CN104531549B - A kind of lactobacillus fermenti Lactobacillus fermentum strain Zhao and application thereof of adjustable intestinal movement, Constipation - Google Patents
A kind of lactobacillus fermenti Lactobacillus fermentum strain Zhao and application thereof of adjustable intestinal movement, Constipation Download PDFInfo
- Publication number
- CN104531549B CN104531549B CN201410578613.3A CN201410578613A CN104531549B CN 104531549 B CN104531549 B CN 104531549B CN 201410578613 A CN201410578613 A CN 201410578613A CN 104531549 B CN104531549 B CN 104531549B
- Authority
- CN
- China
- Prior art keywords
- lactobacillus
- zhao
- constipation
- strain
- fermentum strain
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 241000186840 Lactobacillus fermentum Species 0.000 title claims abstract description 117
- 206010010774 Constipation Diseases 0.000 title claims abstract description 47
- 230000000968 intestinal effect Effects 0.000 title claims abstract description 20
- 229940012969 lactobacillus fermentum Drugs 0.000 claims abstract description 47
- 235000013305 food Nutrition 0.000 claims abstract description 20
- 241000894006 Bacteria Species 0.000 claims description 49
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 claims description 40
- 235000013336 milk Nutrition 0.000 claims description 28
- 210000004080 milk Anatomy 0.000 claims description 28
- 239000008267 milk Substances 0.000 claims description 28
- 235000020167 acidified milk Nutrition 0.000 claims description 21
- 235000014655 lactic acid Nutrition 0.000 claims description 20
- 239000004310 lactic acid Substances 0.000 claims description 20
- 239000000843 powder Substances 0.000 claims description 14
- 238000000855 fermentation Methods 0.000 claims description 10
- 230000004151 fermentation Effects 0.000 claims description 10
- 239000002775 capsule Substances 0.000 claims description 8
- 235000020124 milk-based beverage Nutrition 0.000 claims description 7
- 235000021107 fermented food Nutrition 0.000 claims description 6
- 241000699666 Mus <mouse, genus> Species 0.000 abstract description 23
- 229940099352 cholate Drugs 0.000 abstract description 16
- BHQCQFFYRZLCQQ-OELDTZBJSA-N cholic acid Chemical compound C([C@H]1C[C@H]2O)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(O)=O)C)[C@@]2(C)[C@@H](O)C1 BHQCQFFYRZLCQQ-OELDTZBJSA-N 0.000 abstract description 16
- 230000001580 bacterial effect Effects 0.000 abstract description 15
- 241000699670 Mus sp. Species 0.000 abstract description 12
- 239000012530 fluid Substances 0.000 abstract description 12
- 210000003608 fece Anatomy 0.000 abstract description 11
- 239000003814 drug Substances 0.000 abstract description 10
- 230000013872 defecation Effects 0.000 abstract description 9
- 230000002496 gastric effect Effects 0.000 abstract description 9
- 210000002966 serum Anatomy 0.000 abstract description 9
- 230000004083 survival effect Effects 0.000 abstract description 9
- 239000003651 drinking water Substances 0.000 abstract description 8
- 235000020188 drinking water Nutrition 0.000 abstract description 8
- 239000008187 granular material Substances 0.000 abstract description 7
- 210000001809 melena Anatomy 0.000 abstract description 7
- 238000012360 testing method Methods 0.000 abstract description 7
- 239000002253 acid Substances 0.000 abstract description 6
- 229940079593 drug Drugs 0.000 abstract description 5
- 230000003449 preventive effect Effects 0.000 abstract description 5
- 230000001141 propulsive effect Effects 0.000 abstract description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 abstract description 5
- 238000001727 in vivo Methods 0.000 abstract description 2
- 230000001954 sterilising effect Effects 0.000 description 26
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 22
- 235000020185 raw untreated milk Nutrition 0.000 description 20
- 238000004659 sterilization and disinfection Methods 0.000 description 20
- 241000186660 Lactobacillus Species 0.000 description 14
- 229940039696 lactobacillus Drugs 0.000 description 14
- 238000002474 experimental method Methods 0.000 description 11
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 10
- 238000003304 gavage Methods 0.000 description 9
- 239000000047 product Substances 0.000 description 9
- 239000000725 suspension Substances 0.000 description 9
- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 description 8
- 244000199885 Lactobacillus bulgaricus Species 0.000 description 8
- 235000013960 Lactobacillus bulgaricus Nutrition 0.000 description 8
- 210000000481 breast Anatomy 0.000 description 8
- 235000013365 dairy product Nutrition 0.000 description 8
- 229940004208 lactobacillus bulgaricus Drugs 0.000 description 8
- 239000006041 probiotic Substances 0.000 description 8
- 235000018291 probiotics Nutrition 0.000 description 8
- 210000000813 small intestine Anatomy 0.000 description 8
- 230000001965 increasing effect Effects 0.000 description 7
- CTQNGGLPUBDAKN-UHFFFAOYSA-N O-Xylene Chemical compound CC1=CC=CC=C1C CTQNGGLPUBDAKN-UHFFFAOYSA-N 0.000 description 6
- 210000000941 bile Anatomy 0.000 description 6
- 201000010099 disease Diseases 0.000 description 6
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 6
- 239000000243 solution Substances 0.000 description 6
- 238000001694 spray drying Methods 0.000 description 6
- 235000011187 glycerol Nutrition 0.000 description 5
- 238000000034 method Methods 0.000 description 5
- 238000002360 preparation method Methods 0.000 description 5
- 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 description 4
- 102000012440 Acetylcholinesterase Human genes 0.000 description 4
- 108010022752 Acetylcholinesterase Proteins 0.000 description 4
- 241000283690 Bos taurus Species 0.000 description 4
- 102000016938 Catalase Human genes 0.000 description 4
- 108010053835 Catalase Proteins 0.000 description 4
- 101800002372 Motilin Proteins 0.000 description 4
- 102400001357 Motilin Human genes 0.000 description 4
- 241000194020 Streptococcus thermophilus Species 0.000 description 4
- 108010003205 Vasoactive Intestinal Peptide Proteins 0.000 description 4
- 102400000015 Vasoactive intestinal peptide Human genes 0.000 description 4
- 229940022698 acetylcholinesterase Drugs 0.000 description 4
- 230000004913 activation Effects 0.000 description 4
- 239000007853 buffer solution Substances 0.000 description 4
- VBUWHHLIZKOSMS-RIWXPGAOSA-N invicorp Chemical compound C([C@@H](C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(O)=O)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CCCCN)NC(=O)[C@@H](NC(=O)[C@H](C)NC(=O)[C@H](CCSC)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@@H](NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC(O)=O)NC(=O)[C@@H](NC(=O)[C@H](CC=1C=CC=CC=1)NC(=O)[C@@H](NC(=O)[C@H](C)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC=1NC=NC=1)C(C)C)[C@@H](C)O)[C@@H](C)O)C(C)C)C1=CC=C(O)C=C1 VBUWHHLIZKOSMS-RIWXPGAOSA-N 0.000 description 4
- 238000002156 mixing Methods 0.000 description 4
- 235000015097 nutrients Nutrition 0.000 description 4
- 238000001556 precipitation Methods 0.000 description 4
- 230000031068 symbiosis, encompassing mutualism through parasitism Effects 0.000 description 4
- 230000004580 weight loss Effects 0.000 description 4
- 241000193830 Bacillus <bacterium> Species 0.000 description 3
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 3
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 3
- 239000003795 chemical substances by application Substances 0.000 description 3
- 238000011161 development Methods 0.000 description 3
- 238000010790 dilution Methods 0.000 description 3
- 239000012895 dilution Substances 0.000 description 3
- 239000001963 growth medium Substances 0.000 description 3
- 230000036541 health Effects 0.000 description 3
- 238000011081 inoculation Methods 0.000 description 3
- 235000015141 kefir Nutrition 0.000 description 3
- 238000005259 measurement Methods 0.000 description 3
- 244000005700 microbiome Species 0.000 description 3
- 238000000386 microscopy Methods 0.000 description 3
- 238000011084 recovery Methods 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 235000002639 sodium chloride Nutrition 0.000 description 3
- 238000002560 therapeutic procedure Methods 0.000 description 3
- 241000186000 Bifidobacterium Species 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 2
- 108050009340 Endothelin Proteins 0.000 description 2
- 102000002045 Endothelin Human genes 0.000 description 2
- 241000233866 Fungi Species 0.000 description 2
- 241000590002 Helicobacter pylori Species 0.000 description 2
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical compound C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 description 2
- 239000002250 absorbent Substances 0.000 description 2
- 230000002745 absorbent Effects 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 238000005119 centrifugation Methods 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
- ZUBDGKVDJUIMQQ-UBFCDGJISA-N endothelin-1 Chemical compound C([C@@H](C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(O)=O)NC(=O)[C@H]1NC(=O)[C@H](CC=2C=CC=CC=2)NC(=O)[C@@H](CC=2C=CC(O)=CC=2)NC(=O)[C@H](C(C)C)NC(=O)[C@H]2CSSC[C@@H](C(N[C@H](CO)C(=O)N[C@@H](CO)C(=O)N[C@H](CC(C)C)C(=O)N[C@@H](CCSC)C(=O)N[C@H](CC(O)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCC(O)=O)C(=O)N2)=O)NC(=O)[C@@H](CO)NC(=O)[C@H](N)CSSC1)C1=CNC=N1 ZUBDGKVDJUIMQQ-UBFCDGJISA-N 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 210000000232 gallbladder Anatomy 0.000 description 2
- 235000021472 generally recognized as safe Nutrition 0.000 description 2
- 238000010438 heat treatment Methods 0.000 description 2
- 229940037467 helicobacter pylori Drugs 0.000 description 2
- 230000002209 hydrophobic effect Effects 0.000 description 2
- 239000005457 ice water Substances 0.000 description 2
- 230000000877 morphologic effect Effects 0.000 description 2
- 235000016709 nutrition Nutrition 0.000 description 2
- 230000035764 nutrition Effects 0.000 description 2
- 239000008194 pharmaceutical composition Substances 0.000 description 2
- 230000000529 probiotic effect Effects 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 230000002829 reductive effect Effects 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 235000020183 skimmed milk Nutrition 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 230000002269 spontaneous effect Effects 0.000 description 2
- 239000007921 spray Substances 0.000 description 2
- 238000010186 staining Methods 0.000 description 2
- 210000002784 stomach Anatomy 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- 235000013618 yogurt Nutrition 0.000 description 2
- 229920001817 Agar Polymers 0.000 description 1
- KHOITXIGCFIULA-UHFFFAOYSA-N Alophen Chemical compound C1=CC(OC(=O)C)=CC=C1C(C=1N=CC=CC=1)C1=CC=C(OC(C)=O)C=C1 KHOITXIGCFIULA-UHFFFAOYSA-N 0.000 description 1
- 244000153158 Ammi visnaga Species 0.000 description 1
- 235000010585 Ammi visnaga Nutrition 0.000 description 1
- 206010002153 Anal fissure Diseases 0.000 description 1
- 208000016583 Anus disease Diseases 0.000 description 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- 208000024172 Cardiovascular disease Diseases 0.000 description 1
- 241001478240 Coccus Species 0.000 description 1
- 241000588724 Escherichia coli Species 0.000 description 1
- 208000009531 Fissure in Ano Diseases 0.000 description 1
- 102400000921 Gastrin Human genes 0.000 description 1
- 108010052343 Gastrins Proteins 0.000 description 1
- 241000726221 Gemma Species 0.000 description 1
- 108010051696 Growth Hormone Proteins 0.000 description 1
- 206010053759 Growth retardation Diseases 0.000 description 1
- 206010019375 Helicobacter infections Diseases 0.000 description 1
- 208000031226 Hyperlipidaemia Diseases 0.000 description 1
- 206010020772 Hypertension Diseases 0.000 description 1
- 238000012449 Kunming mouse Methods 0.000 description 1
- 240000001929 Lactobacillus brevis Species 0.000 description 1
- 235000013957 Lactobacillus brevis Nutrition 0.000 description 1
- 244000199866 Lactobacillus casei Species 0.000 description 1
- 235000013958 Lactobacillus casei Nutrition 0.000 description 1
- 240000006024 Lactobacillus plantarum Species 0.000 description 1
- 235000013965 Lactobacillus plantarum Nutrition 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 102000057297 Pepsin A Human genes 0.000 description 1
- 108090000284 Pepsin A Proteins 0.000 description 1
- 208000034189 Sclerosis Diseases 0.000 description 1
- 102100038803 Somatotropin Human genes 0.000 description 1
- 241000194017 Streptococcus Species 0.000 description 1
- 241001052560 Thallis Species 0.000 description 1
- 210000000683 abdominal cavity Anatomy 0.000 description 1
- 230000003187 abdominal effect Effects 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 235000019730 animal feed additive Nutrition 0.000 description 1
- 230000003064 anti-oxidating effect Effects 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 210000000709 aorta Anatomy 0.000 description 1
- 210000001367 artery Anatomy 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 229960000503 bisacodyl Drugs 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 230000017531 blood circulation Effects 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- VTYYLEPIZMXCLO-UHFFFAOYSA-L calcium carbonate Substances [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 1
- 229910000019 calcium carbonate Inorganic materials 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 208000026106 cerebrovascular disease Diseases 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- AOXOCDRNSPFDPE-UKEONUMOSA-N chembl413654 Chemical compound C([C@H](C(=O)NCC(=O)N[C@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@H](CCSC)C(=O)N[C@H](CC(O)=O)C(=O)N[C@H](CC=1C=CC=CC=1)C(N)=O)NC(=O)[C@@H](C)NC(=O)[C@@H](CCC(O)=O)NC(=O)[C@@H](CCC(O)=O)NC(=O)[C@@H](CCC(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC=1C2=CC=CC=C2NC=1)NC(=O)[C@H]1N(CCC1)C(=O)CNC(=O)[C@@H](N)CCC(O)=O)C1=CC=C(O)C=C1 AOXOCDRNSPFDPE-UKEONUMOSA-N 0.000 description 1
- 235000012000 cholesterol Nutrition 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 235000005911 diet Nutrition 0.000 description 1
- 230000037213 diet Effects 0.000 description 1
- 235000019621 digestibility Nutrition 0.000 description 1
- 238000002224 dissection Methods 0.000 description 1
- 230000002526 effect on cardiovascular system Effects 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 230000002550 fecal effect Effects 0.000 description 1
- 235000021001 fermented dairy product Nutrition 0.000 description 1
- 239000000796 flavoring agent Substances 0.000 description 1
- 235000019634 flavors Nutrition 0.000 description 1
- 235000011389 fruit/vegetable juice Nutrition 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- 210000004051 gastric juice Anatomy 0.000 description 1
- 235000021474 generally recognized As safe (food) Nutrition 0.000 description 1
- 235000021473 generally recognized as safe (food ingredients) Nutrition 0.000 description 1
- 239000003292 glue Substances 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 239000000122 growth hormone Substances 0.000 description 1
- 231100000001 growth retardation Toxicity 0.000 description 1
- 230000002650 habitual effect Effects 0.000 description 1
- 235000001497 healthy food Nutrition 0.000 description 1
- 208000014617 hemorrhoid Diseases 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 238000005286 illumination Methods 0.000 description 1
- 230000036737 immune function Effects 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- 210000000936 intestine Anatomy 0.000 description 1
- 230000002427 irreversible effect Effects 0.000 description 1
- 229940017800 lactobacillus casei Drugs 0.000 description 1
- 229940072205 lactobacillus plantarum Drugs 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 230000002906 microbiologic effect Effects 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000036651 mood Effects 0.000 description 1
- 210000004400 mucous membrane Anatomy 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 229940111202 pepsin Drugs 0.000 description 1
- 230000001766 physiological effect Effects 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 238000007747 plating Methods 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 230000002035 prolonged effect Effects 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 210000001187 pylorus Anatomy 0.000 description 1
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Natural products COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 description 1
- 150000003254 radicals Chemical class 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 239000004576 sand Substances 0.000 description 1
- 239000002893 slag Substances 0.000 description 1
- GNBVPFITFYNRCN-UHFFFAOYSA-M sodium thioglycolate Chemical compound [Na+].[O-]C(=O)CS GNBVPFITFYNRCN-UHFFFAOYSA-M 0.000 description 1
- 229940046307 sodium thioglycolate Drugs 0.000 description 1
- 229910001220 stainless steel Inorganic materials 0.000 description 1
- 239000010935 stainless steel Substances 0.000 description 1
- ADNPLDHMAVUMIW-CUZNLEPHSA-N substance P Chemical compound C([C@@H](C(=O)NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCSC)C(N)=O)NC(=O)[C@H](CC=1C=CC=CC=1)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H]1N(CCC1)C(=O)[C@H](CCCCN)NC(=O)[C@H]1N(CCC1)C(=O)[C@@H](N)CCCN=C(N)N)C1=CC=CC=C1 ADNPLDHMAVUMIW-CUZNLEPHSA-N 0.000 description 1
- 238000000967 suction filtration Methods 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 239000003053 toxin Substances 0.000 description 1
- 231100000765 toxin Toxicity 0.000 description 1
- 238000012549 training Methods 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 150000003722 vitamin derivatives Chemical class 0.000 description 1
- 230000005186 women's health 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
- A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
- A23C9/00—Milk preparations; Milk powder or milk powder preparations
- A23C9/12—Fermented milk preparations; Treatment using microorganisms or enzymes
- A23C9/123—Fermented milk preparations; Treatment using microorganisms or enzymes using only microorganisms of the genus lactobacteriaceae; Yoghurt
-
- 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
- A23V2400/143—Fermentum
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/225—Lactobacillus
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Microbiology (AREA)
- Organic Chemistry (AREA)
- Biotechnology (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Genetics & Genomics (AREA)
- Polymers & Plastics (AREA)
- Biomedical Technology (AREA)
- Virology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Medicinal Chemistry (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Food Science & Technology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
Abstract
It is CCTCC NO the invention discloses lactobacillus fermenti (Lactobacillus fermentum strain Zhao) deposit number of a kind of adjustable intestinal movement, Constipation：M 2013513 and application thereof, the acid resistance of the bacterial strain is stronger, survival rate has reached 85.12 ± 5.18% after 3h under the simulated gastric fluid of pH3.0, it can slowly be grown under 1.0% concentration cholate, the hydrophobicity that growth efficiency reaches 18.33 ± 0.52, the Lactobacillus fermentum strain Zhao cells of no cholate culture has also reached 70.80 ± 0.61%.Can in the enteron aisle of people normal growth.In-vivo test in mice also shows, lactobacillus fermenti (Lactobacillus fermentum strain Zhao) can slow down mouse weight caused by constipation and mitigate trend, accelerate row's melena time, improve Intestinal propulsive rate, MTL, Gas, ET, AchE, SP and VIP factor level of different degrees of increase serum, reduce the SS factors, increase food ration and amount of drinking water, reduce feces volume, defecation granule number and excrement water content downward trend, play the role of with constipation drug similar, shows certain constipation preventive effect.
Description
Technical field
The present invention relates to microorganisms technical field more particularly to the acidified milks of a kind of adjustable intestinal movement, Constipation
Bacillus Lactobacillus fermentum strain Zhao and application thereof.
Background technology
Lactobacillus is in close relations with human lives, is to be widely used in food fermentation, industrial lactic fermentation and health care
One of beneficial microbe in field.Many kinds in the lactobacillus and Bifidobacterium that have recognized all have it is specific excellent
Benign matter and inhereditary feature show extremely strong probiotic properties.Lactobacillus can not only improve food value, improve food
Product flavor extends the holding time, can also improve the functional character of food.It can be by decomposing protein, sugar in food
Class, synthesis vitamin, improve the digestibility and biological value of food, promote to digest and assimilate.Can also reduce human cholesterol content,
Enhance immune function, anti-allergenicity, antitumor, anti-hypertension etc..In recent years, lactobacillus is with its special physiological activity and battalion
Support function and as probiotics main source, just have attracted increasing attention.
Constipation is a kind of very universal disease, and the illness rate in crowd is up to 27%, it can influence each age group
People.Habitual constipation is to induce middle-aging male hyperlipidemia, cardiovascular and cerebrovascular disease because firmly increasing abdominal pressure, defecation with strength of holding one's breath
The arch-criminal of the diseases such as disease, artery sclerosis, tumour and its complication, drastically so as to cause old man's cardio cerebrovascular affection death rate
Increase, the problems such as hemorrhoid, anal fissure occurs, it is painful to can't bear.In addition prolonged constipation, largely just slag can be trapped in hair in enteron aisle
Ferment rots, and a large amount of toxin and harmful bacteria are absorbed repeatedly by enteron aisle, reaches partes corporis humani position by blood circulation, it is strong to become women
Health and beautiful arch-criminal.Constipation can also make child's gastrointestinal bacterial flora unbalance, cause energy not concentrated, thought slowness, nutrition
It is bad, mood is irritable, becomes thin, immunity degradation, easy catching a cold, growth retardation, such as improve not in time, irreversible disease may be caused
Become.Because constipation incidence is high, the cause of disease is complicated, patient often has manifold vexations, and quality of life can be influenced when constipation is serious.Therefore, it seeks
Look for it is a kind of can alternatives to medication carry out the biological products of Constipation to have prodigious realistic meaning, Tiny ecosystem therapy be just fine with one
New means solve various problems existing for traditional remedies.
There is a large amount of research report to the physiological function of lactic acid bacteria both at home and abroad, but the functionality of lactic acid bacteria Constipation is ground
It is seldom to study carefully report.Research about lactic acid bacteria Constipation to many subjects such as Food Science, preventive medicine and microecology all
There is very big value.Fermented dairy product is the important channel that people take in lactic acid bacteria, screen it is excellent have Constipation and
The lactic acid bacteria strains of safety will have development functionality dairy products, the added value enriched existing dairy products type, improve dairy products
It is significant.Therefore, the lactic acid bacteria with Constipation has prodigious research and application value.
In the document and patent that have disclosed at present or patent application, such as CN101144064A discloses one kind and has
The Lactobacillus brevis and application thereof of antimutagenicity, extracellular polysaccharide, for, to improve food quality, increasing it in fermented food
Nutrition function.CN1796540A discloses the Bifidobacterium and application thereof with anti-enterovirus and anti-oxidation characteristics, uses
In preparing food compositions or pharmaceutical composition, helicobacter pylori or Escherichia coli are killed or inhibited, treats and be induced by it
Various diseases.CN102174450A discloses a kind of lactobacillus plantarum of Helicobacter pylori Infection and application thereof, is used to prepare
Prevent or mitigate the infection of helicobacter pylori in pharmaceutical composition, leavening, animal feed additive and health products.
The oxidation resistant Lactobacillus casei and application thereof that CN101144065A discloses a kind of resisting hydrogen peroxide, removes free radical, for sending out
With development functionality dairy products, the added value enriched existing dairy products type, improve dairy products in ferment food and dairy products capsule product.
But these patents or patent application are not related to completely with the microorganism for adjusting the performances such as intestinal movement, Constipation.Cause
This, also needs to a kind of food with the microorganism for adjusting intestinal movement, Constipation and its containing them at present.
Invention content
To solve the above-mentioned problems, the present invention provides a kind of adjustable intestinal movement, the lactobacillus fermentis of Constipation
Lactobacillus fermentum strain Zhao and application thereof mainly provide a kind of adjustable intestinal movement, prevent just
Purposes of the secret L fermentum strain and a kind of lactobacillus fermenti of offer in fermented food.
Lactobacillus fermenti (the Lactobacillus of a kind of adjustable intestinal movement provided by the invention, Constipation
Fermentum strain Zhao) on October 30th, 2013 it is preserved in China typical culture collection center (CCTCC), it should
Collection is located at：The Chinese Wuhan Wuhan Universitys, deposit number are CCTCC NO：M2013513.
The present inventor filters out one plant of lactobacillus fermenti (Lactobacillus from spontaneous fermentation yak yogurt
Fermentum strain Zhao), using Microbiological Characteristics such as morphological feature, cultural colony and physiological and biochemical properties to this
Bacillus acidi lactici Lactobacillus fermentum strain Zhao are accredited as lactobacillus fermenti, and the bacterial strain is in 2013
It was preserved in China typical culture collection center (CCTCC) on October 30, deposit number is CCTCC NO：M2013513.
The morphological feature of the lactobacillus fermenti is specially：
Thalline feature：In Gram-positive, cell is rod-shaped, and thalline is about 0.5-1.0 μm wide, 1.5-4 μm long, Cheng Dan,
Pairs of or chaining, does not form gemma, and both ends are round.
Colony characteristics：Apparent bacterium colony is formed on MRS culture mediums, diameter is round between 0.5-2.0mm, and edge is whole
Together, milky, transparent, surface wettability is smooth, not chromogenesis.
Tolerance is characterized as in the lactobacillus fermenti body：
The bacterial strain acid resistance of lactobacillus fermenti is stronger, under the simulated gastric fluid of pH3.0 after 3h survival rate reached 85.12 ±
5.18%；It can slowly be grown under 1.0% concentration cholate, growth efficiency reaches the 18.33 ± 0.52% of no cholate culture；Hair
The hydrophobicity of kefir milk bacilli-cell has reached 70.80 ± 0.61%.
The lactobacillus fermenti derives from traditional fermented food, belongs to generally recognized as safe (Generally Recognized As
Safe, GRAS) strain, in fermented food.
The fermented food is in lactic acid bacteria milk beverage, milk powder, capsule product or acidified milk containing lactobacillus fermenti
It is one or more.
The lactic acid bacteria milk beverage containing lactobacillus fermenti is prepared as steps described below：
The original strain of the lactobacillus fermenti is preserved at -75 DEG C of temperature with 30% (weight) glycerine suspensions, or
Person is saved backup at 4 DEG C of temperature in the form of being freeze-dried bacterium powder；
Lactobacillus fermenti working stock culture is prepared using the original strain of the lactobacillus fermenti；
Raw milk heats the sterilization 20min or high temperature thermal sterilization 2s at 140 DEG C at 95 DEG C, is cooled to 4 DEG C, described in addition
Lactobacillus fermenti working stock culture makes its concentration reach 106Cfu/mL or more, it is stored refrigerated at 4 DEG C；
The raw milk is one or more raw milks selected from defatted milk, fresh milk, recovery milk.
The milk powder containing lactobacillus fermenti is prepared as steps described below：
The original strain of the lactobacillus fermenti is preserved at -75 DEG C of temperature with 30% (weight) glycerine suspensions, or
Person is saved backup at 4 DEG C of temperature in the form of being freeze-dried bacterium powder；
Lactobacillus fermenti working stock culture is prepared using the original strain of the lactobacillus fermenti；
Raw milk heats the sterilization 20min or high temperature thermal sterilization 2s at 140 DEG C at 95 DEG C, is then cooled to 37 DEG C, then
The bacterium amount inoculation lactobacillus fermenti working stock culture is connect with the 4% of raw milk volume, ferment 16h at 37 DEG C, is fermented
Lactobacillus ferment breast；Raw milk after the lactobacillus fermenti acidified milk and sterilizing is matched according to 1: 3 (V: V) to be added, and is carried out equal
Matter is concentrated in vacuo, spray drying obtains the milk powder containing lactobacillus fermenti.
The capsule product containing lactobacillus fermenti is prepared as steps described below：
The original strain of the lactobacillus fermenti is preserved at -75 DEG C of temperature with 30% (weight) glycerine suspensions, or
Person is saved backup at 4 DEG C of temperature in the form of being freeze-dried bacterium powder；
Lactobacillus fermenti working stock culture is prepared using the original strain of the lactobacillus fermenti；
By raw milk in 140 DEG C of high temperature thermal sterilization 2s, 37 DEG C are subsequently cooled to, connecing bacterium amount with the 4% of raw milk volume connects
The kind lactobacillus fermenti working stock culture, ferment 16h at 37 DEG C, obtains lactobacillus fermenti acidified milk；By the acidified milk
Raw milk after bacillus fermentation breast and sterilizing is according to 1: 3 (V: V)) proportioning addition, homogeneous, vacuum concentration, spray drying treatment
After obtain milk powder, the capsule product containing lactobacillus fermenti will be obtained.
The acidified milk containing lactobacillus fermenti is prepared as steps described below：
The original strain of the lactobacillus fermenti is preserved at -75 DEG C of temperature with 30% (weight) glycerine suspensions, or
Person is saved backup at 4 DEG C of temperature in the form of being freeze-dried bacterium powder；
Lactobacillus fermenti working stock culture is prepared using the original strain of the lactobacillus fermenti；
Raw milk heats the sterilization 20min or high temperature thermal sterilization 2s at 140 DEG C at 95 DEG C, is then cooled to 37 DEG C, then
Lactobacillus fermenti working stock culture is added according to 3-5% (volume), add 3-5% (volume) can symbiosis be used to prepare fermentation
The leavening of dairy products, mixed fungus fermentation is subsequently cooled to 4 to titratable acidity 0.6-0.7% in terms of lactic acid at 37 DEG C after mixing
DEG C, then carry out stored refrigerated obtaining the acidified milk containing lactobacillus fermenti
The preparation method of the lactobacillus fermenti working stock culture includes：
The original strain of the lactobacillus fermenti is inoculated in the skimmed milk of 12% (weight), 110 DEG C of sterilizing 10min,
14-16h is cultivated under the conditions of 37 DEG C to curdled milk, in continuous culture two generations of activation, is used as mother culture；The mother culture is pressed into 3-
5% (volume) is inoculated in sterile milk, and 14-16h is to curdled milk for culture, until the viable count about 10 in the curdled milk9Cfu/mL obtains work
Make leavening, directly this working stock culture can be added in food, or with can symbiosis the business for preparing acidified milk
Leavening such as lactobacillus bulgaricus is used together with streptococcus thermophilus prepares acidified milk；Or
The original strain of the lactobacillus fermenti is inoculated in MRS fluid nutrient mediums, 12- is cultivated under the conditions of 37 DEG C
16h is activated, continuous to activate for two generations, is then inoculated in activation culture object in MRS culture mediums by 2-4% (volume), is cultivated
16-18h, 4000r/min centrifuges 15min under the conditions of 4 DEG C, removes supernatant, cell precipitation is obtained, by precipitation sterile absorbent
Suspension is made in breast, obtains working stock culture.
The heating sterilization is the TW10D1000 for example sold using Shanghai Triowin Automation Machinery Limited
What type tubular sterilization machine carried out.
The high temperature thermal sterilization is the YC-104 types for example sold using Beijing Yong Chuanjiaye mechanical equipments Co., Ltd
What board-like superhigh temperature sterilization machine carried out.
The homogeneous be the small-sized homogenizers of GJB500-40 for example sold using Changzhou homogeneous Machinery Co., Ltd. into
Capable.
The concentration is that the vacuum concentration pan for example sold using Shanghai Wei Zhou light industry and machinery Co., Ltd is carried out.
The spray drying be the experiment type spray drier for example sold using Shanghai Triowin Tech. Co., Ltd. into
Capable.
The commodity leavening is preferably lactobacillus bulgaricus or streptococcus thermophilus.
In the sense of the present invention, the raw milk is one or more raw materials selected from defatted milk, fresh milk, recovery milk
Breast.
What the technical solution of the embodiment of the present invention was brought has the beneficial effect that：
The present invention is a kind of adjustable enteron aisle fortune being isolated from Sichuan Province Hongyuan County herdsman man spontaneous fermentation yak yogurt
The dynamic, lactobacillus fermenti of Constipation and using the bacterial strain as the leavening or acidified milk of addition, healthy food and animal health
The acid resistance of application in product, lactobacillus fermenti (Lactobacillus fermentum strain Zhao) bacterial strain is stronger,
Survival rate has reached 85.12 ± 5.18% after 3h under the simulated gastric fluid of pH3.0, can slowly be given birth under 1.0% concentration cholate
Long, growth efficiency reaches 18.33 ± 0.52%, the Lactobacillus fermentum strain Zhao of no cholate culture
The hydrophobicity of cell has also reached 70.80 ± 0.61%.This shows lactobacillus fermenti (Lactobacillus fermentum
Strain Zhao) can in the enteron aisle of people normal growth.In-vivo test in mice is also shown that lactobacillus fermenti
(Lactobacilius fermentum strain Zhao) can slow down mouse weight caused by constipation and mitigate trend, accelerate
The melena time is arranged, Intestinal propulsive rate, MTL, Gas, ET, AchE, SP and VIP factor water of different degrees of increase serum are improved
It is flat, the SS factors are reduced, food ration and amount of drinking water are increased, reduce feces volume, defecation granule number and excrement water content downward trend,
Play the role of with constipation drug similar, shows certain constipation preventive effect.
Description of the drawings
For the clearer technical solution for illustrating the embodiment of the present invention, required use in being described below to embodiment
Attached drawing be briefly described, it is clear that, drawings in the following description are only some embodiments of the invention, for this
For the those of ordinary skill of field, without creative efforts, others are can also be obtained according to these attached drawings
Attached drawing.
Fig. 1 lactobacillus fermentis (Lactobacillus fermentum strain Zhao) colonial morphology
The thalli morphology (1000x) of Fig. 2 lactobacillus fermentis (Lactobacillus fermentum strain Zhao)
Fig. 3 mouse weights with experimental period variation
The time required to Fig. 4 separate groups of mice arranges melena for the first time
The total small intestinal length of Fig. 5 separate groups of mice
The Intestinal propulsive rate of Fig. 6 separate groups of mice
Each factor level in Fig. 7 separate groups of mice serum
The lactobacillus fermenti (Lactobacillus fermentum strain Zhao) of the present invention was October 30 in 2013
Day is preserved in China typical culture collection center, which is located at：The Chinese Wuhan Wuhan Universitys, number CCTCC
NO：M2013513.
Specific implementation mode
Following will be combined with the drawings in the embodiments of the present invention, and technical solution in the embodiment of the present invention carries out clear, complete
Whole description, it is clear that the described embodiments are merely a part of the embodiments of the present invention, the embodiment being not all of, and is based on this hair
Embodiment in bright, the every other implementation that those of ordinary skill in the art are obtained under the premise of not making the creative labor
Example, shall fall within the protection scope of the present invention.
Embodiment 1：The separation of lactobacillus fermenti (Lactobacillus fermentum strain Zhao), purifying and
Preliminary Identification.
The embodiment carries out as steps described below：
(1) separation, purifying of Lactobacillus fermentum strain Zhao.
It is added to 5mL sterile salines with 500 μ L of sterile working pipette samples and mixes the uniform dilution for being made 1: 10,
And continue to make a certain proportion of dilution.The dilution for selecting suitable gradient is respectively drawn 100 μ L with sterile pipette tips and is respectively coated
In MRS solid plate culture mediums, 30 DEG C of culture 48-72h observe and record colonial morphology, as shown in Figure 1.With oese (or sterilizing
Toothpick) from planar surface and internal picking difference colony inoculation in MRS fluid nutrient mediums, set 30 DEG C, the training of 300r/min shaking tables
Support 24-48h；It repeats to carry out gram stain smear microscopy after above step continuously activated for 2 generations, as shown in Fig. 2, being determined as G+ bacterium
Continue activate, until obtain pure bacterium colony (microscopy is without miscellaneous bacteria), and carry out catalase experiment.G+ and catalase is cloudy
Sporeless bacterium, coccus and the streptococcus of property are fixed tentatively as lactic acid bacteria, and are preserved, concrete operations reference《Microbiology Experiment skill
Art》.
Experimental result is shown, 56 plants of lactic acid bacteria is isolated from ten samples of Ruoergai country, and viable count of lactobacillus is flat
108The cfu/mL orders of magnitude.
(2) Preliminary Identification of Lactobacillus fermentum strain Zhao.
Gram's staining：Colonies typical smear is taken, Gram's staining is carried out, in microbioscope oil microscopic observation thalline
Form and its arrangement mode.Picking gets a clear view, the typical bacterial strain of form, takes pictures.
Catalase is tested：One oese of bacterium colony, is placed in clean tube on picking solid medium, and 3% mistake is added dropwise
Hydrogen peroxide solution 2mL, observation result (being the positive in gassing person in half a minute, gassing person is not feminine gender).
The result shows that the bacterial strain isolated and purified out from sample can be containing 5 ‰ CaCO3MRS plating mediums on it is raw
It is long, and form molten calcium circle, Gram-positive, catalase test feminine gender in periphery of bacterial colonies.
Embodiment 2：The in-vitro screening of lactobacillus fermenti Lactobacillus fermentum strain Zhao.
(1) screening of probiotics tolerance pH3.0 simulated gastric fluids.
The preparation of simulated gastric fluid：NaCl 0.2%, pepsin 0.35%, after being 3.0 with the HCl of 1M adjustment pH value,
With spare after vacuum pump suction filtration degerming in aseptic operating platform.
Measurement of the probiotics to artificial gastric juice resistance's property：5mL activated strain cultured solutions are taken, in aseptic operating platform
In pour into and sterilized 10mL centrifuge tubes, collect thalline through 3000r/min centrifugations 10min, 5mL sterile saline mixings be added
Bacteria suspension is made, takes 1mL bacteria suspensions and the simulated gastric fluid of 9mL pH3.0 to mix, shakes up, is placed in constant temperature oscillator and cultivates (37
DEG C, 300r/min), and sampled respectively in 0h and 3h, pour into 37 DEG C of culture 48h with MRS agar mediums.It is surveyed with colony counting method
Determine viable count, calculates its survival rate (%)：Viable count × 100% of viable count/0h of survival rate (%)=3h.
(2) measurement of probiotics tolerance various concentration cholate.
The survival rate in pH3.0 simulated gastric fluids is selected to do the survey of the growth under different gallbladder salinities in 10% or more strain
Examination.Activated strain 5mL is inoculated in liquid-transfering gun by 2% inoculum concentration (100 μ L) containing 0.0% bovine bile (i.e. sky respectively
In vain), 0.3% bovine bile, 0.5% bovine bile, 1.0% bovine bile (W/V) MRS-THIO culture mediums (in MRS culture mediums plus
0.2% sodium thioglycolate).It is that control is (nonvaccinated with blank cultures in constant temperature oscillator after 37 DEG C of cultures for 24 hours
MRS-THIO culture mediums), the OD values of above-mentioned various concentration culture medium are measured respectively, calculate tolerance of the bacterial strain to cholate.Cholate
OD value × 100% of OD values/blank cultures of tolerance=culture medium containing cholate.
(3) bacterial concentration adjusts.
Selection survival rate strain stronger in 10% or more, bile tolerance in pH3.0 simulated gastric fluids observes bacterium through microscopy
It is tested after body growthform is good, by activated bacterial cultures 5mL, the 10mL that sterilized is poured into aseptic operating platform
In centrifuge tube, thalline is collected with 3000r/min centrifugations 10min.With 5mL PBS (50mM, pH6.5) buffer solution washing thalline,
3000r/min centrifuges 10min, and repeated washing is twice.Using PBS buffer solution as blank control, strain subject bacterium is adjusted with buffer solution
Bulk concentration, it is about 1.00 to make its A0 value under 560nm wavelength.
It takes the bacterium solution that 4mL adjusts turbidity in the 10mL centrifuge tubes that sterilized, 0.8mL dimethylbenzene is added, control group is not added with
Dimethylbenzene, vibrate 30s, pause 10s, after vibrate 30s again, on rack for test tube stand 5~10min layering, remove a layer water phase, with
PBS buffer solution is blank control, and A values are measured at 560nm and are recorded.Hydrophobic rate H%=[(A0-A)/A0] × 100 its
Middle A0 and A is the A values measured at 560nm with the forward and backward bacterium solution of dimethylbenzene mixing respectively.
Lactic acid bacteria is a part for normal bowel bacterium, one of the most important condition as probiotics, and being can be before stomach and small intestine
Section survival, the bacterial strain being screened should be with the characteristics of growth and development under enough acid resistance and acidic environment.
The selection result of Lactobacillus fermentum strain Zhao is as shown in table 1 below：
Table 1
The selection result shows that Lactobacillus fermentum strain Zhao bacterial strains are resistant to the ring of pH3.0
Border, and survival rate is 80% or more.
It will be contacted with the cholate in small intestine by the thalline survived after stomach, this experiment is lactic acid bacteria to the resistivity of cholate
For a selection criteria as potential probiotics.The preferable Lactobacillus fermentum strain of acid resistance
Zhao bacterium be resistant to various concentration cholate the result is that the cholate to 0.3% has certain tolerance, when gallbladder salinity is increased to
When 0.5% and 1.0%, Lactobacillus fermentum strain Zhao thalline have revealed good cholate tolerance.
While except having certain resistivity to cholate in small intestine, lactic acid bacteria need to also have good adherency on mucous membrane of small intestine
Property, therefore by the hydrophobic ability of lactic acid bacteria alternately standard, from table 1 it follows that Lactobacillus
The hydrophobicity of fermentum strain Zhao bacterium has reached 70.80 ± 0.61%.
Acid resistance, bile tolerance and the preferable Lactobacillus of hydrophobicity have been filtered out by testing us above
Fermentum strain Zhao bacterium, the bacterial strain are preserved in China typical culture collection center on October 30th, 2013
(CCTCC), deposit number CCTCC NO：M2013513.
Embodiment 3：Activated carbon inducing constipation is tested.
Feeding of healthy kunming mice, 6 week old, 23 ± 2g of weight, female, 50.It, will be small after feeding basal feed adapts to 5d
Mouse is randomly divided into 5 groups according to weight, respectively：Normal group, constipation control group, Bisacody group, lactobacillus bulgaricus group and
The Lactobacillus fermentum strain Zhao group groups of high and low given low, every group 10.Raising is in stainless steel
In cage, 24 ± 2 DEG C of room temperature holding, relative humidity 50 ± 10%, 12h light and shades rotation (8：00~20：00 illumination).Entirely testing
Period, normal group and the equal feeding basal feed of constipation control group, experimental mice press 1.0 × 10 daily respectively9CFU/kg gavages
Probiotics.Start every morning 9 after 2 weeks：00 gives 2 DEG C of activated carbons of other 5 groups of intragastric administration on mice a concentration of 10% in addition to normal group
Ice water, only, daily gavage is primary, continuous gavage 3 days by 0.2mL/.Until experiment last day, all mouse fasting for 24 hours, then
Gavage activated carbon ice water again, 0.2mL/ is only.5 mouse are for measuring changes of weight, food ration, amount of drinking water, first row in every group
Go out melena required time, defecation just quantity and moisture in feces.Other 5 mouse of mouse are after gavage activated carbon 30min
Dead dissection.
With reference to as shown in Figure 3 and Figure 4 the time required to the mouse weight variation of experiment periods Nei Ge groups, first discharge melena.
Normal group (Normal) mouse weight is put on weight increase with experimental period, control group (Control) after 6 days by
There is constipation weight loss in gavage activated carbon, Bisacody (bisacodyl) constipation medicine group slows down weight loss,
Lactobacillus fermentum strain Zhao groups also slow down weight loss, and high concentration group weight loss is less.
Compared with normal group, constipation control group mice arranges the melena time for the first time pole significant difference (P ＜ 0.05).It is high and low
The average time that the Lactobacillus fermentum strain Zhao groups mouse of given low arranges melena for the first time is low
In constipation control group certain constipation preventive effect is shown higher than normal group and constipation therapy drug Bisacody group.
Variation such as the following table 2 of experiment periods Nei Ge groups mouse diet intake：
Table 2
Every group of mouse n=10ICR, Bisacody：100mg/kg b.w., lactobacillus bulgaricus：1.0×109CFU/kg
B.w., LF-Zhao：Lactobacillus fermentum strain Zhao.
Normal group (Normal) mouse weight is slightly increased with experimental period food ration, and control group is after 6 days due to gavage
There is the reduction of constipation food ration in activated carbon, and Bisacody constipation medicine group food ration is less than normal group higher than control group,
Also there is the situation similar with Bisacody in Lactobacillus fermentum strain Zhao groups, and high concentration group is ingested
Amount reduces less.
Variation such as the following table 3 of experiment periods Nei Ge groups mouse amount of drinking water：
Table 3
Every group of mouse n=10ICR, Bisacody：100mg/kgb.w., lactobacillus bulgaricus：1.0×109CFU/kg
B.w., LF-Zhao：Lactobacillus fermentum strain Zhao.
Normal group mouse weight is slightly increased with experimental period amount of drinking water, and control group is after 6 days since gavage activated carbon goes out
Existing constipation amount of drinking water is reduced, and Bisacody constipation medicine group amount of drinking water is less than normal group higher than control group, Lactobacillus
Also there is the situation similar with Bisacody in fermentum strain Zhao groups, and high concentration group amount of drinking water reduces less.
The feces volume of experimental period Nei Ge group mouse, variation such as the following table 4 of defecation granule number and moisture in feces：
Table 4
Every group of mouse n=10ICR, Bisacody：100mg/kg b.w., lactobacillus bulgaricus：1.0×109CFU/kg
B.w., LF-Zhao：Lactobacillus fermentum strain Zhao.
The feces volume of preceding 6 days each groups, defecation granule number and moisture in feces difference are little.After 6 days after constipation induction, just
Normal pack variable is decreased obviously without significant change, control group feces volume, defecation granule number and more normal group of moisture in feces, than
Sand can pyridine and each probiotic group feces volume, defecation granule number and fecal water are higher than control group, Lactobacillus
The defecation granule number and moisture in feces of fermentum strain Zhao high concentrations are close to medicine group and normal group.
Embodiment 4：Small intestine activated carbon Promoting Experiment.
Mouse cervical dislocation is put to death, splits the mesenterium of abdominal cavity and separating mouse, from pylorus, down toward ileocecus on clip
Whole intestinal tube, is put on blotting paper, and whole small intestine is pulled into straight line, to measure Length of intestine, as " total small intestinal length ", from deep and remote
Door to activated carbon juice forward position is " activated carbon propulsion length ".By measuring the propulsion length of activated carbon in small intestine, it is calculated as follows
Propulsion rate.Propulsion rate (%)=(activated carbon advance distance)/(total small intestinal length) × 100.
Total small intestinal length, the Intestinal propulsive rate of experimental period Nei Ge group mouse are as shown in Figure 5 and Figure 6.
Compared with normal group, constipation control group mice Intestinal propulsive rate has pole significant difference (P ＜ 0.05), high and low gavage
The Lactobacillus fermentum strain Zhao group mouse small intestine propulsion rates of dosage are higher than constipation control group, are less than
Normal group and constipation therapy drug Bisacody group, show certain constipation preventive effect.
Embodiment 5：The measurement of each factor level in mice serum.
0.2mL rat aorta blood is taken, at 4 DEG C, 3000r/min centrifuges 10min, takes upper serum.According to motilin
(MTL), gastrin (Gas), Endothelin (ET), growth hormone release inhibiting hormone (SS), serum acetylcholinesterase (AchE), Arg-Pro-Lys-Pro-Gln-Gln-Phe-Phe-Gly-Leu-Met-NH2 (SP) and
The method of vasoactive intestinal peptide (VIP) test agent box specification measures above-mentioned factor level in serum.
Each factor level is as shown in Figure 7 in experiment periods Nei Ge groups mice serum.
Relative to constipation control group, Lactobacillus fermentum strain Zhao height is organized can different journeys
MTL, Gas, ET, AchE, SP and VIP factor level of the increase serum of degree, reduce SS factor levels.So
Lactobacillus fermentum strain Zhao have prevention effect to constipation.
Application Example 1：Breast is manufactured using lactobacillus fermenti (Lactobacillus fermentum strain Zhao)
Sour bacterium milk drink.
First, lactobacillus fermenti (the Lactobacillus fermentum strain Zhao) original strain is in temperature
It is preserved, or is preserved in the form of being freeze-dried bacterium powder at 4 DEG C of temperature standby with 30 weight % glycerine suspensions at -75 DEG C of degree
With.
It is then possible to prepare lactobacillus fermenti (the Lactobacillus fermentum of the present invention using two methods
Strain Zhao) working stock culture：
First method is that above-mentioned lactobacillus fermenti (Lactobacillus fermentum strain Zhao) is original
Strain be inoculated in 12 weight % sterilize at 110 DEG C 10min skimmed milk in, 14-16h is cultivated under the conditions of 37 DEG C to curdled milk, even
In continuous culture two generations of activation, it is used as mother culture；The mother culture is inoculated in by 3-5% (volume) in sterile milk, 14- is cultivated
16h is to curdled milk, the viable count about 10 in the curdled milk at this time9Cfu/mL obtains the working stock culture, can directly will be this
Working stock culture is added in food, or with can symbiosis the commercial fermentation agent such as lactobacillus bulgaricus for preparing acidified milk and
Streptococcus thermophilus, which is used together, prepares acidified milk.
Second method is that above-mentioned lactobacillus fermenti (Lactobacillus fermentum strain Zhao) is original
Strain is inoculated in MRS fluid nutrient mediums, and 12-16h is cultivated under the conditions of 37 DEG C and is activated, continuous to activate for two generations, then will
Activation culture object is inoculated in by 2-4% (volume) in MRS culture mediums, cultivates 16-18h, and 4000r/min is centrifuged under the conditions of 4 DEG C
15min removes supernatant, obtains cell precipitation, and suspension is made with a certain amount of sterile absorbent breast in precipitation, obtains work hair
Ferment agent is spare.
Then, raw milk heats the sterilization 20min or high temperature thermal sterilization 2s at 140 DEG C at 95 DEG C, is then cooled to 4
DEG C, foregoing lactobacillus fermenti (Lactobacillus fermentum strain Zhao) working stock culture is added,
Its concentration is set to reach 106Cfu/ml or more, 4 DEG C it is stored refrigerated i.e. obtain the (Lactobacillus containing lactobacillus fermenti
Fermentum strain Zhao) viable bacteria lactic acid bacteria milk beverage.
In the present invention, the MRS fluid nutrient mediums are known to those skilled in the art, are Suo Laibao public affairs
Take charge of the culture medium for lactobacillus culture of sale.
The heating sterilization is the TW10D1000 for example sold using Shanghai Triowin Automation Machinery Limited
What type tubular sterilization machine carried out.
The high temperature thermal sterilization is the YC-104 types for example sold using Beijing Yong Chuanjiaye mechanical equipments Co., Ltd
What board-like superhigh temperature sterilization machine carried out.
Application Example 2：Breast is manufactured using lactobacillus fermenti (Lactobacillus fermentum strain Zhao)
Powder.
According to above-mentioned lactic acid bacteria milk beverage preparation method prepare, only raw milk heat at 95 DEG C sterilize 20min or
The high temperature thermal sterilization 2s at 140 DEG C, is then cooled to 37 DEG C, then with the 4% of raw milk volume to connect bacterium amount inoculation foregoing
Lactobacillus fermenti (Lactobacillus fermentum strain Zhao) working stock culture, then the 16h that ferments at 37 DEG C,
Obtain lactobacillus fermenti (Lactobacillus fermentum strain Zhao) acidified milk；Then the acidified milk bar
Bacterium (Lactobacillus fermentum strain Zhao) acidified milk is added to above-mentioned sterilizing raw milk according to 1: 3 (V: V)
In, homogeneous is carried out, is concentrated in vacuo, spray drying obtains (the Lactobacillus fermentum strain containing lactobacillus fermenti
Zhao milk powder).
The homogeneous be the small-sized homogenizers of GJB500-40 for example sold using Changzhou homogeneous Machinery Co., Ltd. into
Capable.
The concentration is that the vacuum concentration pan for example sold using Shanghai Wei Zhou light industry and machinery Co., Ltd is carried out.
The spray drying be the experiment type spray drier for example sold using Shanghai Triowin Tech. Co., Ltd. into
Capable.
Application Example 3：Glue is manufactured using lactobacillus fermenti (Lactobacillus fermentum strain Zhao)
Capsule product.
By raw milk in 140 DEG C of high temperature thermal sterilization 2s, 37 DEG C are subsequently cooled to, connecing bacterium amount with the 4% of raw milk volume connects
Lactobacillus fermenti (Lactobacillus fermentum strain Zhao) working stock culture of the kind present invention, at 37 DEG C
Ferment 16h, obtains lactobacillus fermenti (Lactobacillus fermentum strain Zhao) acidified milk.By acidified milk bar
Bacterium (Lactobacillus fermentum strain Zhao) acidified milk is added in the raw milk after sterilizing with 1: 3 (V: V)
Matter obtains milk powder through being concentrated in vacuo, after spray drying treatment, obtained milk powder is attached in capsule, capsule product is made.
Application Example 4：It is prepared and is sent out using lactobacillus fermenti (Lactobacillus fermentum strain Zhao)
Kefir milk.
According to above-mentioned lactic acid bacteria milk beverage preparation method prepare, only raw milk heat at 95 DEG C sterilize 20min or
The high temperature thermal sterilization 2s at 140 DEG C, is then cooled to 37 DEG C, and foregoing acidified milk bar is added according still further to 3-5% (volume)
Bacterium (Lactobacillus fermentum strain Zhao) working stock culture, add 3-5% (volume) can symbiosis system
Preparation kefir milk commodity leavening, at 37 DEG C then mixed fungus fermentation is cooled down to titratable acidity 0.6-0.7% in terms of lactic acid after mixing
To 4 DEG C, then carry out the acidified milk described in stored refrigerated obtain.
The commodity leavening is preferably lactobacillus bulgaricus or streptococcus thermophilus.
In the sense of the present invention, the raw milk is one or more raw materials selected from defatted milk, fresh milk, recovery milk
Breast.
The above, only specific embodiments of the present invention, but the feature of the present invention is not limited thereto, it is any to be familiar with being somebody's turn to do
In the field of the present invention, the changes or modifications that can be readily occurred in should all cover special in application of the invention below the people of item technology
In sharp range.
Claims (2)
1. the lactobacillus fermenti of a kind of adjustable intestinal movement, Constipation, the lactobacillus fermenti is Lactobacillus
Fermentum strain zhao were preserved in China typical culture collection center (CCTCC) on October 30th, 2013, protected
It is CCTCC NO to hide number：M2013513.
2. purposes of the lactobacillus fermenti as described in claim 1 in preparing fermented food, which is characterized in that the fermentation food
Product are lactic acid bacteria milk beverage, milk powder, capsule containing lactobacillus fermenti Lactobacillus fermentum strain zhao
It is one or more in product or acidified milk.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410578613.3A CN104531549B (en) | 2014-05-28 | 2014-10-27 | A kind of lactobacillus fermenti Lactobacillus fermentum strain Zhao and application thereof of adjustable intestinal movement, Constipation |
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2014102277625 | 2014-05-28 | ||
| CN201410227762 | 2014-05-28 | ||
| CN201410578613.3A CN104531549B (en) | 2014-05-28 | 2014-10-27 | A kind of lactobacillus fermenti Lactobacillus fermentum strain Zhao and application thereof of adjustable intestinal movement, Constipation |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104531549A CN104531549A (en) | 2015-04-22 |
| CN104531549B true CN104531549B (en) | 2018-08-10 |
Family
ID=52847184
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410578613.3A Active CN104531549B (en) | 2014-05-28 | 2014-10-27 | A kind of lactobacillus fermenti Lactobacillus fermentum strain Zhao and application thereof of adjustable intestinal movement, Constipation |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104531549B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110938563A (en) * | 2019-12-05 | 2020-03-31 | 南京伯恩世纪信息科技有限公司 | Lactobacillus BJ-REBORN001 and application thereof in preparation of helicobacter pylori inhibiting fermentation broth |
Families Citing this family (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105132330A (en) * | 2015-09-14 | 2015-12-09 | 西南大学 | Lactobacillus fermentum strain Lee capable of preventing gastric ulcers and application of lactobacillus fermentum strain Lee |
| CN105132328B (en) * | 2015-09-14 | 2019-01-11 | 西南大学 | It is a kind of can preventing gastric ulcer lactobacillus fermenti Lactobacillus fermentum strain suo and application thereof |
| JP6847577B2 (en) * | 2015-12-28 | 2021-03-24 | 森永乳業株式会社 | Bifidobacterium and / or lactic acid bacteria growth promoter and / or reduction inhibitor |
| CN107227275A (en) * | 2017-07-06 | 2017-10-03 | 西南大学 | A kind of lactobacillus fermenti HY01 and application thereof |
| CN109628346B (en) * | 2018-12-29 | 2022-07-12 | 善恩康生物科技（苏州）有限公司 | Lactobacillus fermentum CQPC04 and application thereof in preparing fermented food |
| KR20220118438A (en) * | 2019-11-26 | 2022-08-25 | 쿼럼 이노베이션즈 엘엘씨 | Novel protective barrier compositions and uses thereof |
| CN112535700B (en) * | 2020-12-09 | 2022-07-22 | 杭州娃哈哈科技有限公司 | Weight-losing powder based on lactobacillus and preparation method thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1701116A (en) * | 2002-09-06 | 2005-11-23 | Vri生物医学有限公司 | Probiotic bacterium: lactobacillus fermentum |
| CN101144065A (en) * | 2007-09-03 | 2008-03-19 | 江南大学 | Oxidation resistant Lactobacillus casei capable of resisting hydrogen peroxide and eliminating free radical, and use thereof |
| EP2520644A1 (en) * | 2011-05-06 | 2012-11-07 | Biotmicrogen, S.L. | Lactobacillus fermentum CECT 7472 strain with probiotic properties |
-
2014
- 2014-10-27 CN CN201410578613.3A patent/CN104531549B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1701116A (en) * | 2002-09-06 | 2005-11-23 | Vri生物医学有限公司 | Probiotic bacterium: lactobacillus fermentum |
| CN101144065A (en) * | 2007-09-03 | 2008-03-19 | 江南大学 | Oxidation resistant Lactobacillus casei capable of resisting hydrogen peroxide and eliminating free radical, and use thereof |
| EP2520644A1 (en) * | 2011-05-06 | 2012-11-07 | Biotmicrogen, S.L. | Lactobacillus fermentum CECT 7472 strain with probiotic properties |
Non-Patent Citations (1)
| Title |
|---|
| Synbiotic administration of canine-derived strain Lactobacillus fermentum CCM 7421 and inulin to healthy dogs;Viola Strompfová等;《Can. J. Microbiol》;20130319;347-352 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110938563A (en) * | 2019-12-05 | 2020-03-31 | 南京伯恩世纪信息科技有限公司 | Lactobacillus BJ-REBORN001 and application thereof in preparation of helicobacter pylori inhibiting fermentation broth |
Also Published As
| Publication number | Publication date |
|---|---|
| CN104531549A (en) | 2015-04-22 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104531549B (en) | A kind of lactobacillus fermenti Lactobacillus fermentum strain Zhao and application thereof of adjustable intestinal movement, Constipation | |
| CN104498383B (en) | A kind of lactobacillus fermenti Lactobacillus fermentum strain suo and application thereof of adjustable intestinal movement, Constipation | |
| CN111484957B (en) | Bifidobacterium animalis subsp lactis i797, and separation and purification method and application thereof | |
| CN104430847B (en) | Lactobacillus fermenti Lactobacillus fermentum Lee working stock cultures products and its Constipation health purpose | |
| CN104381440B (en) | Lactobacillus fermenti Lactobacillus fermentum strain Lee working stock cultures products and its Constipation health purpose | |
| CN106834187B (en) | Bifidobacterium bifidum and application thereof | |
| CN112175864B (en) | Bifidobacterium animalis and breeding method and application thereof | |
| CN109182207B (en) | Lactobacillus acidophilus La-SJLH001 with probiotic functions of regulating blood sugar level, cholesterol level and the like and application thereof | |
| CN110106122B (en) | Lactobacillus plantarum capable of improving sleep and application thereof | |
| CN107893044A (en) | One plant of bifidobacterium longum and its application | |
| CN107164263A (en) | A kind of adjustable function of intestinal canal, the lactobacillus fermenti HY01 for preventing colitis and application thereof | |
| CN105132328B (en) | It is a kind of can preventing gastric ulcer lactobacillus fermenti Lactobacillus fermentum strain suo and application thereof | |
| CN114085792B (en) | Lactobacillus paracasei for preventing and treating colon cancer and application thereof | |
| CN107227275A (en) | A kind of lactobacillus fermenti HY01 and application thereof | |
| CN114381411B (en) | Lactococcus lactis JYLL-60 and application thereof in preparation of product for improving immunity | |
| CN108486000A (en) | A kind of Bifidobacterium single bacterium acidified milk preparation method and applications | |
| CN107475162A (en) | Lactobacillus rhamnosus and its application with the high inhibitory activity of DPP IV | |
| JP2023544655A (en) | Compositions that promote defecation and their uses | |
| CN109628346A (en) | Lactobacillus fermenti CQPC04 and its application in the food that preparation improves constipation | |
| CN105420150A (en) | Lactobacillus acidophilus and application thereof | |
| CN104651271B (en) | Lactobacillus casei Lactobacillus casei strain Qian working stock cultures products and its enteron aisle adjust health purpose | |
| CN108018248B (en) | Lactobacillus casei capable of regulating flora structural disorder caused by antibiotics | |
| CN116970539B (en) | Lactobacillus murine complex, composition and application thereof | |
| CN109207389B (en) | Thrombolytic lipid-lowering probiotic compound bacteria traditional Chinese medicine oral liquid and preparation method thereof | |
| CN116445356B (en) | Bifidobacterium animalis subspecies BA67 for regulating intestinal flora and enhancing immunity and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |