KR100865363B1 - Bifidobacterium infantis MAEIL-K9 strain and health-promoting food containing the same - Google Patents
Bifidobacterium infantis MAEIL-K9 strain and health-promoting food containing the same Download PDFInfo
- Publication number
- KR100865363B1 KR100865363B1 KR1020020056172A KR20020056172A KR100865363B1 KR 100865363 B1 KR100865363 B1 KR 100865363B1 KR 1020020056172 A KR1020020056172 A KR 1020020056172A KR 20020056172 A KR20020056172 A KR 20020056172A KR 100865363 B1 KR100865363 B1 KR 100865363B1
- Authority
- KR
- South Korea
- Prior art keywords
- maeil
- bifidobacterium
- lactic acid
- bifidus
- acid bacteria
- Prior art date
Links
- 241000186015 Bifidobacterium longum subsp. infantis Species 0.000 title claims abstract description 25
- 229940004120 bifidobacterium infantis Drugs 0.000 title claims abstract description 25
- 235000013305 food Nutrition 0.000 title description 6
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 claims abstract description 132
- 241000894006 Bacteria Species 0.000 claims abstract description 75
- 239000004310 lactic acid Substances 0.000 claims abstract description 66
- 235000014655 lactic acid Nutrition 0.000 claims abstract description 66
- 235000013336 milk Nutrition 0.000 claims abstract description 19
- 239000008267 milk Substances 0.000 claims abstract description 19
- 210000004080 milk Anatomy 0.000 claims abstract description 19
- 235000013376 functional food Nutrition 0.000 claims description 12
- 230000036541 health Effects 0.000 claims description 11
- 238000000034 method Methods 0.000 claims description 7
- 235000014048 cultured milk product Nutrition 0.000 claims description 2
- 235000020124 milk-based beverage Nutrition 0.000 claims description 2
- 239000000843 powder Substances 0.000 claims description 2
- 210000004251 human milk Anatomy 0.000 claims 1
- 235000020256 human milk Nutrition 0.000 claims 1
- 230000000968 intestinal effect Effects 0.000 abstract description 30
- 241000186660 Lactobacillus Species 0.000 abstract description 25
- 229940039696 lactobacillus Drugs 0.000 abstract description 25
- 239000002253 acid Substances 0.000 abstract description 15
- 230000000529 probiotic effect Effects 0.000 abstract description 14
- 239000006041 probiotic Substances 0.000 abstract description 13
- 235000018291 probiotics Nutrition 0.000 abstract description 13
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 abstract description 12
- 239000001301 oxygen Substances 0.000 abstract description 12
- 229910052760 oxygen Inorganic materials 0.000 abstract description 12
- 210000000941 bile Anatomy 0.000 abstract description 11
- 230000001766 physiological effect Effects 0.000 abstract description 10
- 210000004027 cell Anatomy 0.000 description 88
- 241000186000 Bifidobacterium Species 0.000 description 53
- 210000000936 intestine Anatomy 0.000 description 17
- 230000035899 viability Effects 0.000 description 11
- 230000002776 aggregation Effects 0.000 description 9
- 238000004220 aggregation Methods 0.000 description 9
- 210000002490 intestinal epithelial cell Anatomy 0.000 description 8
- 230000009471 action Effects 0.000 description 6
- 230000006870 function Effects 0.000 description 6
- 229920001817 Agar Polymers 0.000 description 5
- 238000002835 absorbance Methods 0.000 description 5
- 239000008272 agar Substances 0.000 description 5
- 210000000110 microvilli Anatomy 0.000 description 5
- 108090000623 proteins and genes Proteins 0.000 description 5
- 238000010998 test method Methods 0.000 description 5
- 238000012360 testing method Methods 0.000 description 5
- 241000186016 Bifidobacterium bifidum Species 0.000 description 4
- 244000052616 bacterial pathogen Species 0.000 description 4
- 229940002008 bifidobacterium bifidum Drugs 0.000 description 4
- 150000001720 carbohydrates Chemical class 0.000 description 4
- 235000014633 carbohydrates Nutrition 0.000 description 4
- 238000002474 experimental method Methods 0.000 description 4
- HEBKCHPVOIAQTA-UHFFFAOYSA-N meso ribitol Natural products OCC(O)C(O)C(O)CO HEBKCHPVOIAQTA-UHFFFAOYSA-N 0.000 description 4
- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Chemical compound [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 description 4
- 230000008569 process Effects 0.000 description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- 241000186012 Bifidobacterium breve Species 0.000 description 3
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 3
- 238000003794 Gram staining Methods 0.000 description 3
- 230000032683 aging Effects 0.000 description 3
- 230000004888 barrier function Effects 0.000 description 3
- SRBFZHDQGSBBOR-UHFFFAOYSA-N beta-D-Pyranose-Lyxose Natural products OC1COC(O)C(O)C1O SRBFZHDQGSBBOR-UHFFFAOYSA-N 0.000 description 3
- 239000008280 blood Substances 0.000 description 3
- 210000004369 blood Anatomy 0.000 description 3
- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical compound P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 238000011534 incubation Methods 0.000 description 3
- 230000002401 inhibitory effect Effects 0.000 description 3
- 230000003993 interaction Effects 0.000 description 3
- 239000010410 layer Substances 0.000 description 3
- 230000007246 mechanism Effects 0.000 description 3
- 238000000386 microscopy Methods 0.000 description 3
- 230000000877 morphologic effect Effects 0.000 description 3
- 239000002953 phosphate buffered saline Substances 0.000 description 3
- 235000018102 proteins Nutrition 0.000 description 3
- 102000004169 proteins and genes Human genes 0.000 description 3
- 239000000725 suspension Substances 0.000 description 3
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 2
- 241001134770 Bifidobacterium animalis Species 0.000 description 2
- SHZGCJCMOBCMKK-UHFFFAOYSA-N D-mannomethylose Natural products CC1OC(O)C(O)C(O)C1O SHZGCJCMOBCMKK-UHFFFAOYSA-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
- IFQSXNOEEPCSLW-DKWTVANSSA-N L-cysteine hydrochloride Chemical compound Cl.SC[C@H](N)C(O)=O IFQSXNOEEPCSLW-DKWTVANSSA-N 0.000 description 2
- PNNNRSAQSRJVSB-UHFFFAOYSA-N L-rhamnose Natural products CC(O)C(O)C(O)C(O)C=O PNNNRSAQSRJVSB-UHFFFAOYSA-N 0.000 description 2
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 230000002378 acidificating effect Effects 0.000 description 2
- 229940118852 bifidobacterium animalis Drugs 0.000 description 2
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 description 2
- 235000015872 dietary supplement Nutrition 0.000 description 2
- 210000002249 digestive system Anatomy 0.000 description 2
- 239000003623 enhancer Substances 0.000 description 2
- 210000002919 epithelial cell Anatomy 0.000 description 2
- 210000003608 fece Anatomy 0.000 description 2
- 230000037406 food intake Effects 0.000 description 2
- 150000004676 glycans Chemical class 0.000 description 2
- DCAYPVUWAIABOU-UHFFFAOYSA-N hexadecane Chemical compound CCCCCCCCCCCCCCCC DCAYPVUWAIABOU-UHFFFAOYSA-N 0.000 description 2
- 208000015181 infectious disease Diseases 0.000 description 2
- 238000011081 inoculation Methods 0.000 description 2
- 230000009545 invasion Effects 0.000 description 2
- 230000000813 microbial effect Effects 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- 239000008363 phosphate buffer Substances 0.000 description 2
- 229910052697 platinum Inorganic materials 0.000 description 2
- 229920001282 polysaccharide Polymers 0.000 description 2
- 239000005017 polysaccharide Substances 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 230000002062 proliferating effect Effects 0.000 description 2
- 230000035755 proliferation Effects 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 239000002356 single layer Substances 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- 241000894007 species Species 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 239000003053 toxin Substances 0.000 description 2
- 231100000765 toxin Toxicity 0.000 description 2
- 108700012359 toxins Proteins 0.000 description 2
- 229940088594 vitamin Drugs 0.000 description 2
- 235000013343 vitamin Nutrition 0.000 description 2
- 229930003231 vitamin Natural products 0.000 description 2
- 239000011782 vitamin Substances 0.000 description 2
- HSINOMROUCMIEA-FGVHQWLLSA-N (2s,4r)-4-[(3r,5s,6r,7r,8s,9s,10s,13r,14s,17r)-6-ethyl-3,7-dihydroxy-10,13-dimethyl-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1h-cyclopenta[a]phenanthren-17-yl]-2-methylpentanoic acid Chemical compound C([C@@]12C)C[C@@H](O)C[C@H]1[C@@H](CC)[C@@H](O)[C@@H]1[C@@H]2CC[C@]2(C)[C@@H]([C@H](C)C[C@H](C)C(O)=O)CC[C@H]21 HSINOMROUCMIEA-FGVHQWLLSA-N 0.000 description 1
- 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 description 1
- IZSRJDGCGRAUAR-MROZADKFSA-M 5-dehydro-D-gluconate Chemical compound OCC(=O)[C@@H](O)[C@H](O)[C@@H](O)C([O-])=O IZSRJDGCGRAUAR-MROZADKFSA-M 0.000 description 1
- PLXMOAALOJOTIY-FPTXNFDTSA-N Aesculin Natural products OC[C@@H]1[C@@H](O)[C@H](O)[C@@H](O)[C@H](O)[C@H]1Oc2cc3C=CC(=O)Oc3cc2O PLXMOAALOJOTIY-FPTXNFDTSA-N 0.000 description 1
- ZHGNHOOVYPHPNJ-UHFFFAOYSA-N Amigdalin Natural products FC(F)(F)C(=O)OCC1OC(OCC2OC(OC(C#N)C3=CC=CC=C3)C(OC(=O)C(F)(F)F)C(OC(=O)C(F)(F)F)C2OC(=O)C(F)(F)F)C(OC(=O)C(F)(F)F)C(OC(=O)C(F)(F)F)C1OC(=O)C(F)(F)F ZHGNHOOVYPHPNJ-UHFFFAOYSA-N 0.000 description 1
- BJRNKVDFDLYUGJ-RMPHRYRLSA-N Arbutin Natural products O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1=CC=C(O)C=C1 BJRNKVDFDLYUGJ-RMPHRYRLSA-N 0.000 description 1
- 241000901050 Bifidobacterium animalis subsp. lactis Species 0.000 description 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- WNBCMONIPIJTSB-BGNCJLHMSA-N Cichoriin Natural products O([C@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1)c1c(O)cc2c(OC(=O)C=C2)c1 WNBCMONIPIJTSB-BGNCJLHMSA-N 0.000 description 1
- 206010010774 Constipation Diseases 0.000 description 1
- HEBKCHPVOIAQTA-NGQZWQHPSA-N D-Arabitol Natural products OC[C@H](O)C(O)[C@H](O)CO HEBKCHPVOIAQTA-NGQZWQHPSA-N 0.000 description 1
- RFSUNEUAIZKAJO-VRPWFDPXSA-N D-Fructose Natural products OC[C@H]1OC(O)(CO)[C@@H](O)[C@@H]1O RFSUNEUAIZKAJO-VRPWFDPXSA-N 0.000 description 1
- RGHNJXZEOKUKBD-SQOUGZDYSA-M D-gluconate Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C([O-])=O RGHNJXZEOKUKBD-SQOUGZDYSA-M 0.000 description 1
- WQZGKKKJIJFFOK-QTVWNMPRSA-N D-mannopyranose Chemical compound OC[C@H]1OC(O)[C@@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-QTVWNMPRSA-N 0.000 description 1
- HMFHBZSHGGEWLO-SOOFDHNKSA-N D-ribofuranose Chemical compound OC[C@H]1OC(O)[C@H](O)[C@@H]1O HMFHBZSHGGEWLO-SOOFDHNKSA-N 0.000 description 1
- SRBFZHDQGSBBOR-IOVATXLUSA-N D-xylopyranose Chemical compound O[C@@H]1COC(O)[C@H](O)[C@H]1O SRBFZHDQGSBBOR-IOVATXLUSA-N 0.000 description 1
- 206010012735 Diarrhoea Diseases 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 239000004386 Erythritol Substances 0.000 description 1
- UNXHWFMMPAWVPI-UHFFFAOYSA-N Erythritol Natural products OCC(O)C(O)CO UNXHWFMMPAWVPI-UHFFFAOYSA-N 0.000 description 1
- RFSUNEUAIZKAJO-ARQDHWQXSA-N Fructose Chemical compound OC[C@H]1O[C@](O)(CO)[C@@H](O)[C@@H]1O RFSUNEUAIZKAJO-ARQDHWQXSA-N 0.000 description 1
- 229920002527 Glycogen Polymers 0.000 description 1
- SQUHHTBVTRBESD-UHFFFAOYSA-N Hexa-Ac-myo-Inositol Natural products CC(=O)OC1C(OC(C)=O)C(OC(C)=O)C(OC(C)=O)C(OC(C)=O)C1OC(C)=O SQUHHTBVTRBESD-UHFFFAOYSA-N 0.000 description 1
- 241000282412 Homo Species 0.000 description 1
- 229920001202 Inulin Polymers 0.000 description 1
- SHZGCJCMOBCMKK-PQMKYFCFSA-N L-Fucose Natural products C[C@H]1O[C@H](O)[C@@H](O)[C@@H](O)[C@@H]1O SHZGCJCMOBCMKK-PQMKYFCFSA-N 0.000 description 1
- HEBKCHPVOIAQTA-IMJSIDKUSA-N L-arabinitol Chemical compound OC[C@H](O)C(O)[C@@H](O)CO HEBKCHPVOIAQTA-IMJSIDKUSA-N 0.000 description 1
- SHZGCJCMOBCMKK-JFNONXLTSA-N L-rhamnopyranose Chemical compound C[C@@H]1OC(O)[C@H](O)[C@H](O)[C@H]1O SHZGCJCMOBCMKK-JFNONXLTSA-N 0.000 description 1
- 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 description 1
- OVRNDRQMDRJTHS-UHFFFAOYSA-N N-acelyl-D-glucosamine Natural products CC(=O)NC1C(O)OC(CO)C(O)C1O OVRNDRQMDRJTHS-UHFFFAOYSA-N 0.000 description 1
- MBLBDJOUHNCFQT-LXGUWJNJSA-N N-acetylglucosamine Natural products CC(=O)N[C@@H](C=O)[C@@H](O)[C@H](O)[C@H](O)CO MBLBDJOUHNCFQT-LXGUWJNJSA-N 0.000 description 1
- HAUHVTUBJKXLLM-SZPZTJOGSA-N OC[C@H](O)[C@H](O)[C@@H](O)C=O.O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1O[C@H]1[C@H](O)[C@@H](O)C(O)O[C@@H]1CO Chemical compound OC[C@H](O)[C@H](O)[C@@H](O)C=O.O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1O[C@H]1[C@H](O)[C@@H](O)C(O)O[C@@H]1CO HAUHVTUBJKXLLM-SZPZTJOGSA-N 0.000 description 1
- PYMYPHUHKUWMLA-LMVFSUKVSA-N Ribose Natural products OC[C@@H](O)[C@@H](O)[C@@H](O)C=O PYMYPHUHKUWMLA-LMVFSUKVSA-N 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- MUPFEKGTMRGPLJ-UHFFFAOYSA-N UNPD196149 Natural products OC1C(O)C(CO)OC1(CO)OC1C(O)C(O)C(O)C(COC2C(C(O)C(O)C(CO)O2)O)O1 MUPFEKGTMRGPLJ-UHFFFAOYSA-N 0.000 description 1
- TVXBFESIOXBWNM-UHFFFAOYSA-N Xylitol Natural products OCCC(O)C(O)C(O)CCO TVXBFESIOXBWNM-UHFFFAOYSA-N 0.000 description 1
- 230000004520 agglutination Effects 0.000 description 1
- PYMYPHUHKUWMLA-WISUUJSJSA-N aldehydo-L-xylose Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)C=O PYMYPHUHKUWMLA-WISUUJSJSA-N 0.000 description 1
- 229930195726 aldehydo-L-xylose Natural products 0.000 description 1
- HMFHBZSHGGEWLO-UHFFFAOYSA-N alpha-D-Furanose-Ribose Natural products OCC1OC(O)C(O)C1O HMFHBZSHGGEWLO-UHFFFAOYSA-N 0.000 description 1
- 229940024606 amino acid Drugs 0.000 description 1
- 235000001014 amino acid Nutrition 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 229910021529 ammonia Inorganic materials 0.000 description 1
- 230000003712 anti-aging effect Effects 0.000 description 1
- 230000001093 anti-cancer Effects 0.000 description 1
- PYMYPHUHKUWMLA-UHFFFAOYSA-N arabinose Natural products OCC(O)C(O)C(O)C=O PYMYPHUHKUWMLA-UHFFFAOYSA-N 0.000 description 1
- 229960000271 arbutin Drugs 0.000 description 1
- 230000000621 autoagglutination Effects 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 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 description 1
- DLRVVLDZNNYCBX-ZZFZYMBESA-N beta-melibiose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1OC[C@@H]1[C@@H](O)[C@H](O)[C@@H](O)[C@H](O)O1 DLRVVLDZNNYCBX-ZZFZYMBESA-N 0.000 description 1
- 229940009289 bifidobacterium lactis Drugs 0.000 description 1
- 239000003613 bile acid Substances 0.000 description 1
- 230000000975 bioactive effect Effects 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 230000011748 cell maturation Effects 0.000 description 1
- 239000006285 cell suspension Substances 0.000 description 1
- 210000002421 cell wall Anatomy 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 235000013681 dietary sucrose Nutrition 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 230000007515 enzymatic degradation Effects 0.000 description 1
- 235000019414 erythritol Nutrition 0.000 description 1
- 229940009714 erythritol Drugs 0.000 description 1
- UNXHWFMMPAWVPI-ZXZARUISSA-N erythritol Chemical compound OC[C@H](O)[C@H](O)CO UNXHWFMMPAWVPI-ZXZARUISSA-N 0.000 description 1
- 229940093496 esculin Drugs 0.000 description 1
- AWRMZKLXZLNBBK-UHFFFAOYSA-N esculin Natural products OC1OC(COc2cc3C=CC(=O)Oc3cc2O)C(O)C(O)C1O AWRMZKLXZLNBBK-UHFFFAOYSA-N 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- 239000000834 fixative Substances 0.000 description 1
- 235000013350 formula milk Nutrition 0.000 description 1
- 229930182830 galactose Natural products 0.000 description 1
- 210000001035 gastrointestinal tract Anatomy 0.000 description 1
- 229940050410 gluconate Drugs 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 229940096919 glycogen Drugs 0.000 description 1
- 230000035931 haemagglutination Effects 0.000 description 1
- 210000000987 immune system Anatomy 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 229960000367 inositol Drugs 0.000 description 1
- CDAISMWEOUEBRE-GPIVLXJGSA-N inositol Chemical compound O[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@H](O)[C@@H]1O CDAISMWEOUEBRE-GPIVLXJGSA-N 0.000 description 1
- 210000004692 intercellular junction Anatomy 0.000 description 1
- 230000007413 intestinal health Effects 0.000 description 1
- 229940029339 inulin Drugs 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 210000002429 large intestine Anatomy 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- HOVAGTYPODGVJG-ZFYZTMLRSA-N methyl alpha-D-glucopyranoside Chemical compound CO[C@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O HOVAGTYPODGVJG-ZFYZTMLRSA-N 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 210000003097 mucus Anatomy 0.000 description 1
- 229950006780 n-acetylglucosamine Drugs 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 238000000879 optical micrograph Methods 0.000 description 1
- 238000000399 optical microscopy Methods 0.000 description 1
- BJRNKVDFDLYUGJ-UHFFFAOYSA-N p-hydroxyphenyl beta-D-alloside Natural products OC1C(O)C(O)C(CO)OC1OC1=CC=C(O)C=C1 BJRNKVDFDLYUGJ-UHFFFAOYSA-N 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 230000035479 physiological effects, processes and functions Effects 0.000 description 1
- 230000010287 polarization Effects 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 235000004252 protein component Nutrition 0.000 description 1
- MUPFEKGTMRGPLJ-ZQSKZDJDSA-N raffinose 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[C@@H]2[C@@H]([C@@H](O)[C@@H](O)[C@@H](CO)O2)O)O1 MUPFEKGTMRGPLJ-ZQSKZDJDSA-N 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- CDAISMWEOUEBRE-UHFFFAOYSA-N scyllo-inosotol Natural products OC1C(O)C(O)C(O)C(O)C1O CDAISMWEOUEBRE-UHFFFAOYSA-N 0.000 description 1
- 210000002955 secretory cell Anatomy 0.000 description 1
- 235000020183 skimmed milk Nutrition 0.000 description 1
- 238000004611 spectroscopical analysis Methods 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 230000000638 stimulation Effects 0.000 description 1
- 229960004793 sucrose Drugs 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
- 241001148471 unidentified anaerobic bacterium Species 0.000 description 1
- 150000003722 vitamin derivatives Chemical class 0.000 description 1
- 239000000811 xylitol Substances 0.000 description 1
- 235000010447 xylitol Nutrition 0.000 description 1
- HEBKCHPVOIAQTA-SCDXWVJYSA-N xylitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)CO HEBKCHPVOIAQTA-SCDXWVJYSA-N 0.000 description 1
- 229960002675 xylitol Drugs 0.000 description 1
Images
Classifications
-
- 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
-
- 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
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Biotechnology (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Wood Science & Technology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Medicinal Chemistry (AREA)
- Biomedical Technology (AREA)
- Virology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Nutrition Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
본 발명은, 종래의 프로바이오틱 유산균의 기본적인 생리활성인 내산소성, 내산성, 내담즙성 및 우유 배양성 등이 뛰어나면서도, 특히 장 정착성이 우수하여, 장에서의 높은 증식능을 갖는 신규 비피더스 유산균 균주로서 비피도박테리움 인팬티스 MAEIL-K9 비피더스 유산균 KCTC 10294BP를 제공한다.The present invention is a novel bifidus lactic acid bacterium having excellent intestinal fixation ability and excellent intestinal fixation ability while being excellent in oxygen resistance, acid resistance, bile resistance and milk cultureability, which are basic physiological activities of conventional probiotic lactic acid bacteria. As strain, Bifidobacterium infantis MAEIL-K9 bifidus lactic acid bacteria KCTC 10294BP is provided.
장 정착성, 비피더스 유산균, 비피도박테리움 인팬티스, 프로바이오틱 유산균Intestinal Settlement, Bifidus Lactobacillus, Bifidobacterium Infantis, Probiotic Lactobacillus
Description
도 1은 실험에 이용된 각 비피더스 유산균의 세포표면 소수성을 비교한 도면이고,1 is a diagram comparing the cell surface hydrophobicity of each bifidus lactic acid bacteria used in the experiment,
도 2는 실험에 이용된 각 비피더스 유산균의 세포 응집능을 비교한 결과이며,Figure 2 is a result of comparing the cell aggregation capacity of each bifidus lactic acid bacteria used in the experiment,
도 3은 직접검경법을 이용한 비피더스 유산균의 Caco-2 세포 정착능을 확인한 결과이며,3 is a result of confirming the Caco-2 cell fixation ability of bifidus lactic acid bacteria using direct microscopy,
도 4는 본 발명에서 선발한 비피더스 유산균인 MAEIL-K9이 Caco-2 세포에 정착된 광학현미경 사진이다(도 4에서, (A)는 비피도박테리움 인팬티스 MAEIL-K9의 형태(×1500)를 나타내며,(B)는 Caco-2 세포의 형태(×200)를 나타내고, (C)는 그람(Gram) 염색 후의 Caco-2 세포의 형태(×1500)를 나타내며, (D)는 Caco-2 세포에 정착된 비피더스 유산균의 형태(×1500)를 나타냄).Figure 4 is an optical micrograph of the bifidus lactic acid bacteria MAEIL-K9 selected in the present invention is fixed to Caco-2 cells (Fig. 4, (A) is the form of Bifidobacterium infantis MAEIL-K9 (x 1500) (B) shows the morphology of Caco-2 cells (× 200), (C) shows the morphology of Caco-2 cells after Gram staining (× 1500), and (D) shows Caco- Form of bifidus lactic acid bacteria (* 1500) settled in 2 cells).
본 발명은 신규 비피더스 유산균 균주 및 그를 함유하는 기능성 식품에 관한 것이며, 더욱 상세하게는 종래의 프로바이오틱 유산균의 기본적인 생리활성인 내산소성, 내산성, 내담즙성 및 우유 배양성 등이 뛰어나면서도, 특히 장 정착성이 우수하여, 장에서의 높은 증식능을 갖는 신규 비피더스 유산균 균주에 관한 것이다.The present invention relates to a novel bifidus lactobacillus strain and a functional food containing the same, and more particularly, in spite of being excellent in oxygen resistance, acid resistance, bile resistance and milk culture ability, which are basic physiological activities of conventional probiotic lactic acid bacteria, It relates to a novel bifidus lactic acid bacteria strain having excellent intestinal fixability and having a high proliferative capacity in the intestine.
사람의 장내에서 유용균으로서 가장 많이 알려진 유산균은 포도당 또는 탄수화물을 분해하여 생체에 유용한 유산을 생성하는 세균으로서 정의되고 있으며, 이들은 단백질을 분해하지만 부패시키지는 않으며, 인체에 해로운 물질들을 생성하지 않고 유익한 작용을 한다.Lactobacillus, best known as a useful bacterium in the human intestine, is defined as a bacterium that breaks down glucose or carbohydrates to produce a useful lactic acid for the living body, which breaks down proteins but does not rot and does not produce harmful substances to the human body. do.
메치니코프는 장내 부패세균에 의해서 만들어지는 독소가 노화를 일으킨다는 것을 찾아냈으며, 부패세균의 작용을 억제함으로써 노화가 억제될 수 있다고도 주장하였다. 그는 이러한 작용이 비피더스균과 같은 유산균에 의해서 수행될 수 있음을 확인하였다. 즉, 비피더스 유산균은 대장 내에서 유산과 초산 등을 생성하여 장내의 pH를 산성으로 유지시키므로, 장내 부패균의 증식을 억제하여 이러한 노화억제 기능을 수행하는 것으로 보여졌다. 또한, 비피더스 유산균은 노화 억제작용 뿐만 아니라, 혈중 콜레스테롤 저하작용, 비타민 합성 기능, 변비 치료작용, 항암 작용, 면역 작용 및 설사 개선 작용 등의 기능도 갖고 있다고 알려졌다.Mezzinikov found that toxins produced by intestinal rot bacteria cause aging and argue that aging can be suppressed by inhibiting the action of rot bacteria. He confirmed that this action could be performed by lactic acid bacteria such as Bifidus. In other words, bifidus lactic acid bacteria to produce lactic acid and acetic acid in the intestine to maintain the pH in the intestinal acidic acid, inhibiting the growth of intestinal rot bacteria has been shown to perform this anti-aging function. In addition, bifidus lactic acid bacteria, as well as inhibiting aging, blood cholesterol lowering action, vitamin synthesis function, constipation treatment action, anti-cancer action, immune action and diarrhea improvement effect is known to have a function.
이러한 비피더스 유산균의 기능을 이용한 기능성 식품이 많이 개발되었으며 현재 시판되고 있다. 그러나, 현재까지 개발되거나 시판되고 있는 비피더스 유산균을 함유하고 있는 기능성 식품들은 비피더스 유산균이 장까지 살아서 들어가는 것에 대해서만 관심이 집중되었다. 즉, 소화기관을 통과하여 장까지 도달시킬 때, 소화기관에서의 산성 환경 및 효소분해 환경에 저항성을 갖는 비피더스 유산균 균주의 개발에만 관심이 집중되었다.Many functional foods using the functions of these bifidus lactic acid bacteria have been developed and are now commercially available. However, functional foods containing bifidus lactic acid bacteria that have been developed or marketed so far have focused only on the entry of bifidus lactic acid bacteria into the intestines. That is, when reaching through the digestive system to the intestine, attention was focused only on the development of the bifidus lactic acid bacteria strain resistant to the acidic environment and the enzymatic degradation environment in the digestive system.
그러나, 사람의 대장에서는 인체에 필요한 비타민, 아미노산 및 단백질을 합성하며 감염 방어하는 등의 유익한 작용을 하는 유용균과, 암모니아, 페놀, 독소 등을 생성하여 건강을 해치는 유해균이 경쟁하고 있으며, 양자의 균형에 의하여 건강상태가 조절된다고 알려져 있다. 특히, 장에서 유산균의 점유상태가 생체 건강의 중요한 요인으로 작용한다고 알려져 있다.However, in the large intestine of humans, useful bacteria that have beneficial effects such as synthesizing vitamins, amino acids and proteins necessary for the human body and defending against infection, and harmful bacteria that produce ammonia, phenol, and toxins and harm health, are competing. It is known that the state of health is controlled by. In particular, the occupied state of lactic acid bacteria in the intestine is known to act as an important factor of the health of the body.
따라서, 유산균이 장에서 충분한 기능을 발휘하기 위해서는 장까지의 생존성이 우수해야 할 뿐만이 아니라, 유해균과의 경쟁에서 더욱 많은 군집을 형성할 수 있기 위해서 장에서의 정착성(장 정착능)이 우수해야만 한다. 현재 식품에 이용되는 비피더스 유산균은 단지 장까지의 생존성만을 주로 고려하여 개발된 것이며, 아직까지는 장 정착능을 고려하여 균주를 개발하려는 시도는 거의 없었다.Therefore, in order for lactic acid bacteria to function sufficiently in the intestine, not only the viability to the intestine should be excellent, but also the intestinal fixability (intestinal fixation ability) is excellent in order to form more colonies in competition with harmful bacteria. must do it. Bifidus lactic acid bacteria currently used in food have been developed mainly considering only viability to the intestine, and so far there has been little attempt to develop strains considering the intestinal fixation ability.
이에, 본 발명자는 병원성 세균에 대한 장내에서의 1차적인 보호 장벽을 형성하는데 있어서, 유산균이 가져야 하는 가장 중요한 요소 중 하나인 장 정착성이 우수한 균주를 선발하고자 많은 연구를 수행한 결과, 모유 영양아(2개월)로부터 분리한 비피도박테리움 인팬티스 MAEIL-K9 비피더스 유산균(KCTC 10294BP)이 프로바이오틱 유산균이 가져야 하는 기본적인 생리활성인 내산소성, 내산성, 내담즙성, 우유 배양능이 우수하면서도, 공지된 균주들에 비해서 장 정착능이 매우 우수함을 발견하고, 본 발명을 완성하기에 이르렀다. 따라서, 본 발명의 목적은 장 정착능이 우수한 신규 비피더스 유산균을 제공하는 것이다. 본 발명의 또 다른 목적은 상기 비피더스 유산균을 함유하는 건강기능성 식품을 제공하는 것이다.Accordingly, the present inventors conducted a number of studies to select strains excellent in intestinal fixability, which is one of the most important factors that lactic acid bacteria should have in forming the first protective barrier in the gut against pathogenic bacteria. Bifidobacterium infantis MAEIL-K9 bifidus lactic acid bacterium (KCTC 10294BP) isolated from 2 months old baby has excellent physiological activities such as oxygen resistance, acid resistance, bile resistance, and milk culture ability. , It was found that the intestinal fixation ability is very superior to the known strains, and came to complete the present invention. Accordingly, it is an object of the present invention to provide a novel bifidus lactic acid bacterium excellent in intestinal fixation ability. Still another object of the present invention is to provide a health functional food containing the bifidus lactic acid bacteria.
상기한 목적을 달성하기 위해서, 본 발명은 종래의 프로바이오틱 유산균의 기본적인 생리활성인 내산소성, 내산성, 내담즙성 및 우유 배양성 등이 뛰어나면서도, 특히 장 정착성이 우수하여 장에서의 높은 증식능을 갖는 신규 비피더스 유산균 균주로서 비피도박테리움 인팬티스 MAEIL-K9 비피더스 유산균(KCTC 10294BP)을 제공한다.In order to achieve the above object, the present invention is excellent in the intestinal fixability and excellent in the intestinal fixability, but also excellent in oxygen resistance, acid resistance, bile resistance and milk culture ability, which are basic physiological activities of the conventional probiotic lactic acid bacteria As a novel bifidus lactobacillus strain having proliferative capacity, Bifidobacterium infantis MAEIL-K9 bifidus lactic acid bacteria (KCTC 10294BP) is provided.
상기 균주는 2개월된 모유영양아로부터 분리한 비피도박테리움 인팬티스 균주로서, 다음 방법에 의하여 분리하였다. 즉, 상기 영양아로부터 채집된 분변을 0.85% 멸균 생리식염수에 10% 희석하여 잘 용해한 후 화염 멸균된 백금이를 이용하여 BL agar(5% horse blood 함유)에 스트리킹(streaking)하였다. 이것을 37℃/72시간 혐기배양한 다음 비피더스균으로 인정되는 집락을 선정하여 그람 염색, 현미경하에서의 형태 비교, F6PPK 테스트에 의해 최종 비피더스균임을 확인하였다. 이를 2002년 6월 25일자로 생명공학연구소 유전자 은행에 기탁하여, 수탁번호 KCTC 10294BP를 수여받았다.The strain was a Bifidobacterium infantis strain isolated from 2 months old breast-feeding infants and was isolated by the following method. That is, feces collected from the nutrients were diluted 10% in 0.85% sterile saline and dissolved well, and then streaked in BL agar (containing 5% horse blood) using flame sterilized platinum. After anaerobic incubation at 37 ° C./72 hours, colonies recognized as bifidus were selected and identified as final bifidus by gram staining, morphological comparison under a microscope, and F6PPK test. It was deposited on June 25, 2002 to the Biotechnology Research Institute Gene Bank, and received accession number KCTC 10294BP.
이하, 본 발명을 더욱 구체적으로 설명한다.Hereinafter, the present invention will be described in more detail.
본 발명에서 선발된 비피도박테리움 인팬티스 MAEIL-K9 비피더스 유산균과 종래에 공지된 비피더스균 균주 및 그 외의 임상균주들의 세포표면 소수성, 장 정착성, 우유 배양능 및 프로바이오틱 유산균의 기초적 생리활성인 내산소성, 내산성 및 내답즙성 등의 내성을 평가한 결과를 하기 표 1에 나타냈다.Basic Physiology of Cell Surface Hydrophobicity, Intestinal Fixation, Milk Culture Capacity, and Probiotic Lactobacillus of Bifidobacterium Infantis MAEIL-K9 Bifidobacterium Lactobacillus and Other Known Bifidobacterium Strains and Other Clinical Strains Selected in the Present Invention The results of evaluating the resistance such as active oxygen resistance, acid resistance and bile resistance are shown in Table 1 below.
상기 표 1에서 +++ 는 아주 좋음을, ++ 는 좋음을, + 는 약간 좋음을, 그리고 - 는 좋지 않음을 나타낸다.In Table 1, +++ is very good, ++ is good, + is slightly good, and-is not good.
상기 표 1에서 보는 바와 같이, 본 발명에서 선발된 비피도박테리움 인팬티스 MAEIL-K9은 종래의 공지된 균주 및 여러 임상균주에 비해서 세포표면 소수성, 정착성, 우유 배양능 및 프로바이오틱 유산균의 기초적 생리활성인 여러 내성이 균 형있게 모두 우수하였다.As shown in Table 1, Bifidobacterium infantis MAEIL-K9 selected in the present invention is cell surface hydrophobicity, fixation, milk culture ability and probiotic lactic acid bacteria compared to the known strains and various clinical strains All of the basic physiological activities of were well balanced.
본 발명에서 선발된 비피도박테리움 인팬티스 MAEIL-K9은, 건강기능성 식품에 포함시키기 위한 비피더스 유산균 균주의 선발기준과는 다른 기준에 의해서 선발된 균주이다. 즉, 현재까지 건강기능성 식품용 유산균은 장까지 살아서 들어가는 것에 대해서만 그의 선발기준으로서 삼아왔지만, 비피더스 유산균을 포함한 모든 프로바이오틱 유산균은 장내에 정착하여 증식 및 서식할 수 있어야만 장내에서 기능을 발휘할 수 있으므로, 본 발명의 균주는 장 정착능을 충분히 고려하여 선발되었다. 따라서, 본 발명에서 선발된 균주는 장에서의 생존성, 정착성 및 증식성이 우수하므로, 이는 일차적으로 병원성 세균의 장벽 침입을 억제하고, 유용한 미생물이 생성하는 부산물에 의해서 병원성 세균의 성장을 억제시키며, 장 정착에 의한 면역체계 자극으로 인한 면역증강 등의 효과를 유발할 수 있다.The Bifidobacterium infantis MAEIL-K9 selected in the present invention is a strain selected by a criterion different from the selection criteria of the bifidus lactic acid bacteria strain for inclusion in a health functional food. That is, until now, lactic acid bacteria for health functional foods have been used as the selection criteria only for living in the intestine, but all probiotic lactic acid bacteria including bifidus lactic acid bacteria can function in the intestine only if they can settle, proliferate and inhabit the intestines. , The strain of the present invention was selected in consideration of the intestinal fixation ability. Therefore, since the strains selected in the present invention are excellent in viability, fixability and proliferation in the intestine, this primarily inhibits barrier invasion of pathogenic bacteria and inhibits growth of pathogenic bacteria by by-products produced by useful microorganisms. In addition, it may cause effects such as immunity enhancement due to stimulation of the immune system by intestinal settlement.
이러한 선별기준에 의한 균주의 선발은 종래에 그다지 많이 행하여 지지 않았고, 단지 몇몇 유산간균, 예를 들어 락토바실러스 GG 유산균, 락토바실러스 가세리 ADH, 락토바실러스 애시도필러스 BG2FO4 균주만이 장내 서식능에 대해서 연구되어 왔다. 현재까지 우수한 장 정착능력을 가진 것으로 알려진 프로바이오틱 유산간균으로서는 락토바실러스 GG 유산균, 락토바실러스 애시도필러스 LA1, 락토바실러스 애시도필러스 NCFB 1748, 락토바실러스 케이시아이 시로타 등의 유산간균을 들 수 있다. 이외에 일부 비피더스 유산균도 프로바이오틱 활성을 나타내는데, 실제로 비피도박테리움 랙티스 Bb-12의 점막 결합력은 락토바실러스 GG 유산균과 락토바실러스 델부뤼에키 불가리커스가 존재할 때 더욱 높아지는 것으로 밝혀졌다. Selection of strains based on these selection criteria has not been carried out in the past, and only a few lactobacillus strains, for example, Lactobacillus GG lactobacillus, Lactobacillus gaseri ADH, and Lactobacillus ashdophyllus BG2FO4 strains, have an intestinal viability. Has been studied. Probiotic lactic acid bacteria known to have excellent intestinal fixation ability to date include Lactobacillus GG lactic acid bacteria, Lactobacillus ashdophyllus LA1, Lactobacillus ashdophyllus NCFB 1748, Lactobacillus Keshiai Shirota, etc. have. In addition, some of the bifidus lactic acid bacteria also show probiotic activity, and in fact, the mucosal binding capacity of the Bifidobacterium lactis Bb-12 was found to be higher in the presence of Lactobacillus GG lactic acid bacteria and Lactobacillus delburueki vulgaris.
비피더스 유산균이 세포벽에 정착하는 기작은 현재 구체적으로 알려진 바는 없지만, 일반적으로는 단백질, 다당류, 이온전하, 리포테이콘산(lipoteichoic acid) 등의 정착인자의 농도와 균주간의 생리활성의 차이에 기인하는 것으로 알려져 있다. 즉, 유산균의 정착능은 장 상피세포인 Caco-2 세포의 기능성쇄자연(brush border)과 HT-29-MTX 점질물 분비 세포에 의해서 분비되는 점액질에 의해서 나타난다고 알려져 있다. 또한, 비피더스 유산균은 Caco-2 세포에 아무런 손상도 미치지 않으면서 첨단의 미세융모와 상호작용하며, 비피더스 유산균이 Caco-2 세포에 정착하는 과정에는 칼슘을 필요로 하지 않고, 비피더스 유산균 전체 세포와 배양 상등액에 존재하는 단백질성 정착능 증강인자에 의해서 매개된다. 또한, 이러한 정착능 증강 인자는 유산균의 정착능 촉진인자와 마찬가지로 균주 특이성을 나타낸다고 알려져 있다.The mechanism by which bifidus lactic acid bacteria settle in the cell wall is currently unknown, but is generally due to the difference in the concentrations of fixation factors such as proteins, polysaccharides, ionic charges, and lipoteichoic acid and the physiological activity between strains. It is known. In other words, the lactic acid bacteria's ability to settle is known to be caused by the brush border of Caco-2 cells, the intestinal epithelial cells, and the mucus secreted by the HT-29-MTX stromal secretory cells. In addition, bifidus lactobacillus interacts with the microvilli of the tip without damaging Caco-2 cells, and does not require calcium in the process of the bifidus lactic acid bacteria settling into Caco-2 cells, and cultures with whole cells of bifidus lactobacillus. Mediated by proteinaceous fixation enhancers present in the supernatant. In addition, these fixation enhancers are known to exhibit strain specificity similarly to the fixation promoters of lactic acid bacteria.
한편, 정착능이 우수한 비피더스 유산균은, 세포표면 소수성 검사, 세포 응집능 검사, 직접 검경법에 의한 비피더스 유산균의 장 상피세포 Caco-2에의 정착능 등을 조사함으로써 선발될 수 있다.On the other hand, bifidus lactic acid bacteria having excellent fixation ability can be selected by examining cell surface hydrophobicity test, cell aggregation ability test, and ability to fix bifidus lactic acid bacteria intestinal epithelial cell Caco-2 by direct spectroscopy.
일반적으로, 유산균의 장 상피세포에의 정착능은 유산균과 상피세포 사이의 흡인력과 반발력의 상호작용의 결과로 볼 수 있으며, 이러한 상호작용에는 세포표면 소수성이 요구된다. 당업계에서는 일반적으로 정착능을 갖기 위해서는 85% 이상의 세포표면 소수성이 요구된다고 알려져 있으며, 세포표면 소수성은 균주 특이성에 의존한다는 것으로 당업계에 알려져 있다. 본 발명에 의하면, 본원 발명에서 선발된 비피도박테리움 인팬티스 MAEIL-K9은 매우 우수한 세포표면 소수성을 갖는 다.In general, the ability of the lactic acid bacteria to settle into intestinal epithelial cells is the result of the interaction between the aspiratory and repulsive forces between the lactic acid bacteria and the epithelial cells, which requires cell surface hydrophobicity. It is generally known in the art that cell surface hydrophobicity of 85% or more is required to have fixation capacity, and cell surface hydrophobicity is known in the art to depend on strain specificity. According to the present invention, the Bifidobacterium infantis MAEIL-K9 selected in the present invention has very good cell surface hydrophobicity.
본 발명에서는, 장 정착능을 조사하기 위한 세포 모델로서 Caco-2 세포를 이용하여 장 정착성 균주를 선발하였다. 인체 장내 상피세포 계열인 Caco-2 세포는 실험실 조건에서도 형태학적, 기능적 분화를 일으킬 수 있으며, 또한 극성화(polarization), 기능성 쇄자연(brush border) 및 장융모 첨단(microvilli)에서 분비하는 가수분해효소(hydrolase) 등의 성숙한 장 상피세포의 특성을 나타낸다. 또한, Caco-2 세포는 정상적인 소장 융모세포의 다양한 특성을 나타내므로, 여러가지 병원성 세균의 정착과 침입 기작에 대한 연구에 있어서 중요한 모델로서 제공될 수 있다. 즉, Caco-2 세포는 정상적인 장내균총 중에서 어떤 균종이 장에 정착하며, 장내균총이 병원성 세균 및 비피더스 유산균과 상호 경쟁하는 작용 기작을 연구하는데 훌륭한 세포 모델을 제공할 수 있기 때문에, 비피더스 유산균 등의 감염 및 정착의 연구에도 이용될 수 있다.In the present invention, enteric fixative strains were selected using Caco-2 cells as a cell model for investigating enteric fixation ability. Caco-2 cells of human intestinal epithelial cells can cause morphological and functional differentiation even under laboratory conditions, and also hydrolyzed by polarization, functional brush border, and microvilli. Mature intestinal epithelial cells such as enzyme (hydrolase). In addition, since Caco-2 cells exhibit various characteristics of normal small intestinal chollocytes, Caco-2 cells may serve as an important model in the study of the settlement and invasion mechanism of various pathogenic bacteria. That is, Caco-2 cells can provide an excellent cell model for studying the mechanism of interaction among pathogens and bifidus lactobacillus, in which certain strains of the normal intestinal flora are established in the intestine. It can also be used to study infection and settlement.
본 발명에서는 Caco-2 세포의 단일층에 비피더스 유산균 현탁액을 접종하여 그람 염색한 후, 현미경 하에서 계수하는 직접 검경법을 통해서 Caco-2 세포에 대한 정착능을 조사하여, 장 정착능이 우수한 비피더스 유산균 균주를 선발하였다. 그 결과, 본 발명에서 선발된 비피도박테리움 인팬티스 MAEIL-K9가 매우 우수한 Caco-2 세포에 대한 정착능, 즉 우수한 장 정착능을 갖는 균주로서 선발되었다.In the present invention, a single layer of Caco-2 cells were inoculated with Gram stained by inoculating bifidus lactobacillus suspension, and then examined for fixing ability against Caco-2 cells by direct microscopy counting under a microscope, and showed excellent bifidus lactic acid bacteria strains. Was selected. As a result, the Bifidobacterium infantis MAEIL-K9 selected in the present invention was selected as a strain having a very good Caco-2 cell fixation ability, that is, an excellent intestinal fixation ability.
한편, 본 발명에서 선발한 비피도박테리움 인팬티스 MAEIL-K9 균주는 내산소성, 내산성 및 내담즙성의 기초적인 프로바이오틱 유산균의 내성 특성 및 우유 배양능도 매우 우수하였다. 이러한 과정에서 선발된 프로바이오틱 유산균은 식품 내에서의 생존성, 섭취 후 장까지의 생존성, 장벽에의 정착능에서 우수한 특성을 가지게 되며, 따라서 프로바이오틱 유산균이 기본적으로 가져야 하는 모든 조건에 적합한 것으로 인정된다.On the other hand, the Bifidobacterium infantis MAEIL-K9 strain selected in the present invention was also excellent in the resistance characteristics and milk culture ability of the basic probiotic lactic acid bacteria, acid resistance and bile resistance. The probiotic lactic acid bacteria selected in this process have excellent characteristics in viability in food, viability to the intestine after ingestion, and ability to settle to the barrier. It is considered appropriate.
따라서, 본 발명에서 선발된 균주는 기존에 공지된 비피도박테리움 균주들 보다 더욱 우수한 장 정착능을 가지면서도, 기존의 균주들과 비슷하거나 더욱 우수한 프로바이오틱 생리활성을 가지고 있기 때문에, 건강기능성 식품에 포함되어 장에서 그의 활성을 충분히 발휘할 수 있다. 예를 들어, 비피도박테리움 인팬티스 MAEIL-K9 균주는 본 발명의 유음료, 발효유 제품, 분유 제품, 정장제, 생식 등에 함유되어 매우 유용한 건강기능성 식품을 제공할 수 있다. Therefore, the strains selected in the present invention have better intestinal fixation ability than the known Bifidobacterium strains, and have similar or better probiotic physiological activity than the existing strains, and thus have a health functional property. It is included in foods and can fully exhibit its activity in the intestine. For example, the Bifidobacterium infantis MAEIL-K9 strain may be contained in the milk beverage, fermented milk product, milk powder product, formal preparation, raw food, etc. of the present invention to provide a very useful health functional food.
한편, 본 발명에 따른 비피도박테리움 인팬티스 MAEIL-K9를 함유하는 건강 기능성 식품의 제조 및 그의 함량은 당업계에 공지된 통상적인 방법으로부터 용이하게 파악될 수 있으며, 당업자에게 자명하게 알려져 있는 섭취 후 장까지의 생존성 및 식품 내에서의 생존성을 유지하기 위한 다른 물질 등도 함유할 수도 있다.On the other hand, the preparation of the health functional food containing Bifidobacterium infantis MAEIL-K9 and its content according to the present invention can be easily understood from conventional methods known in the art, it is obvious to those skilled in the art It may also contain other substances for maintaining viability to the intestine after ingestion and viability in food.
[실시예]EXAMPLE
이하, 실시예에 의거하여 본 발명을 보다 상세히 설명한다. 하기 실시예는 본 발명의 이해를 보다 용이하게 하기 위하여 제공되는 것이지만, 본 발명의 범위가 이들 실시예로 한정되는 것은 아니다. Hereinafter, the present invention will be described in more detail based on Examples. The following examples are provided to facilitate the understanding of the present invention, but the scope of the present invention is not limited to these examples.
상기 표 1에 기재된 20종의 비피더스 유산균을 이용하여 먼저 세포표면 소수성과 직접검경법(direct count)을 이용한 Caco-2 세포 정착능을 조사하였으며, 이때 비피도박테리움 애돌센티스 MAEIL-K8과 비피도박테리움 인팬티스 MAEIL-K9를 선 발하였다. 또한, 이 두 균주에 대한 내산소성, 내산성, 내담즙성, 우유 배양성 등을 조사하여 비피도박테리움 인팬티스 MAEIL-K9를 최종 선발하였다. 따라서, 본 발명에 의한 비피도박테리움 인팬티스 MAEIL-K9는 식품과 건강보조제 등에 다양하게 활용될 수 있다.Using the 20 kinds of bifidus lactic acid bacteria described in Table 1, first, cell surface hydrophobicity and Caco-2 cell fixation ability using direct count were examined, wherein Bifidobacterium adolescents MAEIL-K8 and Bifidobacterium were used. Therium Infantis MAEIL-K9 was selected. In addition, by examining the oxygen resistance, acid resistance, bile resistance, milk culture, etc. for these two strains, Bifidobacterium infantis MAEIL-K9 was finally selected. Therefore, Bifidobacterium infantis MAEIL-K9 according to the present invention can be used in various ways such as food and health supplements.
상기 비피도박테리움 인팬티스 MAEIL-K9의 선발과정은 다음과 같다. 즉, 2개월된 모유영양아로부터 채집된 분변을 0.85% 멸균 생리식염수에 10% 희석하여 잘 용해한 후 화염 멸균된 백금이를 이용하여 BL agar(5% horse blood 함유)에 스트리킹(streaking)하였다. 이것을 37℃/72시간 혐기배양한 다음 비피더스균으로 인정되는 집락을 선정하여 그람 염색, 현미경하에서의 형태 비교, F6PPK 테스트에 의해 최종 비피더스균임을 확인하였다.The selection process of the Bifidobacterium infant MAEIL-K9 is as follows. That is, feces collected from two-month-old breastfeeding infants were diluted 10% in 0.85% sterile saline, and dissolved well, and then streaked in BL agar (containing 5% horse blood) using flame sterilized platinum. After anaerobic incubation at 37 ° C./72 hours, colonies recognized as bifidus were selected and identified as final bifidus by gram staining, morphological comparison under a microscope, and F6PPK test.
한편, 상기 선발된 균주가 비피도박테리움 인팬티스 종에 속한다는 것을 보여주는 분류학적 실험 자료는 표 2에서 보는 바와 같다. 표 2는 동정을 위한 선별된 비피도박테리움 인팬티스 MAEIL-K9의 탄수화물 발효 패턴을 보여주고 있다.On the other hand, the taxonomic experimental data showing that the selected strain belongs to the Bifidobacterium inpantis species is shown in Table 2. Table 2 shows the carbohydrate fermentation patterns of selected Bifidobacterium infantis MAEIL-K9 for identification.
한편, 상기 균주의 배양적 특성은 일반적인 비피더스균의 그것과 동일하며, 본 발명의 균주를 2002년 6월 25일자로 한국생명과학연구소 유전자 은행에 기탁하여 수탁번호 KCTC 10294BP를 수여받았다.On the other hand, the culture characteristics of the strain is the same as that of the general bifidus bacteria, the strain of the present invention was deposited on June 25, 2002 to the Korea Life Science Research Institute Gene Bank and received the accession number KCTC 10294BP.
실시예 1. 비피더스 유산균의 세포표면 소수성(cell surface hydrophobicity, CSH %)Example 1 Cell Surface Hydrophobicity (CSH%) of Bifidus Lactic Acid Bacteria
세포표면 소수성 검사 방법에 대해서 간략히 설명하면, 먼저 비피더스 균주 를 modified MRS broth에 2.0% 접종하여 혐기배양(37℃/15hr)시킨 후, 원심분리(×3,000rpm/10min)하였다. 그후 세포 덩어리를 50mM 인산염 완충액(22.2g K2HPO4·H2O, 7.26g KH2PO4, pH 7.0)으로 2회 수세 반복 후, 세포 현탁액 3ml에 동량의 hexadecane(Sima Co., USA)을 첨가하여 60초간 혼합한 후, 20분간 방치하여 층이 분리되도록 하였다. 분리된 층 중에서 하층의 수용성 층의 흡광도를 600nm에서 측정하여 세포표면 소수성(CHS)을 아래와 같이 계산하였다. Briefly, the cell surface hydrophobicity test method, the bifidus strain was inoculated 2.0% in modified MRS broth anaerobic culture (37 ℃ / 15hr), and then centrifuged (x 3,000rpm / 10min). The cell mass was then washed twice with 50 mM phosphate buffer (22.2 g K 2 HPO 4 H 2 O, 7.26 g KH 2 PO 4 , pH 7.0), followed by the same amount of hexadecane (Sima Co., USA) in 3 ml of cell suspension. After adding for 60 seconds, the mixture was allowed to stand for 20 minutes to separate the layers. The cell surface hydrophobicity (CHS) was calculated as follows by measuring the absorbance of the lower water-soluble layer at 600 nm among the separated layers.
CHS(%)=(Ai-Af)×100/AiCHS (%) = (Ai-Af) × 100 / Ai
여기서 Ai=초기흡광도(OD=1.0±0.05), Af=최종 흡광도이다.
Where Ai = initial absorbance (OD = 1.0 ± 0.05) and Af = final absorbance.
각 비피더스 유산균의 세포표면 소수성에 대한 결과는 도 1에 나타냈다. 일반적으로 모든 미생물의 장 상피세포 정착능은 유산균과 상피세포 사이의 흡인력 및 반발력과의 상호작용의 결과라고 할 수 있는데, 세포표면 소수성이 85% 이상이 되어야 정착능과 기타 생리활성 기능이 우수하다고 할 수 있다. 도 1의 결과에서는 20 균종의 비피더스 유산균 중에서 비피도박테리움 비피덤 MAEIL-M3, 비피도박테리움 애돌센티스 MAEIL-K8, 비피도박테리움 인팬티스 MAEIL-K9 만이 85% 이상의 높은 세포표면 소수성을 나타냈다. 이러한 세포표면 소수성은 아종(subspecies)에 의한 것이라기보다는 균주 특이성에 더욱 의존하는 것으로 보여졌다.Results of cell surface hydrophobicity of each bifidus lactic acid bacteria are shown in FIG. 1. In general, the intestinal epithelial cell settling capacity of all microorganisms is a result of the interaction between the aspiratory and repulsive forces between lactic acid bacteria and epithelial cells. can do. In the results of FIG. 1, only 20 Bifidobacterium bifidum MAEIL-M3, Bifidobacterium adolescents MAEIL-K8, and Bifidobacterium inpantis MAEIL-K9 showed high cell surface hydrophobicity of 85% or more. Indicated. This cell surface hydrophobicity has been shown to be more dependent on strain specificity than by subspecies.
실시예 2. 비피더스 유산균의 세포 응집능(cell agglutination, CA) Example 2 Cell Agglutination of Bifidus Lactic Acid Bacteria
세포 응집능은 세포표면 소수성과 유사한 세포표면 특성을 측정하는 방법으 로 인정되고 있다. 실험에 이용된 비피더스 유산균의 세포 응집능은 도 2에 나타냈다. 모든 실험균주에서의 세포 응집능은 31.45±0.54 건물중㎎/㎖로서 균주간에 커다란 차이를 나타내지 않았다. 따라서, 세포 응집능으로 비피더스 유산균의 세포표면 특성에 따른 균주간의 특이성을 구분하기는 힘든 것으로 밝혀졌다. Cell cohesion is recognized as a method of measuring cell surface properties similar to cell surface hydrophobicity. Cell aggregation ability of the bifidus lactic acid bacteria used in the experiment is shown in FIG. Cell aggregation capacity in all experimental strains was 31.45 ± 0.54 mg / ml of dry matter, showing no significant difference among the strains. Therefore, it was found that it is difficult to distinguish the specificity between the strains according to the cell surface characteristics of bifidus lactic acid bacteria by cell aggregation ability.
실시예 3. 비피더스 유산균의 Caco-2 세포 정착능Example 3 Caco-2 Cell Fixation Capacity of Bifidus Lactic Acid Bacteria
장 상피세포에 대한 프로바이오틱 유산균의 정착능은 생리활성 증진 효과의 기본적인 조건으로 알려져 있다. 본 실시예에서는 Caco-2 세포에 대한 각 비피더스 유산균의 정착능을 직접 검경법으로 확인함으로써 각 균주의 장 정착능을 확인하였다.The ability of the probiotic lactic acid bacteria to intestinal epithelial cells is known as a basic condition of the biological activity enhancing effect. In this example, the intestinal fixation ability of each strain was confirmed by directly confirming the fixation ability of each bifidus lactic acid bacteria to Caco-2 cells by microscopy.
간략히 설명하면, 원형 플라스틱 플레이트(round plastic plate)에 형성된 Caco-2 세포의 단일층(monolayer)에 610nm에서의 흡광도값이 1.0인 비피더스 유산균 현탁액을 100㎕ 접종하여 2시간 동안 반응시킨 후, 그람(Gram) 염색법으로 염색한 다음, 현미경 하에서 직접적으로 계수하였다. Caco-2 세포에 대한 20종의 각 비피더스 유산균의 장 상피세포에 대한 정착능을 도 3에 나타냈다. 도 3에서 보는 바와 같이, Caco-2 세포 100개당 100개 이상의 균수가 측정된 장 정착능이 우수한 비피더스 유산균은 비피도박테리움 비피덤 MAEIL-M3, 비피도박테리움 애니말리스 MAEIL-M10, 비피도박테리움 롱검 MAEIL-K1, 비피도박테리움 롱검 MAEIL-K2, 비피도박테리움 애돌센티스 MAEIL-K8, 비피도박테리움 인팬티스 MAEIL-K9, 비피도박테리움 인팬티스 MAEIL-K10 균주이었다.Briefly, a monolayer of Caco-2 cells formed on a round plastic plate was inoculated with 100 μl of a bifidus lactic acid suspension having an absorbance value of 1.0 at 610 nm, and reacted for 2 hours. Gram) staining was followed by direct counting under a microscope. The settling capacity of 20 bifidus lactic acid bacteria against intestinal epithelial cells with respect to Caco-2 cells is shown in FIG. 3. As shown in FIG. 3, the Bifidobacterium lactic acid bacteria having excellent intestinal fixation ability, in which 100 or more bacteria were measured per 100 Caco-2 cells, Bifidobacterium bifidomum MAEIL-M3, Bifidobacterium animalis MAEIL-M10, and Bifidobacterium Leeum Longgum MAEIL-K1, Bifidobacterium longgum MAEIL-K2, Bifidobacterium adolescents MAEIL-K8, Bifidobacterium inpantis MAEIL-K9, Bifidobacterium inpantis MAEIL-K10 strain.
도 3에서, 장 정착능은 균종에 따라서 일관성 없이 차이를 나타났다. 따라 서, 비피더스 유산균 균주마다 Caco-2 세포 정착능이 서로 상이하게 나타나는 것은 일반적으로 단백질 성분, 다당류, 이온전하, 리포테이콘산(lipoteichoic acid) 등의 정착인자의 농도와 균주 자체의 생리활성 특성 차이에 기인하는 것으로 보여졌다.In FIG. 3, the intestinal fixation ability was inconsistently different depending on the species. Therefore, the Caco-2 cell fixation ability of the bifidus lactic acid bacteria is different from each other in terms of the concentrations of fixation factors such as protein components, polysaccharides, ion charges, and lipoteichoic acid and the physiological activity characteristics of the strain itself. It was shown to be due.
한편, 세포표면 소수성(CSH), 세포 응집능(CA) 및 정착능의 결과를 비교하면 비피도박테리움 비피덤 MAEIL-M3, 비피도박테리움 애돌센티스 MAEIL-K8, 비피도박테리움 인팬티스 MAEIL-K9가 세포 정착성에 관련된 모든 특성이 우수한 것으로 나타났다. 이러한 선발 과정에서, 비피도박테리움 인팬티스 MAEIL-M10의 경우에 정착능은 우수하였지만 세포표면 소수성이 매우 낮기 때문에 선발에서 제외하였다. 또한, 비피도박테리움 롱검 MAEIL-K1은 세포표면 특성과 정착능이 우수하였지만, 균주 자체의 활력(일상적으로 균주의 성장 조건에 대한 민감성, 예를 들어 배양이 어렵거나 성장이 매우 안되는 경우에는 균주 자체의 활력이 낮다고 표현함)이 매우 낮기 때문에 선발에서 제외하였다.On the other hand, when comparing the results of cell surface hydrophobicity (CSH), cell aggregation ability (CA) and fixation capacity, Bifidobacterium bifidem MAEIL-M3, Bifidobacterium adolescents MAEIL-K8, Bifidobacterium infantis MAEIL-K9 has been shown to be superior in all properties related to cell fixation. In the selection process, Bifidobacterium infantis MAEIL-M10 had excellent fixation ability but was excluded from selection because of its very low cell surface hydrophobicity. In addition, Bifidobacterium longgum MAEIL-K1 was excellent in cell surface properties and fixation ability, but the strain itself had a vitality (usually sensitive to growth conditions of the strain, for example, if the culture is difficult or very difficult to grow, the strain itself. Was not selected because of its very low level.
도 4는 비피도박테리움 인팬티스 MAEIL-K9(A), Caco-2 세포(B), 염색된 Caco-2 세포(C), 비피도박테리움 인팬티스 MAEIL-K9가 정착된 Caco-2 세포(D)의 모습을 광학현미경으로 촬영한 사진을 나타낸 것이다. 광학현미경으로 확인한 결과, 비피더스 유산균의 Caco-2 세포 정착부위는 매우 다양하지만, 여기서는 세포 상단의 기능성쇄자연(brush border, microvilli)과 세포와 세포 사이의 세포 분지점(tight junction)에 비피더스 유산균이 정착하는 것을 확인할 수 있었다. 4 shows Caco- with Bifidobacterium infantism MAEIL-K9 (A), Caco-2 cells (B), stained Caco-2 cells (C), and Bifidobacterium infantism MAEIL-K9. 2 cells (D) is a photograph taken with an optical microscope. As a result of optical microscopy, Caco-2 cell fixation sites of Bifidobacterium Lactobacillus are very diverse, but here, Bifidobacteric Lactobacillus is found in the functional border of the cell (microvilli) and the cell junction between the cells. It was confirmed that it settled.
실시예 4. 정착능이 우수한 비피더스 유산균의 선발Example 4. Selection of bifidus lactic acid bacteria having excellent fixation ability
미생물의 세포표면 특성인 세포표면 소수성(CSH)과 세포 응집능(CA) 및 직접검경법(direct count)에 의한 Caco-2 세포에 대한 정착능을 기준으로 하여 장 정착능이 우수한 것으로 선발된 균주는 비피도박테리움 비피덤 MAEIL-M3, 비피도박테리움 애돌센티스 MAEIL-K8, 비피도박테리움 인팬티스 MAEIL-K9이었다. 이들 균주의 세포표면 소수성(CSH)과 세포 응집능(CA) 및 Caco-2 세포에 대한 정착능에 대한 결과를 하기 표 3에 나타냈다. 대조군으로는 비피도박테리움 롱검 MAEIL-M8을 이용하였다.Based on the cell surface hydrophobicity (CSH), cell aggregation ability (CA), and direct counting ability of Caco-2 cells by microbial cell surface characteristics, the strains selected for superior intestinal fixation ability Bifidobacterium bifidum MAEIL-M3, Bifidobacterium adolescents MAEIL-K8, Bifidobacterium inpantis MAEIL-K9. The results of cell surface hydrophobicity (CSH) and cell aggregation ability (CA) of these strains and the fixing ability to Caco-2 cells are shown in Table 3 below. Bifidobacterium long gum MAEIL-M8 was used as a control.
상기 네 균주의 균 현탁액(흡광도 1.0, 610nm) 내의 균수는 1.2~2.3×108 cfu/㎖로서 거의 유사하였으며, 세포표면 소수성은 비피도박테리움 애돌센티스 MAEIL-M8을 제외하고는 90% 이상의 높은 결과를 나타냈다. 정착능에서도 비피도박테리움 애돌센티스 MAEIL-M8을 제외하고는 모두 100개 이상의 높은 결과를 나타냈고, 세포 응집능은 유의적인 차이를 보이지 않았다.The bacterial counts in the suspensions of the four strains (absorbance 1.0, 610 nm) were almost similar as 1.2-2.3 × 10 8 cfu / ml, and the cell surface hydrophobicity was higher than 90% except for Bifidobacterium adolescents MAEIL-M8. The result was shown. Except for Bifidobacterium adolescents MAEIL-M8, all of the results showed a high fixation ability of 100 or more, and cell cohesion was not significantly different.
상기 표 3의 결과로부터, 자동응집반응(autoagglutination)과 혈구응집반응 이 측정되지 않고 세포표면 소수성이 높은 균주라고 할지라도 Caco-2 세포에 대한 정착능이 항상 우수한 것은 아니라는 것을 알 수 있다. 그러나, 일반적으로 장 세포에 대한 유산간균의 정착은 다양한 미생물 세포 표면 특성에 의존한다.From the results in Table 3, it can be seen that even if the autoagglutination and hemagglutination reaction were not measured and the strain having high cell surface hydrophobicity, the fixation capacity for Caco-2 cells was not always excellent. However, the settlement of lactobacillus to enteric cells generally depends on various microbial cell surface properties.
실시예 5. 우유 배양성 검사Example 5. Milk Culture Test
우유 배양성 특성을 선발 기준에 포함시킨 이유는, 통상적으로 비피더스균은 혐기성 균이므로 우유 배양을 호기로 할 경우 균주 특성에 따라 성장이 되지 않는 경우가 매우 많으며, 따라서 우유 배양성 실험은 이러한 비피더스 균주를 차후에 상용화하는데 매우 필수적인 배양특성이라 할 수 있다.The reason why the milk culture characteristics are included in the selection criteria is that the bifidus bacteria are usually anaerobic bacteria, and thus, when the milk culture is aerobic, the growth is not very likely depending on the strain characteristics. Can be said to be a very essential culture characteristics for future commercialization.
우유 배양성 검사 방법에 대해서 간략히 설명하면, 다음과 같다.A brief description of the milk culture test method is as follows.
정착능이 우수하다고 인정되는 비피도박테리움 비피덤 MAEIL-M3, 비피도박테리움 롱검 MAEIL-M8, 비피도박테리움 애돌센티스 MAEIL-K8 및 비피도박테리움 인팬티스 MAEIL-K9를 mMRS 액체배지에서 3회 계대배양하여 활력을 높인 후에, 그 배양물을 원심분리하여 인산완충생리식염수(phosphate-buffered saline, PBS)로 2회 세척후 동량의 PBS로 치환하였다. 또한, 실험에 이용된 우유로는 11% 멸균환원탈지유를 이용하였으며, 접종률은 1%, 배양조건은 호기와 혐기배양, 배양 조건은 37℃에서 24시간으로 하였다. 우유 배양성에 대한 결과는 표 4에 나타냈다. 비피도박테리움 비피덤 MAEIL-M3는 우유 배양성이 매우 열악하였지만 비피도박테리움 롱검 MAEIL-M8, 비피도박테리움 애돌센티스 MAEIL-K8 및 비피도박테리움 인팬티스 MAEIL-K9는 배양성이 우수하였다.Bifidobacterium bifiderm MAEIL-M3, Bifidobacterium longgum MAEIL-M8, Bifidobacterium adolescents MAEIL-K8 and Bifidobacterium infantis MAEIL-K9, which are considered to have excellent fixation ability, were After three passages to increase vitality, the culture was centrifuged, washed twice with phosphate-buffered saline (PBS), and replaced with the same amount of PBS. In addition, 11% sterilized reduced skim milk was used as the milk used in the experiment, the inoculation rate was 1%, the culture conditions were the aerobic and anaerobic culture, and the culture conditions were set to 37 hours at 37 ℃. Results for milk cultures are shown in Table 4. Bifidobacterium Bifidum MAEIL-M3 was very poor in milk culture, but Bifidobacterium longgum MAEIL-M8, Bifidobacterium adolescents MAEIL-K8 and Bifidobacterium inpantis MAEIL-K9 were cultured Excellent.
실시예 6. 비피도박테리움 애돌센티스 MAEIL-K8과 비피도박테리움 인팬티스 MAEIL-K9의 기초적인 생리활성 특성 비교Example 6 Comparison of Basic Bioactive Properties of Bifidobacterium Adolescents MAEIL-K8 and Bifidobacterium Infantis MAEIL-K9
비피도박테리움 애돌센티스 MAEIL-K8과 비피도박테리움 인팬티스 MAEIL-K9의 기초적인 생리활성을 조사하기 위하여 내산소성, 내산성 및 내담즙성을 조사하였으며, 그 결과를 하기 표 5에 나타냈다. In order to investigate the basic physiological activities of Bifidobacterium adolescents MAEIL-K8 and Bifidobacterium infantis MAEIL-K9, oxygen resistance, acid resistance and bile resistance were examined, and the results are shown in Table 5 below.
내산소성 실험 방법은, 균체를 1×108-9 cfu/ml 수준으로 BL agar에 도말하여 37℃ 호기배양기에서 6, 15, 24시간 전 배양시킨 후, 혐기배양기에서 37℃/7시간 배양하여 생존력을 측정하였다.Oxygen resistance test method, the cells were plated in BL agar at the level of 1 × 10 8-9 cfu / ml and incubated 6, 15, 24 hours in aerobic incubator at 37 ° C., and then incubated at 37 ° C./7 hours in anaerobic incubator Viability was measured.
내산성 실험 방법은, 원심분리한 균체를 1N-HCl을 이용하여 pH를 조정한 0.05% L-cysteine-HCl이 함유된 50mM 인상염 완충액(pH 7.0, 3.0)에 1×107-8 cfu/ml 수준으로 접종하여 37℃ 혐기배양기에서 12시간 처리 후, 생존성을 BL agar에서 확인하였다.Acid resistance test method, centrifuged cells in 1 × 10 7-8 cfu / ml in 50 mM impression salt buffer (pH 7.0, 3.0) containing 0.05% L-cysteine-HCl pH adjusted using 1N-HCl After incubation at the level for 12 hours in a anaerobic incubator at 37 ℃, the viability was confirmed in BL agar.
내담즙성 실험 방법은, 0.05% L-cysteine-HCl이 함유된 pH 7.0의 50mM 인산 염 완충액에 0, 1.0%의 oxgall을 첨가한 후, 균체를 1×107-8 cfu/ml 수준으로 접종하여 37℃ 혐기배양기에 두고 12시간 처리 후, 생존성을 BL agar에서 확인하였다.The test method for biliary resistance was that 0, 1.0% of oxgall was added to 50 mM phosphate buffer at pH 7.0 containing 0.05% L-cysteine-HCl, and then the cells were inoculated at a level of 1 × 10 7-8 cfu / ml. After the treatment for 12 hours at 37 ℃ anaerobic incubator, the viability was confirmed in BL agar.
내산소성의 결과에서는 두 균주간에 차이가 없는 것으로 인정되었지만, 내산성과 내담즙성에서는 비피도박테리움 애돌센티스 MAEIL-K8보다 비피도박테리움 인팬티스 MAEIL-K9이 매우 우수한 것으로 나타났다. In the results of oxygen resistance, it was recognized that there was no difference between the two strains, but it was found that Bifidobacterium infantis MAEIL-K9 was superior to Bifidobacterium adolescents MAEIL-K8 in acid resistance and bile resistance.
이상에서 살펴본 바와 같이, 본 발명에서 선발한 비피더스 유산균인 비피도 박테리움 인팬티스 MAEIL-K9(기탁번호: KCTC 10294BP)은 종래의 프로바이오틱 유산균의 기본적인 생리활성인 내산소성, 내산성, 내담즙성 및 우유 배양성 등이 뛰어나면서도 특히 장 정착성이 우수하고, 장에서의 높은 증식능을 갖고 있기 때문에 건강기능성 식품에 포함시킬 경우 매우 우수한 장 건강촉진 효과를 얻을 수 있다. As described above, the Bifidobacterium inpantis MAEIL-K9 (Accession No .: KCTC 10294BP), which is selected from the present invention, is an oxygen-resistant, acid-resistant, bile biliary which is a basic physiological activity of a conventional probiotic lactic acid bacterium. In addition to excellent sex and milk culture, but also excellent intestinal fixability, and high intestinal proliferation ability, when included in the functional foods can be very good intestinal health promoting effect.
Claims (3)
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| KR1020020056172A KR100865363B1 (en) | 2002-09-16 | 2002-09-16 | Bifidobacterium infantis MAEIL-K9 strain and health-promoting food containing the same |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| KR1020020056172A KR100865363B1 (en) | 2002-09-16 | 2002-09-16 | Bifidobacterium infantis MAEIL-K9 strain and health-promoting food containing the same |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| KR20040024739A KR20040024739A (en) | 2004-03-22 |
| KR100865363B1 true KR100865363B1 (en) | 2008-10-24 |
Family
ID=37327783
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| KR1020020056172A KR100865363B1 (en) | 2002-09-16 | 2002-09-16 | Bifidobacterium infantis MAEIL-K9 strain and health-promoting food containing the same |
Country Status (1)
| Country | Link |
|---|---|
| KR (1) | KR100865363B1 (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR101589464B1 (en) * | 2015-05-21 | 2016-02-01 | 주식회사 쎌바이오텍 | New bifidobacterium strain and nutraceutical composition for improving growth comprising the same |
| WO2016186246A1 (en) * | 2015-05-21 | 2016-11-24 | 주식회사 쎌바이오텍 | Bifidobacterium bifidum cbt bf3 strain for promoting growth and functional food composition containing same for promoting growth |
| WO2016186245A1 (en) * | 2015-05-21 | 2016-11-24 | 주식회사 쎌바이오텍 | Bifidobacterium longum cbt bg7 strain and functional food composition containing same for promoting growth |
| WO2016186244A1 (en) * | 2015-05-21 | 2016-11-24 | 주식회사 쎌바이오텍 | Bifidobacterium breve cbt br3 strain for promoting growth and functional food composition containing same for promoting growth |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR102164198B1 (en) * | 2020-01-16 | 2020-10-12 | 한국생명공학연구원 | Bifidobacterium longum subsp. infantis IN02 and Food composition comprising thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4870020A (en) * | 1985-02-28 | 1989-09-26 | Nestec S.A. | Preparation of compositions including acid-resistant bifidobacteria |
| KR0142615B1 (en) * | 1995-09-16 | 1998-07-01 | 이은선 | New Lactic Acid Bacteria with Excellent Acid Resistance and Cholesterol Degradation Isolated from Feces of Korean Infant and Their Use |
| KR100472865B1 (en) * | 2002-08-28 | 2005-03-10 | (주) 피엘바이오 | Korean Isolate Bifidobacterium infantis PL9506 with High Nutraceutical Activity |
-
2002
- 2002-09-16 KR KR1020020056172A patent/KR100865363B1/en active IP Right Grant
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4870020A (en) * | 1985-02-28 | 1989-09-26 | Nestec S.A. | Preparation of compositions including acid-resistant bifidobacteria |
| KR0142615B1 (en) * | 1995-09-16 | 1998-07-01 | 이은선 | New Lactic Acid Bacteria with Excellent Acid Resistance and Cholesterol Degradation Isolated from Feces of Korean Infant and Their Use |
| KR100472865B1 (en) * | 2002-08-28 | 2005-03-10 | (주) 피엘바이오 | Korean Isolate Bifidobacterium infantis PL9506 with High Nutraceutical Activity |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR101589464B1 (en) * | 2015-05-21 | 2016-02-01 | 주식회사 쎌바이오텍 | New bifidobacterium strain and nutraceutical composition for improving growth comprising the same |
| WO2016186246A1 (en) * | 2015-05-21 | 2016-11-24 | 주식회사 쎌바이오텍 | Bifidobacterium bifidum cbt bf3 strain for promoting growth and functional food composition containing same for promoting growth |
| WO2016186245A1 (en) * | 2015-05-21 | 2016-11-24 | 주식회사 쎌바이오텍 | Bifidobacterium longum cbt bg7 strain and functional food composition containing same for promoting growth |
| WO2016186244A1 (en) * | 2015-05-21 | 2016-11-24 | 주식회사 쎌바이오텍 | Bifidobacterium breve cbt br3 strain for promoting growth and functional food composition containing same for promoting growth |
| US10597740B2 (en) | 2015-05-21 | 2020-03-24 | Cell Biotech Co., Ltd. | Bifidobacterium longum CBT BG7 strain for promotion of growth and nutraceutical composition for promotion of growth containing the same |
| US11026983B2 (en) | 2015-05-21 | 2021-06-08 | Cell Biotech Co., Ltd. | Bifidobacterium breve CBT BR3 strain for promotion of growth and nutraceutical composition for promotion of growth containing the same |
Also Published As
| Publication number | Publication date |
|---|---|
| KR20040024739A (en) | 2004-03-22 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US20230034193A1 (en) | Bifidobacterium animalis subsp. lactis i797, method for separation and purification thereof, and use thereof | |
| CN109929773B (en) | Bifidobacterium capable of being used for selenium-rich culture and active protein and application thereof | |
| US11026983B2 (en) | Bifidobacterium breve CBT BR3 strain for promotion of growth and nutraceutical composition for promotion of growth containing the same | |
| CN110106119B (en) | Lactobacillus rhamnosus M9 separated from breast milk and application thereof | |
| CN116676225B (en) | Lactobacillus rhamnosus LR-28 strain with nerve soothing and sleep aiding effects, fermentation product, hypnotic fungus group mixture and application | |
| CN111528283B (en) | Application of lactobacillus rhamnosus X253 with anti-fatigue effect and capability of improving body fatigue tolerance | |
| CN116103208A (en) | Application of lactobacillus salivarius in antioxidation | |
| CN113249244B (en) | Lactobacillus paracasei for antagonizing pharyngitis pathogenic bacteria beta hemolytic streptococcus | |
| KR100865363B1 (en) | Bifidobacterium infantis MAEIL-K9 strain and health-promoting food containing the same | |
| KR19990079232A (en) | Lactobacillus plantarum pmeo 08 (KF-11028) microbe having cholesterol lowering ability | |
| JP2019517810A (en) | Bacillus licheniformis NY1505 strain producing a large amount of α-glucosidase inhibitor | |
| KR100609779B1 (en) | Lactic acid bacteria degrading alcohol and acetaldehyde | |
| CN113881592B (en) | Lactobacillus reuteri and application thereof | |
| CN112080449B (en) | Enterococcus faecium R40 and application thereof in cholesterol reduction, exopolysaccharide production and antioxidation | |
| KR100725012B1 (en) | The new lactobacillus acidophilus HY 7036 isolated from korean adult, having high constipation improvement effect, high relieving intestinal disorders and high organic acid producibility, and products containing it | |
| KR20020072807A (en) | Novel lactobacillus sp. strain having the effect of immune enhancement | |
| KR100299306B1 (en) | Lactic acid bacteria with high acid resistance and growth rate and excellent blood cholesterol lowering ability | |
| RU2506308C1 (en) | CONSORTIUM OF PROBIOTIC STRAINS OF Lactobacillus rhamnosus AND Lactobacillus plantarum FOR PRODUCING BACTERIAL PREPARATION AND DIRECT ADMINISTRATION FERMENT FOR PRODUCING FERMENTED MILK AND FERMENTED BEET JUICE | |
| CN112322531A (en) | Production method and application of high-activity lactobacillus acidophilus freeze-dried powder | |
| KR101726207B1 (en) | --glucosidase LT19-2 [KCTC18497P]Novel Bifidobacterium animalis LT19-2 strain[KCTC18497P] having excellent -glucosidase activity | |
| KR100218233B1 (en) | Medium for high concentration culture of bifidus | |
| WO2024090413A1 (en) | Lactic acid bacterium, natural immunoactivator derived from said lactic acid bacterium, and food containing said lactic acid bacterium | |
| WO2024090415A1 (en) | Lactic acid bacteria, innate immunity activator derived from said lactic acid bacteria, and food containing said lactic acid bacteria | |
| KR0142615B1 (en) | New Lactic Acid Bacteria with Excellent Acid Resistance and Cholesterol Degradation Isolated from Feces of Korean Infant and Their Use | |
| WO2024090414A1 (en) | Lactic acid bacterium, natural immunoactivating agent derived from said lactic acid bacterium, and food containing said lactic acid bacterium |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| A201 | Request for examination | ||
| E902 | Notification of reason for refusal | ||
| E701 | Decision to grant or registration of patent right | ||
| GRNT | Written decision to grant | ||
| FPAY | Annual fee payment |
Payment date: 20121016 Year of fee payment: 5 |
|
| FPAY | Annual fee payment |
Payment date: 20131011 Year of fee payment: 6 |
|
| FPAY | Annual fee payment |
Payment date: 20141002 Year of fee payment: 7 |
|
| FPAY | Annual fee payment |
Payment date: 20151006 Year of fee payment: 8 |
|
| FPAY | Annual fee payment |
Payment date: 20161005 Year of fee payment: 9 |
|
| FPAY | Annual fee payment |
Payment date: 20171012 Year of fee payment: 10 |
|
| FPAY | Annual fee payment |
Payment date: 20181017 Year of fee payment: 11 |
|
| FPAY | Annual fee payment |
Payment date: 20191008 Year of fee payment: 12 |