CN116024130B - Lactobacillus fermentum A21215 for reducing blood uric acid and application thereof - Google Patents
Lactobacillus fermentum A21215 for reducing blood uric acid and application thereof Download PDFInfo
- Publication number
- CN116024130B CN116024130B CN202211600780.4A CN202211600780A CN116024130B CN 116024130 B CN116024130 B CN 116024130B CN 202211600780 A CN202211600780 A CN 202211600780A CN 116024130 B CN116024130 B CN 116024130B
- Authority
- CN
- China
- Prior art keywords
- lactobacillus fermentum
- uric acid
- purine
- strain
- lactobacillus
- 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 79
- 229940012969 lactobacillus fermentum Drugs 0.000 title claims abstract description 77
- LEHOTFFKMJEONL-UHFFFAOYSA-N Uric Acid Chemical compound N1C(=O)NC(=O)C2=C1NC(=O)N2 LEHOTFFKMJEONL-UHFFFAOYSA-N 0.000 title abstract description 59
- TVWHNULVHGKJHS-UHFFFAOYSA-N Uric acid Natural products N1C(=O)NC(=O)C2NC(=O)NC21 TVWHNULVHGKJHS-UHFFFAOYSA-N 0.000 title abstract description 57
- 229940116269 uric acid Drugs 0.000 title abstract description 57
- 210000004369 blood Anatomy 0.000 title abstract description 33
- 239000008280 blood Substances 0.000 title abstract description 33
- 230000001603 reducing effect Effects 0.000 title description 22
- KDCGOANMDULRCW-UHFFFAOYSA-N 7H-purine Chemical compound N1=CNC2=NC=NC2=C1 KDCGOANMDULRCW-UHFFFAOYSA-N 0.000 claims abstract description 32
- 235000013305 food Nutrition 0.000 claims abstract description 9
- 244000005700 microbiome Species 0.000 claims abstract description 8
- 238000004519 manufacturing process Methods 0.000 claims abstract description 5
- 235000021107 fermented food Nutrition 0.000 claims abstract description 3
- NYHBQMYGNKIUIF-UUOKFMHZSA-N Guanosine Chemical compound C1=NC=2C(=O)NC(N)=NC=2N1[C@@H]1O[C@H](CO)[C@@H](O)[C@H]1O NYHBQMYGNKIUIF-UUOKFMHZSA-N 0.000 claims description 28
- 229940029575 guanosine Drugs 0.000 claims description 17
- MIKUYHXYGGJMLM-GIMIYPNGSA-N Crotonoside Natural products C1=NC2=C(N)NC(=O)N=C2N1[C@H]1O[C@@H](CO)[C@H](O)[C@@H]1O MIKUYHXYGGJMLM-GIMIYPNGSA-N 0.000 claims description 14
- NYHBQMYGNKIUIF-UHFFFAOYSA-N D-guanosine Natural products C1=2NC(N)=NC(=O)C=2N=CN1C1OC(CO)C(O)C1O NYHBQMYGNKIUIF-UHFFFAOYSA-N 0.000 claims description 14
- UGQMRVRMYYASKQ-KQYNXXCUSA-N Inosine Chemical compound O[C@@H]1[C@H](O)[C@@H](CO)O[C@H]1N1C2=NC=NC(O)=C2N=C1 UGQMRVRMYYASKQ-KQYNXXCUSA-N 0.000 claims description 11
- 229930010555 Inosine Natural products 0.000 claims description 11
- 229960003786 inosine Drugs 0.000 claims description 11
- 239000002126 C01EB10 - Adenosine Substances 0.000 claims description 9
- 229960005305 adenosine Drugs 0.000 claims description 9
- OIRDTQYFTABQOQ-KQYNXXCUSA-N adenosine Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](CO)[C@@H](O)[C@H]1O OIRDTQYFTABQOQ-KQYNXXCUSA-N 0.000 claims description 8
- 230000000593 degrading effect Effects 0.000 claims description 8
- 238000002360 preparation method Methods 0.000 claims description 8
- 235000015140 cultured milk Nutrition 0.000 claims description 5
- 239000002775 capsule Substances 0.000 claims description 3
- 239000008176 lyophilized powder Substances 0.000 claims description 3
- 239000002777 nucleoside Substances 0.000 abstract description 28
- 230000000694 effects Effects 0.000 abstract description 27
- 150000003833 nucleoside derivatives Chemical class 0.000 abstract description 22
- 108010093894 Xanthine oxidase Proteins 0.000 abstract description 20
- 102100033220 Xanthine oxidase Human genes 0.000 abstract description 20
- 201000001431 Hyperuricemia Diseases 0.000 abstract description 19
- 244000052616 bacterial pathogen Species 0.000 abstract description 8
- 210000001072 colon Anatomy 0.000 abstract description 6
- 210000002919 epithelial cell Anatomy 0.000 abstract description 6
- 230000012010 growth Effects 0.000 abstract description 5
- 238000010172 mouse model Methods 0.000 abstract description 2
- 229940030225 antihemorrhagics Drugs 0.000 abstract 1
- 230000000025 haemostatic effect Effects 0.000 abstract 1
- 238000004321 preservation Methods 0.000 abstract 1
- 238000006731 degradation reaction Methods 0.000 description 28
- 230000015556 catabolic process Effects 0.000 description 27
- 230000001580 bacterial effect Effects 0.000 description 23
- 210000004027 cell Anatomy 0.000 description 22
- 239000006228 supernatant Substances 0.000 description 18
- 241000699670 Mus sp. Species 0.000 description 15
- 239000007788 liquid Substances 0.000 description 15
- 239000000243 solution Substances 0.000 description 15
- 239000001963 growth medium Substances 0.000 description 14
- 241000894006 Bacteria Species 0.000 description 11
- 239000003833 bile salt Substances 0.000 description 11
- 230000000968 intestinal effect Effects 0.000 description 10
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 10
- 238000009630 liquid culture Methods 0.000 description 10
- 108020004465 16S ribosomal RNA Proteins 0.000 description 9
- 239000000843 powder Substances 0.000 description 9
- 239000006041 probiotic Substances 0.000 description 9
- 235000018291 probiotics Nutrition 0.000 description 9
- 230000002829 reductive effect Effects 0.000 description 9
- 230000002401 inhibitory effect Effects 0.000 description 8
- 239000002212 purine nucleoside Substances 0.000 description 8
- 241000588724 Escherichia coli Species 0.000 description 7
- 241000186605 Lactobacillus paracasei Species 0.000 description 7
- 241001052560 Thallis Species 0.000 description 7
- 238000000855 fermentation Methods 0.000 description 7
- 230000004151 fermentation Effects 0.000 description 7
- 238000000034 method Methods 0.000 description 7
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 6
- 201000005569 Gout Diseases 0.000 description 6
- 241000186660 Lactobacillus Species 0.000 description 6
- 230000003321 amplification Effects 0.000 description 6
- 238000012258 culturing Methods 0.000 description 6
- 230000006870 function Effects 0.000 description 6
- 210000004051 gastric juice Anatomy 0.000 description 6
- 229940039696 lactobacillus Drugs 0.000 description 6
- 239000000463 material Substances 0.000 description 6
- 238000003199 nucleic acid amplification method Methods 0.000 description 6
- 125000003835 nucleoside group Chemical group 0.000 description 6
- 235000020183 skimmed milk Nutrition 0.000 description 6
- 239000000725 suspension Substances 0.000 description 6
- 241000699666 Mus <mouse, genus> Species 0.000 description 5
- OFCNXPDARWKPPY-UHFFFAOYSA-N allopurinol Chemical compound OC1=NC=NC2=C1C=NN2 OFCNXPDARWKPPY-UHFFFAOYSA-N 0.000 description 5
- 229960003459 allopurinol Drugs 0.000 description 5
- 210000001035 gastrointestinal tract Anatomy 0.000 description 5
- 239000002054 inoculum Substances 0.000 description 5
- 239000004310 lactic acid Substances 0.000 description 5
- 235000014655 lactic acid Nutrition 0.000 description 5
- 239000002504 physiological saline solution Substances 0.000 description 5
- 239000000047 product Substances 0.000 description 5
- 238000000746 purification Methods 0.000 description 5
- 210000002966 serum Anatomy 0.000 description 5
- 239000000126 substance Substances 0.000 description 5
- 230000004083 survival effect Effects 0.000 description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 5
- LRFVTYWOQMYALW-UHFFFAOYSA-N 9H-xanthine Chemical compound O=C1NC(=O)NC2=C1NC=N2 LRFVTYWOQMYALW-UHFFFAOYSA-N 0.000 description 4
- 206010021131 Hypouricaemia Diseases 0.000 description 4
- 241000191967 Staphylococcus aureus Species 0.000 description 4
- 239000002253 acid Substances 0.000 description 4
- 229940093761 bile salts Drugs 0.000 description 4
- 238000006243 chemical reaction Methods 0.000 description 4
- 238000001514 detection method Methods 0.000 description 4
- 239000003814 drug Substances 0.000 description 4
- 238000002474 experimental method Methods 0.000 description 4
- 230000002496 gastric effect Effects 0.000 description 4
- 238000004128 high performance liquid chromatography Methods 0.000 description 4
- 238000011534 incubation Methods 0.000 description 4
- 230000005764 inhibitory process Effects 0.000 description 4
- 230000004060 metabolic process Effects 0.000 description 4
- 239000000203 mixture Substances 0.000 description 4
- MRWXACSTFXYYMV-FDDDBJFASA-N nebularine Chemical compound O[C@@H]1[C@H](O)[C@@H](CO)O[C@H]1N1C2=NC=NC=C2N=C1 MRWXACSTFXYYMV-FDDDBJFASA-N 0.000 description 4
- 108020004707 nucleic acids Proteins 0.000 description 4
- 102000039446 nucleic acids Human genes 0.000 description 4
- 230000000529 probiotic effect Effects 0.000 description 4
- 230000001105 regulatory effect Effects 0.000 description 4
- 210000002700 urine Anatomy 0.000 description 4
- 239000003643 water by type Substances 0.000 description 4
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- 238000010521 absorption reaction Methods 0.000 description 3
- 230000009286 beneficial effect Effects 0.000 description 3
- 235000013361 beverage Nutrition 0.000 description 3
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 3
- 235000005911 diet Nutrition 0.000 description 3
- 238000010790 dilution Methods 0.000 description 3
- 239000012895 dilution Substances 0.000 description 3
- 229940079593 drug Drugs 0.000 description 3
- 238000001035 drying Methods 0.000 description 3
- 238000000338 in vitro Methods 0.000 description 3
- 230000007774 longterm Effects 0.000 description 3
- 239000002609 medium Substances 0.000 description 3
- 238000002156 mixing Methods 0.000 description 3
- 230000002265 prevention Effects 0.000 description 3
- -1 purine nucleic acid Chemical class 0.000 description 3
- 230000000284 resting effect Effects 0.000 description 3
- 238000000926 separation method Methods 0.000 description 3
- 239000007787 solid Substances 0.000 description 3
- 238000005406 washing Methods 0.000 description 3
- 239000012224 working solution Substances 0.000 description 3
- 235000013618 yogurt Nutrition 0.000 description 3
- IXPNQXFRVYWDDI-UHFFFAOYSA-N 1-methyl-2,4-dioxo-1,3-diazinane-5-carboximidamide Chemical compound CN1CC(C(N)=N)C(=O)NC1=O IXPNQXFRVYWDDI-UHFFFAOYSA-N 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 108090000790 Enzymes 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
- 241001465754 Metazoa Species 0.000 description 2
- 108010046377 Whey Proteins Proteins 0.000 description 2
- 102000007544 Whey Proteins Human genes 0.000 description 2
- 238000000246 agarose gel electrophoresis Methods 0.000 description 2
- 229960002529 benzbromarone Drugs 0.000 description 2
- WHQCHUCQKNIQEC-UHFFFAOYSA-N benzbromarone Chemical compound CCC=1OC2=CC=CC=C2C=1C(=O)C1=CC(Br)=C(O)C(Br)=C1 WHQCHUCQKNIQEC-UHFFFAOYSA-N 0.000 description 2
- 230000033228 biological regulation Effects 0.000 description 2
- 238000009835 boiling Methods 0.000 description 2
- 229940099352 cholate Drugs 0.000 description 2
- 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 description 2
- 230000001276 controlling effect Effects 0.000 description 2
- 235000020247 cow milk Nutrition 0.000 description 2
- 230000037213 diet Effects 0.000 description 2
- 238000007865 diluting Methods 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 229940088598 enzyme Drugs 0.000 description 2
- 230000029142 excretion Effects 0.000 description 2
- 238000000605 extraction Methods 0.000 description 2
- 238000011049 filling Methods 0.000 description 2
- 238000007710 freezing Methods 0.000 description 2
- 230000008014 freezing Effects 0.000 description 2
- 230000002068 genetic effect Effects 0.000 description 2
- FDGQSTZJBFJUBT-UHFFFAOYSA-N hypoxanthine Chemical compound O=C1NC=NC2=C1NC=N2 FDGQSTZJBFJUBT-UHFFFAOYSA-N 0.000 description 2
- 230000002779 inactivation Effects 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 230000002503 metabolic effect Effects 0.000 description 2
- 239000003094 microcapsule Substances 0.000 description 2
- 239000011259 mixed solution Substances 0.000 description 2
- NAFSTSRULRIERK-UHFFFAOYSA-M monosodium urate Chemical class [Na+].N1C([O-])=NC(=O)C2=C1NC(=O)N2 NAFSTSRULRIERK-UHFFFAOYSA-M 0.000 description 2
- 150000007523 nucleic acids Chemical class 0.000 description 2
- 229960003081 probenecid Drugs 0.000 description 2
- DBABZHXKTCFAPX-UHFFFAOYSA-N probenecid Chemical compound CCCN(CCC)S(=O)(=O)C1=CC=C(C(O)=O)C=C1 DBABZHXKTCFAPX-UHFFFAOYSA-N 0.000 description 2
- 239000003223 protective agent Substances 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 238000007789 sealing Methods 0.000 description 2
- 238000012163 sequencing technique Methods 0.000 description 2
- 235000010413 sodium alginate Nutrition 0.000 description 2
- 239000000661 sodium alginate Substances 0.000 description 2
- 229940005550 sodium alginate Drugs 0.000 description 2
- HEMHJVSKTPXQMS-UHFFFAOYSA-M sodium hydroxide Inorganic materials [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 2
- 230000001954 sterilising effect Effects 0.000 description 2
- UCSJYZPVAKXKNQ-HZYVHMACSA-N streptomycin Chemical compound CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](NC(N)=N)[C@H](O)[C@@H](NC(N)=N)[C@H](O)[C@H]1O UCSJYZPVAKXKNQ-HZYVHMACSA-N 0.000 description 2
- 229940075420 xanthine Drugs 0.000 description 2
- HDTRYLNUVZCQOY-UHFFFAOYSA-N α-D-glucopyranosyl-α-D-glucopyranoside Natural products OC1C(O)C(O)C(CO)OC1OC1C(O)C(O)C(O)C(CO)O1 HDTRYLNUVZCQOY-UHFFFAOYSA-N 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- 238000012371 Aseptic Filling Methods 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- 241001260012 Bursa Species 0.000 description 1
- BHPQYMZQTOCNFJ-UHFFFAOYSA-N Calcium cation Chemical compound [Ca+2] BHPQYMZQTOCNFJ-UHFFFAOYSA-N 0.000 description 1
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 description 1
- 241001333951 Escherichia coli O157 Species 0.000 description 1
- RFSUNEUAIZKAJO-ARQDHWQXSA-N Fructose Chemical class OC[C@H]1O[C@](O)(CO)[C@@H](O)[C@@H]1O RFSUNEUAIZKAJO-ARQDHWQXSA-N 0.000 description 1
- 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 1
- 244000068988 Glycine max Species 0.000 description 1
- 235000010469 Glycine max Nutrition 0.000 description 1
- 238000003794 Gram staining Methods 0.000 description 1
- 241000282412 Homo Species 0.000 description 1
- UGQMRVRMYYASKQ-UHFFFAOYSA-N Hypoxanthine nucleoside Natural products OC1C(O)C(CO)OC1N1C(NC=NC2=O)=C2N=C1 UGQMRVRMYYASKQ-UHFFFAOYSA-N 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
- 241000186606 Lactobacillus gasseri Species 0.000 description 1
- 240000006024 Lactobacillus plantarum Species 0.000 description 1
- 235000013965 Lactobacillus plantarum Nutrition 0.000 description 1
- 241000186604 Lactobacillus reuteri Species 0.000 description 1
- 229920002774 Maltodextrin Polymers 0.000 description 1
- 239000005913 Maltodextrin Substances 0.000 description 1
- 229920000881 Modified starch Polymers 0.000 description 1
- 239000004368 Modified starch Substances 0.000 description 1
- 102000019055 Nucleoside Transport Proteins Human genes 0.000 description 1
- 229930182555 Penicillin Natural products 0.000 description 1
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 description 1
- 239000001888 Peptone Substances 0.000 description 1
- 108010080698 Peptones Proteins 0.000 description 1
- 241000700159 Rattus Species 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- HDTRYLNUVZCQOY-WSWWMNSNSA-N Trehalose Natural products O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-WSWWMNSNSA-N 0.000 description 1
- 108010092464 Urate Oxidase Proteins 0.000 description 1
- 239000005862 Whey Substances 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 230000001154 acute effect Effects 0.000 description 1
- 230000006978 adaptation Effects 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 230000002776 aggregation Effects 0.000 description 1
- 238000004220 aggregation Methods 0.000 description 1
- HDTRYLNUVZCQOY-LIZSDCNHSA-N alpha,alpha-trehalose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-LIZSDCNHSA-N 0.000 description 1
- 239000003708 ampul Substances 0.000 description 1
- 238000000137 annealing Methods 0.000 description 1
- 235000015278 beef Nutrition 0.000 description 1
- 235000013405 beer Nutrition 0.000 description 1
- 239000003613 bile acid Substances 0.000 description 1
- 230000037396 body weight Effects 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 239000001110 calcium chloride Substances 0.000 description 1
- 229910001628 calcium chloride Inorganic materials 0.000 description 1
- 229910001424 calcium ion Inorganic materials 0.000 description 1
- 239000001768 carboxy methyl cellulose Substances 0.000 description 1
- 235000010418 carrageenan Nutrition 0.000 description 1
- 239000000679 carrageenan Substances 0.000 description 1
- 229920001525 carrageenan Polymers 0.000 description 1
- 229940113118 carrageenan Drugs 0.000 description 1
- 210000000845 cartilage Anatomy 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 230000019522 cellular metabolic process Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 235000013351 cheese Nutrition 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 230000001684 chronic effect Effects 0.000 description 1
- 230000009194 climbing Effects 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 230000000112 colonic effect Effects 0.000 description 1
- 210000004922 colonic epithelial cell Anatomy 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 238000004132 cross linking Methods 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- YXVFQADLFFNVDS-UHFFFAOYSA-N diammonium citrate Chemical compound [NH4+].[NH4+].[O-]C(=O)CC(O)(C(=O)O)CC([O-])=O YXVFQADLFFNVDS-UHFFFAOYSA-N 0.000 description 1
- 230000000378 dietary effect Effects 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
- 201000010099 disease Diseases 0.000 description 1
- 208000016097 disease of metabolism Diseases 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 239000012154 double-distilled water Substances 0.000 description 1
- 239000003651 drinking water Substances 0.000 description 1
- 235000020188 drinking water Nutrition 0.000 description 1
- 210000003722 extracellular fluid Anatomy 0.000 description 1
- 230000002550 fecal effect Effects 0.000 description 1
- 210000003608 fece Anatomy 0.000 description 1
- 239000012091 fetal bovine serum Substances 0.000 description 1
- 239000012467 final product Substances 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 235000013376 functional food Nutrition 0.000 description 1
- 238000003304 gavage Methods 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 230000036449 good health Effects 0.000 description 1
- 230000003394 haemopoietic effect Effects 0.000 description 1
- 230000015784 hyperosmotic salinity response Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 208000027866 inflammatory disease Diseases 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 230000003870 intestinal permeability Effects 0.000 description 1
- 230000006799 invasive growth in response to glucose limitation Effects 0.000 description 1
- 210000003734 kidney Anatomy 0.000 description 1
- 229940072205 lactobacillus plantarum Drugs 0.000 description 1
- 229940001882 lactobacillus reuteri Drugs 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 1
- 235000019341 magnesium sulphate Nutrition 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 229940035034 maltodextrin Drugs 0.000 description 1
- 229940099596 manganese sulfate Drugs 0.000 description 1
- 239000011702 manganese sulphate Substances 0.000 description 1
- 235000007079 manganese sulphate Nutrition 0.000 description 1
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 208000030159 metabolic disease Diseases 0.000 description 1
- 230000008558 metabolic pathway by substance Effects 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- 235000013336 milk Nutrition 0.000 description 1
- 239000008267 milk Substances 0.000 description 1
- 210000004080 milk Anatomy 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 235000019426 modified starch Nutrition 0.000 description 1
- 230000000877 morphologic effect Effects 0.000 description 1
- 239000006872 mrs medium Substances 0.000 description 1
- 210000003205 muscle Anatomy 0.000 description 1
- 230000035772 mutation Effects 0.000 description 1
- 201000001119 neuropathy Diseases 0.000 description 1
- 230000007823 neuropathy Effects 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 108091006527 nucleoside transporters Proteins 0.000 description 1
- 239000002773 nucleotide Substances 0.000 description 1
- 125000003729 nucleotide group Chemical group 0.000 description 1
- 238000004806 packaging method and process Methods 0.000 description 1
- 235000019629 palatability Nutrition 0.000 description 1
- 229940049954 penicillin Drugs 0.000 description 1
- 235000019319 peptone Nutrition 0.000 description 1
- 208000033808 peripheral neuropathy Diseases 0.000 description 1
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 description 1
- 229920000053 polysorbate 80 Polymers 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- 238000012257 pre-denaturation Methods 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 235000018102 proteins Nutrition 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 150000003212 purines Chemical class 0.000 description 1
- 230000000306 recurrent effect Effects 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 235000014102 seafood Nutrition 0.000 description 1
- 239000001632 sodium acetate Substances 0.000 description 1
- 235000017281 sodium acetate Nutrition 0.000 description 1
- 229920001027 sodium carboxymethylcellulose Polymers 0.000 description 1
- 239000012064 sodium phosphate buffer Substances 0.000 description 1
- 238000005507 spraying Methods 0.000 description 1
- 239000012086 standard solution Substances 0.000 description 1
- 210000002784 stomach Anatomy 0.000 description 1
- 229960005322 streptomycin Drugs 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 210000001258 synovial membrane Anatomy 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 235000018648 unbalanced nutrition Nutrition 0.000 description 1
- 229940005267 urate oxidase Drugs 0.000 description 1
- 238000009777 vacuum freeze-drying Methods 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
- 210000001835 viscera Anatomy 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
- 235000021119 whey protein Nutrition 0.000 description 1
- 235000008939 whole milk Nutrition 0.000 description 1
- UHVMMEOXYDMDKI-JKYCWFKZSA-L zinc;1-(5-cyanopyridin-2-yl)-3-[(1s,2s)-2-(6-fluoro-2-hydroxy-3-propanoylphenyl)cyclopropyl]urea;diacetate Chemical compound [Zn+2].CC([O-])=O.CC([O-])=O.CCC(=O)C1=CC=C(F)C([C@H]2[C@H](C2)NC(=O)NC=2N=CC(=CC=2)C#N)=C1O UHVMMEOXYDMDKI-JKYCWFKZSA-L 0.000 description 1
Classifications
-
- 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
Abstract
The invention relates to the field of microorganisms, and discloses a haemostatic acid-reducing lactobacillus fermentum A21215 and application thereof, wherein the lactobacillus fermentum A21215 is preserved in the microorganism strain preservation center of Guangdong province at 11/2022, and the preservation number is GDMCCNO 62963. The microorganism classification was named Limosilactobacillus fermentum. The strain can degrade nucleoside purine, inhibit xanthine oxidase activity, has higher adhesiveness to colon epithelial cells, inhibits pathogenic bacteria growth, effectively reduces blood uric acid level of hyperuricemia model mice, and is suitable for production of low-purine foods and fermented foods with purine-reducing function.
Description
Technical Field
The invention relates to the field of intestinal microorganisms, in particular to lactobacillus fermentum A21215 with purine nucleoside reducing and hyperuricemia model mouse blood uric acid reducing functions and application thereof.
Background
Hyperuricemia (HUA) is a metabolic disease characterized by a significant higher than normal serum uric acid content in humans, and most studies define hyperuricemia as being defined as two times of measurement of > 420. Mu. Mol/L for male fasting uric acid and > 360. Mu. Mol/L for female fasting uric acid in a non-daily normal dietary state. The current trend of hyperuricemia in China has the characteristics of younger and high epidemic, male is higher than female, the incidence rate in coastal areas is higher than that in inland areas, and the incidence rate of genetic aggregation is from 5% to 23%, which is close to western developed countries. Long-term hyperuricemia can cause monosodium urate crystals (MSU) or uric acid to form a supersaturated state in extracellular fluid, so that the urate crystals are deposited in a large amount in joint synovium, bursa, cartilage and other tissues, and cause acute, chronic and recurrent inflammatory diseases, namely gout.
Uric acid is produced by purine nucleic acid substance metabolism, and in the long-term genetic evolution process, the gene of human encoding urate oxidase is subjected to inactivation mutation, and uric acid becomes a final product of purine nucleoside metabolism in vivo. The source of uric acid in human body can be divided into endogenous and exogenous, wherein 2/3 of uric acid is produced by metabolism of nucleic acid and other purine compounds decomposed by endogenous cell metabolism, 1/3 of uric acid is decomposed by enzyme action of purine in exogenous food, exogenous purine can be derived from animal viscera rich in purine nucleic acid, seafood, beer and the like, and the intake of purine components directly influences uric acid level in blood, and the current method for controlling uric acid level to prevent gout is mainly drug treatment and food prevention. Common drugs include allopurinol, benzbromarone, probenecid, and the like. Among them, allopurinol mainly inhibits xanthine oxidase which promotes uric acid production, benzbromarone, probenecid and the like promote uric acid excretion, however, long-term drug treatment is accompanied with various side effects such as liver and kidney damage, gastrointestinal symptoms, muscle and neuropathy and the like. Food prevention is one of the main methods for reducing uric acid production by reducing the intake of exogenous purine substances, but has the problems of inaccurate purine substance content in various foods, poor palatability of low-purine foods, easy unbalanced nutrition of patients and the like. The intestinal probiotics can reduce the absorption of purine nucleoside substances by taking and degrading the purine substances in the digestive tract; directly degrading uric acid or promoting excretion of uric acid; changing intestinal permeability, regulating balance of intestinal flora, etc. to achieve the purposes of controlling blood uric acid level, preventing and treating hyperuricemia and gout.
Lactic acid bacteria are safe food-grade microorganisms and play an important role in preventing and treating metabolic regulation diseases. In recent years, some lactic acid bacteria strains with reduced purine nucleosides and reduced uric acid are discovered: lactobacillus gasseri 0LL2922,1 ×10 strain of Ming Zhi Co., ltd 9 The degradation rates of CFU/mL bacterial suspension to 1.25mM guanosine and 1.25mM inosine were 90% and 70%, respectively. Lactobacillus brevis DM9218 is reported by university of Dalian medical science, and 1.26mM inosine and 1.26mM guanosine are degraded simultaneously, wherein the degradation rates are 99.31 percent and 99.64 percent respectively. The institute of agricultural products processing of Jilin province institute reported that Lactobacillus reuteri TSR332 and Lactobacillus fermentum TSF331 with in vitro nucleoside-reducing ability, which are found by Fenghua Biotechnology Co., ltd, can degrade 1.25mM inosine and guanosine degradation rate of about 60% by Lactobacillus plantarum UA149The degradation rate of 1.26mM inosine and guanosine reaches the highest (CN 111388509A), wang Chenghua and the like, and the degradation rate of 1mM inosine and guanosine is about 60% for Lactobacillus fermentum Lactobacillus fermentum-4 (CN 110684685A). In addition, zhu et al isolated and identified a strain of Lactobacillus fermentum from Sinkiang traditional yogurt and demonstrated its ability to reduce guanosine in vitro, with guanosine degradation rate of 63.06% at a guanosine concentration of 1mM, and animal experiments indicated that it had a blood uric acid reducing function (CN 110079476A). These strains with in vitro degradation of nucleosides also show a blood uric acid lowering effect in the intervention of mice in hyperuricemia models. However, the nucleic acid and purine substances contained in the daily diet are high, and thus it is required to develop a strain having a higher nucleotide degrading ability. In addition, it has been found that there are various ways of affecting blood uric acid levels by the strain, whether or not xanthine oxidase activity is inhibited in addition to reduction of the intake of nucleoside purine, the tolerance of the strain to gastrointestinal acidity, bile salts, adhesion to intestinal cells, and the balance of intestinal flora regulation by the strain also affect the effect of probiotics on the function of lowering blood uric acid, so that the development of the strain with higher nucleoside degradation capability and the strain with multiple functions of inhibiting xanthine oxidase, facilitating colonization in the intestinal tract, inhibiting harmful bacteria and the like has important significance in preventing and treating gout and hyperuricemia.
Disclosure of Invention
The invention aims to provide a lactobacillus fermentum A21215 for reducing blood uric acid and application thereof, and the strain has the capability of reducing purine nucleosides and reducing the blood uric acid of mice with hyperuricemia models.
In order to achieve the above purpose, the present invention adopts the following technical scheme: a strain of Lactobacillus fermentum A21215 with the blood uric acid reducing effect, wherein the Lactobacillus fermentum A21215 is deposited with the microorganism strain collection of Guangdong province at 11/2022 under the deposit number GDMCCNO 62963, and has the capabilities of degrading purine nucleosides, inhibiting xanthine oxidase and reducing the blood uric acid of mice with hyperuricemia models.
The 16S rDNA sequence of the strain is shown as SEQ ID NO. 1.
The colony characteristics of the lactobacillus fermentum A21215 are as follows: the colony surface is smooth, white or milky single colony is round or oval under microscope, no spore exists, and gram staining is positive. The Lactobacillus fermentum A21215 with reduced blood uric acid grows well on MRS culture medium, and the colony is milky white and has smooth surface. The bacterial forms were examined microscopically and gram-stained purple and rod-like.
The haemopoietic acid reducing lactobacillus fermentum A21215 is obtained by separating, purifying and identifying faeces samples of old people with good health and long life in Guangxi Hepu county at 98 years.
The application of the lactobacillus fermentum A21215 in preparing fermented food.
The application of the lactobacillus fermentum A21215 in preparing low-purine foods.
The application of the blood uric acid reducing lactobacillus fermentum A21215 in preparing freeze-dried powder, capsules, fermented milk and products thereof.
The application of the lactobacillus fermentum A21215 in degrading nucleosides is to add the lactobacillus fermentum A21215 cells or the culture and the metabolite thereof into a purine-containing system.
The screening method of the lactobacillus fermentum A21215 for reducing blood uric acid comprises the following steps:
(1) Diluting and coating a sample on an MRS plate, selecting a characteristic colony, separating and purifying to obtain a purified candidate strain;
(2) Performing 16S rDNA sequencing, and identifying and storing lactobacillus strains through NCBI and RDP database Blast comparison;
(3) The method comprises the steps of using resting cell nucleoside metabolism capability as a detection index, and using an enzyme-labeled instrument to perform preliminary screening on nucleoside degradation capability of lactic acid bacteria, so as to screen out strains with higher nucleoside metabolism capability;
(4) The degradation of nucleosides by resting cells was determined using a water e2695 HPLC system equipped with a 2998PDA uv detector.
The steps (1) - (4) are carried out to obtain the lactobacillus fermentum A21215 with high nucleoside metabolic capacity, wherein the degradation rate of 3mM inosine-guanosine-adenosine is 89.91 +/-1.27%, 95.78+/-3.09%, 100+/-0.00% and the degradation rate of 5mM inosine-guanosine-adenosine is 85.29 +/-1.18%, 65.68+/-4.12% and 89.94 +/-0.86%.
The acid-resistant and cholate-resistant test of the hypouricemia lactobacillus fermentum A21215 shows that the hypouricemia lactobacillus fermentum has certain tolerance to acid and cholate. After 4 hours of artificial gastric juice with pH of 2.0 and pH of 2.5, the survival rate of the lactobacillus fermentum A21215 is 192.86 +/-21.43 percent and 257.14 +/-14.29 percent respectively; can survive 8 hours under 0.3% high concentration bile salt.
The genomic DNA of Lactobacillus paracasei A21215 was PCR amplified and sequenced using the 27F/1492R of bacterial universal primer 16S rDNA to obtain a 1467bp SEQUENCE as shown in SEQUENCE Listing No. 1. The sequence was subjected to BLAST alignment in NCBI database and phylogenetic tree was constructed, and A21215 was identified as Lactobacillus fermentum.
Compared with the prior art, the invention has the beneficial effects that:
the lactobacillus fermentum A21215 for reducing blood uric acid provided by the invention has the capability of degrading purine nucleosides and inhibiting xanthine oxidase activity more efficiently. Fermentation broth OD of A21215 600 When the ratio is adjusted to 2, the degradation rate of resting cells prepared from 2mL of bacterial liquid to 3mM inosine-guanosine-adenosine solution is 89.91 +/-1.27% and 95.78+/-3.09%, 100+/-0.00% respectively in 1h, and the degradation rate to 5mM inosine-guanosine-adenosine solution is 85.29 +/-1.18%, 65.68+/-4.12% and 89.94 +/-0.86% respectively. Through degrading nucleoside, the absorption of intestinal nucleoside transporter to purine is reduced, so that the generation and accumulation of uric acid are reduced, and the blood uric acid of mice with hyperuricemia models is effectively reduced.
In addition, the lactobacillus fermentum A21215 for reducing blood uric acid provided by the invention has the capabilities of tolerating gastrointestinal tract environment, ensuring good adhesion of intestinal cells and inhibiting the growth of pathogenic bacteria. The strain affects blood uric acid level through various ways, whether xanthine oxidase activity is inhibited or not besides reducing the intake of nucleoside purine, the strain tolerance to gastrointestinal acidity and bile salts, the adhesion to intestinal cells and the effect of regulating intestinal flora balance of the strain also affect probiotics to exert the function of reducing blood uric acid, so that the strain with higher nucleoside degradation capability needs to be developed, and the strain is resistant to gastrointestinal environment and has better viscosity to intestinal cellsAnd a strain with multiple functions of attaching, inhibiting pathogenic bacteria and the like. After 4 hours of artificial gastric juice with pH of 2.0 and pH of 2.5, the survival rate of the lactobacillus fermentum A21215 is 192.86 +/-21.43 percent and 257.14 +/-14.29 percent respectively; can survive for 8h under 0.3% high concentration bile salt, and lactobacillus fermentum A21215 can adhere to colon epithelial cells NCM460 in large quantity, gram stain count shows that the number of adhered bacteria is 3.6X10 on 100cells 5 100cells, lactobacillus fermentum A21215 can form transparent ring around pathogenic bacteria, and inhibit pathogenic bacteria growth. The good performance of the hypouricemia lactobacillus fermentum A21215 makes the hypouricemia lactobacillus fermentum A21215 more beneficial to develop into a probiotic strain, is applied to the field of functional foods, is used for producing the hypopurine probiotic bacteria and the low-purine foods, and has important significance for preventing, improving and treating gout and hyperuricemia diseases.
Compared with the existing purine-reducing lactobacillus fermentum, the blood uric acid-reducing lactobacillus fermentum A21215 provided by the invention has higher nucleoside degradation capability, can inhibit xanthine oxidase activity, can resist gastrointestinal tract environment, has good adhesion to colon epithelial cells, can effectively inhibit pathogenic bacteria from growing, can play a role in reducing blood uric acid from multiple ways, and can effectively reduce the blood uric acid level of a model mouse in the intervention of the model mouse by the lactobacillus fermentum A21215.
Drawings
FIG. 1 is a colony morphology of Lactobacillus fermentum A21215.
FIG. 2 shows the results of genome extraction (a) and 16S rDNA amplification (b) of Lactobacillus fermentum A21215.
FIG. 3 is a phylogenetic tree of Lactobacillus fermentum A21215 with reduced blood uric acid.
FIG. 4 is an HPLC chart of A21215 for degradation of 3mM inosine-Guanosine (Inisine-Guanosine) within 30min and 60min.
FIG. 5 shows the effect of different volumes of incubation of Lactobacillus paracasei A21215 on xanthine oxidase activity.
FIG. 6 shows the results of experiments on the bile salt tolerance (a) and the acid tolerance (b) of Lactobacillus paracasei A21215.
FIG. 7 shows adhesion of Lactobacillus paracasei A21215 to NCM460, a colonic epithelial cell.
FIG. 8 shows the inhibitory effects of A21215 on Staphylococcus aureus (a), escherichia coli (b) and Escherichia coli (c).
FIG. 9 shows uric acid lowering effect of Lactobacillus fermentum A21215 on serum and urine of mice with hyperuricemia model.
Detailed Description
The technical scheme of the invention is further described in detail below with reference to specific embodiments.
Example 1
The invention discloses separation, purification and identification of a Lactobacillus fermentum A21215 strain for Ming-Jiang blood uric acid, which comprises the following steps:
the Mingjiang blood uric acid fermenting lactobacillus A21215 (hereinafter referred to as fermenting lactobacillus A21215) is obtained by separating a fecal sample from old people with a healthy life and a long life of 98 years in Guangxi Hepu county, and specifically, the separation, purification and identification method of the blood uric acid fermenting lactobacillus A21215 is as follows:
preparation of MRS culture medium: 20g of glucose, 10g of beef extract, 10g of peptone, 5g of yeast powder, 2g of dipotassium hydrogen phosphate, 2g of diammonium citrate, 5g of sodium acetate, 0.58g of magnesium sulfate, 0.25g of manganese sulfate, 1mL of Tween 80, 1000mL of distilled water, pH of 6.6-6.8, and when preparing a solid culture medium, 20g of agar is added and the mixture is autoclaved at 115 ℃ for 20min.
Separating and purifying strains: adding 0.1mL or 0.1g sample to be separated into 0.9mL sterile physiological saline, shaking and mixing uniformly to obtain sample suspension, and diluting 10 times with physiological saline to obtain 10 -1 ～10 -5 Dilution gradient series, dilution was 10 -3 ～10 -5 0.1mL of the dilution of (C) was applied to MRS solid medium, and the plate was placed in a constant temperature incubator at 37℃for 1-2 days. After the culture is finished, the bacterial colonies are selected according to morphological characteristics of the bacterial colonies, and are subjected to plate streak purification. The growth morphology of Lactobacillus paracasei A21215 on MRS plates is shown in FIG. 1. Single colony on the purification plate is selected and inoculated in MRS liquid culture medium, after standing culture for 18 hours at 37 ℃, bacterial liquid and 40% glycerol are mixed according to the proportion of 1:1, and the bacterial liquid is preserved at-80 ℃.
Strain16S rDNA identification: after separation and purification, lactobacillus fermentum A21215 is cultured for 16 hours by using MRS liquid culture medium according to the following stepsThe operation of the bacterial genome extraction kit showed that the genome of lactobacillus fermentum a21215 was extracted and verified by agarose gel electrophoresis, and the result is shown in fig. 2 (a). 1. Mu.L of genome diluted to 1 ng/. Mu.L is taken as a template and added into a 16S rDNA amplification system, and 50. Mu.L of amplification system is as follows: 1. Mu.L of genome as template, 1. Mu.L of each of primer 27F (SEQ ID No. 2) and primer 1492R (SEQ ID No. 3), 25. Mu.L of 2 XEs Taq Mix was added, and ddH2O 22. Mu.L was amplified by 5min pre-denaturation at 94 ℃, 30s annealing at 55 ℃, 1min extension at 72℃and 30 cycles of final extension at 72℃were performed. After the amplification was completed, 1% agarose gel electrophoresis was performed, the 16S rDNA amplification product of the A21215 strain was shown in FIG. 2 (b), 16S rDNA of about 1500bp was obtained, sequencing was performed by the engineering and bioengineering Co., ltd, the 16S rDNA SEQUENCE of the A21215 strain was obtained as shown in SEQUENCE LISTING (SEQ ID No. 1), the obtained 16S rDNA SEQUENCE was aligned and a phylogenetic tree was constructed using database NCBI (www.ncbi.nlm.nih.gov), the phylogenetic relationship between strains was analyzed using MEGA-X, the SEQUENCE was aligned using ClustalW, and the phylogenetic tree boottrap was set to 1000 and shown in percentage form, as shown in FIG. 3. The A21215 strain has the highest similarity with the sequence (GenBank: MT 510461.) of Lactobacillus fermentum Lactobacillus fermentum strain 5638, which is the closest relation to Lactobacillus fermentum, and was initially identified as Lactobacillus fermentum. The strain has been deposited at 11.11.2022 with the collection of microorganism strains, guangdong province, under the accession number GDMCC NO. 62963.
Example 2
HPLC determination of degradation of nucleosides by Lactobacillus fermentum A21215
In a Waters alliance acquity e2695 liquid chromatograph equipped with a Waters 2998PDA photodiode matrix chromatograph detector, the nucleoside content was analyzed at 30℃using a Waters symmetry shield RP chromatographic column (5 μm, 4.6X250 mm, waters, USA), detection wavelength 254nm, flowMobile phase 20mM H at pH 4.1 3 PO 4 -KH 2 PO 4 Buffer and methanol (95:5) at a flow rate of 0.5mL/min. After a 10mmol/L PBS was used to prepare a 1mmol/L standard solution of inosine, guanosine and adenosine, a 0.22 μm aqueous filter membrane was used to filter the solution, standards (0, 0.25,0.5,0.75,1 mmol/L) of different concentrations were analyzed according to the set chromatographic conditions, retention time and peak area of the standards were analyzed, linear fitting was performed on peak areas with the concentration of each nucleoside purine, and a standard curve between inosine and guanosine contents and peak areas was established, wherein the inosine standard curve was: y= 2.47275 ×10 7 x，R 2 =0.9999; the guanosine standard curve is: y= 3.08853 ×10 7 x，R 2 =0.9999; the adenosine standard curve is: y= 3.19242 ×10 7 x，R 2 ＝0.9999。
Lactobacillus fermentum a21215 was inoculated in an inoculum size of 1% in MRS liquid, and after 16h stationary culture at 37 ℃ the OD600 of the broth was adjusted to 2.00 using a blank medium, 2mL broth was taken, centrifuged at 4 ℃,8000rpm for 2min, the supernatant was discarded, and the cells were collected. To the cells, 750. Mu.L of physiological saline was added to wash the cells, and the washing was repeated 1 time to collect the washed cells. Using PBS to prepare 1, 2, 3 and 5mmol/L inosine-guanosine mixed solution or inosine-guanosine-adenosine working solution, adding 750 mu L of nucleoside working solution into the collected bacterial cells, taking the nucleoside working solution without adding bacterial cells as blank control of a nucleoside degradation system under the same operation, and setting bacterial cells inactivated by boiling water bath for 10min as heat inactivation control. And (3) placing the prepared nucleoside degradation reaction system at 37 ℃, and incubating for 30min and 60min under constant-temperature shaking at 120 rpm. After the reaction was completed, the system was inactivated in a boiling water bath for 10min, centrifuged at 10000rpm for 2min, and the supernatant was filtered using a PES filter membrane of 0.22 μm and placed in a refrigerator at 4℃for use. And quantifying the nucleoside degradation capacity of the strain according to the chromatographic conditions, wherein the sample injection amount is 5 mu L, and calculating the residual nucleoside in the degraded system according to a standard curve of the corresponding nucleoside. And calculating the degradation rate of the thalli to the nucleoside according to the following formula:
α＝[(n-X)/n]×100％
alpha: degradation rate (%), n: content (mmol) of degraded inosine, guanosine or adenosine, X: inosine or guanosine content (mmol) after degradation.
Degradation rates of lactobacillus paracasei a21215 after incubation with three different concentrations of nucleosides for 60min are shown in table 1, and HPLC detection of 3mM nucleoside degradation is shown in fig. 4.
TABLE 1 degradation Rate of Lactobacillus fermentum A21215 on nucleosides of different concentrations
Example 3
Inhibition of Xanthine Oxidase (XOD) by lactobacillus fermentum a21215 incubation supernatant
Xanthine Oxidase (XOD) catalyzes hypoxanthine and xanthine to generate uric acid, and inhibiting the activity of xanthine oxidase can reduce the generation of uric acid, and allopurinol is one of the main medicaments used in the current clinical treatment of gout, and the principle is that allopurinol with a structure similar to xanthine is used for competitively binding to the active site of XOD, so that the generation of uric acid is reduced. Inoculating a target strain to MRS liquid culture medium with an inoculum size of 1%, standing and culturing for 24 hours at a constant temperature of 37 ℃, centrifuging for 2min, collecting thalli, using sodium phosphate buffer PBS with pH of 7.4 to resuspend and wash the thalli twice, finally adding PBS according to the volume of 20% of initial fermentation liquor, placing at 37 ℃,160rpm, taking out the centrifugally reserved supernatant after 4 hours, adding supernatants with different volumes into a reaction system shown in table 2, taking sterile PBS as a blank control, measuring and repeating for three times, measuring the change of the absorbance value of uric acid of a product at a characteristic absorption peak wavelength of 293nm, calculating the activity of XOD, taking the activity of XOD as 100% when the supernatant is not added, calculating the relative activity of XOD when the supernatant is added and the supernatant is incubated by fermenting lactobacillus A21215 with different pH values, and the micromolar activity of uric acid generated per milliliter of the protein is shown in a graph 5a, taking the relative activity of XOD as 47.17%, the relative activity of the supernatant is 62.66% when the supernatant is added, and taking the relative activity of the supernatant is the supernatant as a graph of 20.83% when the supernatant is incubated with the same pH value as the supernatant, and the relative activity of the supernatant is the same as the pH value of 62.62.83, and the activity of the supernatant is inhibited by the same, and the activity of the supernatant is shown in a graph is about the trend of increasing to be about the activity of the same.
TABLE 2 xanthine oxidase 200. Mu.L reaction system
Example 4
Bile salt and acid resistance of Lactobacillus fermentum A21215
The bile salt mass fractions of MRS liquid culture medium are respectively regulated to 0.1%,0.2% and 0.3% by using ox gall salt, lactobacillus fermentum A21215 is inoculated into the MRS liquid culture medium, after the culture is carried out for 18 hours at 37 ℃, 1mL of bacterial liquid is respectively taken and added into 9mL of bile salt culture medium containing different mass fractions, the viable count is respectively counted at 0 hours, 2 hours, 4 hours and 8 hours, and as a result, the viable count of lactobacillus fermentum A21215 is relatively stably kept at 10 in a 1% bile salt environment as shown in figure 6a 9 About CFU/mL, the number of viable bacteria A21215 is kept at 10 when the bile salts with different concentrations are tolerated for 2 hours 5 Above CFU/mL, lactobacillus fermentum a21215 was shown to survive in high concentrations of bile salts.
Will be purchased from 1mol/L sterile HCl and 1mol/L NaOHThe pH of the artificial gastric juice is respectively regulated to 2.0 and 2.5, lactobacillus fermentum A21215 is inoculated in MRS liquid culture medium, after the culture is carried out for 18 hours at 37 ℃, 1mL of bacterial liquid is respectively taken and added into 9mL of the artificial gastric juice with the pH of 2.0 and the pH of 2.5, 100 mu L of the bacterial liquid is taken for viable count after the bacterial liquid is fully and uniformly mixed, the rest of the bacterial liquid is cultured at the constant temperature of 37 ℃ for 4 hours, sampling is carried out for viable count after 4 hours, the result is shown in Table 3, the viable count of 0 hour is taken as 100%, and the relative survival rate is calculated, and the result is shown in Table 6 b. After 4 hours of artificial gastric juice with pH of 2.0 and pH of 2.5, the survival rate of the Lactobacillus fermentum A21215 is 192.86 +/-21.43% and 257.14 +/-14.29%, respectively, which shows that the Lactobacillus fermentum A21215 corresponds to pH2.0 and pH 2.5 has stronger tolerance.
TABLE 3 viable count and survival rate of Lactobacillus fermentum A21215 in artificial gastric juice at different pH for 4h
Example 5
Adhesion of Lactobacillus fermentum A21215 to colonic epithelial NCM460
The 9 th generation colon epithelial cells NCM460 stored in liquid nitrogen are resuscitated, and the cells are cultured by using 10% fetal bovine serum, penicillin and streptomycin and DEME high sugar culture medium, and after passage to 11 generations, the cells are subjected to 2.5X10 5 Inoculating the cells into a six-hole plate with a cell climbing sheet, culturing overnight, adding cell culture solution, and re-suspending to OD 600 After 2.0 incubation for 2h with A21215 bacterial solution, the cell slide was removed and gram stained with sterile PBS for 4-6 washes. As a result, as shown in FIG. 7, a large amount of cells of A21215 were adhered around each cell, and the statistics revealed that 3.6X10 were adhered to 100 colon epithelial cells NCM460 5 The individual thalli show that the lactobacillus fermentum A21215 has better adhesiveness to colon epithelial cells, and is beneficial to the colonization of the lactobacillus fermentum A21215 in the intestinal tract.
Example 6
Inhibition of Lactobacillus fermentum A21215 on Escherichia coli, staphylococcus aureus and Escherichia coli
Staphylococcus aureus (ATCC 43300), escherichia coli O157 were cultured overnight at 37 ℃, 220rpm using LB liquid culture: h7 After (ATCC 35150) and Escherichia coli (ATCC 25922), 100. Mu.L of the bacterial liquid was applied to an LB plate, 200. Mu.L of the fermentation liquid of A21215 was added to an oxford cup, and the LB plate was placed in a constant temperature incubator at 37℃for stationary culture with MRS medium as a control. The inhibition conditions of A21215 on escherichia coli, staphylococcus aureus and escherichia coli are shown in fig. 8a, 8b and 8c respectively, wherein blank MRS culture solution is used as a control at the upper left part of a flat plate, a bacteriostasis ring is not formed around the blank MRS, a transparent ring without pathogenic bacteria growth is formed around the fermentation liquor of A21215, and the inhibition effect of the fermentation liquor of lactobacillus fermentum A21215 on three different pathogenic bacteria is shown.
Example 7
Effect of lactobacillus fermentum a21215 on hyperuricemia model mice
SPF-grade Kunming mice were purchased 24, male, 4-5 weeks old, and weighing 16-20g. Before the experiment, the mice are adapted for one week, and after 7 days of adapted feeding, the mice are randomly divided into a normal control group, a hyperuricemia model group, a lactobacillus fermentum A21215 intervention group and an allopurinol treatment group, and each group is 6. Prevention with probiotics after the end of the adaptation period: probiotics gavage of 11 points per day for the probiotic groups except for the normal control group, model group, and positive control group 0.2mL containing 1×10 10 Skim milk of CFU lactobacillus fermentum a21215, and the remaining group mice were gavaged at 11 points daily with 0.2mL skim milk. All rats were normally fed and freely drinking water for 14 days. Starting on day 21, mice except the normal control group were fed with high purine custom-made mice and were intraperitoneally injected with 0.2ml 350 mg/(kg·bw) of potassium oxazinate-sodium carboxymethyl cellulose suspension at 10 points daily according to body weight standard; normal control group provided normal mouse diet, and mice treated with probiotic group had 1×10 stomach lavage at 11 points per day 10 Skim milk of CFU. All mice were free to drink water for 7 days up to 28 days. And collecting blood and urine of all mice at the end of the experiment, centrifuging the blood at 3500r/min for 15min, taking the supernatant as serum, and storing in a refrigerator at-80 ℃ for later use. The uric acid content was determined using uric acid detection kit purchased from the institute of bioengineering, built in south kyo. The effect of lactobacillus fermentum a21215 on uric acid content in serum and urine of mice with hyperuricemia model is shown in fig. 9, and the results show that the lactobacillus fermentum a21215 has the capability of reducing uric acid in serum and urine of mice with hyperuricemia model.
Example 8
Preparation of lyophilized powder
And (3) preparing a protective agent: preparing 12% skimmed milk solution with skimmed milk powder, sterilizing at 115deg.C for 15min, and cooling.
Strain activation and culture: inoculating Lactobacillus paracasei A21215 stored in an glycerol pipe at-80 ℃ to an MRS flat plate by using an inoculating loop, streaking, culturing at 37 ℃ for 24 hours, streaking the activated single colony again, inoculating the single colony subjected to secondary streaking to the MRS liquid culture medium, culturing at 37 ℃ for 12 hours, inoculating 4.5mL of culture solution to 150mL of MRS liquid culture medium, inoculating 10.5mL of seed solution to 350mL of MRS liquid culture medium according to 3% inoculum size after 12 hours, continuously culturing for 20 hours, centrifuging at 4 ℃ for 5 minutes at 4000rpm, collecting the thalli, re-suspending and washing the thalli by using 350mL of sterile physiological saline at 4 ℃ for 5 minutes at 4000rpm, and repeatedly washing once according to the operation. The collected thalli are weighed, and 1:1 (W: V) skim milk is added as a protective agent, and the mixture is uniformly mixed to prepare bacterial suspension and split charging is carried out.
Vacuum freeze drying: filling the split-packed bacterial suspension into a material bottle, sealing the mouth, placing the material bottle in a refrigerating chamber at 4 ℃ for refrigerating for 30min, then freezing for 1.5h in a refrigerator at-20 ℃, then freezing for 20min in a refrigerator at-80 ℃, freeze-drying the sample by using a four-ring Foring vacuum freeze dryer, inserting the material bottle into a T-shaped frame of the vacuum freeze dryer, opening a switch to communicate an ampoule tube with the vacuum freeze dryer, drying the bacterial agent for 8-20h, stopping drying and vacuum sealing after judging that the material is dried, placing the material in a drying place for standby, preparing the freeze-dried powder into bacterial suspension, performing specific amplification verification and viable bacteria counting.
Example 9
Preparation of solid beverages
Lactobacillus fermentum a21215 was prepared as in example 5 to obtain a freeze-dried powder of lactobacillus fermentum a21215 with reduced blood uric acid, 40% whole milk powder, 10% fructo-oligosaccharides, 10% trehalose, 10% strain freeze-dried powder, 10% whey protein powder, 5% citric acid, 5% maltodextrin, 5% carrageenan and 5% edible essence were weighed, stirred with a stirrer for 20min, and the mixture was packaged and sealed with a powder packaging machine.
Example 10
Preparation of microcapsules
Lactobacillus fermentum a21215 was inoculated into MRS liquid medium, cultured at 37 ℃ for 18 hours, and then centrifuged to collect the cells and washed twice with physiological saline. Preparing 2% sodium alginate solution and 2% modified starch respectively, sterilizing, and mixing the two solutions according to a ratio of 1:1 volume ratio, as wall material solution, then mixing with centrifugal washed pure thallus, spraying into calcium chloride (2%, m/V) solution after atomizing by sprayer nozzle, making calcium ion and sodium alginate cross-linking reaction to form microcapsule, and standing and solidifying for 120min.
Example 11
Preparation of fermented milk
The lyophilized powder of Lactobacillus fermentum A21215 frozen at-20deg.C was taken out, inoculated into 10mL of previously prepared, homogenized and sterilized pure cow milk at an inoculum size of 1%, fermented at 40deg.C for 24h, and then refrigerated at 4deg.C. And the fermented milk is taken as seed liquid for subsequent yogurt fermentation, added into pure cow milk according to the addition amount of 2 percent, fermented for 24 hours at 40 ℃, and then refrigerated for 24 hours at 4 ℃. Counting viable bacteria of fermented milk, wherein the viable bacteria number is greater than 1.0X10 8 CFU/mL。
Example 12
Lactic acid bacteria beverage
Inoculating Lactobacillus fermentum A21215 with 5% inoculum size into sterile mixed solution containing 100L soybean milk, cheese whey 2kg and sucrose 8kg, culturing at 40deg.C for 20 hr, filling into paper container by aseptic filling method to obtain lactobacillus beverage, standing at 5deg.C for 7 days, and keeping flowing state with lactic acid content of 1.2% and viable count greater than 10 7 CFU/mL。
The above examples prove the purine nucleoside degradation capability of the strain, the capability of reducing blood uric acid of a model mouse and the application of the strain in the production of freeze-dried preparations, yogurt fermentation, capsules and other products. The invention is not limited to the above-mentioned technical solutions, and any improvement of the invention, including making various simple modifications to the technical solutions, falls within the scope of the invention.
In addition, the specific features described in the above embodiments may be combined in any suitable manner, and in order to avoid unnecessary repetition, various possible combinations are not described further.
In addition, various embodiments of the present invention may be arbitrarily combined, and should also be regarded as the disclosure of the present invention, as long as the idea of the present invention is not deviated.
Claims (5)
1. Lactobacillus fermentum（Lactobacillus fermentum）A21215, wherein the Lactobacillus fermentum A21215 was deposited with the microorganism culture Collection, canon province, at 11/2022, under accession number GDMCC NO 62963.
2. Use of lactobacillus fermentum a21215 as claimed in claim 1 in the manufacture of a fermented food product.
3. Use of lactobacillus fermentum a21215 as claimed in claim 1 in the preparation of low purine foods.
4. Use of lactobacillus fermentum a21215 according to claim 1 for the preparation of a lyophilized powder, a capsule, a fermented milk.
5. Use of lactobacillus fermentum a21215 according to claim 1 for degrading inosine, guanosine, adenosine.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211600780.4A CN116024130B (en) | 2022-12-13 | 2022-12-13 | Lactobacillus fermentum A21215 for reducing blood uric acid and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211600780.4A CN116024130B (en) | 2022-12-13 | 2022-12-13 | Lactobacillus fermentum A21215 for reducing blood uric acid and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN116024130A CN116024130A (en) | 2023-04-28 |
| CN116024130B true CN116024130B (en) | 2023-11-24 |
Family
ID=86071428
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202211600780.4A Active CN116024130B (en) | 2022-12-13 | 2022-12-13 | Lactobacillus fermentum A21215 for reducing blood uric acid and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN116024130B (en) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116694530B (en) * | 2023-06-28 | 2023-11-07 | 北京量化健康科技有限公司 | Lactobacillus composition and application thereof |
| CN116656578B (en) * | 2023-07-26 | 2023-11-28 | 杭州微致生物科技有限公司 | Lactobacillus mucilaginosus VB216 and application thereof |
| CN117025456B (en) * | 2023-07-31 | 2024-02-06 | 广西爱生生命科技有限公司 | Lactobacillus paracasei A21151 with blood uric acid reducing and anti-inflammatory effects and application thereof |
Citations (19)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2008005834A (en) * | 2006-05-31 | 2008-01-17 | Meiji Milk Prod Co Ltd | Lactobacillus having blood uric acid level-reducing activity |
| CA2851018A1 (en) * | 2007-11-29 | 2009-06-04 | Hiroshi Tsuboi | Lactic acid bacteria having action of lowering blood uric acid level |
| CN101932697A (en) * | 2007-11-29 | 2010-12-29 | 明治乳业株式会社 | Lactic acid bacteria having action of lowering blood uric acid level |
| CN102747004A (en) * | 2007-11-29 | 2012-10-24 | 株式会社明治 | Lactic acid bacteria with effect of uric acid level reducing in blood |
| JP2013048636A (en) * | 2006-05-31 | 2013-03-14 | Meiji Co Ltd | Lactobacillus having blood uric acid level-reducing activity |
| CN107208029A (en) * | 2014-11-18 | 2017-09-26 | 株式会社明治 | Lactic acid bacteria of ability and application thereof is absorbed with purine |
| CN110079476A (en) * | 2019-04-24 | 2019-08-02 | 杭州娃哈哈科技有限公司 | One plant of lactobacillus fermenti that can reduce blood uric acid |
| WO2019157585A1 (en) * | 2018-02-13 | 2019-08-22 | Proviva Pharma Inc. | Probiotic formulations for the treatment and alleviation of metabolic and oxidative stress, inflammation and neurodegeneration |
| CN110684685A (en) * | 2019-10-14 | 2020-01-14 | 广西大学 | Lactobacillus fermentum 9-4 and application thereof |
| CN113215012A (en) * | 2020-01-21 | 2021-08-06 | 宁波倍益嘉生物科技有限公司 | Probiotic strain for reducing blood uric acid, composition and application thereof |
| CN113999805A (en) * | 2021-12-06 | 2022-02-01 | 四川高福记生物科技有限公司 | Lactobacillus fermentum for preventing and treating hyperuricemia, and composition and application thereof |
| CN114657095A (en) * | 2022-03-22 | 2022-06-24 | 南昌大学 | Lactobacillus fermentum NCU001464 |
| CN115287239A (en) * | 2022-09-14 | 2022-11-04 | 天津科技大学 | Lactobacillus plantarum capable of degrading nucleosides and purines in vitro and reducing uric acid and application thereof |
| CN115927116A (en) * | 2022-12-30 | 2023-04-07 | 广西爱生生命科技有限公司 | Lactobacillus fermentum strain capable of reducing triglyceride, cholesterol and fat and application thereof |
| CN116004455A (en) * | 2022-12-28 | 2023-04-25 | 广西爱生生命科技有限公司 | Lactobacillus fermentum strain A21196 with function of secreting 5-hydroxytryptamine and application thereof |
| CN116144541A (en) * | 2022-12-30 | 2023-05-23 | 广西爱生生命科技有限公司 | Lactobacillus plantarum strain with triglyceride reducing, cholesterol reducing and lipid reducing functions and application thereof |
| CN116445321A (en) * | 2022-12-13 | 2023-07-18 | 广西爱生生命科技有限公司 | Lactobacillus reuteri A21160 capable of lowering nucleoside and blood uric acid and application thereof |
| CN116925946A (en) * | 2022-04-08 | 2023-10-24 | 江苏省农业科学院 | Lactobacillus fermentum FM-LF-SR6 with high adhesion performance and uric acid reducing function and application thereof |
| CN116925951A (en) * | 2023-02-21 | 2023-10-24 | 熊涛 | Lactobacillus fermentum NCU326 with uric acid reducing effect and application thereof |
-
2022
- 2022-12-13 CN CN202211600780.4A patent/CN116024130B/en active Active
Patent Citations (19)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2008005834A (en) * | 2006-05-31 | 2008-01-17 | Meiji Milk Prod Co Ltd | Lactobacillus having blood uric acid level-reducing activity |
| JP2013048636A (en) * | 2006-05-31 | 2013-03-14 | Meiji Co Ltd | Lactobacillus having blood uric acid level-reducing activity |
| CA2851018A1 (en) * | 2007-11-29 | 2009-06-04 | Hiroshi Tsuboi | Lactic acid bacteria having action of lowering blood uric acid level |
| CN101932697A (en) * | 2007-11-29 | 2010-12-29 | 明治乳业株式会社 | Lactic acid bacteria having action of lowering blood uric acid level |
| CN102747004A (en) * | 2007-11-29 | 2012-10-24 | 株式会社明治 | Lactic acid bacteria with effect of uric acid level reducing in blood |
| CN107208029A (en) * | 2014-11-18 | 2017-09-26 | 株式会社明治 | Lactic acid bacteria of ability and application thereof is absorbed with purine |
| WO2019157585A1 (en) * | 2018-02-13 | 2019-08-22 | Proviva Pharma Inc. | Probiotic formulations for the treatment and alleviation of metabolic and oxidative stress, inflammation and neurodegeneration |
| CN110079476A (en) * | 2019-04-24 | 2019-08-02 | 杭州娃哈哈科技有限公司 | One plant of lactobacillus fermenti that can reduce blood uric acid |
| CN110684685A (en) * | 2019-10-14 | 2020-01-14 | 广西大学 | Lactobacillus fermentum 9-4 and application thereof |
| CN113215012A (en) * | 2020-01-21 | 2021-08-06 | 宁波倍益嘉生物科技有限公司 | Probiotic strain for reducing blood uric acid, composition and application thereof |
| CN113999805A (en) * | 2021-12-06 | 2022-02-01 | 四川高福记生物科技有限公司 | Lactobacillus fermentum for preventing and treating hyperuricemia, and composition and application thereof |
| CN114657095A (en) * | 2022-03-22 | 2022-06-24 | 南昌大学 | Lactobacillus fermentum NCU001464 |
| CN116925946A (en) * | 2022-04-08 | 2023-10-24 | 江苏省农业科学院 | Lactobacillus fermentum FM-LF-SR6 with high adhesion performance and uric acid reducing function and application thereof |
| CN115287239A (en) * | 2022-09-14 | 2022-11-04 | 天津科技大学 | Lactobacillus plantarum capable of degrading nucleosides and purines in vitro and reducing uric acid and application thereof |
| CN116445321A (en) * | 2022-12-13 | 2023-07-18 | 广西爱生生命科技有限公司 | Lactobacillus reuteri A21160 capable of lowering nucleoside and blood uric acid and application thereof |
| CN116004455A (en) * | 2022-12-28 | 2023-04-25 | 广西爱生生命科技有限公司 | Lactobacillus fermentum strain A21196 with function of secreting 5-hydroxytryptamine and application thereof |
| CN115927116A (en) * | 2022-12-30 | 2023-04-07 | 广西爱生生命科技有限公司 | Lactobacillus fermentum strain capable of reducing triglyceride, cholesterol and fat and application thereof |
| CN116144541A (en) * | 2022-12-30 | 2023-05-23 | 广西爱生生命科技有限公司 | Lactobacillus plantarum strain with triglyceride reducing, cholesterol reducing and lipid reducing functions and application thereof |
| CN116925951A (en) * | 2023-02-21 | 2023-10-24 | 熊涛 | Lactobacillus fermentum NCU326 with uric acid reducing effect and application thereof |
Non-Patent Citations (6)
| Title |
|---|
| 上海人民出版社.《辞海 生物分册》.上海人民出版社,1975,第79页. * |
| 乳酸菌的抗氧化活性及其在水产品加工中的应用;王悦齐;吴燕燕;李来好;王锡昌;蔡秋杏;杨贤庆;;食品工业科技(08);全文 * |
| 益生菌抗衰老作用机制研究进展;余洁;刘昕;张驰;赵吉春;李富华;明建;;生物技术通报(01);全文 * |
| 豆酱微生物宏蛋白质组提取及分析;乌日娜;薛亚婷;张平;唐筱扬;陶冬冰;岳媛媛;张鹏飞;陈卫;;食品科学(14);全文 * |
| 降尿酸乳酸菌的筛选及其对高尿酸血症小鼠的影响;张沙沙 等;《生物技术》;全文 * |
| 降血尿酸乳酸菌筛选及其对高尿酸血症模型大鼠作用研究;杨殿斌 等;《中国微生态学杂志》;全文 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN116024130A (en) | 2023-04-28 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN116024130B (en) | Lactobacillus fermentum A21215 for reducing blood uric acid and application thereof | |
| CN116445321B (en) | Lactobacillus reuteri A21160 capable of lowering nucleoside and blood uric acid and application thereof | |
| CN111607538A (en) | Lactobacillus rhamnosus and application thereof in inhibiting helicobacter pylori | |
| CN110218681B (en) | Lactobacillus fermentum KP101 and application thereof | |
| CN110106119B (en) | Lactobacillus rhamnosus M9 separated from breast milk and application thereof | |
| CN114990011B (en) | Lactobacillus reuteri capable of reducing cholesterol and inhibiting gardnerella and application | |
| CN109749957B (en) | Preparation and application of lactobacillus gasseri preparation with aquatic pathogenic bacteria antagonistic property | |
| CN112625979B (en) | Lactobacillus casei for resisting helicobacter pylori and application thereof | |
| CN114634901A (en) | Lactobacillus casei LC16 for promoting bone health and culture method and application thereof | |
| CN114181864A (en) | Lactobacillus rhamnosus HF01 and application thereof | |
| CN116200290A (en) | Lactobacillus paracasei capable of inhibiting proliferation of colorectal cancer cells and application thereof | |
| CN114214230B (en) | Lactobacillus North with helicobacter pylori copolymerization capability and application thereof | |
| CN113797232B (en) | Composition with insulin resistance relieving function and application thereof | |
| CN116970539B (en) | Lactobacillus murine complex, composition and application thereof | |
| CN116445356B (en) | Bifidobacterium animalis subspecies BA67 for regulating intestinal flora and enhancing immunity and application thereof | |
| CN116083325B (en) | Lactobacillus rhamnosus for improving helicobacter pylori related gastrointestinal diseases and application thereof | |
| CN115895966B (en) | Bifidobacterium bifidum BL002 for assisting in relieving gout and application thereof | |
| CN113913334B (en) | Enterococcus faecalis EF-ZA1107-06 and application thereof | |
| CN110897166B (en) | Edible composition containing probiotics and casein phosphopeptide with digestion promoting effect | |
| CN108330082B (en) | Lactobacillus paracasei and application thereof | |
| CN117025456B (en) | Lactobacillus paracasei A21151 with blood uric acid reducing and anti-inflammatory effects and application thereof | |
| CN114231449B (en) | Lactobacillus acidophilus with helicobacter pylori copolymerization capability and application thereof | |
| CN116019842B (en) | Antibacterial new application of lactobacillus acidophilus LA85 and application of lactobacillus acidophilus LA85 in preparation of medicines for relieving EIEC diarrhea | |
| WO2024056081A1 (en) | Bifidobacterium breve b51 and b280, product thereof, and use thereof | |
| CN114854628A (en) | Lactobacillus fermentum TY-S07 and application thereof |
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 |