CN117535176A - Bifidobacterium animalis subspecies lactis TA-2688 and application thereof in constipation improvement, intestinal mucosa barrier repair and immunity adjustment - Google Patents
Bifidobacterium animalis subspecies lactis TA-2688 and application thereof in constipation improvement, intestinal mucosa barrier repair and immunity adjustment Download PDFInfo
- Publication number
- CN117535176A CN117535176A CN202311341637.2A CN202311341637A CN117535176A CN 117535176 A CN117535176 A CN 117535176A CN 202311341637 A CN202311341637 A CN 202311341637A CN 117535176 A CN117535176 A CN 117535176A
- Authority
- CN
- China
- Prior art keywords
- product
- bifidobacterium animalis
- animalis subspecies
- intestinal
- lactis
- 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.)
- Pending
Links
- 241001134770 Bifidobacterium animalis Species 0.000 title claims abstract description 46
- 229940118852 bifidobacterium animalis Drugs 0.000 title claims abstract description 46
- 206010010774 Constipation Diseases 0.000 title claims abstract description 24
- 230000036039 immunity Effects 0.000 title claims abstract description 11
- 230000004888 barrier function Effects 0.000 title abstract description 11
- 210000004347 intestinal mucosa Anatomy 0.000 title abstract description 10
- 230000006872 improvement Effects 0.000 title description 4
- 230000008439 repair process Effects 0.000 title description 3
- QZAYGJVTTNCVMB-UHFFFAOYSA-N serotonin Chemical compound C1=C(O)C=C2C(CCN)=CNC2=C1 QZAYGJVTTNCVMB-UHFFFAOYSA-N 0.000 claims abstract description 54
- 230000000968 intestinal effect Effects 0.000 claims abstract description 28
- 230000008855 peristalsis Effects 0.000 claims abstract description 16
- 229940076279 serotonin Drugs 0.000 claims abstract description 13
- 210000005027 intestinal barrier Anatomy 0.000 claims abstract description 3
- 239000000047 product Substances 0.000 claims description 70
- 230000014509 gene expression Effects 0.000 claims description 28
- 239000001963 growth medium Substances 0.000 claims description 16
- 239000002207 metabolite Substances 0.000 claims description 15
- 108050009302 Claudin Proteins 0.000 claims description 14
- LVRVABPNVHYXRT-BQWXUCBYSA-N 52906-92-0 Chemical compound C([C@H](N)C(=O)N[C@H](C(=O)N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)NCC(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCCCN)C(=O)NCC(=O)N[C@@H](CCC(N)=O)C(O)=O)C(C)C)C1=CC=CC=C1 LVRVABPNVHYXRT-BQWXUCBYSA-N 0.000 claims description 13
- 102000002419 Motilin Human genes 0.000 claims description 13
- 101800002372 Motilin Proteins 0.000 claims description 13
- 102000002029 Claudin Human genes 0.000 claims description 12
- 238000000034 method Methods 0.000 claims description 12
- 230000001976 improved effect Effects 0.000 claims description 11
- 239000006228 supernatant Substances 0.000 claims description 11
- 102000003940 Occludin Human genes 0.000 claims description 10
- 108090000304 Occludin Proteins 0.000 claims description 10
- 241000901050 Bifidobacterium animalis subsp. lactis Species 0.000 claims description 9
- 102000003777 Interleukin-1 beta Human genes 0.000 claims description 9
- 108090000193 Interleukin-1 beta Proteins 0.000 claims description 9
- 238000004519 manufacturing process Methods 0.000 claims description 8
- 230000001737 promoting effect Effects 0.000 claims description 7
- 241000186660 Lactobacillus Species 0.000 claims description 6
- 210000003608 fece Anatomy 0.000 claims description 6
- 230000000770 proinflammatory effect Effects 0.000 claims description 6
- 102000011632 Caseins Human genes 0.000 claims description 5
- 108010076119 Caseins Proteins 0.000 claims description 5
- 239000005018 casein Substances 0.000 claims description 5
- BECPQYXYKAMYBN-UHFFFAOYSA-N casein, tech. Chemical compound NCCCCC(C(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(CC(C)C)N=C(O)C(CCC(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(C(C)O)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(COP(O)(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(N)CC1=CC=CC=C1 BECPQYXYKAMYBN-UHFFFAOYSA-N 0.000 claims description 5
- 235000021240 caseins Nutrition 0.000 claims description 5
- 108090000765 processed proteins & peptides Proteins 0.000 claims description 5
- 238000009630 liquid culture Methods 0.000 claims description 4
- 238000004321 preservation Methods 0.000 claims description 4
- 230000004913 activation Effects 0.000 claims description 3
- 239000012530 fluid Substances 0.000 claims description 3
- 229940039696 lactobacillus Drugs 0.000 claims description 3
- 230000004673 intestinal mucosal barrier function Effects 0.000 claims 1
- 230000006870 function Effects 0.000 abstract description 11
- 230000001105 regulatory effect Effects 0.000 abstract description 10
- 238000002360 preparation method Methods 0.000 abstract description 9
- 210000004369 blood Anatomy 0.000 abstract description 8
- 239000008280 blood Substances 0.000 abstract description 8
- 206010020772 Hypertension Diseases 0.000 abstract description 7
- 239000002253 acid Substances 0.000 abstract description 6
- 244000005700 microbiome Species 0.000 abstract description 5
- 208000024891 symptom Diseases 0.000 abstract description 5
- 239000003629 gastrointestinal hormone Substances 0.000 abstract description 3
- 230000007358 intestinal barrier function Effects 0.000 abstract description 3
- 239000002243 precursor Substances 0.000 abstract description 3
- 230000028327 secretion Effects 0.000 abstract description 3
- 239000000843 powder Substances 0.000 description 46
- 241001465754 Metazoa Species 0.000 description 44
- 241000186000 Bifidobacterium Species 0.000 description 34
- 241000699670 Mus sp. Species 0.000 description 21
- 239000007788 liquid Substances 0.000 description 16
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 16
- 238000002156 mixing Methods 0.000 description 15
- 239000002775 capsule Substances 0.000 description 14
- 210000001035 gastrointestinal tract Anatomy 0.000 description 14
- 235000013305 food Nutrition 0.000 description 11
- 241000894006 Bacteria Species 0.000 description 10
- 241000699666 Mus <mouse, genus> Species 0.000 description 10
- RDOIQAHITMMDAJ-UHFFFAOYSA-N loperamide Chemical compound C=1C=CC=CC=1C(C=1C=CC=CC=1)(C(=O)N(C)C)CCN(CC1)CCC1(O)C1=CC=C(Cl)C=C1 RDOIQAHITMMDAJ-UHFFFAOYSA-N 0.000 description 10
- 229960001571 loperamide Drugs 0.000 description 10
- 239000013642 negative control Substances 0.000 description 10
- 239000003826 tablet Substances 0.000 description 10
- 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 9
- 239000008103 glucose Substances 0.000 description 9
- 235000013361 beverage Nutrition 0.000 description 8
- 239000002552 dosage form Substances 0.000 description 8
- 239000008187 granular material Substances 0.000 description 8
- 235000013402 health food Nutrition 0.000 description 8
- 229910052500 inorganic mineral Inorganic materials 0.000 description 8
- HQKMJHAJHXVSDF-UHFFFAOYSA-L magnesium stearate Chemical compound [Mg+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O HQKMJHAJHXVSDF-UHFFFAOYSA-L 0.000 description 8
- 235000010755 mineral Nutrition 0.000 description 8
- 239000011707 mineral Substances 0.000 description 8
- 229930003231 vitamin Natural products 0.000 description 8
- 239000011782 vitamin Substances 0.000 description 8
- 235000013343 vitamin Nutrition 0.000 description 8
- 229940088594 vitamin Drugs 0.000 description 8
- 210000001072 colon Anatomy 0.000 description 7
- 230000029087 digestion Effects 0.000 description 7
- 239000000243 solution Substances 0.000 description 7
- 206010016100 Faeces discoloured Diseases 0.000 description 6
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 6
- XUJNEKJLAYXESH-REOHCLBHSA-N L-Cysteine Chemical compound SC[C@H](N)C(O)=O XUJNEKJLAYXESH-REOHCLBHSA-N 0.000 description 6
- 229940009289 bifidobacterium lactis Drugs 0.000 description 6
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 6
- 230000000694 effects Effects 0.000 description 6
- 238000004806 packaging method and process Methods 0.000 description 6
- 210000001519 tissue Anatomy 0.000 description 6
- 238000005303 weighing Methods 0.000 description 6
- 230000001580 bacterial effect Effects 0.000 description 5
- 239000003833 bile salt Substances 0.000 description 5
- 229960002433 cysteine Drugs 0.000 description 5
- 235000013365 dairy product Nutrition 0.000 description 5
- 238000001514 detection method Methods 0.000 description 5
- 238000002474 experimental method Methods 0.000 description 5
- 238000000855 fermentation Methods 0.000 description 5
- FTSSQIKWUOOEGC-RULYVFMPSA-N fructooligosaccharide Chemical compound OC[C@H]1O[C@@](CO)(OC[C@@]2(OC[C@@]3(OC[C@@]4(OC[C@@]5(OC[C@@]6(OC[C@@]7(OC[C@@]8(OC[C@@]9(OC[C@@]%10(OC[C@@]%11(O[C@H]%12O[C@H](CO)[C@@H](O)[C@H](O)[C@H]%12O)O[C@H](CO)[C@@H](O)[C@@H]%11O)O[C@H](CO)[C@@H](O)[C@@H]%10O)O[C@H](CO)[C@@H](O)[C@@H]9O)O[C@H](CO)[C@@H](O)[C@@H]8O)O[C@H](CO)[C@@H](O)[C@@H]7O)O[C@H](CO)[C@@H](O)[C@@H]6O)O[C@H](CO)[C@@H](O)[C@@H]5O)O[C@H](CO)[C@@H](O)[C@@H]4O)O[C@H](CO)[C@@H](O)[C@@H]3O)O[C@H](CO)[C@@H](O)[C@@H]2O)[C@@H](O)[C@@H]1O FTSSQIKWUOOEGC-RULYVFMPSA-N 0.000 description 5
- 229940107187 fructooligosaccharide Drugs 0.000 description 5
- 235000021255 galacto-oligosaccharides Nutrition 0.000 description 5
- 150000003271 galactooligosaccharides Chemical class 0.000 description 5
- 235000013336 milk Nutrition 0.000 description 5
- 239000008267 milk Substances 0.000 description 5
- 210000004080 milk Anatomy 0.000 description 5
- 239000000203 mixture Substances 0.000 description 5
- 239000000523 sample Substances 0.000 description 5
- 239000007787 solid Substances 0.000 description 5
- LDCYZAJDBXYCGN-VIFPVBQESA-N 5-hydroxy-L-tryptophan Chemical compound C1=C(O)C=C2C(C[C@H](N)C(O)=O)=CNC2=C1 LDCYZAJDBXYCGN-VIFPVBQESA-N 0.000 description 4
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical group OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 4
- 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 description 4
- 208000031226 Hyperlipidaemia Diseases 0.000 description 4
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 4
- 229930195725 Mannitol Natural products 0.000 description 4
- 239000001888 Peptone Substances 0.000 description 4
- 108010080698 Peptones Proteins 0.000 description 4
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 4
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 4
- VLSOAXRVHARBEQ-UHFFFAOYSA-N [4-fluoro-2-(hydroxymethyl)phenyl]methanol Chemical compound OCC1=CC=C(F)C=C1CO VLSOAXRVHARBEQ-UHFFFAOYSA-N 0.000 description 4
- 235000015278 beef Nutrition 0.000 description 4
- 229940041514 candida albicans extract Drugs 0.000 description 4
- 230000001684 chronic effect Effects 0.000 description 4
- -1 compound vitamin Chemical class 0.000 description 4
- 238000001816 cooling Methods 0.000 description 4
- XUJNEKJLAYXESH-UHFFFAOYSA-N cysteine Natural products SCC(N)C(O)=O XUJNEKJLAYXESH-UHFFFAOYSA-N 0.000 description 4
- 235000018417 cysteine Nutrition 0.000 description 4
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 description 4
- 239000012153 distilled water Substances 0.000 description 4
- 229940079593 drug Drugs 0.000 description 4
- 239000003814 drug Substances 0.000 description 4
- 239000003623 enhancer Substances 0.000 description 4
- 230000002550 fecal effect Effects 0.000 description 4
- 238000011049 filling Methods 0.000 description 4
- 238000010438 heat treatment Methods 0.000 description 4
- 201000001421 hyperglycemia Diseases 0.000 description 4
- 235000019359 magnesium stearate Nutrition 0.000 description 4
- 239000000594 mannitol Substances 0.000 description 4
- 235000010355 mannitol Nutrition 0.000 description 4
- 108020004999 messenger RNA Proteins 0.000 description 4
- 235000016709 nutrition Nutrition 0.000 description 4
- 230000035764 nutrition Effects 0.000 description 4
- 235000019319 peptone Nutrition 0.000 description 4
- 239000000825 pharmaceutical preparation Substances 0.000 description 4
- 229940127557 pharmaceutical product Drugs 0.000 description 4
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 description 4
- 229920000053 polysorbate 80 Polymers 0.000 description 4
- 235000013406 prebiotics Nutrition 0.000 description 4
- 239000006041 probiotic Substances 0.000 description 4
- 230000000529 probiotic effect Effects 0.000 description 4
- 235000018291 probiotics Nutrition 0.000 description 4
- 210000002966 serum Anatomy 0.000 description 4
- 235000020183 skimmed milk Nutrition 0.000 description 4
- 239000007901 soft capsule Substances 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- 239000012138 yeast extract Substances 0.000 description 4
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- 229920000168 Microcrystalline cellulose Polymers 0.000 description 3
- 229920002472 Starch Polymers 0.000 description 3
- 238000007664 blowing Methods 0.000 description 3
- 239000011575 calcium Substances 0.000 description 3
- 239000006285 cell suspension Substances 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 235000020247 cow milk Nutrition 0.000 description 3
- 230000013872 defecation Effects 0.000 description 3
- 239000000796 flavoring agent Substances 0.000 description 3
- 235000013355 food flavoring agent Nutrition 0.000 description 3
- 239000007902 hard capsule Substances 0.000 description 3
- 238000011534 incubation Methods 0.000 description 3
- XFIKMPRBSORKQP-JATHGWPISA-M lithium;9-[(e)-4-[(2s,3r,4r,5s)-3,4-dihydroxy-5-[[(2s,3s)-3-[(2s,3s)-3-hydroxybutan-2-yl]oxiran-2-yl]methyl]oxan-2-yl]-3-methylbut-2-enoyl]oxynonanoate Chemical class [Li+].C[C@H](O)[C@H](C)[C@@H]1O[C@H]1C[C@@H]1[C@@H](O)[C@@H](O)[C@H](C\C(C)=C\C(=O)OCCCCCCCCC([O-])=O)OC1 XFIKMPRBSORKQP-JATHGWPISA-M 0.000 description 3
- 239000002609 medium Substances 0.000 description 3
- 239000008108 microcrystalline cellulose Substances 0.000 description 3
- 235000019813 microcrystalline cellulose Nutrition 0.000 description 3
- 229940016286 microcrystalline cellulose Drugs 0.000 description 3
- 210000000813 small intestine Anatomy 0.000 description 3
- 239000001632 sodium acetate Substances 0.000 description 3
- 235000017281 sodium acetate Nutrition 0.000 description 3
- 239000008107 starch Substances 0.000 description 3
- 235000019698 starch Nutrition 0.000 description 3
- 239000000725 suspension Substances 0.000 description 3
- 238000005406 washing Methods 0.000 description 3
- 235000013618 yogurt Nutrition 0.000 description 3
- PVXPPJIGRGXGCY-TZLCEDOOSA-N 6-O-alpha-D-glucopyranosyl-D-fructofuranose 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(O)(CO)O1 PVXPPJIGRGXGCY-TZLCEDOOSA-N 0.000 description 2
- 244000215068 Acacia senegal Species 0.000 description 2
- 229920001817 Agar Polymers 0.000 description 2
- 102100028292 Aladin Human genes 0.000 description 2
- 101710065039 Aladin Proteins 0.000 description 2
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical class [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 2
- ZZZCUOFIHGPKAK-UHFFFAOYSA-N D-erythro-ascorbic acid Natural products OCC1OC(=O)C(O)=C1O ZZZCUOFIHGPKAK-UHFFFAOYSA-N 0.000 description 2
- 229920000084 Gum arabic Polymers 0.000 description 2
- FYYHWMGAXLPEAU-UHFFFAOYSA-N Magnesium Chemical compound [Mg] FYYHWMGAXLPEAU-UHFFFAOYSA-N 0.000 description 2
- 229920002774 Maltodextrin Polymers 0.000 description 2
- 239000005913 Maltodextrin Substances 0.000 description 2
- 229920000057 Mannan Polymers 0.000 description 2
- BUGBHKTXTAQXES-UHFFFAOYSA-N Selenium Chemical compound [Se] BUGBHKTXTAQXES-UHFFFAOYSA-N 0.000 description 2
- 229930006000 Sucrose Natural products 0.000 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 description 2
- 229930003270 Vitamin B Chemical group 0.000 description 2
- 229930003268 Vitamin C Natural products 0.000 description 2
- 229930003316 Vitamin D Natural products 0.000 description 2
- QYSXJUFSXHHAJI-XFEUOLMDSA-N Vitamin D3 Natural products C1(/[C@@H]2CC[C@@H]([C@]2(CCC1)C)[C@H](C)CCCC(C)C)=C/C=C1\C[C@@H](O)CCC1=C QYSXJUFSXHHAJI-XFEUOLMDSA-N 0.000 description 2
- HCHKCACWOHOZIP-UHFFFAOYSA-N Zinc Chemical compound [Zn] HCHKCACWOHOZIP-UHFFFAOYSA-N 0.000 description 2
- 239000000205 acacia gum Substances 0.000 description 2
- 235000010489 acacia gum Nutrition 0.000 description 2
- 239000008272 agar Substances 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 229910052791 calcium Inorganic materials 0.000 description 2
- 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 2
- RQFQJYYMBWVMQG-IXDPLRRUSA-N chitotriose Chemical compound O[C@@H]1[C@@H](N)[C@H](O)O[C@H](CO)[C@H]1O[C@H]1[C@H](N)[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O2)N)[C@@H](CO)O1 RQFQJYYMBWVMQG-IXDPLRRUSA-N 0.000 description 2
- 238000007796 conventional method Methods 0.000 description 2
- YXVFQADLFFNVDS-UHFFFAOYSA-N diammonium citrate Chemical compound [NH4+].[NH4+].[O-]C(=O)CC(O)(C(=O)O)CC([O-])=O YXVFQADLFFNVDS-UHFFFAOYSA-N 0.000 description 2
- 235000013325 dietary fiber Nutrition 0.000 description 2
- 230000001079 digestive effect Effects 0.000 description 2
- 230000004151 fermentation Effects 0.000 description 2
- 239000000945 filler Substances 0.000 description 2
- 238000001914 filtration Methods 0.000 description 2
- 238000009472 formulation Methods 0.000 description 2
- 238000007710 freezing Methods 0.000 description 2
- 230000008014 freezing Effects 0.000 description 2
- 238000003304 gavage Methods 0.000 description 2
- 238000003306 harvesting Methods 0.000 description 2
- 239000002054 inoculum Substances 0.000 description 2
- 210000000936 intestine Anatomy 0.000 description 2
- 229910052742 iron Inorganic materials 0.000 description 2
- 238000002955 isolation Methods 0.000 description 2
- 230000007774 longterm Effects 0.000 description 2
- 239000011777 magnesium Substances 0.000 description 2
- 229910052749 magnesium Inorganic materials 0.000 description 2
- 229940035034 maltodextrin Drugs 0.000 description 2
- LUEWUZLMQUOBSB-GFVSVBBRSA-N mannan Chemical class 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]2[C@H](O[C@@H](O[C@H]3[C@H](O[C@@H](O)[C@@H](O)[C@H]3O)CO)[C@@H](O)[C@H]2O)CO)[C@H](O)[C@H]1O LUEWUZLMQUOBSB-GFVSVBBRSA-N 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 238000009629 microbiological culture Methods 0.000 description 2
- 235000020124 milk-based beverage Nutrition 0.000 description 2
- 239000006872 mrs medium Substances 0.000 description 2
- 239000002245 particle Substances 0.000 description 2
- 239000001814 pectin Substances 0.000 description 2
- 235000010987 pectin Nutrition 0.000 description 2
- 229920001277 pectin Polymers 0.000 description 2
- 230000002035 prolonged effect Effects 0.000 description 2
- 239000003223 protective agent Substances 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- 210000001187 pylorus Anatomy 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 230000002040 relaxant effect Effects 0.000 description 2
- 229910052711 selenium Inorganic materials 0.000 description 2
- 239000011669 selenium Substances 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- 210000002784 stomach Anatomy 0.000 description 2
- 238000003860 storage Methods 0.000 description 2
- 239000005720 sucrose Substances 0.000 description 2
- 235000019156 vitamin B Nutrition 0.000 description 2
- 239000011720 vitamin B Chemical group 0.000 description 2
- 235000019154 vitamin C Nutrition 0.000 description 2
- 239000011718 vitamin C Substances 0.000 description 2
- 235000019166 vitamin D Nutrition 0.000 description 2
- 239000011710 vitamin D Substances 0.000 description 2
- 150000003710 vitamin D derivatives Chemical class 0.000 description 2
- 229940046008 vitamin d Drugs 0.000 description 2
- 150000003722 vitamin derivatives Chemical class 0.000 description 2
- 239000011701 zinc Substances 0.000 description 2
- 229910052725 zinc Inorganic materials 0.000 description 2
- HDTRYLNUVZCQOY-UHFFFAOYSA-N α-D-glucopyranosyl-α-D-glucopyranoside Natural products OC1C(O)C(O)C(CO)OC1OC1C(O)C(O)C(O)C(CO)O1 HDTRYLNUVZCQOY-UHFFFAOYSA-N 0.000 description 1
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 1
- IIZPXYDJLKNOIY-JXPKJXOSSA-N 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCC\C=C/C\C=C/C\C=C/C\C=C/CCCCC IIZPXYDJLKNOIY-JXPKJXOSSA-N 0.000 description 1
- 108020004465 16S ribosomal RNA Proteins 0.000 description 1
- 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 description 1
- 241000283690 Bos taurus Species 0.000 description 1
- YNXLOPYTAAFMTN-SBUIBGKBSA-N C([C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCCCN)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(=O)N1[C@@H](CCC1)C(=O)NCC(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CO)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H](C)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=1NC=NC=1)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(N)=O)C1=CC=C(O)C=C1 Chemical compound C([C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCCCN)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(=O)N1[C@@H](CCC1)C(=O)NCC(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CO)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H](C)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=1NC=NC=1)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(N)=O)C1=CC=C(O)C=C1 YNXLOPYTAAFMTN-SBUIBGKBSA-N 0.000 description 1
- 235000005976 Citrus sinensis Nutrition 0.000 description 1
- 240000002319 Citrus sinensis Species 0.000 description 1
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- 238000002965 ELISA Methods 0.000 description 1
- 238000008157 ELISA kit Methods 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- 229920002488 Hemicellulose Polymers 0.000 description 1
- 229920001202 Inulin Polymers 0.000 description 1
- 239000004201 L-cysteine Substances 0.000 description 1
- 235000013878 L-cysteine Nutrition 0.000 description 1
- 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 description 1
- JLVVSXFLKOJNIY-UHFFFAOYSA-N Magnesium ion Chemical compound [Mg+2] JLVVSXFLKOJNIY-UHFFFAOYSA-N 0.000 description 1
- 229920000881 Modified starch Polymers 0.000 description 1
- 238000010802 RNA extraction kit Methods 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- HDTRYLNUVZCQOY-WSWWMNSNSA-N Trehalose Natural products 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@@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-WSWWMNSNSA-N 0.000 description 1
- 206010047139 Vasoconstriction Diseases 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 210000000577 adipose tissue Anatomy 0.000 description 1
- 239000002671 adjuvant Substances 0.000 description 1
- HDTRYLNUVZCQOY-LIZSDCNHSA-N alpha,alpha-trehalose Chemical compound 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@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-LIZSDCNHSA-N 0.000 description 1
- 229940040526 anhydrous sodium acetate Drugs 0.000 description 1
- 229940093761 bile salts Drugs 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 230000017531 blood circulation Effects 0.000 description 1
- 230000037396 body weight Effects 0.000 description 1
- 229910001424 calcium ion Inorganic materials 0.000 description 1
- OKTJSMMVPCPJKN-YPZZEJLDSA-N carbon-10 atom Chemical compound [10C] OKTJSMMVPCPJKN-YPZZEJLDSA-N 0.000 description 1
- 230000020411 cell activation Effects 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000007910 chewable tablet Substances 0.000 description 1
- 229940068682 chewable tablet Drugs 0.000 description 1
- 239000002299 complementary DNA Substances 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 230000001276 controlling effect Effects 0.000 description 1
- 239000012228 culture supernatant Substances 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 239000012470 diluted sample Substances 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- PXEDJBXQKAGXNJ-QTNFYWBSSA-L disodium L-glutamate Chemical compound [Na+].[Na+].[O-]C(=O)[C@@H](N)CCC([O-])=O PXEDJBXQKAGXNJ-QTNFYWBSSA-L 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 239000003651 drinking water Substances 0.000 description 1
- 235000020188 drinking water Nutrition 0.000 description 1
- 230000005284 excitation Effects 0.000 description 1
- 235000021001 fermented dairy product Nutrition 0.000 description 1
- 239000000706 filtrate Substances 0.000 description 1
- 239000010794 food waste Substances 0.000 description 1
- 235000021433 fructose syrup Nutrition 0.000 description 1
- 235000011389 fruit/vegetable juice Nutrition 0.000 description 1
- 230000005176 gastrointestinal motility Effects 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 229920000591 gum Polymers 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 230000004957 immunoregulator effect Effects 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 206010022498 insulinoma Diseases 0.000 description 1
- 230000003871 intestinal function Effects 0.000 description 1
- JYJIGFIDKWBXDU-MNNPPOADSA-N inulin Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)OC[C@]1(OC[C@]2(OC[C@]3(OC[C@]4(OC[C@]5(OC[C@]6(OC[C@]7(OC[C@]8(OC[C@]9(OC[C@]%10(OC[C@]%11(OC[C@]%12(OC[C@]%13(OC[C@]%14(OC[C@]%15(OC[C@]%16(OC[C@]%17(OC[C@]%18(OC[C@]%19(OC[C@]%20(OC[C@]%21(OC[C@]%22(OC[C@]%23(OC[C@]%24(OC[C@]%25(OC[C@]%26(OC[C@]%27(OC[C@]%28(OC[C@]%29(OC[C@]%30(OC[C@]%31(OC[C@]%32(OC[C@]%33(OC[C@]%34(OC[C@]%35(OC[C@]%36(O[C@@H]%37[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O%37)O)[C@H]([C@H](O)[C@@H](CO)O%36)O)[C@H]([C@H](O)[C@@H](CO)O%35)O)[C@H]([C@H](O)[C@@H](CO)O%34)O)[C@H]([C@H](O)[C@@H](CO)O%33)O)[C@H]([C@H](O)[C@@H](CO)O%32)O)[C@H]([C@H](O)[C@@H](CO)O%31)O)[C@H]([C@H](O)[C@@H](CO)O%30)O)[C@H]([C@H](O)[C@@H](CO)O%29)O)[C@H]([C@H](O)[C@@H](CO)O%28)O)[C@H]([C@H](O)[C@@H](CO)O%27)O)[C@H]([C@H](O)[C@@H](CO)O%26)O)[C@H]([C@H](O)[C@@H](CO)O%25)O)[C@H]([C@H](O)[C@@H](CO)O%24)O)[C@H]([C@H](O)[C@@H](CO)O%23)O)[C@H]([C@H](O)[C@@H](CO)O%22)O)[C@H]([C@H](O)[C@@H](CO)O%21)O)[C@H]([C@H](O)[C@@H](CO)O%20)O)[C@H]([C@H](O)[C@@H](CO)O%19)O)[C@H]([C@H](O)[C@@H](CO)O%18)O)[C@H]([C@H](O)[C@@H](CO)O%17)O)[C@H]([C@H](O)[C@@H](CO)O%16)O)[C@H]([C@H](O)[C@@H](CO)O%15)O)[C@H]([C@H](O)[C@@H](CO)O%14)O)[C@H]([C@H](O)[C@@H](CO)O%13)O)[C@H]([C@H](O)[C@@H](CO)O%12)O)[C@H]([C@H](O)[C@@H](CO)O%11)O)[C@H]([C@H](O)[C@@H](CO)O%10)O)[C@H]([C@H](O)[C@@H](CO)O9)O)[C@H]([C@H](O)[C@@H](CO)O8)O)[C@H]([C@H](O)[C@@H](CO)O7)O)[C@H]([C@H](O)[C@@H](CO)O6)O)[C@H]([C@H](O)[C@@H](CO)O5)O)[C@H]([C@H](O)[C@@H](CO)O4)O)[C@H]([C@H](O)[C@@H](CO)O3)O)[C@H]([C@H](O)[C@@H](CO)O2)O)[C@@H](O)[C@H](O)[C@@H](CO)O1 JYJIGFIDKWBXDU-MNNPPOADSA-N 0.000 description 1
- 229940029339 inulin Drugs 0.000 description 1
- DLRVVLDZNNYCBX-RTPHMHGBSA-N isomaltose 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(O)O1 DLRVVLDZNNYCBX-RTPHMHGBSA-N 0.000 description 1
- 239000008101 lactose Substances 0.000 description 1
- 235000010445 lecithin Nutrition 0.000 description 1
- 239000000787 lecithin Substances 0.000 description 1
- 229940067606 lecithin Drugs 0.000 description 1
- 239000008176 lyophilized powder Substances 0.000 description 1
- 229910001425 magnesium ion Inorganic materials 0.000 description 1
- WRUGWIBCXHJTDG-UHFFFAOYSA-L magnesium sulfate heptahydrate Chemical compound O.O.O.O.O.O.O.[Mg+2].[O-]S([O-])(=O)=O WRUGWIBCXHJTDG-UHFFFAOYSA-L 0.000 description 1
- 229940061634 magnesium sulfate heptahydrate Drugs 0.000 description 1
- 229940099596 manganese sulfate Drugs 0.000 description 1
- 239000011702 manganese sulphate Substances 0.000 description 1
- 235000007079 manganese sulphate Nutrition 0.000 description 1
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 description 1
- ISPYRSDWRDQNSW-UHFFFAOYSA-L manganese(II) sulfate monohydrate Chemical compound O.[Mn+2].[O-]S([O-])(=O)=O ISPYRSDWRDQNSW-UHFFFAOYSA-L 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 230000004630 mental health Effects 0.000 description 1
- 210000000713 mesentery Anatomy 0.000 description 1
- 208000030159 metabolic disease Diseases 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 235000013923 monosodium glutamate Nutrition 0.000 description 1
- 230000008621 organismal health Effects 0.000 description 1
- 230000003204 osmotic effect Effects 0.000 description 1
- 239000012057 packaged powder Substances 0.000 description 1
- 208000021255 pancreatic insulinoma Diseases 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 238000003825 pressing Methods 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000010079 rubber tapping Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 239000007974 sodium acetate buffer Substances 0.000 description 1
- 229940073490 sodium glutamate Drugs 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 230000004936 stimulating effect Effects 0.000 description 1
- 208000011580 syndromic disease Diseases 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 239000006188 syrup Substances 0.000 description 1
- 235000020357 syrup Nutrition 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 239000003053 toxin Substances 0.000 description 1
- 231100000765 toxin Toxicity 0.000 description 1
- 238000009777 vacuum freeze-drying Methods 0.000 description 1
- 230000025033 vasoconstriction Effects 0.000 description 1
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
- C12N1/205—Bacterial isolates
-
- 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
- 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
- A61K35/745—Bifidobacteria
-
- 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
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/10—Laxatives
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/02—Immunomodulators
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P1/00—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes
- C12P1/04—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes by using bacteria
-
- 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
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Medicinal Chemistry (AREA)
- General Chemical & Material Sciences (AREA)
- Mycology (AREA)
- Veterinary Medicine (AREA)
- Pharmacology & Pharmacy (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- Microbiology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Genetics & Genomics (AREA)
- Biotechnology (AREA)
- Immunology (AREA)
- General Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- Virology (AREA)
- Epidemiology (AREA)
- Molecular Biology (AREA)
- Biomedical Technology (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
- Nutrition Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Tropical Medicine & Parasitology (AREA)
Abstract
The invention relates to the technical field of microorganisms, in particular to bifidobacterium animalis subspecies lactis TA-2688 and application thereof in preparation of products for improving constipation, repairing intestinal mucosa barriers and regulating immunity. The strain can promote the generation of 5-hydroxytryptamine precursor substance-serotonin acid, regulate and control the secretion of gastrointestinal hormone, the intestinal peristalsis is mildly promoted, so that constipation symptoms of middle-aged and elderly people with high blood pressure, high blood sugar and high blood sugar are relieved, and meanwhile, the functions of improving intestinal barrier and regulating immunity are also realized.
Description
Technical Field
The invention relates to the technical field of microorganisms, in particular to an animal bifidobacterium subspecies lactis TA-2688 and application thereof in constipation improvement, intestinal mucosa barrier repair and immunity adjustment.
Background
Chronic constipation is a common middle-aged and elderly syndrome, and is mainly manifested by reduced defecation times, dry stool and/or difficult defecation. The prevalence of chronic constipation increases year by year with age in the middle-aged and elderly people. Hyperglycemia, hypertension and hyperlipidemia are common chronic metabolic diseases of middle-aged and elderly people, and research shows that the people suffering from hypertension, hyperglycemia and hyperlipidemia are more prone to constipation, because hyperglycemia can cause osmotic pressure in intestinal tracts to be increased, moisture in the intestinal tracts is absorbed, and excrement is dry and difficult to discharge. In addition, hyperglycemia affects the peristalsis and digestive functions of the intestines, so that food residues stay in the intestines for a prolonged period of time, and the risk of constipation is increased. Hypertension can cause vasoconstriction, affect the blood circulation of the intestinal tract, and slow down intestinal peristalsis. In addition, hypertension can also cause damage to intestinal mucosa, so that the number of bacteria in the intestinal tract is increased, and normal peristalsis and digestion functions of the intestinal tract are affected. Hyperlipidemia can cause fat accumulation in the intestinal tract, affecting the normal peristalsis of the intestinal tract. In addition, hyperlipidemia can cause damage to intestinal mucosa, so that the number of bacteria in the intestinal tract is increased, and normal peristalsis and digestion functions of the intestinal tract are affected. Chronic constipation not only seriously affects the quality of life and physical and mental health of middle-aged and elderly people, but also consumes a great deal of medical expenses.
The use of the cathartic is a main treatment mode for relieving constipation of middle-aged and elderly people at present, but long-term use of the cathartic can influence barrier function and intestinal microecological balance of intestinal tracts, and long-term use of the cathartic can cause colon blackening disease, so that constipation is more serious. Therefore, there is a need to develop a gentle, healthy and effective method to fundamentally improve the intestinal function of the middle aged and elderly people, thereby alleviating the constipation problem.
Disclosure of Invention
Aiming at the technical problems, the invention provides an animal bifidobacterium subspecies TA-2688 strain and application thereof in preparing products for improving constipation, repairing intestinal mucosa barrier and regulating immunity, the strain can promote the generation of 5-hydroxytryptamine precursor substance-serotonin, regulate and control secretion of gastrointestinal hormone, and mildly promote intestinal peristalsis, thereby relieving constipation symptoms of middle-aged and elderly people with high blood pressure, high blood sugar and high blood sugar, and simultaneously has the functions of improving intestinal barrier and regulating immunity.
In order to achieve the aim of the invention, the invention adopts the following technical scheme:
a bifidobacterium animalis subspecies lactis TA-2688, which is classified under the name of bifidobacterium animalis subsp.lactis, is preserved in China general microbiological culture collection center (CGMCC) No.27908 at the date of 7.14 in 2023; the preservation address is North Chen West Lu No.1 and 3 of the Korean area of Beijing, and the institute of microorganisms of the national academy of sciences.
Compared with other bifidobacterium animalis subspecies, the bifidobacterium animalis subspecies TA-2688 with the preservation number of CGMCC No.27908 has the following advantages:
(1) The animal bifidobacterium lactosub-species TA-2688 provided by the invention can promote the levels of Motilin (MTL) and 5-hydroxytryptamine (5-HT) and reduce the level of casein Peptide (PYY), thereby promoting intestinal peristalsis;
(2) Animal experiments prove that the animal bifidobacterium subspecies lactis TA-2688 provided by the invention can reduce the water content of excrement, promote intestinal peristalsis, keep the stool smooth and moist, and has good bowel relaxing effect;
(3) The animal bifidobacterium subspecies of the invention TA-2688 can improve the relative expression rate of mRNA of the intestinal canal compact connecting protein Occludin, claudin and improve the barrier function of the intestinal canal
(4) The animal bifidobacterium lactosub-species TA-2688 provided by the invention can also reduce the expression of the proinflammatory factor IL-1 beta, thereby generating an immunoregulatory function.
(5) The animal bifidobacterium lactosub-species TA-2688 provided by the invention has good acid resistance and bile salt resistance, and can still keep higher activity after being digested by gastrointestinal tracts.
In a second aspect, the invention also provides a metabolite of the bifidobacterium animalis subspecies lactobacilli TA-2688.
In a third aspect, the invention also provides a preparation method of the animal bifidobacterium subspecies lactis TA-2688 metabolite, which specifically comprises the following operations: inoculating the bifidobacterium animalis subspecies of the Lactobacillus TA-2688 into an improved MRS liquid culture medium for activation, centrifuging and taking supernatant fluid to obtain the bifidobacterium animalis strain.
In combination with the third aspect, the components of the improved MRS liquid medium include: 18.0 to 22.0g/L of glucose, 9.0 to 11.0g/L of peptone, 6.0 to 7.0g/L of beef powder, 4.5 to 5.5g/L of yeast extract powder, 4.5 to 5.5g/L of sodium acetate, 1.8 to 2.2g/L of diammonium hydrogen citrate, 1.8 to 2.2g/L of dipotassium hydrogen phosphate and MgSO 4 ·7H 2 O0.52～0.64g/L，MnSO 4 ·H 2 0.23-0.27 g/L of O, 0.45-0.55 g/L of L-cysteine hydrochloride and 0.9-1.1 ml/L of Tween 80.
Preferably, the components of the improved MRS liquid culture medium include: glucose 20.0g/L, peptone 10.0g/L, beef powder 6.5g/L, yeast extract powder 5.0g/L, sodium acetate 5.0g/L, diammonium citrate 2.0g/L, dipotassium hydrogen phosphate 2.0g/L, mgSO 4 ·7H 2 O 0.58g/L，MnSO 4 ·H 2 O0.25 g/L, L-cysteine hydrochloride 0.5g/L and Tween 80 1.0ml/L.
In a fourth aspect, the invention also provides the use of the bifidobacterium animalis subspecies lactis TA-2688 in the preparation of a product for improving constipation.
With reference to the fourth aspect, the product comprises at least one of a viable cell or a metabolite of the bifidobacterium animalis subspecies lactis TA-2688.
With reference to the fourth aspect, the product is a product for increasing the moisture content of feces.
With reference to the fourth aspect, the product is a product that promotes intestinal peristalsis.
Preferably, the product is a product that promotes production of serotonin by cells.
Preferably, the product is a product that increases the expression of intestinal motilin.
Preferably, the product is a product that increases expression of 5 hydroxytryptamine.
Preferably, the product is a product that reduces the expression of casein peptides.
With reference to the fourth aspect, the product comprises a pharmaceutical product, a health food or a food. The dosage forms of the medicines and the health foods comprise conventional dosage forms such as powder, granules, capsules, tablets or oral liquid, and the forms of the foods comprise liquid beverages, solid beverages, dairy products, cold cakes and the like.
In a fifth aspect, the invention also provides application of the bifidobacterium animalis subspecies lactis TA-2688 in preparation of products for repairing intestinal mucosa barriers. Repairing intestinal mucosa barrier is not only beneficial to maintaining normal intestinal peristalsis and digestion, but also can prevent the organism from being affected by endogenous microorganism and toxin thereof.
With reference to the fifth aspect, the product comprises at least one of a viable cell or a metabolite of the bifidobacterium animalis subspecies lactis TA-2688.
With reference to the fifth aspect, the product is a product that promotes expression of intestinal claudin Occludin.
With reference to the fifth aspect, the product is a product that promotes expression of intestinal Claudin.
With reference to the fifth aspect, the product includes a pharmaceutical product, a health food or a food. The dosage forms of the medicines and the health foods comprise conventional dosage forms such as powder, granules, capsules, tablets or oral liquid, and the forms of the foods comprise liquid beverages, solid beverages, dairy products, cold cakes and the like.
In a sixth aspect, the invention also provides the use of the bifidobacterium animalis subspecies lactis TA-2688 in the preparation of products for regulating immunity.
With reference to the sixth aspect, the product comprises at least one of a viable cell or a metabolite of the bifidobacterium animalis subspecies lactis TA-2688.
With reference to the sixth aspect, the product is a product that reduces the expression of the pro-inflammatory factor IL-1 beta.
With reference to the sixth aspect, the product includes a pharmaceutical product, a health food or a food product. The dosage forms of the medicines and the health foods comprise conventional dosage forms such as powder, granules, capsules, tablets or oral liquid, and the forms of the foods comprise liquid beverages, solid beverages, dairy products, cold cakes and the like.
Drawings
FIG. 1 shows the tolerance of bifidobacterium animalis subspecies lactis TA-2688 in example 2 at pH3.0 and 0.3% bile salts;
FIG. 2 shows the content of serotonin produced by RIN14B cells of different groups in example 3.
Detailed Description
The present invention will be described in further detail with reference to the following examples in order to make the objects, technical solutions and advantages of the present invention more apparent. It should be understood that the specific embodiments described herein are for purposes of illustration only and are not intended to limit the scope of the invention.
The animal bifidobacterium subspecies TA-2688 provided by the invention is from intestinal flora of healthy infants, can promote the generation of 5-hydroxytryptamine precursor substance-serotonin, can promote MTL and 5-HT levels, reduce PYY levels, mildly promote intestinal peristalsis and can also improve the water content of excrement, thereby effectively relieving constipation symptoms of middle-aged and elderly people with high blood pressure, high blood sugar and high blood sugar; meanwhile, the strain can also improve the relative expression rate of mRNA of the intestinal canal compact connecting protein Occludin, claudin and reduce the expression of the proinflammatory factor IL-1 beta, thereby having the functions of improving intestinal canal barrier and regulating immunity and being beneficial to maintaining the health of organisms.
Based on the functions, the bifidobacterium animalis subspecies of the TA-2688 strain can be prepared into products for improving constipation, repairing intestinal mucosa barriers and regulating immunity, or products with two or more functions. The forms of the product include, but are not limited to, a pharmaceutical product, a health food or a food product. The dosage forms of the medicines and the health foods comprise conventional dosage forms such as powder, granules, capsules, tablets or oral liquid, and the forms of the foods comprise liquid beverages, solid beverages, dairy products, cold cakes and the like.
When the product is prepared, other auxiliary materials are added according to the requirement, and the product is prepared by a conventional method.
For example, in the preparation of powders, the following methods may be used: weighing animal bifidobacterium lactobionic TA-2688 freeze-dried powder, prebiotics, dietary fibers, nutrition enhancers and the balance of food raw materials, mixing and packaging to obtain a powder product containing animal bifidobacterium lactobionic TA-2688, wherein packaging specifications can be adjusted according to actual requirements, for example, each independently packaged powder can contain 100-300 hundred million live bacteria of animal bifidobacterium lactobionic TA-2688. The prebiotics can be selected from at least one of fructo-oligosaccharide, galacto-oligosaccharide, isomaltulose, chitosan oligosaccharide or mannooligosaccharide, the dietary fiber can be selected from at least one of mannans, inulin, hemicellulose, pectin, gum and agar, the nutrition enhancer can be selected from vitamin powder, mineral powder, etc., the vitamins can be selected from vitamin C, vitamin B group, vitamin D, etc., the minerals can be selected from calcium, iron, magnesium, zinc, selenium, etc., the food raw material can be selected from at least one of milk powder or maltodextrin, and the flavoring agent can be added according to actual requirements.
In preparing the granule, the preparation method comprises the following steps: mixing the animal bifidobacterium lactosub-species TA-2688 freeze-dried powder, prebiotics, filling agent and nutrition enhancer, granulating by adopting a conventional method, adding a glidant, mixing, and sub-packaging to obtain a granular product containing the animal bifidobacterium lactosub-species TA-2688, wherein the sub-packaging specification can be adjusted according to actual requirements, for example, each independently packaged granular product can contain 100-300 hundred million live bacteria of the animal bifidobacterium lactosub-species TA-2688. The prebiotics can be selected from at least one of fructo-oligosaccharide, galacto-oligosaccharide, isomaltulose, chitosan oligosaccharide or mannooligosaccharide, the filler can be selected from at least one of mannitol, milk powder, maltodextrin, soluble starch and sugar powder, the nutrition enhancer can be selected from vitamin powder, mineral powder, etc., the vitamins can be selected from vitamin C, vitamin B group, vitamin D, etc., the minerals can be selected from calcium, iron, magnesium, zinc, selenium, etc., and the flavoring agent can be added according to actual requirements.
In preparing tablets, the following procedure may be used: mixing animal Bifidobacterium lactobionic TA-2688 lyophilized powder with common tablet adjuvants such as sucrose, lactose, pregelatinized starch, microcrystalline cellulose, mannitol, magnesium stearate, correctant, etc., and tabletting. The tablet weight and the tablet shape can be selected according to actual requirements during tabletting.
In preparing the capsule, the preparation method comprises the following steps: the hard capsule can be prepared into powder or granule from animal bifidobacterium lactobionic TA-2688, and then prepared into capsule products by a capsule machine; the soft capsule can be prepared by mixing animal Bifidobacterium lactobionic TA-2688 powder with oil and fat, and making into soft capsule product by capsule mechanism. Both capsules can be specified according to gram weight or viable count.
In preparing a dairy product, the following method can be used:
fermented yogurt: heating pasteurized fresh cow milk or skim milk to 45-55 ℃, adding 2-5% of glucose, 0.2-0.5% of yeast autolysate and 0.04-0.06% of cysteine, completely dissolving, cooling to 37-42 ℃, inoculating 5-10% of animal bifidobacterium lactobionic TA-2688, carrying out anaerobic fermentation at 37-42 ℃ to pH 4.3-4.5, and carrying out cold storage after-ripening at 2-7 ℃ to obtain the coagulated or stirred yogurt.
Milk beverage: heating pasteurized fresh cow milk or skim milk to 45-55 ℃, adding 2-5% of glucose, 0.2-0.5% of yeast autolysate and 0.04-0.06% of cysteine, completely dissolving, cooling to 37-42 ℃, inoculating 5-10% of animal bifidobacterium lactobionic TA-2688, anaerobic fermenting to pH 4.3-4.5 at 37-42 ℃, mixing with water and glucose syrup, regulating acidity to pH 3.5-5.0 with citric acid, adding a flavoring agent, homogenizing under 10MPa pressure, and refrigerating after filling.
In the preparation of the product, the adopted bifidobacterium B11 freeze-dried powder can be prepared according to the following method:
inoculating the bifidobacterium animalis subspecies TA-2688 frozen and preserved at the temperature of minus 70 ℃ into a fresh and improved MRS culture medium, carrying out anaerobic culture at the temperature of 37 ℃ for 24 hours, and harvesting first-stage seeds; transferring the first-level seeds to a fresh improved MRS culture medium in an inoculum size of 5%, carrying out anaerobic culture for 24 hours at 37 ℃, and harvesting second-level seeds; transferring the second-level seed liquid to a fresh improved MRS culture medium according to an inoculum size of 5%, carrying out anaerobic culture for 20-24 h at 37 ℃, centrifuging for 3min at 12000g at 4 ℃, discarding the supernatant, washing bacterial mud for 2 times by using sterile physiological saline, uniformly mixing the bacterial mud with a protective agent according to a ratio of 1:2.5 (m/v), pre-freezing for 12h at-70 ℃, then placing in a freeze dryer for vacuum freeze-drying, and crushing to obtain bifidobacterium lactis subspecies TA-2688 freeze-dried powder. Wherein the protective agent can be prepared by the following method: 10 to 15g of skim milk powder, 4 to 9g of trehalose, 3 to 7g of sucrose, 1 to 2g of glycerol, 1 to 2g of gelatin, 1 to 2g of sodium glutamate, 0.1 to 0.2g of L-cysteine and 0.3 to 0.4g of manganese sulfate are uniformly mixed, and distilled water is added to be added to 100g of the mixture to obtain the compound.
The formulation and formulation of the media used in the following examples were as follows:
improvement of MRS culture medium: 20.0g of glucose, 10.0g of peptone, 6.5g of beef powder, 5.0g of yeast extract powder, 2.0g of diammonium hydrogen citrate, 5.0g of anhydrous sodium acetate, 2.0g of dipotassium hydrogen phosphate, 0.58g of magnesium sulfate heptahydrate, 0.25g of manganese sulfate monohydrate, 0.5g of L-cysteine hydrochloride, 1.0mL of Tween 80, 1000mL of water are fully dissolved, the pH is adjusted to 6.8 (15 g of agar powder is added to a solid culture medium), and the mixture is autoclaved at 115 ℃/20min after subpackaging.
MRS medium supplemented with mupirocin lithium salt: to every 100ml of MRS medium, 1 piece of mupirocin lithium salt (5 mg) was added.
Example 1
This example provides isolation, purification and identification of strains.
1.1 isolation and purification of Strain
Placing faeces sample of healthy infant in sterile container, adding normal saline, diluting and mixing, 10 times gradient dilution, selecting 10 -3 -10 -5 The diluted samples of (2) are respectively coated on an MRS culture medium and an MRS culture medium added with mupirocin lithium salt, anaerobically cultured for 48 hours at 37 ℃, and respectively spotted on the MRS culture medium for at least 3 times according to different colony forms until strains with uniform colony forms are obtained.
1.2 identification of strains
The isolated strain is subjected to 16SrRNA identification, the strain conforming to a food list is frozen, and the specific operation is that 600ul of seed liquid and 600ul of 30% glycerol (V: V=1:1) are uniformly mixed in a sterilized freezing tube, and then the strain is preserved at-70 ℃. Through examining the performance of each strain, one strain is found to have the performance of promoting cells to produce serotonin, regulating and controlling secretion of gastrointestinal hormone, promoting intestinal peristalsis and the like. The 16S rRNA sequence of the strain (shown as SEQ ID NO. 1) is as follows:
AGTTCCTGCGCGTGCTTACCATGCAAGTCGAACGGGATCCCTGGCAGCTTGCTGTCGGGGTGAGAGTGGCGAACGGGTGAGTAATGCGTGACCAACCTGCCCTGTGCACCGGAATAGCTCCTGGAAACGGGTGGTAATACCGGATGCTCCGCTCCATCGCATGGTGGGGTGGGAAATGCTTTTGCGGCATGGGATGGGGTCGCGTCCTATCAGCTTGTTGGCGGGGTGATGGCCCACCAAGGCGTTGACGGGTAGCCGGCCTGAGAGGGTGACCGGCCACATTGGGACTGAGATACGGCCCAGACTCCTACGGGAGGCAGCAGTGGGGAATATTGCACAATGGGCGCAAGCCTGATGCAGCGACGCCGCGTGCGGGATGGAGGCCTTCGGGTTGTAAACCGCTTTTGTTCAAGGGCAAGGCACGGTTTCGGCCGTGTTGAGTGGATTGTTCGAATAAGCACCGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGGTGCGAGCGTTATCCGGATTTATTGGGCGTAAAGGGCTCGTAGGCGGTTCGTCGCGTCCGGTGTGAAAGTCCATCGCCTAACGGTGGATCTGCGCCGGGTACGGGCGGGCTGGAGTGCGGTAGGGGAGACTGGAATTCCCGGTGTAACGGTGGAATGTGTAGATATCGGGAAGAACACCAATGGCGAAGGCAGGTCTCTGGGCCGTCACTGACGCTGAGGAGCGAAAGCGTGGGGAGCGAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGGTGGATGCTGGATGTGGGGCCCTTTCCACGGGTCCCGTGTCGGAGCCAACGCGTTAAGCATCCCGCCTGGGGAGTACGGCCGCAAGGCTAAAACTCAAAGAAATTGACGGGGGCCCGCACAAGCGGCGGAGCATGCGGATTAATTCGATGCAACGCGAAGAACCTTACCTGGGCTTGACATGTGCCGGATCGCCGTGGAGACACGGTTTCCCTTCGGGGCCGGTTCACAGGTGGTGCATGGTCGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTCGCCGCATGTTGCCAGCGGGTGATGCCGGGAACTCATGTGGGACCGCCGGGGTCAACTCGGAGGAAGGTGGGGATGACGTCAGATCATCATGCCCCTTACGTCCAGGGCTTCACGCATGCTACAATGGCCGGTACAACGCGGTGCGACACGGTGACGTGGGGCGGATCGCTGAAAACCGGTCTCAGTTCGGATCGCAGTCTGCAACTCGACTGCGTGAAGGCGGAGTCGCTAGTAATCGCGGATCAGCAACGCCGCGGTGAATGCGTTCCCGGGCCTTGTACACACCGCCCGTCAAGTCATGAAAGTGGGTAGCACCCGAAGCCGGTGGCCCGACCCTTGTGGGGGGAGCCGTCTAAGGGTAGACTTCGT
through BLAST comparison in NCBI, the strain has 99.86% similarity with Bifidobacterium animalis subsp.lacti strain 912, so the strain belongs to bifidobacterium animalis subspecies lactis, which is named as bifidobacterium animalis subspecies TA-2688, and is preserved in China general microbiological culture Collection center (CGMCC) No.27908 at 7.14 of 2023; the preservation address is North Chen West Lu No.1 and 3 of the Korean area of Beijing, and the institute of microorganisms of the national academy of sciences.
Example 2
This example provides the ability of bifidobacterium animalis subspecies lactis TA-2688 to tolerate gastrointestinal tract.
1. Test method
1.1 acid resistance test
The third generation animal bifidobacterium lactobacillus subspecies TA-2688 culture solution (culture time 18 h) is taken and inoculated with 10 percent of the inoculation amount of the modified liquid MRS culture medium with the pH of 3.0. Viable counts were performed at 0h,2h, and 4h of incubation, respectively. Anaerobic culture at 37 ℃.
1.2 bile salt resistance experiments
Taking an activated third-generation animal bifidobacterium subspecies lactis TA-2688 culture solution (culture time is 18 h), and inoculating 10% of the culture solution into a liquid modified liquid MRS culture medium containing 0.3% of bovine bile salt. Viable counts were performed at 0h,2h, and 4h of incubation, respectively. Anaerobic culture at 37 ℃.
2. Experimental results
The detection results are shown in FIG. 1.
The results show that the bifidobacterium animalis subspecies lactis TA-2688 has good acid resistance and bile salt resistance, and can still keep higher activity after being digested by gastrointestinal tracts.
Example 3
This example provides the effect of bifidobacterium animalis subspecies lactis TA-2688 on promoting serotonin acid production function.
1. Experimental method
1.1 cultivation of bifidobacterium animalis subspecies lactis TA-2688
(1) Strain activation
Inoculating isolated animal Bifidobacterium lactis subspecies TA-2688 into modified MRS liquid culture medium (glucose 20.0g, peptone 10.0g, beef powder 6.5g, yeast extract 5.0g, sodium acetate 5.0g, diammonium citrate 2.0g, dipotassium hydrogen phosphate 2.0g, mgSO) 4 ·7H 2 O 0.58g，MnSO 4 ·H 2 O0.25 g, L-cysteine hydrochloride 0.5g, tween 80 1.0mL and distilled water 1000 mL), shaking and mixing, anaerobic culturing at 37 ℃ for 24h, and sequentially activating for three generations.
(2) Strain treatment
Centrifuging activated animal Bifidobacterium lactis subspecies TA-2688 at 4deg.C and 12000rpm/min for 3min, collecting thallus, re-suspending with 50mM sterile PBS and washing twice, and adjusting cell concentration to 1×10 8 CFU/mL was ready for use. Filtering and sterilizing the supernatant obtained by centrifugation through a 0.22 mu m organic filter membrane, and refrigerating for later use.
1.2RIN14B cell activation
(1) Resuscitates cells: the frozen tube containing 1mL of cell suspension was thawed by shaking in a 37 ℃ water bath and 5mL of medium was added to mix well. Centrifuging at 1000RPM for 5min, discarding supernatant, adding 4-6mL of complete medium, and blowing uniformly. All cell suspensions were then added to the flask and incubated overnight. The next day the fluid was changed and the cell density was checked.
(2) Cell passage: after the cells grow to 80%, the culture supernatant is discarded, and the cells are rinsed 1-2 times with PBS without calcium and magnesium ions. Adding 1-2ml of digestive juice (0.25% Trypsin-0.53mM EDTA) into a culture flask, placing into a culture flask for digestion at 37 ℃, observing the digestion condition of cells under a microscope every 1-2min, quickly taking back to an operating table when the cells are mostly rounded and fall off, tapping the culture flask, adding more than 5ml of complete culture medium containing 10% serum, stopping digestion (if the culture flask is not rounded and fall off, continuing digestion until the cells are mostly rounded and fall off). Gently blowing the cells, completely removing the cells, sucking out, centrifuging at 1000RPM for 8-10min, removing supernatant, adding 1-2mL culture solution, and uniformly blowing. The culture solution is added according to 5-6 ml/bottle, and the cell suspension is added according to 1:2 to 1: the ratio of 5 was divided into new dishes or flasks containing 5-6ml of culture medium. The culture solution is replaced every 2-3 days for 4 times.
1.3 Co-incubation of Bifidobacterium animalis subspecies lactobacilli TA-2688 with RIN14B cells (rat insulinoma cells)
Collecting activated RIN14B cells, and adjusting cell concentration to 1×10 4 CFU/mL are plated, and after 72h of culture, three groups are separated, 20 mu L of TA-2688 bacterial suspension with the concentration adjusted by washing in the step (2) in the treatment group 1 is added, 20 mu L of sterile supernatant obtained by filtering in the step (2) in the treatment group 2 is added in the step 1.1, 20 mu L of PBS is added in the control group, and culture is continued for 12h.
1.4 detection of the content of serotonin (5-HTP)
The cell supernatants of each treatment group of 1.3 were filtered through a 0.22 μm filter, and the filtrate was measured for 5-HTP content by high performance liquid chromatography equipped with a fluorescence detector (RF-20A, shimadzu). The column was a Agilent Zorbax XDB-C18 column (250 mM. Times.4.6 mM,5 μm) and the mobile phase was 0.1M sodium acetate buffer (containing 0.1mM EDTA-2 Na) and methanol (85:15, v/v). The column temperature was 30℃and the flow rate was 1.0mL/min. Fluorescence intensities were detected at 330nm (emission) and 290nm (excitation) wavelengths.
2. Experimental results
The results of the detection are shown in Table 1 and FIG. 2.
TABLE 1 content of serotonin (5-HTP)
| Group of | 5-HTP(ng/mL) |
| Control group | 1.07±0.21 |
| Treatment group 1 | 23.56±2.87 |
| Treatment group 2 | 62.39±7.41 |
The results show that the bifidobacterium animalis subspecies TA-2688 and the fermentation supernatant thereof can promote RIN14B cells to produce serotonin acid, wherein the bifidobacterium animalis subspecies TA-2688 fermentation supernatant has the best effect. Namely, the TA-2688 cells and the metabolites thereof can stimulate the production of the serotonin, and the metabolites have the best effect of stimulating the production of the serotonin.
Example 4
The present example provides the effect of bifidobacterium animalis subspecies lactis TA-2688 on relieving constipation in mice.
1. Test animals and reagents
Male mice (Beijing Wanfukang), 6-8 weeks, 18-22g body weight, 12 animals per group; activated carbon powder (aladin, a 2213554); gum arabic (Coolaber, CG 31163150); loperamide (aladin, F2117174); ink (5% activated carbon powder and 10% gum arabic).
2. Method of
2.1 first grain black stool time and determination of the moisture content of the mouse feces
The test sample was given for 15 days (200. Mu.L of the liquid containing TA-2688 viable bacteria 1X10 per day by stomach lavage) 8 CFU bacterial suspension), each group of mice fasted for 16 hours without water. Loperamide (4 mg/kg BW) was given by gavage to the model control and probiotic groups, and the negative control group was gavaged with equal volumes of distilled water. After 0.5 hour of loperamide administration, mice in the negative control and model control groups were perfused with 200. Mu.L of ink and the probiotic group was given 200. Mu.L of TA-2688 containing ink (TA-2688 containing live bacteria 1X 10) 8 CFU), animals were kept in single cages and fed with normal drinking water. From the ink filling, the first black stool time and the first black stool weight of each animal were recorded. Weighing to obtain wet weight, lyophilizing to obtain dry weight, and calculating faeces water content according to the following formula:
fecal moisture = (fecal wet weight-fecal dry weight)/fecal wet weight.
2.2 ink Propulsion
The test sample was given for 15 days (200. Mu.L of the liquid containing TA-2688 viable bacteria 1X10 per day by stomach lavage) 8 CFU bacterial suspension), each group of mice fasted for 16 hours without water. The negative control group was given distilled water and the model control group and the probiotic group were given loperamide (4 mg/kg BW) by gavage. After 0.5 hours from loperamide administration, the probiotic group was given 200. Mu.L of TA-2688 containing ink (TA-2688 containing live bacteria 1X 10) 8 CFU), negative control and model control mice were lavaged with 200 μl of ink. Immediately after 25 minutes, the animals were sacrificed by cervical vertebra removal, the celiac separation mesentery was opened, the upper end from the pylorus, the lower end to the ileocecum were cut off, the intestinal canal was placed on a tray, the small intestine was gently pulled into a straight line, the length of the intestinal canal was measured as the "total length of the small intestine", and the front edge from the pylorus to the ink was the "push length of the ink". The ink push rate was calculated as follows:
ink advance (%) =ink advance length (cm)/total length of small intestine (cm) ×100%
2.3 determination of colon Occludin, claudin Gene expression in mice
Taking 20mg colon tissue of 2.2 mice, extracting RNA according to the specification of animal tissue total RNA extraction kit (Tiangen, DP 431), reversely transcribing the first strand according to the specification of FastKing cDNA first strand synthesis kit (Tiangen, KR 116-2), and setting a PCR reaction strip after the sample is added to a PCR reaction systemThe PCR primers are shown in Table 2, after the PCR reaction was performed on the machine (Roche, light Cycle480 ll). Ct value was measured using 2 -△△Ct The (Livak) method (the negative group gene expression rate was assigned to 1, and the comparison was made as a reference, with the final relative expression rate being a multiple of that of the negative control group). The expression level of each group of the claudin Occludin, claudin gene was calculated.
TABLE 2 primer sequences
2.4 measurement of levels of mouse Casein Peptide (PYY), motilin (MTL), 5-hydroxytryptamine (5-HT), IL-1 beta
Serum samples: after killing 2.2 mice, the collected blood of mice was allowed to stand for 2 hours and centrifuged at 3000 Xg for 15min to obtain serum, and experiments were performed with reference to the instructions of the mouse casein Peptide (PYY), motilin (MTL), 5-hydroxytryptamine (5-HT) enzyme-linked immunosorbent assay kit (mlbrio ELISA), and the concentration of MTL in serum was calculated according to a standard curve.
Colon sample: the colon tissue was rinsed with pre-chilled PBS, residual blood was removed, surrounding adipose tissue was removed, and weighed and minced. The sheared tissues and PBS with corresponding volumes (according to the weight-to-volume ratio of 1:9) are crushed on a high-flux tissue crusher, and finally homogenate and 5000 Xg are centrifuged for 5-10 minutes, supernatant is taken for detection, experiments are carried out by referring to the corresponding kit instruction, and the contents of 5-HT and pro-inflammatory factors IL-1 beta in the tissues are calculated according to a standard curve.
3. Experimental results
3.1 measurement of first-grain black stool time, water content of mouse feces and ink push rate
TABLE 3 defecation index of mice of different groups
Note that: the same index data is marked with a letter to indicate significant differences between groups (P < 0.05).
The results show that compared with the negative control group, the black stool time of the mice fed with loperamide is prolonged, the ink propelling rate of the fecal water content is obviously reduced, which indicates that the loperamide can cause constipation symptoms of the mice fed with loperamide. Meanwhile, the first grain black stool time can be obviously shortened by feeding the animal bifidobacterium lactobionic TA-2688, the water content of the mouse stool can be improved, the ink propulsion rate of the intestinal tract of the mouse can be improved, and the animal bifidobacterium lactobionic TA-2688 can promote intestinal peristalsis, keep stool smooth and moist, has a good bowel relaxing effect, and can relieve constipation symptoms of the mouse.
3.2 determination of colon Occludin, claudin Gene expression in mice
TABLE 4 relative expression Rate of the colon Occludin, claudin Gene in mice of different groups
Note that: 1. * the expression level of the negative control group was set to 1, and the relative expression rate was a multiple of that of the negative control group. 2. The same index data is marked with letters to indicate significant differences between groups (P<0.05)。
The results showed that the mice fed loperamide had a reduced relative expression of mRNA of the intestinal claudin Occludin, claudin compared to the negative control group. Meanwhile, the relative expression rate of mRNA of the mouse intestinal canal compact junction protein Occludin, claudin can be obviously improved by feeding bifidobacterium lactis subspecies TA-2688 to animals. TA-2688 was demonstrated to improve intestinal barrier function in constipation mice.
3.3 determination of PYY, MTL, 5-HT, IL-1 beta levels in mice
TABLE 5 PYY, MTL, 5-HT and IL-1 beta levels detection results for different groups of mice
Note that: the same index data is marked with a letter to indicate significant differences between groups (P < 0.05).
The results show that the MTL, 5-HT expression was reduced and PYY and IL-1β levels were increased in loperamide-fed mice compared to the negative control group. Meanwhile, the bifidobacterium lactis subspecies TA-2688 of the animal is fed at the same time, so that the MTL and 5-HT levels of mice can be obviously improved, the PYY level is reduced, and intestinal peristalsis is promoted; and can reduce the expression of the proinflammatory factor IL-1 beta. The animal bifidobacterium lactis TA-2688 not only can improve gastrointestinal motility function of a constipation mouse, but also can regulate and control immune level of the mouse.
Example 5
This example provides a powder containing bifidobacterium animalis subspecies lactis TA-2688.
Weighing 30% of animal bifidobacterium lactosub-species TA-2688 freeze-dried powder, 5% of fructo-oligosaccharide, 7% of galacto-oligosaccharide, 12% of mannans, 10% of pectin, 1.5% of compound vitamin powder, 1.5% of mineral powder and 33% of milk powder according to mass percentage, mixing, preparing the product specification according to the viable count, and packaging to obtain the powder product containing animal bifidobacterium lactosub-species TA-2688.
Example 6
The present example provides a granule comprising bifidobacterium animalis subspecies lactis TA-2688.
Weighing 35% of animal bifidobacterium subspecies TA-2688 freeze-dried powder, 7% of fructo-oligosaccharide, 9% of galacto-oligosaccharide, 25% of sugar powder, 10% of soluble starch, 11% of mannitol, 1.5% of compound vitamin powder and 1.5% of mineral powder according to mass percentage, mixing, granulating by a dry method, adding magnesium stearate accounting for 0.5% of the total particle weight after granulating, preparing product specifications according to viable count, and packaging to obtain the granules containing animal bifidobacterium subspecies TA-2688.
Example 7
The present example provides a chewable tablet comprising bifidobacterium animalis subspecies lactis TA-2688.
Weighing 35% of animal bifidobacterium subspecies TA-2688 freeze-dried powder, 9% of galactooligosaccharide, 6% of mannooligosaccharide, 23% of microcrystalline cellulose, 10% of mannitol, 13% of sugar powder, 1.5% of compound vitamin powder, 1.5% of mineral powder, 0.5% of sweet orange essence and 0.5% of magnesium stearate according to mass percentage, uniformly mixing, tabletting, and selecting tablet weight and tablet type according to the number of viable bacteria and actual requirements. The pressure is adjusted according to the hardness of the product in the tabletting process.
Example 8
The present example provides a bifidobacteria capsule product.
Hard capsule: weighing 35% of animal bifidobacterium subspecies TA-2688 freeze-dried powder, 8% of fructo-oligosaccharide, 8% of isomaltooligosaccharide, 30% of soluble starch, 16% of microcrystalline cellulose, 1.5% of compound vitamin powder and 1.5% of mineral powder according to mass percentage, mixing, granulating by a dry method, adding magnesium stearate accounting for 0.5% of total particle weight after granulating, selecting a capsule shell model according to gram weight or viable count, and filling by a capsule machine to obtain the hard capsule containing animal bifidobacterium subspecies TA-2688.
Soft capsule: mixing animal bifidobacterium lactobionic TA-2688 powder with lecithin, selecting capsule specification according to gram weight or viable bacteria number, and pressing into soft capsule product by a capsule machine.
Example 9
The present example provides a fermented dairy product comprising bifidobacterium animalis subspecies lactis TA-2688.
Fermented yogurt: heating pasteurized fresh cow milk to 50 ℃, adding 5% glucose, 0.3% yeast autolysate and 0.05% cysteine, stirring until the mixture is completely dissolved, cooling to 40 ℃, inoculating 8% bifidobacterium animalis subspecies of milk TA-2688, carrying out anaerobic fermentation at 40-42 ℃ until the pH is 4.4, and carrying out cold storage and after-ripening at 4-6 ℃ to obtain the milk.
Milk beverage: heating pasteurized skim milk to 50 ℃, adding 5% glucose, 0.3% yeast autolysate and 0.05% cysteine, stirring until complete dissolution, cooling to 40 ℃, carrying out anaerobic fermentation at 40-42 ℃ to pH 4.4 with 8% bifidobacterium animalis subspecies TA-2688, refrigerating at 4-6 ℃, after-ripening, mixing with water and high fructose syrup according to a mass ratio of 1.5:2:0.8, regulating acidity to pH 4.2 with citric acid, dripping flavoring essence, homogenizing under 10MPa pressure, filling, and refrigerating to obtain the finished product.
The foregoing description of the preferred embodiments of the invention is not intended to be limiting, but rather is intended to cover all modifications, equivalents, or alternatives falling within the spirit and principles of the invention.
Claims (13)
1. A bifidobacterium animalis subspecies lactis TA-2688 is classified as bifidobacterium animalis subspecies lactis (Bifidobacterium animalis subsp.lactis), and the strain preservation number is CGMCC No.27908.
2. A metabolite of bifidobacterium animalis subspecies lactis TA-2688 as claimed in claim 1.
3. The method for preparing the metabolite of the bifidobacterium animalis subspecies lactobacilli TA-2688 according to claim 2, comprising the following operations: inoculating the bifidobacterium animalis subspecies of the Lactobacillus TA-2688 into an improved MRS liquid culture medium for activation, centrifuging and taking supernatant fluid to obtain the bifidobacterium animalis strain.
4. Use of a metabolite of bifidobacterium animalis subspecies lactis TA-2688 as claimed in claim 1 in the manufacture of a product for improving constipation.
5. The use according to claim 4, wherein the product comprises at least one of viable cells or metabolites of the bifidobacterium animalis subspecies lacto TA-2688.
6. Use according to claim 4 or 5, characterized in that the product is a product for increasing the moisture content of faeces; and/or
The product is a product for promoting intestinal peristalsis.
7. The use according to claim 6, wherein the product is a product that promotes the production of serotonin by cells; and/or
The product is a product for improving the expression of intestinal motilin; and/or
The product is a product for improving the expression of 5-hydroxytryptamine; and/or
The product is a product for reducing the expression of casein peptide.
8. Use of a metabolite of bifidobacterium animalis subspecies lactis TA-2688 as claimed in claim 1 in the manufacture of a product for repairing intestinal mucosal barriers.
9. The use according to claim 8, wherein said product comprises at least one of viable cells or metabolites of said bifidobacterium animalis subspecies lactis TA-2688.
10. The use according to claim 8 or 9, wherein the product is a product promoting expression of intestinal claudin Occludin; and/or
The product is a product for promoting the expression of the intestinal canal compact junction protein Claudin.
11. Use of a metabolite of bifidobacterium animalis subspecies lactis TA-2688 as claimed in claim 1 in the manufacture of a product for modulating immunity.
12. The use according to claim 11, wherein said product comprises at least one of viable cells or metabolites of said bifidobacterium animalis subspecies lactis TA-2688.
13. The use according to claim 11 or 12, wherein the product is a product that reduces the expression of the pro-inflammatory factor IL-1 β.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311341637.2A CN117535176A (en) | 2023-10-16 | 2023-10-16 | Bifidobacterium animalis subspecies lactis TA-2688 and application thereof in constipation improvement, intestinal mucosa barrier repair and immunity adjustment |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311341637.2A CN117535176A (en) | 2023-10-16 | 2023-10-16 | Bifidobacterium animalis subspecies lactis TA-2688 and application thereof in constipation improvement, intestinal mucosa barrier repair and immunity adjustment |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN117535176A true CN117535176A (en) | 2024-02-09 |
Family
ID=89794677
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202311341637.2A Pending CN117535176A (en) | 2023-10-16 | 2023-10-16 | Bifidobacterium animalis subspecies lactis TA-2688 and application thereof in constipation improvement, intestinal mucosa barrier repair and immunity adjustment |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN117535176A (en) |
-
2023
- 2023-10-16 CN CN202311341637.2A patent/CN117535176A/en active Pending
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US11116806B2 (en) | Composite probiotic lactic acid bacteria powder and preparation method and use thereof | |
| CN103908585B (en) | For the probiotics fermention compositions of prevention and therapy constipation | |
| CN111996153B (en) | Bifidobacterium breve and application thereof | |
| CN108782758B (en) | Fermented synbiotic goat milk powder and preparation method thereof | |
| CN108477617A (en) | A kind of probiotic powder and preparation method thereof to relax bowel | |
| WO2022206300A1 (en) | Composition capable of facilitating defecation and use thereof | |
| CN110643524B (en) | Composite probiotic preparation with gastrointestinal tract mucosa protection effect and application thereof | |
| CN115353988B (en) | Lactobacillus paracasei LC-37 with digestion promoting effect and application | |
| CN112980734B (en) | Bifidobacterium bifidum for relieving constipation and regulating intestinal flora disorder and application thereof | |
| CN113943681B (en) | Bifidobacterium longum capable of reducing inflammatory reaction and relieving constipation | |
| EP3808357A1 (en) | Composition and uses thereof | |
| CN114146101A (en) | Application of bifidobacterium animalis subsp lactis BLA80 in preparing medicine or food for regulating intestinal motility | |
| CN114651984A (en) | Application of lactobacillus casei LC89 and microbial preparation containing same in preparation of product for relieving functional constipation | |
| CN114774328B (en) | Bifidobacterium breve capable of down-regulating IL-17 and relieving constipation and application thereof | |
| CN103228143A (en) | Compositions and methods for augmenting kidney function | |
| CN117143765A (en) | Bifidobacterium longum subspecies capable of regulating intestinal canal steady state and relieving intractable constipation and application thereof | |
| CN114686405B (en) | Bifidobacterium bifidum with functions of reducing fat, relieving hyperglycemia and regulating intestinal immunity and application thereof | |
| CN113943683B (en) | Bifidobacterium longum subspecies capable of relieving constipation and increasing total bile acid content of feces and application thereof | |
| CN115747121A (en) | Bifidobacterium lactis HSLA-009, microbial inoculum and preparation method and application thereof | |
| CN113943682B (en) | Bifidobacterium longum subspecies longum for relieving constipation and fermented food and probiotic preparation prepared from same | |
| CN117535176A (en) | Bifidobacterium animalis subspecies lactis TA-2688 and application thereof in constipation improvement, intestinal mucosa barrier repair and immunity adjustment | |
| US11969447B2 (en) | Use of spray-dried powder derived from lactic acid bacterial strains and herbal extracts for promoting defecation | |
| CA3197953A1 (en) | Compositions and methods using at least one strain of staphylococcus carnosus therapeutically or prophylactically | |
| CN115996736A (en) | Novel lactobacillus reuteri strain and use thereof | |
| CN117603841A (en) | Bifidobacterium longum subspecies THRK-7782 and application thereof in improving constipation, repairing intestinal mucosa barrier and regulating immunity |
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 |