile and Clostridium perfringens", BMC MICROBIOLOGY, vol. 17, pages 1 - 9 * |
| 刘洁等: "两种乳酸菌及其发酵液对单核细胞增生李斯特菌生长的抑制作用", 上海交通大学学报(农业科学版), vol. 24, no. 03, pages 221 - 225 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN117143782B (zh) | 2024-03-22 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Bin Masalam et al. | Isolation, molecular characterization and probiotic potential of lactic acid bacteria in Saudi raw and fermented milk | |
| DK1859022T3 (en) | New acid tolerant Lactobacillus sakei Probio-65 with the ability to inhibit growth of pathogenic microorganisms and antiallergic effect | |
| KR100911115B1 (ko) | 내산성, 내담즙산성 및 항균 효과를 가진 신규한 유산균 및 이를 포함하는 조성물 | |
| CN113170842B (zh) | 一种防治家禽坏死性肠炎的复合微生态制剂及其应用 | |
| CN109749957B (zh) | 一种具有水产病原菌拮抗特性的格氏乳杆菌制剂的制备及应用 | |
| CN108148789B (zh) | 一种鼠李糖乳杆菌及其在制备细菌素中的应用 | |
| CN108753650B (zh) | 屎肠球菌以及由其制备的复合微生态制剂 | |
| CN113832077A (zh) | 鼠李糖乳杆菌及其应用 | |
| WO2017071346A1 (zh) | 脆弱拟杆菌在抗水产病原菌中的应用 | |
| CN114480231A (zh) | 一种抗幽门螺杆菌感染的罗伊氏乳杆菌及其应用 | |
| KR20080109585A (ko) | 내산성, 내담즙산성, 내염성 및 항균 효과를 가진 신규한식물성 유산균 및 이를 포함하는 조성물 | |
| CN114891675B (zh) | 胃源性植物乳杆菌lpf-01及其应用 | |
| CN112940985A (zh) | 一种增强人体免疫力的鼠李糖乳杆菌制剂及其制备方法 | |
| CN114317334B (zh) | 一株能够与幽门螺杆菌共聚集的清酒乳杆菌及其应用 | |
| CN108432996B (zh) | 一种猴头菇乳酸菌发酵饮料及其制备方法 | |
| CN112940984B (zh) | 抗幽门螺杆菌、降血糖、调理肠胃和增加免疫力的复合乳酸杆菌制剂及其制备方法 | |
| CN112940983A (zh) | 一种能增加抗幽门螺杆菌效果的植物乳杆菌制剂及其制备方法 | |
| KR100585391B1 (ko) | 돼지 전염성위장염 (TGE) 코로나바이러스 및 유해 미생물 억제 활성을 갖는 신규 락토바실러스 프란타럼 Probio-38 및 이를 함유하는 생균활성제 | |
| CN116064324B (zh) | 一株鼠李糖乳杆菌及其培养方法和在防治腹泻和肠炎方面的应用 | |
| CN112251380A (zh) | 一株促进机体健康长寿的植物乳杆菌flpl05 | |
| CN117143782B (zh) | 一种唾液链球菌嗜热亚种vb331及其应用 | |
| KR20050025002A (ko) | 유해미생물 억제 활성을 가지는 신규 내산성 락토바실러스루테리 Probio-054 및 이를 함유하는 생균활성제 | |
| CN102399728B (zh) | 一种乳酸杆菌及其发酵方法 | |
| TW201016847A (en) | A strain of Lactobacillus plantarum and its use for inhibiting Helicobacter pylori growth | |
| EP2040723A1 (en) | Method and use of lactic acid bacteria with increased acid tolerance |
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 |