CN114908020B - 一种抗幽门螺旋杆菌感染的植物乳杆菌及其在食用本草酵素产品中的应用 - Google Patents
一种抗幽门螺旋杆菌感染的植物乳杆菌及其在食用本草酵素产品中的应用 Download PDFInfo
- Publication number
- CN114908020B CN114908020B CN202210639283.9A CN202210639283A CN114908020B CN 114908020 B CN114908020 B CN 114908020B CN 202210639283 A CN202210639283 A CN 202210639283A CN 114908020 B CN114908020 B CN 114908020B
- Authority
- CN
- China
- Prior art keywords
- lactobacillus plantarum
- parts
- herbal
- helicobacter pylori
- ferment
- 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
- 240000006024 Lactobacillus plantarum Species 0.000 title claims abstract description 102
- 235000013965 Lactobacillus plantarum Nutrition 0.000 title claims abstract description 102
- 229940072205 lactobacillus plantarum Drugs 0.000 title claims abstract description 102
- 206010019375 Helicobacter infections Diseases 0.000 title claims abstract description 12
- 108090000790 Enzymes Proteins 0.000 title claims description 41
- 102000004190 Enzymes Human genes 0.000 title claims description 41
- 238000004321 preservation Methods 0.000 claims abstract description 10
- 244000005700 microbiome Species 0.000 claims abstract description 5
- 238000009629 microbiological culture Methods 0.000 claims abstract description 3
- 239000002131 composite material Substances 0.000 claims description 38
- 241001076416 Dendrobium tosaense Species 0.000 claims description 24
- 238000000855 fermentation Methods 0.000 claims description 24
- 230000004151 fermentation Effects 0.000 claims description 24
- 235000013361 beverage Nutrition 0.000 claims description 23
- 239000000843 powder Substances 0.000 claims description 20
- 241000222336 Ganoderma Species 0.000 claims description 16
- 150000001875 compounds Chemical class 0.000 claims description 15
- 239000007788 liquid Substances 0.000 claims description 13
- 240000000588 Hericium erinaceus Species 0.000 claims description 9
- 235000007328 Hericium erinaceus Nutrition 0.000 claims description 9
- 239000002994 raw material Substances 0.000 claims description 9
- 235000017784 Mespilus germanica Nutrition 0.000 claims description 7
- 235000000560 Mimusops elengi Nutrition 0.000 claims description 7
- 235000007837 Vangueria infausta Nutrition 0.000 claims description 7
- 238000002360 preparation method Methods 0.000 claims description 7
- 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 claims description 6
- 229940107187 fructooligosaccharide Drugs 0.000 claims description 6
- 235000009917 Crataegus X brevipes Nutrition 0.000 claims description 4
- 235000013204 Crataegus X haemacarpa Nutrition 0.000 claims description 4
- 235000009685 Crataegus X maligna Nutrition 0.000 claims description 4
- 235000009444 Crataegus X rubrocarnea Nutrition 0.000 claims description 4
- 235000009486 Crataegus bullatus Nutrition 0.000 claims description 4
- 235000017181 Crataegus chrysocarpa Nutrition 0.000 claims description 4
- 235000009682 Crataegus limnophila Nutrition 0.000 claims description 4
- 240000000171 Crataegus monogyna Species 0.000 claims description 4
- 235000004423 Crataegus monogyna Nutrition 0.000 claims description 4
- 235000002313 Crataegus paludosa Nutrition 0.000 claims description 4
- 235000009840 Crataegus x incaedua Nutrition 0.000 claims description 4
- 235000002722 Dioscorea batatas Nutrition 0.000 claims description 4
- 235000006536 Dioscorea esculenta Nutrition 0.000 claims description 4
- 240000001811 Dioscorea oppositifolia Species 0.000 claims description 4
- 235000003416 Dioscorea oppositifolia Nutrition 0.000 claims description 4
- 244000303040 Glycyrrhiza glabra Species 0.000 claims description 4
- 235000006200 Glycyrrhiza glabra Nutrition 0.000 claims description 4
- LPLVUJXQOOQHMX-QWBHMCJMSA-N glycyrrhizinic acid Chemical compound O([C@@H]1[C@@H](O)[C@H](O)[C@H](O[C@@H]1O[C@@H]1C([C@H]2[C@]([C@@H]3[C@@]([C@@]4(CC[C@@]5(C)CC[C@@](C)(C[C@H]5C4=CC3=O)C(O)=O)C)(C)CC2)(C)CC1)(C)C)C(O)=O)[C@@H]1O[C@H](C(O)=O)[C@@H](O)[C@H](O)[C@H]1O LPLVUJXQOOQHMX-QWBHMCJMSA-N 0.000 claims description 4
- 235000011477 liquorice Nutrition 0.000 claims description 4
- 210000000582 semen Anatomy 0.000 claims description 4
- 235000005979 Citrus limon Nutrition 0.000 claims description 3
- 244000131522 Citrus pyriformis Species 0.000 claims description 3
- 241000186660 Lactobacillus Species 0.000 claims description 3
- 235000006679 Mentha X verticillata Nutrition 0.000 claims description 3
- 235000002899 Mentha suaveolens Nutrition 0.000 claims description 3
- 235000001636 Mentha x rotundifolia Nutrition 0.000 claims description 3
- 235000011389 fruit/vegetable juice Nutrition 0.000 claims description 3
- 229940039696 lactobacillus Drugs 0.000 claims description 3
- 229940027779 persimmon extract Drugs 0.000 claims description 3
- 108091028043 Nucleic acid sequence Proteins 0.000 claims description 2
- 244000182216 Mimusops elengi Species 0.000 claims 2
- 241000590002 Helicobacter pylori Species 0.000 abstract description 57
- 229940037467 helicobacter pylori Drugs 0.000 abstract description 57
- 238000002474 experimental method Methods 0.000 abstract description 10
- 206010061218 Inflammation Diseases 0.000 abstract description 8
- 230000004054 inflammatory process Effects 0.000 abstract description 8
- 210000002784 stomach Anatomy 0.000 abstract description 7
- 239000002253 acid Substances 0.000 abstract description 5
- 208000015181 infectious disease Diseases 0.000 abstract description 4
- 229940088598 enzyme Drugs 0.000 description 35
- 239000000725 suspension Substances 0.000 description 27
- 239000006228 supernatant Substances 0.000 description 20
- 230000001580 bacterial effect Effects 0.000 description 19
- 239000001963 growth medium Substances 0.000 description 19
- 239000000047 product Substances 0.000 description 16
- 241000699670 Mus sp. Species 0.000 description 14
- 239000000243 solution Substances 0.000 description 14
- 239000011550 stock solution Substances 0.000 description 14
- 238000009630 liquid culture Methods 0.000 description 13
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 12
- 238000012258 culturing Methods 0.000 description 11
- 238000001914 filtration Methods 0.000 description 10
- 240000008397 Ganoderma lucidum Species 0.000 description 9
- 235000001637 Ganoderma lucidum Nutrition 0.000 description 9
- 230000000694 effects Effects 0.000 description 9
- 230000002401 inhibitory effect Effects 0.000 description 9
- 241000894006 Bacteria Species 0.000 description 8
- 210000004051 gastric juice Anatomy 0.000 description 8
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 8
- 230000003385 bacteriostatic effect Effects 0.000 description 7
- 230000002496 gastric effect Effects 0.000 description 7
- 239000002504 physiological saline solution Substances 0.000 description 7
- 241001494479 Pecora Species 0.000 description 6
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 6
- 239000008280 blood Substances 0.000 description 6
- 210000004369 blood Anatomy 0.000 description 6
- 239000006781 columbia blood agar Substances 0.000 description 6
- 235000011194 food seasoning agent Nutrition 0.000 description 6
- 230000005764 inhibitory process Effects 0.000 description 6
- 239000004310 lactic acid Substances 0.000 description 6
- 235000014655 lactic acid Nutrition 0.000 description 6
- 239000012528 membrane Substances 0.000 description 6
- 239000013642 negative control Substances 0.000 description 6
- 239000006041 probiotic Substances 0.000 description 6
- 235000018291 probiotics Nutrition 0.000 description 6
- 238000012360 testing method Methods 0.000 description 6
- 240000002624 Mespilus germanica Species 0.000 description 5
- 238000001816 cooling Methods 0.000 description 5
- 238000011049 filling Methods 0.000 description 5
- 239000000203 mixture Substances 0.000 description 5
- 238000009928 pasteurization Methods 0.000 description 5
- 239000013641 positive control Substances 0.000 description 5
- 238000012163 sequencing technique Methods 0.000 description 5
- 238000005520 cutting process Methods 0.000 description 4
- 239000003814 drug Substances 0.000 description 4
- 230000002255 enzymatic effect Effects 0.000 description 4
- 235000021552 granulated sugar Nutrition 0.000 description 4
- 239000000413 hydrolysate Substances 0.000 description 4
- 229960000282 metronidazole Drugs 0.000 description 4
- VAOCPAMSLUNLGC-UHFFFAOYSA-N metronidazole Chemical compound CC1=NC=C([N+]([O-])=O)N1CCO VAOCPAMSLUNLGC-UHFFFAOYSA-N 0.000 description 4
- 238000000746 purification Methods 0.000 description 4
- 230000004083 survival effect Effects 0.000 description 4
- 238000002560 therapeutic procedure Methods 0.000 description 4
- MZOFCQQQCNRIBI-VMXHOPILSA-N (3s)-4-[[(2s)-1-[[(2s)-1-[[(1s)-1-carboxy-2-hydroxyethyl]amino]-4-methyl-1-oxopentan-2-yl]amino]-5-(diaminomethylideneamino)-1-oxopentan-2-yl]amino]-3-[[2-[[(2s)-2,6-diaminohexanoyl]amino]acetyl]amino]-4-oxobutanoic acid Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CC(O)=O)NC(=O)CNC(=O)[C@@H](N)CCCCN MZOFCQQQCNRIBI-VMXHOPILSA-N 0.000 description 3
- 108010059892 Cellulase Proteins 0.000 description 3
- 108020004414 DNA Proteins 0.000 description 3
- 208000007882 Gastritis Diseases 0.000 description 3
- 241001465754 Metazoa Species 0.000 description 3
- 241000699666 Mus <mouse, genus> Species 0.000 description 3
- 108010059820 Polygalacturonase Proteins 0.000 description 3
- 108060008682 Tumor Necrosis Factor Proteins 0.000 description 3
- 102000000852 Tumor Necrosis Factor-alpha Human genes 0.000 description 3
- 238000009924 canning Methods 0.000 description 3
- 229940106157 cellulase Drugs 0.000 description 3
- 238000004140 cleaning Methods 0.000 description 3
- 239000011248 coating agent Substances 0.000 description 3
- 238000000576 coating method Methods 0.000 description 3
- 108010093305 exopolygalacturonase Proteins 0.000 description 3
- 239000004744 fabric Substances 0.000 description 3
- 210000001156 gastric mucosa Anatomy 0.000 description 3
- 238000011081 inoculation Methods 0.000 description 3
- 238000000034 method Methods 0.000 description 3
- 230000009467 reduction Effects 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 238000012216 screening Methods 0.000 description 3
- 239000011780 sodium chloride Substances 0.000 description 3
- 210000001519 tissue Anatomy 0.000 description 3
- 241000283690 Bos taurus Species 0.000 description 2
- 244000077995 Coix lacryma jobi Species 0.000 description 2
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 2
- 102000003777 Interleukin-1 beta Human genes 0.000 description 2
- 108090000193 Interleukin-1 beta Proteins 0.000 description 2
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 2
- 239000001888 Peptone Substances 0.000 description 2
- 108010080698 Peptones Proteins 0.000 description 2
- 244000088415 Raphanus sativus Species 0.000 description 2
- 235000006140 Raphanus sativus var sativus Nutrition 0.000 description 2
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 239000003242 anti bacterial agent Substances 0.000 description 2
- 239000006285 cell suspension Substances 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
- 208000023652 chronic gastritis Diseases 0.000 description 2
- 230000008029 eradication Effects 0.000 description 2
- 210000004211 gastric acid Anatomy 0.000 description 2
- 230000002068 genetic effect Effects 0.000 description 2
- 239000008103 glucose Substances 0.000 description 2
- 230000036541 health Effects 0.000 description 2
- 230000007407 health benefit Effects 0.000 description 2
- 239000002054 inoculum Substances 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 210000004877 mucosa Anatomy 0.000 description 2
- 235000019319 peptone Nutrition 0.000 description 2
- 229940126409 proton pump inhibitor Drugs 0.000 description 2
- 239000000612 proton pump inhibitor Substances 0.000 description 2
- 230000002829 reductive effect Effects 0.000 description 2
- 239000010802 sludge Substances 0.000 description 2
- 230000001954 sterilising effect Effects 0.000 description 2
- 238000004659 sterilization and disinfection Methods 0.000 description 2
- 239000000758 substrate Substances 0.000 description 2
- 238000012795 verification Methods 0.000 description 2
- 108020004465 16S ribosomal RNA Proteins 0.000 description 1
- 208000002109 Argyria Diseases 0.000 description 1
- 241000186016 Bifidobacterium bifidum Species 0.000 description 1
- 238000009007 Diagnostic Kit Methods 0.000 description 1
- 206010012735 Diarrhoea Diseases 0.000 description 1
- 206010059866 Drug resistance Diseases 0.000 description 1
- 238000002965 ELISA Methods 0.000 description 1
- 229930182566 Gentamicin Natural products 0.000 description 1
- CEAZRRDELHUEMR-URQXQFDESA-N Gentamicin Chemical compound O1[C@H](C(C)NC)CC[C@@H](N)[C@H]1O[C@H]1[C@H](O)[C@@H](O[C@@H]2[C@@H]([C@@H](NC)[C@@](C)(O)CO2)O)[C@H](N)C[C@@H]1N CEAZRRDELHUEMR-URQXQFDESA-N 0.000 description 1
- 102000000589 Interleukin-1 Human genes 0.000 description 1
- 108010002352 Interleukin-1 Proteins 0.000 description 1
- 238000012449 Kunming mouse Methods 0.000 description 1
- 201000010538 Lactose Intolerance Diseases 0.000 description 1
- 206010025323 Lymphomas Diseases 0.000 description 1
- 240000000249 Morus alba Species 0.000 description 1
- 235000008708 Morus alba Nutrition 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 238000012408 PCR amplification Methods 0.000 description 1
- 102000057297 Pepsin A Human genes 0.000 description 1
- 108090000284 Pepsin A Proteins 0.000 description 1
- 208000008469 Peptic Ulcer Diseases 0.000 description 1
- 244000197580 Poria cocos Species 0.000 description 1
- 235000008599 Poria cocos Nutrition 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 240000004534 Scutellaria baicalensis Species 0.000 description 1
- 235000017089 Scutellaria baicalensis Nutrition 0.000 description 1
- 241000589970 Spirochaetales Species 0.000 description 1
- 208000005718 Stomach Neoplasms Diseases 0.000 description 1
- 208000007107 Stomach Ulcer Diseases 0.000 description 1
- 239000004098 Tetracycline Substances 0.000 description 1
- 244000273928 Zingiber officinale Species 0.000 description 1
- 235000006886 Zingiber officinale Nutrition 0.000 description 1
- 230000001154 acute effect Effects 0.000 description 1
- 229960003022 amoxicillin Drugs 0.000 description 1
- LSQZJLSUYDQPKJ-NJBDSQKTSA-N amoxicillin Chemical compound C1([C@@H](N)C(=O)N[C@H]2[C@H]3SC([C@@H](N3C2=O)C(O)=O)(C)C)=CC=C(O)C=C1 LSQZJLSUYDQPKJ-NJBDSQKTSA-N 0.000 description 1
- 229960000723 ampicillin Drugs 0.000 description 1
- AVKUERGKIZMTKX-NJBDSQKTSA-N ampicillin Chemical compound C1([C@@H](N)C(=O)N[C@H]2[C@H]3SC([C@@H](N3C2=O)C(O)=O)(C)C)=CC=CC=C1 AVKUERGKIZMTKX-NJBDSQKTSA-N 0.000 description 1
- 229940088710 antibiotic agent Drugs 0.000 description 1
- 239000012984 antibiotic solution Substances 0.000 description 1
- 229940006383 azithromycin 10 mg/ml Drugs 0.000 description 1
- 235000015278 beef Nutrition 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 210000004556 brain Anatomy 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 230000000711 cancerogenic effect Effects 0.000 description 1
- 239000002775 capsule Substances 0.000 description 1
- 231100000357 carcinogen Toxicity 0.000 description 1
- 239000003183 carcinogenic agent Substances 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 229960002626 clarithromycin Drugs 0.000 description 1
- AGOYDEPGAOXOCK-KCBOHYOISA-N clarithromycin Chemical compound O([C@@H]1[C@@H](C)C(=O)O[C@@H]([C@@]([C@H](O)[C@@H](C)C(=O)[C@H](C)C[C@](C)([C@H](O[C@H]2[C@@H]([C@H](C[C@@H](C)O2)N(C)C)O)[C@H]1C)OC)(C)O)CC)[C@H]1C[C@@](C)(OC)[C@@H](O)[C@H](C)O1 AGOYDEPGAOXOCK-KCBOHYOISA-N 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 description 1
- 208000035475 disorder Diseases 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 208000000718 duodenal ulcer Diseases 0.000 description 1
- 201000006549 dyspepsia Diseases 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 206010017758 gastric cancer Diseases 0.000 description 1
- 244000000075 gastric pathogen Species 0.000 description 1
- 210000001035 gastrointestinal tract Anatomy 0.000 description 1
- 229960002518 gentamicin Drugs 0.000 description 1
- 235000008397 ginger Nutrition 0.000 description 1
- 238000007490 hematoxylin and eosin (H&E) staining Methods 0.000 description 1
- 210000000987 immune system Anatomy 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 230000002757 inflammatory effect Effects 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 230000000968 intestinal effect Effects 0.000 description 1
- 230000002147 killing effect Effects 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 210000003563 lymphoid tissue Anatomy 0.000 description 1
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 1
- 235000019341 magnesium sulphate Nutrition 0.000 description 1
- 229940099596 manganese sulfate Drugs 0.000 description 1
- 235000007079 manganese sulphate Nutrition 0.000 description 1
- 239000011702 manganese sulphate Substances 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
- 239000000463 material Substances 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 238000000386 microscopy Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 239000006872 mrs medium Substances 0.000 description 1
- 230000017066 negative regulation of growth Effects 0.000 description 1
- 150000007523 nucleic acids Chemical group 0.000 description 1
- 230000008520 organization Effects 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- LSQZJLSUYDQPKJ-UHFFFAOYSA-N p-Hydroxyampicillin Natural products O=C1N2C(C(O)=O)C(C)(C)SC2C1NC(=O)C(N)C1=CC=C(O)C=C1 LSQZJLSUYDQPKJ-UHFFFAOYSA-N 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 229940111202 pepsin Drugs 0.000 description 1
- 229920000136 polysorbate Polymers 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 230000000529 probiotic effect Effects 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 239000001632 sodium acetate Substances 0.000 description 1
- 235000017281 sodium acetate Nutrition 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 230000007480 spreading Effects 0.000 description 1
- 238000003892 spreading Methods 0.000 description 1
- 201000011549 stomach cancer Diseases 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 229960002180 tetracycline Drugs 0.000 description 1
- 229930101283 tetracycline Natural products 0.000 description 1
- 235000019364 tetracycline Nutrition 0.000 description 1
- 150000003522 tetracyclines Chemical class 0.000 description 1
- YWYZEGXAUVWDED-UHFFFAOYSA-N triammonium citrate Chemical compound [NH4+].[NH4+].[NH4+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O YWYZEGXAUVWDED-UHFFFAOYSA-N 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- 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
- A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
- A23L2/38—Other non-alcoholic beverages
- A23L2/382—Other non-alcoholic beverages fermented
-
- 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
- A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
- A23L29/065—Microorganisms
-
- 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
-
- 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
- A61K35/747—Lactobacilli, e.g. L. acidophilus or L. brevis
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- 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
-
- 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
- A23V2400/169—Plantarum
-
- 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
- C12R2001/25—Lactobacillus plantarum
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Microbiology (AREA)
- Nutrition Science (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Genetics & Genomics (AREA)
- General Health & Medical Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biotechnology (AREA)
- Mycology (AREA)
- Polymers & Plastics (AREA)
- Public Health (AREA)
- Biomedical Technology (AREA)
- General Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- Veterinary Medicine (AREA)
- Tropical Medicine & Parasitology (AREA)
- Virology (AREA)
- Epidemiology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Oncology (AREA)
- General Chemical & Material Sciences (AREA)
- Molecular Biology (AREA)
- Communicable Diseases (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
本发明涉及微生物技术领域,尤其是涉及一种抗幽门螺旋杆菌感染的植物乳杆菌(Lactobacillus plantarum)RH JS001及其在食用本草酵素产品中的应用。所述植物乳杆菌(Lactobacillus plantarum)RH JS001保藏于中国微生物菌种保藏管理委员会普通微生物中心,保藏地址为北京市朝阳区北辰西路1号院3号,保藏编号为:CGMCC No.23703。植物乳杆菌(Lactobacillus plantarum)RH JS001具有良好的耐酸性,能有效抑制幽门螺旋杆菌的生长,经实验证实,其具有抗幽门螺旋杆菌感染的能力,还可以显著减轻幽门螺杆菌感染小鼠胃部的炎症反应。
Description
技术领域
本发明涉及微生物技术领域,尤其是涉及一种抗幽门螺旋杆菌感染的植物乳杆菌(
Lactobacillus plantarum)RH JS001及其在食用本草酵素产品中的应用。
背景技术
幽门螺旋杆菌(Helicobacter pylori, HP)是一种革兰氏阴性螺旋状微需氧菌,定植于胃黏膜表面和黏膜层之间,最初由澳大利亚学者Marshall和Warren于1982年发现并提出其与消化性溃疡和慢性胃炎的发病相关。现代大量医学研究表明,幽门螺旋杆菌是急、慢性胃炎以及胃、十二指肠溃疡的主要致病原因,并可能与胃癌和胃粘膜相关性淋巴样组织(MALT)恶性淋巴瘤发病有关。世界卫生组织(WHO)已经将其列为I级致癌因子。
关于治疗幽门螺旋杆菌感染的方案,我国科学家一直在不断探索,目前最流行的方案是同时服用质子泵抑制剂(PPI)加两种抗生素(克拉霉素、阿莫西林、四环素、甲硝唑等选二种)的三联疗法。然而,在实际治疗过程中,随着幽门螺旋杆菌耐药率上升,三联疗法的根除率不断下降,且质子泵抑制剂会导致消化不良,大量的抗菌剂也会破坏人体的正常菌群的平衡。
当前形势,寻求一种治疗幽门螺旋杆菌的新疗法刻不容缓。因此,益生菌疗法应运而生。益生菌通常被定义为活的微生物,当摄入一定的量时,可以通过改善宿主肠道微生物平衡来提供健康益处。这些健康益处包括减轻乳糖不耐症,增强免疫***,预防和治疗癌症,降低胆固醇水平,减少包括抗生素相关性腹泻在内的胃肠道疾病,提高胃病原体幽门螺旋杆菌的根除率等。然而,中国专利CN111118106A和CN113980878A并未结合体内验证菌株的应用效果,中国专利CN111548970A和CN111607538A未从抑制幽门螺旋杆菌能力出发筛选乳酸菌,且上述专利仅将益生菌在产品上的应用局限于抑制剂和胶囊等粉剂。
在人民愈发关注中药养生的今天,由益生菌发酵,药食同源原料作为发酵底物的可食用本草酵素,无疑是一个新的赛道。大量研究表明,猴头菇、甘草、黄芩、桑叶、生姜、茯苓、灵芝、山楂、红枣、黄精、山药、薏苡仁、枸杞和铁皮石斛等药食同源原料皆具有抑制幽门螺旋杆菌生长的作用。考虑到益生菌抗幽门螺旋杆菌感染的功能具有菌株依赖性,持续筛选可食用的,具有抑制幽门螺旋杆菌生长能力的菌株并将其应用于食用本草酵素产品,具有现实意义和广阔前景。
发明内容
本发明的目的在于克服上述现有技术的不足之处而提供一种抗幽门螺旋杆菌感染的植物乳杆菌(
Lactobacillus plantarum)RH JS001及其在食用本草酵素产品中的应用。
为实现上述目的,本发明采取的技术方案为:
第一目的,本发明提供了一种包含如SEQ ID No: 1所示的16rRNA序列的植物乳杆菌(
Lactobacillus plantarum)。
本发明的发明人意外地从四川省的农家自制酸萝卜分离筛选出一株植物乳杆菌(
Lactobacillus plantarum)RH JS001,该菌株经测序分析,其16S rDNA序列如SEQ IDNO.1所示,将测序得到的序列在GeneBank中进行核酸序列比对,结果显示菌株为植物乳杆菌。
基于上述发现,本发明提供了一种植物乳杆菌(
Lactobacillus plantarum)RHJS001,所述植物乳杆菌(
Lactobacillus plantarum)RHJS001保藏于中国微生物菌种保藏管理委员会普通微生物中心,保藏地址为北京市朝阳区北辰西路1号院3号,保藏编号为:CGMCC No.23703,保藏时间为2021年11月1日。
本发明的植物乳杆菌(
Lactobacillus plantarum)RH JS001具有良好的耐酸性,能有效抑制幽门螺旋杆菌的生长,经动物实验证实,其具有抗幽门螺旋杆菌感染的能力,还可以显著减轻幽门螺杆菌感染小鼠胃部的炎症反应。
植物乳杆菌(
Lactobacillus plantarum)RH JS001的菌悬液和上清液对幽门螺旋杆菌的抑菌圈大小分别可达18.50±1.70 mm和16.90±2.10mm。
第二目的,本发明提供了上述植物乳杆菌(
Lactobacillus plantarum)接种原料液形成的发酵液。
优选地,植物乳杆菌(
Lactobacillus plantarum)RH JS001按2-6%的接种量接种入原料液中。
第三目的,本发明提供了上述植物乳杆菌(
Lactobacillus plantarum)或其发酵液在制备抗幽门螺杆菌感染制剂中的应用。
第四目的,本发明提供了上述植物乳杆菌(
Lactobacillus plantarum)或其发酵液在制备食用本草酵素产品中的应用。
作为本发明所述应用的优选实施方式,所述植物乳杆菌(
Lactobacillus
plantarum)或其发酵液在发酵过程中抑制幽门螺杆菌中的应用。
第五目的,本发明提供了一种食用本草酵素,所述食用本草酵素包括以下组分通过植物乳杆菌(Lactobacillus plantarum)发酵而成;所述组分包括铁皮石斛或灵芝。
在某些实施方案中,植物乳杆菌(
Lactobacillus plantarum)RH JS001接种至原料液中,获得发酵液,发酵条件:温度为30-40℃,发酵时间48-72 h,静置培养。
本发明提供了由植物乳杆菌(
Lactobacillus plantarum)发酵得到的食用本草酵素,该食用本草酵素能够抑制幽门螺旋杆菌生长。
在某些实施方案中,铁皮石斛酵素对幽门螺旋杆菌的抑菌圈大小可达12.40±0.90 mm。
在某些实施方案中,灵芝酵素对幽门螺旋杆菌的抑菌圈大小可达16.10±1.40mm。
第六目的,本发明提供了一种复合本草酵素,所述复合本草酵素包括以下组分通过上述植物乳杆菌(
Lactobacillus plantarum)发酵而成;
组分包括铁皮石斛、灵芝、猴头菇、甘草、山楂、黄精、山药、薏苡仁和枸杞中的至少三种。
作为本发明所述复合本草酵素的优选实施方式,所述复合本草酵素包括以下重量份数的组分:
铁皮石斛1-8份、灵芝1-8份、猴头菇5-25份、甘草5-20份、山楂2-16份、黄精2-16份、山药2-16份、薏苡仁1-3份和枸杞1-8份。
第七目的,本发明提供了一种本草酵素产品,所述本草酵素产品包括复合本草酵素饮料A和/或复合本草酵素饮料B。
作为本发明所述本草酵素产品的优选实施方式,所述复合本草酵素饮料A包括以下重量份数的组分:
复合本草酵素3-15份、猴头菇粉0.01-0.2份、红枣粉0.1-2份、枸杞果汁粉0.01-0.2份和低聚果糖0.1-2份。
作为本发明所述本草酵素产品的优选实施方式,所述复合本草酵素饮料B包括以下重量份数的组分:
复合本草酵素3-15份、柠檬粉0.1-2份、薄荷粉0.001-0.01份、柿子提取物0.01-0.2份和低聚果糖0.1-2份。
与现有技术相比,本发明具有以下有益效果:
本发明提供的植物乳杆菌(
Lactobacillus plantarum)RH JS001具有良好的耐酸性,能够抑制幽门螺旋杆菌生长,在抑制幽门螺旋杆菌生长方面具有巨大的应用前景。
附图说明
图1为植物乳杆菌(
Lactobacillusplantarum)RH JS001的16rDNA测序图;
图2为植物乳杆菌(
Lactobacillusplantarum)RH JS001的***发育树。
具体实施方式
为更好的说明本发明的目的、技术方案和优点,下面将结合附图和具体实施例对本发明作进一步说明。
在以下实施例中,所使用的实验方法如无特殊说明,均为常规方法,所用的材料、试剂等,如无特殊说明,均可从商业途径得到。
实施例1、抑制幽门螺杆菌生长的植物乳杆菌(
Lactobacillusplantarum)RHJS001的筛选和鉴定
1、乳酸菌菌株的筛选:
本发明中所述的菌株分离自四川农家自制酸萝卜。在无菌的条件下,称取1 g样品,用0.85%的生理盐水将其制成悬液并稀释至合适的梯度,将稀释液涂布于MRS固体培养基上,37°C厌氧培养48 h后挑取具有乳酸菌特征的若干单菌落,反复进行纯化培养;将挑取的单菌落接种于MRS液体培养基,在37°C培养24 h,以2%的接种量连续活化三次,保证菌株活性,以备后续试验使用。
所述MRS液体培养基的组成成分:蛋白胨10.0g、牛肉粉10.0 g、酵母粉5.0g、葡萄糖(C6H12O6•H2O)20.0 g、硫酸镁(MgSO4•7H2O)0.1 g、醋酸钠(CH3COONa•3H2O)5.0 g、柠檬酸铵(C6H17N3O7)2.0 g、磷酸氢二钾(K2HPO4•7H2O)2.0 g、硫酸锰(MnSO4•4H2O)0.05 g和吐温801.0 g。
2、抑制幽门螺旋杆菌生长的乳酸菌的筛选:
(1)将购自ATCC,菌株编号为ATCCA43504的幽门螺旋杆菌从-80°C取出,涂布于含5%绵羊血(v/v)的哥伦比亚血琼脂培养基中,在微需氧环境中,37℃培养48 h后挑取单菌落,反复进行纯化培养;将挑取的单菌落接种于幽门螺旋杆菌液体培养基中,在37°C培养48h,在4°C下以2600 g离心10 min后,取上清过0 .22 μm无菌滤膜,得到上清液;使用幽门螺旋杆菌液体培养基重悬菌体,得到菌悬液,调整菌悬液的浓度为108CFU/mL。
所述幽门螺旋杆菌液体培养基的组成成分:蛋白胨10.0 g、牛脑浸出粉10.0g、牛心浸出粉9.0、氯化钠(NaCl)5.0 g、葡萄糖(C6H12O6•H2O)2.0 g、磷酸氢二钠(Na2HPO4•12H2O)2.5 g。
(2)将活化好的菌株培养液在4°C下以2600 g离心10min,取上清过0.22 μm无菌滤膜,得到上清液;使用0.85%(w/v)生理盐水重悬菌体,得到菌悬液,调整菌悬液的浓度为108CFU/mL。
(3)抑菌圈实验:
将100 μL幽门螺旋杆菌菌悬液涂布于含5%绵羊血的哥伦比亚血琼脂培养基上,在涂布后的平板上放置四个牛津杯,分别取100 μL上述制备的菌株的菌悬液、菌株的上清液注入牛津杯孔径内,以MRS液体培养基作为阴性对照,以0.025%的甲硝唑溶液作为阳性对照。在37℃的微氧环境下培养48h,用游标卡尺测量抑菌圈直径,实验重复3次。
3、菌种的鉴定:
采用生工基因组DNA试剂盒提取待鉴定菌种的基因组DNA,将提取的基因组送至生工生物工程(上海)股份有限公司进行PCR扩增和测序工作,得到的测序结果在NCBI数据库进行BLAST同源性分析与比对,鉴定菌种并构建***发育树,结果如图1所示。由图1可知,该菌株为植物乳杆菌(
Lactobacillus plantarum),命名为植物乳杆菌RH JS001,保藏于中国微生物菌种保藏管理委员会普通微生物中心(CGMCC),其保藏编号为:CGMCC No.23703,位于北京市朝阳区北辰西路1号院3号,保藏时间为2021年11月1日。
实施例2、植物乳杆菌(
Lactobacillusplantarum)RH JS001的耐酸性评价
(1)人工胃液的配制:
人工胃液中含有0.85%(w/v)的NaCl以及0.30%(w/v)的胃蛋白酶,使用0.1 M HCl分别调节其pH为2.0、2.5和3.0,过0.22 μM滤膜过滤除菌备用。
(2)耐胃酸能力测试:
将植物乳杆菌(
Lactobacillus plantarum)RH JS001以2%的接种量接入MRS液体培养基,37°C培养24 h,在4°C下以2600 g离心10 min后,弃上清液,收集菌泥,用0.85%(w/v)生理盐水洗涤菌泥两次后,重悬于0.85%(w/v)生理盐水中,调整菌悬液的浓度为108CFU/mL。取1.0 mL的菌悬液分别与9.0 mL pH为2.0、2.5和3.0的无菌人工胃液混合,在37°C,50rpm条件下培养2 h,通过平板计数法测定一次活菌数,并计算存活率。
测试结果如表1所示。
表1 植物乳杆菌(
Lactobacillusplantarum)RH JS001的耐酸特性结果
从表1从可以看出,本发明所述植物乳杆菌(
Lactobacillusplantarum)RH JS001具有良好的耐胃酸能力,在pH为2.0人工胃液中孵育2h,存活率可达82.25±0.53%;在pH为2.5的人工胃液中孵育2 h,存活率可达92.44±0.42%;在pH为3.0的人工胃液中孵育2 h,存活率可达95.43±0.35%。
实施例3、植物乳杆菌(
Lactobacillusplantarum)RH JS001抑制幽门螺旋杆菌生长的验证
(1)将购自ATCC,菌株编号为ATCCA43504的幽门螺旋杆菌从-80°C取出,涂布于含5%绵羊血(v/v)的哥伦比亚血琼脂培养基中,在微需氧环境中,37℃培养48 h后挑取单菌落,反复进行纯化培养;将挑取的单菌落接种于幽门螺旋杆菌液体培养基中,在37°C培养48h,在4°C下以2600 g离心10 min后,取上清过0 .22 μm无菌滤膜,得到上清液;使用幽门螺旋杆菌液体培养基重悬菌体,得到菌悬液,调整菌悬液的浓度为108CFU/mL。
(2)将活化好的植物乳杆菌(
Lactobacillusplantarum)RH JS001培养液在4°C下以2600 g离心10 min,取上清过0 .22 μm无菌滤膜,得到上清液;使用0.85%(w/v)生理盐水重悬菌体,得到菌悬液,调整菌悬液的浓度为108CFU/mL。
(3)抑菌圈实验:
将100 μL幽门螺旋杆菌菌悬液涂布于含5%绵羊血的哥伦比亚血琼脂培养基上,在涂布后的平板上放置四个牛津杯,分别取100 μL上述制备的(
Lactobacillusplantarum)RHJS001的菌悬液、植物乳杆菌(
Lactobacillus plantarum)RHJS001的上清液注入牛津杯孔径内,以MRS液体培养基作为阴性对照,以0.025%的甲硝唑溶液作为阳性对照。在37℃的微氧环境下培养48h,用游标卡尺测量抑菌圈直径,实验重复3次。
测试结果如表2所示。
表2植物乳杆菌(
Lactobacillusplantarum)RH JS001的抑菌结果
抑菌圈直径(mm) | |
植物乳杆菌悬液 | 18.50±1.70 |
植物乳杆菌上清液 | 16.90±2.10 |
阳性对照组 | 12.30±1.20 |
阴性对照组 | 0 |
从表2从可以看出,阴性对照即MRS培养基无抑制幽门螺旋杆菌生长的作用,植物乳杆菌(
Lactobacillus plantarum)RH JS001菌悬液和上清液能够明显抑制幽门螺旋杆菌生长。中国专利CN113980878A中所述一株抗幽门螺旋杆菌感染的植物乳杆菌(
Lactobacillus plantarum)LP05,培养48 h时,其菌悬液的抑菌圈直径为15.8±2.2 mm,上清液的抑菌圈直径为14.5±2 mm以上,本发明所述植物乳杆菌(
Lactobacillus
plantarum)RH JS001抑制幽门螺旋杆菌生长的能力更优。
实施例4、动物试验
(1)小鼠的分组及幽门螺旋杆菌感染模型的建立
选择4-6周龄,健康的雄性昆明小鼠38只。所有小鼠先灌胃混合抗生素溶液(阿奇霉素10 mg/mL、氨苄青霉素10mg/mL、庆大霉素1.2mg/mL)1周,用量为每天灌胃0.1 mL。将小鼠分为空白组,模型组和干预组,空白组和干预组各12只,模型组14只。除准备留作空白组的12只小鼠灌胃1mL 0.85%(m/v)生理盐水外,其余26只小鼠灌胃幽门螺杆菌(108CFU/mL)新鲜悬菌液0.5 mL,隔天1次,共8次。同时模型组灌胃0.5 mL0.85%(m/v)生理盐水,干预组灌胃0.5 mL实施例1制备的植物乳杆菌(
Lactobacillus plantarum)RHJS001菌悬液(8.2×108CFU/天)。末次灌胃1周后,随机处死模型组小鼠2只,取胃黏膜,采用胃幽门螺旋杆菌诊断试剂盒进行判断,判为阳性,则感染模型建立成功。
(2)小鼠的处理
最后一次灌胃后1周将小鼠全部处死,剪取小鼠的胃组织,取胃黏膜行组织病理学特殊银染色评估幽门螺旋杆菌分布情况,HE染色镜检观察胃黏膜感染分级;于小鼠眼眶静脉丛取血清,用ELISA的方法检测IL-1β、TNF-α的含量。
测试结果如表3、4所示。
表3 各组小鼠胃组织幽门螺旋杆菌感染情况比较(单位:只)
分组 | 空白组 | 模型组 | 干预组 |
HP(-) | 11 | 0* | 5# |
HP(+) | 1 | 1* | 4# |
HP(++) | 0 | 4* | 2# |
HP(+++) | 0 | 7* | 1# |
注:与空白组比较,*P<0.01;与模型组比较,#P<0.05;-,+,++,+++分别代表幽门螺杆菌炎症程度的阴性/无炎症、轻度阳性/轻度炎症、中度阳性/中度炎症和重度阳性/重度炎症。
从表3中可以看出,与空白组比较,模型组小鼠胃组织幽门螺旋杆菌感染的动物数明显增加;与模型组比较,干预组小鼠的幽门螺旋杆菌感染数量和程度显著减少,差异有统计学意义,说明植物乳杆菌(
Lactobacillus plantarum)RH JS001有减轻幽门螺旋杆菌感染的能力。
表4 各组小鼠血清IL-1β、TNF-α水平比较
分组 | 空白组 | 模型组 | 干预组 |
IL-1β | 103.83±10.47 | 220.81±13.20** | 140.28±10.39*# |
TNF-α | 82.38±4.92 | 267.38±2.81** | 119.39±2.49# |
注:与空白组相比,*P<0.05,**p<0.01;与模型组相比,#P<0.05。
从表4中可以看出,在灌胃植物乳杆菌(
Lactobacillusplantarum)RH JS001菌悬液后,小鼠胃内的炎症因子IL-1β、TNF-α的表达减少,说明植物乳杆菌(
Lactobacillus
plantarum)RH JS001可减轻小鼠胃部的炎症反应。
实施例5、植物乳杆菌(
Lactobacillusplantarum)RH JS001在铁皮石斛酵素中的应用
(1)原料的预处理:
选择新鲜、无霉烂的铁皮石斛,用流动水洗净后去皮,切成小块,待用;
(2)铁皮石斛原液的制备:
将铁皮石斛与水以1:10-20的比例榨汁,添加质量比3-5%白砂糖,得到铁皮石斛原液,待用;
(3)酶解:
在步骤(2)中制备的铁皮石斛原液中加入质量比1-3%纤维素酶(酶活20000 U/g)和1-2%果胶酶(酶活10000 U/g),在45-55℃下酶解30-60 min,得到铁皮石斛酶解液,待用;
(4)巴氏杀菌:
将步骤(3)得到的铁皮石斛酶解液在80-90℃进行巴氏杀菌5-15 min,然后冷却至室温,待用;
(5)乳酸菌发酵:
在无菌环境下,将实施例1制备的植物乳杆菌(
Lactobacillus plantarum)RHJS001菌悬液(8.2×108CFU/mL)以2-6%的接种量接种到无菌铁皮石斛酶解液中,37℃恒温培养48-72 h,得到铁皮石斛发酵初液;
(6)离心过滤:
将铁皮石斛发酵初液在4°C下以2600g离心10 min后,取上清液过300目滤布,得铁皮石斛发酵原液;
(7)调味罐装:
在步骤(6)中制备的铁皮石斛发酵原液中加入质量比1-5%白砂糖进行调味,无菌灌装于已灭菌的容器内,即得一种铁皮石斛酵素。
实施例6、植物乳杆菌(
Lactobacillusplantarum)RH JS001在灵芝酵素中的应用
(1)原料的预处理:
选择新鲜、无霉烂的灵芝,用流动水洗净后去皮,切成小块,待用;
(2)灵芝原液的制备:
将灵芝与水以1:10-20的比例榨汁,添加质量比3-5%白砂糖,得到灵芝原液,待用;
(3)酶解:
在步骤(2)中制备的灵芝原液中加入质量比1-3%纤维素酶(酶活20000 U/g)和1-2%果胶酶(酶活10000 U/g),在45-55℃下酶解30-60 min,得到灵芝酶解液,待用;
(4)巴氏杀菌:
将步骤(3)得到的灵芝酶解液在80-90℃进行巴氏杀菌5-15 min,然后冷却至室温,待用;
(5)乳酸菌发酵:
在无菌环境下,将实施例1制备的植物乳杆菌(
Lactobacillus plantarum)RHJS001菌悬液(8.2×108CFU/mL)以2-6%的接种量接种到无菌灵芝酶解液中,37℃恒温培养48-72 h,得到灵芝发酵初液;
(6)离心过滤:
将灵芝发酵初液在4°C下以2600 g离心10 min后,取上清液过300目滤布,得灵芝发酵原液;
(7)调味罐装:
在步骤(6)中制备的灵芝发酵原液中加入质量比1-5%白砂糖进行调味,无菌灌装于已灭菌的容器内,即得一种灵芝酵素。
实施例7、植物乳杆菌(
Lactobacillusplantarum)RH JS001在复合本草酵素中的应用
(1)原料的预处理:
选择新鲜、无霉烂的铁皮石斛、灵芝、猴头菇、甘草、山楂、黄精、山药、薏苡仁和枸杞,用流动水洗净后去皮,切成小块,待用;
(2)复合本草原液配方一的制备:
将步骤(1)中所述中药组合物与水以1:10-20的比例榨汁,其中中药组合物主要包括铁皮石斛4份、灵芝4份、猴头菇15份、甘草5份、山楂8份、黄精10份、山药6份、薏苡仁2份、枸杞4份,添加质量比3-5%白砂糖,得到复合本草原液,待用;
复合本草原液配方二的制备:
将步骤(1)中所述中药组合物与水以1:10-20的比例榨汁,其中中药组合物主要包括铁皮石斛6份、灵芝6份、猴头菇20份、甘草15份、山楂10份、黄精14份、山药10份、薏苡仁2份、枸杞6份,添加质量比3-5%白砂糖,得到复合本草原液,待用;
复合本草原液配方三的制备:
将步骤(1)中所述中药组合物与水以1:10-20的比例榨汁,其中中药组合物主要包括铁皮石斛2份、灵芝2份、猴头菇8份、甘草8份、山楂6份、黄精4份、山药2份、薏苡仁1份、枸杞4份,添加质量比3-5%白砂糖,得到复合本草原液,待用;
(3)酶解:
在步骤(2)中制备的复合本草原液中加入质量比1-3%纤维素酶(酶活20000 U/g)和1-2%果胶酶(酶活10000 U/g),在45-55℃下酶解30-60 min,得到复合本草酶解液,待用;
(4)巴氏杀菌:
将步骤(3)得到的复合本草酶解液在80-90℃进行巴氏杀菌5-15 min,然后冷却至室温,待用;
(5)乳酸菌发酵:
在无菌环境下,将实施例1制备的植物乳杆菌(
Lactobacillus plantarum)RHJS001菌悬液(8.2×108 CFU/mL)以2-6%的接种量接种到无菌复合本草酶解液中,37℃恒温培养48-72 h,得到复合本草发酵初液;
(6)离心过滤:
将复合本草发酵初液在4°C下以2600g离心10 min后,取上清液过300目滤布,得复合本草发酵原液;
(7)调味罐装:
在步骤(6)中制备的复合本草发酵原液中加入质量比1-5%白砂糖进行调味,无菌灌装于已灭菌的容器内,即得三种配方的复合本草酵素。
实施例8、一种复合本草酵素饮料A
(1)复合本草酵素饮料A的制备:
一种复合本草酵素饮料A,主要由实施例7中制备的复合本草酵素、猴头菇粉、红枣粉、枸杞果汁粉、低聚果糖组成,其中按照重量份复合本草酵素3份、猴头菇粉0.05份、红枣粉0.2份、枸杞果汁粉0.1份、低聚果糖0.3份。
(2)杀菌灌装:
将步骤(1)中制备的复合本草酵素饮料进行分装,在80-90℃进行巴氏杀菌5-15min,然后冷却至室温,得到复合本草酵素饮料成品。
实施例9、一种复合本草酵素饮料B
(1)复合本草酵素饮料B的制备:
一种复合本草酵素饮料B,主要由实施例7中制备的复合本草酵素、柠檬粉、薄荷粉、柿子提取物、低聚果糖组成,其中按照重量份复合本草酵素3份、柠檬粉0.2份、薄荷粉0.01份、柿子提取物0.02份、低聚果糖0.4份。
(2)杀菌灌装:
将步骤(1)中制备的复合本草酵素饮料进行分装,在80-90℃进行巴氏杀菌5-15min,然后冷却至室温,得到复合本草酵素饮料成品。
实施例10、本草酵素及其产品抑制幽门螺旋杆菌生长的验证
(1)将购自ATCC,菌株编号为ATCCA43504的幽门螺旋杆菌从-80°C取出,涂布于含5%绵羊血(v/v)的哥伦比亚血琼脂培养基中,在微需氧环境中,37℃培养48 h后挑取单菌落,反复进行纯化培养;将挑取的单菌落接种于幽门螺旋杆菌液体培养基中,在37°C培养48h,在4°C下以2600 g离心10 min后,取上清过0.22 μm无菌滤膜,得到上清液;使用幽门螺旋杆菌液体培养基重悬菌体,得到菌悬液,调整菌悬液的浓度为108CFU/mL。
(2)抑菌圈实验:
将100 μL幽门螺旋杆菌菌悬液涂布于含5%绵羊血的哥伦比亚血琼脂培养基上,在涂布后的平板上放置牛津杯,分别取100 μL铁皮石斛酵素、灵芝酵素、复合本草酵素、复合本草酵素饮料A和复合本草酵素饮料B注入牛津杯孔径内,以MRS液体培养基作为阴性对照,以0.025%的甲硝唑溶液作为阳性对照。在37℃的微氧环境下培养48h,用游标卡尺测量抑菌圈直径,实验重复3次。
测试结果如表5所示。
表5 本草酵素及其产品的抑菌结果
组别 | 抑菌圈直径(mm) |
铁皮石斛酵素 | 12.40±0.90 |
灵芝酵素 | 14.20±1.70 |
复合本草酵素配方一 | 16.10±1.40 |
复合本草酵素配方二 | 16.89±1.10 |
复合本草酵素配方三 | 15.98±1.20 |
复合本草酵素饮料A | 18.47±2.10 |
复合本草酵素饮料B | 18.19±1.30 |
阳性对照 | 10.40±1.30 |
阴性对照 | 0 |
从表5中可以看出,由植物乳杆菌(
Lactobacillusplantarum)RH JS001发酵的本草酵素对幽门螺杆菌的生长具有抑制作用,培养48 h时,由植物乳杆菌RH JS001发酵的铁皮石斛酵素、灵芝酵素、复合本草酵素配方一、复合本草酵素配方二、复合本草酵素配方三、复合本草酵素饮料A和复合本草酵素饮料B的抑菌圈直径分别可达12.40±0.90 mm、14.20±1.70mm、16.10±1.40 mm、16.89±1.10、15.98±1.20、18.47±2.10mm和18.19±1.30mm。
最后所应当说明的是,以上实施例仅用以说明本发明的技术方案而非对本发明保护范围的限制,尽管参照较佳实施例对本发明作了详细说明,本领域的普通技术人员应当理解,可以对本发明的技术方案进行修改或者等同替换,而不脱离本发明技术方案的实质和范围。
SEQUENCE LISTING
<110> 仁和全域(上海)大健康研究院有限公司
<120> 一种抗幽门螺旋杆菌感染的植物乳杆菌及其在食用本草酵素产品中的应用
<130> 2022-05-19
<160> 1
<170> PatentIn version 3.5
<210> 1
<211> 1029
<212> DNA
<213> 人工合成
<400> 1
ccaaccttca actgtcactt aggcggctgg ttcctaaaag gttaccccac cgactttggg 60
tgttacaaac tctcatggtg tgacgggcgg tgtgtacaag gcccgggaac gtattcaccg 120
cggcatgctg atccgcgatt actagcgatt ccgacttcat gtaggcgagt tgcagcctac 180
aatccgaact gagaatggct ttaagagatt agcttactct cgcgagttcg caactcgttg 240
taccatccat tgtagcacgt gtgtagccca ggtcataagg ggcatgatga tttgacgtca 300
tccccacctt cctccggttt gtcaccggca gtctcaccag agtgcccaac ttaatgctgg 360
caactgataa taagggttgc gctcgttgcg ggacttaacc caacatctca cgacacgagc 420
tgacgacaac catgcaccac ctgtatccat gtccccgaag ggaacgtcta atctcttaga 480
tttgcatagt atgtcaagac ctggtaaggt tcttcgcgta gcttcgaatt aaaccacatg 540
ctccaccgct tgtgcgggcc cccgtcaatt cctttgagtt tcagccttgc ggccgtactc 600
cccaggcgga atgcttaatg cgttagctgc agcactgaag ggcggaaacc ctccaacact 660
tagcattcat cgtttacggt atggactacc agggtatcta atcctgtttg ctacccatac 720
tttcgagcct cagcgtcagt tacagaccag acagccgcct tcgccactgg tgttcttcca 780
tatatctacg catttcaccg ctacacatgg agttccactg tcctcttctg cactcaagtt 840
tcccagtttc cgatgcactt cttcggttga gccgaaaggc tttcacatca gacttaaaaa 900
accgcctgcg ctcgctttac gcccaataaa tccggacaac gcttgccacc tacgtattac 960
cgcggctgct ggcacgtagt tagccgtggc tttctggtta aataccgtca ataacctgaa 1020
acagttact 1029
Claims (10)
1.一种植物乳杆菌(Lactobacillus plantarum),其特征在于,所述植物乳杆菌(Lactobacillus plantarum)的16r RNA序列如SEQID No: 1所示;
所述植物乳杆菌(Lactobacillus plantarum)保藏于中国微生物菌种保藏管理委员会普通微生物中心,保藏编号为:CGMCC No.23703,保藏时间为2021年11月1日。
2.如权利要求1所述的植物乳杆菌(Lactobacillus plantarum)接种原料液形成的发酵液。
3.如权利要求1所述的植物乳杆菌(Lactobacillus plantarum)或如权利要求2所述的发酵液在制备抗幽门螺杆菌感染制剂中的应用。
4.如权利要求1所述的植物乳杆菌(Lactobacillus plantarum)或如权利要求2所述的发酵液在制备食用本草酵素产品中的应用。
5.一种食用本草酵素,其特征在于,所述食用本草酵素包括以下组分通过如权利要求1所述的植物乳杆菌(Lactobacillusplantarum)发酵而成;
所述组分包括铁皮石斛或灵芝。
6.一种复合本草酵素,其特征在于,所述复合本草酵素包括以下组分通过如权利要求1所述的植物乳杆菌(Lactobacillus plantarum)发酵而成;
组分包括铁皮石斛、灵芝、猴头菇、甘草、山楂、黄精、山药、薏苡仁和枸杞。
7.如权利要求6所述的复合本草酵素,其特征在于,所述复合本草酵素包括以下重量份数的组分:
铁皮石斛1-8份、灵芝1-8份、猴头菇5-25份、甘草5-20份、山楂2-16份、黄精2-16份、山药2-16份、薏苡仁1-3份和枸杞1-8份。
8.一种食用本草酵素产品,其特征在于,所述食用本草酵素产品包括含有如权利要求6或7所述的复合本草酵素的复合本草酵素饮料。
9.如权利要求8所述的食用本草酵素产品,其特征在于,所述复合本草酵素饮料包括以下重量份数的组分:
如权利要求6或7所述的复合本草酵素3-15份、猴头菇粉0.01-0.2份、红枣粉0.1-2份、枸杞果汁粉0.01-0.2份和低聚果糖0.1-2份。
10.如权利要求8所述的食用本草酵素产品,其特征在于,所述复合本草酵素饮料包括以下重量份数的组分:
如权利要求6或7所述的复合本草酵素3-15份、柠檬粉0.1-2份、薄荷粉0.001-0.01份、柿子提取物0.01-0.2份和低聚果糖0.1-2份。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202210639283.9A CN114908020B (zh) | 2022-05-31 | 2022-05-31 | 一种抗幽门螺旋杆菌感染的植物乳杆菌及其在食用本草酵素产品中的应用 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202210639283.9A CN114908020B (zh) | 2022-05-31 | 2022-05-31 | 一种抗幽门螺旋杆菌感染的植物乳杆菌及其在食用本草酵素产品中的应用 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN114908020A CN114908020A (zh) | 2022-08-16 |
CN114908020B true CN114908020B (zh) | 2023-04-14 |
Family
ID=82769839
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202210639283.9A Active CN114908020B (zh) | 2022-05-31 | 2022-05-31 | 一种抗幽门螺旋杆菌感染的植物乳杆菌及其在食用本草酵素产品中的应用 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN114908020B (zh) |
Families Citing this family (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN115074298A (zh) * | 2022-07-27 | 2022-09-20 | 广东益可维生物技术有限公司 | 具有抗幽护胃和消食化积功效的益生菌组合物及其应用 |
CN116731913B (zh) * | 2023-05-30 | 2024-03-15 | 江西仁仁健康微生态科技有限公司 | 植物乳植杆菌在制备抗幽门螺杆菌产品中的应用 |
CN116606781B (zh) * | 2023-07-14 | 2024-05-14 | 青岛蔚蓝生物集团有限公司 | 一株具有拮抗幽门螺旋杆菌能力的植物乳植杆菌及其应用 |
Family Cites Families (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN107988115B (zh) * | 2017-12-27 | 2020-06-23 | 北京科拓恒通生物技术股份有限公司 | 植物乳杆菌及其复合益生菌发酵液与制备方法 |
-
2022
- 2022-05-31 CN CN202210639283.9A patent/CN114908020B/zh active Active
Also Published As
Publication number | Publication date |
---|---|
CN114908020A (zh) | 2022-08-16 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN114908020B (zh) | 一种抗幽门螺旋杆菌感染的植物乳杆菌及其在食用本草酵素产品中的应用 | |
CN1297653C (zh) | 包含戊糖乳杆菌菌株的组合物及其用途 | |
CN106282072B (zh) | 一种复合乳酸菌微生态制剂及其制备方法与应用 | |
CN103893214B (zh) | 一种全燕麦固态混菌发酵生产益生菌活菌粉剂及制备方法 | |
CN106754619A (zh) | 一种在谷物培养基中添加中药组分促进益生菌生长的方法 | |
CN113170842B (zh) | 一种防治家禽坏死性肠炎的复合微生态制剂及其应用 | |
CN104293716A (zh) | 一种用大型真菌菌液(质)制备高效益生菌制剂的方法 | |
CN110495522B (zh) | 一种饲料用中药微生态制剂 | |
CN109749957B (zh) | 一种具有水产病原菌拮抗特性的格氏乳杆菌制剂的制备及应用 | |
CN110106119B (zh) | 一株分离自母乳的鼠李糖乳杆菌m9及其应用 | |
CN109161509B (zh) | 一株能防治牛羊腹泻病的菌株 | |
CN110564638A (zh) | 一株具有益生特性的罗伊氏乳杆菌及其用途 | |
CN113913346B (zh) | 一种副干酪乳杆菌jn-1及其应用 | |
CN113832077A (zh) | 鼠李糖乳杆菌及其应用 | |
CN113616715B (zh) | 治疗仔猪腹泻及改善其肠道菌群的发酵中药口服液 | |
KR101396842B1 (ko) | 오미자 발효물 및 이를 포함하는 항균 및 면역 증강 조성물 | |
CN112239739B (zh) | 一株可缓解etec致腹泻的植物乳杆菌及其应用 | |
CN114032190A (zh) | 一株可发酵石斛且其发酵液可有效修复日光皮炎的罗伊氏乳杆菌 | |
CN109363004A (zh) | 大鳞鲃鱼用发酵型中草药免疫增强剂的制备方法与应用 | |
CN113337427A (zh) | 一种植物乳杆菌hnu082、组合物及其应用 | |
CN115895966B (zh) | 一株辅助缓解痛风的两歧双歧杆菌bl002及其应用 | |
CN114540232B (zh) | 有水产病原菌拮抗特性的鼠李糖乳杆菌及其制剂的制备及应用 | |
CN113249244B (zh) | 一种拮抗咽炎致病菌乙型溶血性链球菌的副干酪乳杆菌 | |
CN112391317B (zh) | 一种生产燕窝酸的益生菌株组合物及用途 | |
CN112029676B (zh) | 一种有利于提高免疫力的益生菌组合及其应用 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
GR01 | Patent grant | ||
GR01 | Patent grant |