CN112375696A - Donkey milk source pediococcus pentosaceus and application thereof - Google Patents
Donkey milk source pediococcus pentosaceus and application thereof Download PDFInfo
- Publication number
- CN112375696A CN112375696A CN202010593449.9A CN202010593449A CN112375696A CN 112375696 A CN112375696 A CN 112375696A CN 202010593449 A CN202010593449 A CN 202010593449A CN 112375696 A CN112375696 A CN 112375696A
- Authority
- CN
- China
- Prior art keywords
- pediococcus pentosaceus
- hmtn
- strain
- milk
- fermented milk
- 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.)
- Granted
Links
- 241000191996 Pediococcus pentosaceus Species 0.000 title claims abstract description 102
- 235000020250 donkey milk Nutrition 0.000 title claims abstract description 11
- 230000000968 intestinal effect Effects 0.000 claims abstract description 20
- 235000013618 yogurt Nutrition 0.000 claims abstract description 20
- 238000002360 preparation method Methods 0.000 claims abstract description 13
- 238000000855 fermentation Methods 0.000 claims abstract description 12
- 230000004151 fermentation Effects 0.000 claims abstract description 12
- 235000013305 food Nutrition 0.000 claims abstract description 8
- 235000013373 food additive Nutrition 0.000 claims abstract description 3
- 239000002778 food additive Substances 0.000 claims abstract description 3
- 235000015140 cultured milk Nutrition 0.000 claims description 48
- 241000894006 Bacteria Species 0.000 claims description 43
- 239000007788 liquid Substances 0.000 claims description 24
- 241000588724 Escherichia coli Species 0.000 claims description 21
- 239000001963 growth medium Substances 0.000 claims description 19
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 18
- 239000000725 suspension Substances 0.000 claims description 15
- 235000020183 skimmed milk Nutrition 0.000 claims description 14
- 238000000034 method Methods 0.000 claims description 13
- 239000000203 mixture Substances 0.000 claims description 11
- 238000012258 culturing Methods 0.000 claims description 10
- 230000005764 inhibitory process Effects 0.000 claims description 10
- 239000006228 supernatant Substances 0.000 claims description 10
- 241000191967 Staphylococcus aureus Species 0.000 claims description 8
- 239000000047 product Substances 0.000 claims description 8
- 239000007858 starting material Substances 0.000 claims description 8
- 241000186779 Listeria monocytogenes Species 0.000 claims description 7
- 241001354013 Salmonella enterica subsp. enterica serovar Enteritidis Species 0.000 claims description 7
- 241000293869 Salmonella enterica subsp. enterica serovar Typhimurium Species 0.000 claims description 7
- 235000010855 food raising agent Nutrition 0.000 claims description 7
- 230000003213 activating effect Effects 0.000 claims description 6
- 239000003814 drug Substances 0.000 claims description 6
- 238000005406 washing Methods 0.000 claims description 6
- 230000000688 enterotoxigenic effect Effects 0.000 claims description 4
- 235000021001 fermented dairy product Nutrition 0.000 claims description 4
- 230000036541 health Effects 0.000 claims description 4
- 230000001717 pathogenic effect Effects 0.000 claims description 4
- 238000003756 stirring Methods 0.000 claims description 4
- 230000000369 enteropathogenic effect Effects 0.000 claims description 3
- 239000002054 inoculum Substances 0.000 claims description 3
- 238000009630 liquid culture Methods 0.000 claims description 3
- 238000005057 refrigeration Methods 0.000 claims description 3
- 230000001105 regulatory effect Effects 0.000 claims description 3
- 235000013361 beverage Nutrition 0.000 claims description 2
- 235000013351 cheese Nutrition 0.000 claims description 2
- 235000021552 granulated sugar Nutrition 0.000 claims description 2
- 230000005070 ripening Effects 0.000 claims description 2
- 235000014048 cultured milk product Nutrition 0.000 claims 1
- 230000008021 deposition Effects 0.000 claims 1
- 239000002253 acid Substances 0.000 abstract description 29
- 210000001035 gastrointestinal tract Anatomy 0.000 abstract description 20
- 238000004519 manufacturing process Methods 0.000 abstract description 20
- 244000052616 bacterial pathogen Species 0.000 abstract description 18
- 230000009286 beneficial effect Effects 0.000 abstract description 10
- 238000004220 aggregation Methods 0.000 abstract description 9
- 239000000796 flavoring agent Substances 0.000 abstract description 8
- 235000019634 flavors Nutrition 0.000 abstract description 8
- 238000004321 preservation Methods 0.000 abstract description 7
- 230000000813 microbial effect Effects 0.000 abstract description 4
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 32
- 239000006041 probiotic Substances 0.000 description 21
- 235000018291 probiotics Nutrition 0.000 description 21
- 238000012360 testing method Methods 0.000 description 20
- QSJXEFYPDANLFS-UHFFFAOYSA-N Diacetyl Chemical group CC(=O)C(C)=O QSJXEFYPDANLFS-UHFFFAOYSA-N 0.000 description 19
- 239000000243 solution Substances 0.000 description 18
- IKHGUXGNUITLKF-UHFFFAOYSA-N Acetaldehyde Chemical compound CC=O IKHGUXGNUITLKF-UHFFFAOYSA-N 0.000 description 16
- 230000001580 bacterial effect Effects 0.000 description 16
- 239000004310 lactic acid Substances 0.000 description 16
- 235000014655 lactic acid Nutrition 0.000 description 16
- 230000000694 effects Effects 0.000 description 14
- 230000004083 survival effect Effects 0.000 description 14
- 238000011156 evaluation Methods 0.000 description 12
- 238000011081 inoculation Methods 0.000 description 12
- 229910052740 iodine Inorganic materials 0.000 description 12
- 239000011630 iodine Substances 0.000 description 12
- 230000000529 probiotic effect Effects 0.000 description 12
- ZCYVEMRRCGMTRW-UHFFFAOYSA-N 7553-56-2 Chemical compound [I] ZCYVEMRRCGMTRW-UHFFFAOYSA-N 0.000 description 11
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 11
- 241000194020 Streptococcus thermophilus Species 0.000 description 11
- 239000003833 bile salt Substances 0.000 description 11
- 210000004027 cell Anatomy 0.000 description 11
- 238000005259 measurement Methods 0.000 description 10
- 238000002156 mixing Methods 0.000 description 10
- 239000000523 sample Substances 0.000 description 10
- 230000001953 sensory effect Effects 0.000 description 10
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical group O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 10
- 239000003242 anti bacterial agent Substances 0.000 description 9
- 229940088710 antibiotic agent Drugs 0.000 description 9
- 238000010790 dilution Methods 0.000 description 9
- 239000012895 dilution Substances 0.000 description 9
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 8
- 230000000844 anti-bacterial effect Effects 0.000 description 8
- 238000003860 storage Methods 0.000 description 8
- 238000006243 chemical reaction Methods 0.000 description 7
- 238000002474 experimental method Methods 0.000 description 7
- 238000012216 screening Methods 0.000 description 7
- 239000012086 standard solution Substances 0.000 description 7
- ULGZDMOVFRHVEP-RWJQBGPGSA-N Erythromycin 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)(O)[C@H](O[C@H]2[C@@H]([C@H](C[C@@H](C)O2)N(C)C)O)[C@H]1C)(C)O)CC)[C@H]1C[C@@](C)(OC)[C@@H](O)[C@H](C)O1 ULGZDMOVFRHVEP-RWJQBGPGSA-N 0.000 description 6
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 6
- 241000186660 Lactobacillus Species 0.000 description 6
- MYSWGUAQZAJSOK-UHFFFAOYSA-N ciprofloxacin Chemical compound C12=CC(N3CCNCC3)=C(F)C=C2C(=O)C(C(=O)O)=CN1C1CC1 MYSWGUAQZAJSOK-UHFFFAOYSA-N 0.000 description 6
- 239000012530 fluid Substances 0.000 description 6
- 201000010538 Lactose Intolerance Diseases 0.000 description 5
- 239000011248 coating agent Substances 0.000 description 5
- 238000000576 coating method Methods 0.000 description 5
- 229940079593 drug Drugs 0.000 description 5
- 230000015784 hyperosmotic salinity response Effects 0.000 description 5
- 229940039696 lactobacillus Drugs 0.000 description 5
- 235000013336 milk Nutrition 0.000 description 5
- 210000004080 milk Anatomy 0.000 description 5
- 239000008267 milk Substances 0.000 description 5
- 108090000623 proteins and genes Proteins 0.000 description 5
- 108020004465 16S ribosomal RNA Proteins 0.000 description 4
- 206010059866 Drug resistance Diseases 0.000 description 4
- 241001052560 Thallis Species 0.000 description 4
- 230000008859 change Effects 0.000 description 4
- 238000001816 cooling Methods 0.000 description 4
- 230000002496 gastric effect Effects 0.000 description 4
- 230000036449 good health Effects 0.000 description 4
- 238000005070 sampling Methods 0.000 description 4
- 230000035945 sensitivity Effects 0.000 description 4
- FFTVPQUHLQBXQZ-KVUCHLLUSA-N (4s,4as,5ar,12ar)-4,7-bis(dimethylamino)-1,10,11,12a-tetrahydroxy-3,12-dioxo-4a,5,5a,6-tetrahydro-4h-tetracene-2-carboxamide Chemical compound C1C2=C(N(C)C)C=CC(O)=C2C(O)=C2[C@@H]1C[C@H]1[C@H](N(C)C)C(=O)C(C(N)=O)=C(O)[C@@]1(O)C2=O FFTVPQUHLQBXQZ-KVUCHLLUSA-N 0.000 description 3
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- 206010004016 Bacterial diarrhoea Diseases 0.000 description 3
- 102000016938 Catalase Human genes 0.000 description 3
- 108010053835 Catalase Proteins 0.000 description 3
- 206010012735 Diarrhoea Diseases 0.000 description 3
- 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 3
- 229930182566 Gentamicin Natural products 0.000 description 3
- 229930182555 Penicillin Natural products 0.000 description 3
- 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 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 241000194017 Streptococcus Species 0.000 description 3
- 239000004098 Tetracycline Substances 0.000 description 3
- 229960003022 amoxicillin Drugs 0.000 description 3
- 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 3
- 238000003556 assay Methods 0.000 description 3
- 230000003385 bacteriostatic effect Effects 0.000 description 3
- SRBFZHDQGSBBOR-UHFFFAOYSA-N beta-D-Pyranose-Lyxose Natural products OC1COC(O)C(O)C1O SRBFZHDQGSBBOR-UHFFFAOYSA-N 0.000 description 3
- 230000003115 biocidal effect Effects 0.000 description 3
- 229960004261 cefotaxime Drugs 0.000 description 3
- AZZMGZXNTDTSME-JUZDKLSSSA-M cefotaxime sodium Chemical compound [Na+].N([C@@H]1C(N2C(=C(COC(C)=O)CS[C@@H]21)C([O-])=O)=O)C(=O)\C(=N/OC)C1=CSC(N)=N1 AZZMGZXNTDTSME-JUZDKLSSSA-M 0.000 description 3
- 238000005119 centrifugation Methods 0.000 description 3
- 229960005091 chloramphenicol Drugs 0.000 description 3
- WIIZWVCIJKGZOK-RKDXNWHRSA-N chloramphenicol Chemical compound ClC(Cl)C(=O)N[C@H](CO)[C@H](O)C1=CC=C([N+]([O-])=O)C=C1 WIIZWVCIJKGZOK-RKDXNWHRSA-N 0.000 description 3
- 229960003405 ciprofloxacin Drugs 0.000 description 3
- 229960002227 clindamycin Drugs 0.000 description 3
- KDLRVYVGXIQJDK-AWPVFWJPSA-N clindamycin Chemical compound CN1C[C@H](CCC)C[C@H]1C(=O)N[C@H]([C@H](C)Cl)[C@@H]1[C@H](O)[C@H](O)[C@@H](O)[C@@H](SC)O1 KDLRVYVGXIQJDK-AWPVFWJPSA-N 0.000 description 3
- 229960003326 cloxacillin Drugs 0.000 description 3
- LQOLIRLGBULYKD-JKIFEVAISA-N cloxacillin Chemical compound N([C@@H]1C(N2[C@H](C(C)(C)S[C@@H]21)C(O)=O)=O)C(=O)C1=C(C)ON=C1C1=CC=CC=C1Cl LQOLIRLGBULYKD-JKIFEVAISA-N 0.000 description 3
- 230000000875 corresponding effect Effects 0.000 description 3
- 235000013365 dairy product Nutrition 0.000 description 3
- 238000007865 diluting Methods 0.000 description 3
- 229960003276 erythromycin Drugs 0.000 description 3
- 210000004051 gastric juice Anatomy 0.000 description 3
- 229960002518 gentamicin Drugs 0.000 description 3
- 239000011521 glass Substances 0.000 description 3
- 230000036039 immunity Effects 0.000 description 3
- 230000002401 inhibitory effect Effects 0.000 description 3
- 229960000318 kanamycin Drugs 0.000 description 3
- 229930027917 kanamycin Natural products 0.000 description 3
- SBUJHOSQTJFQJX-NOAMYHISSA-N kanamycin Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CN)O[C@@H]1O[C@H]1[C@H](O)[C@@H](O[C@@H]2[C@@H]([C@@H](N)[C@H](O)[C@@H](CO)O2)O)[C@H](N)C[C@@H]1N SBUJHOSQTJFQJX-NOAMYHISSA-N 0.000 description 3
- 229930182823 kanamycin A Natural products 0.000 description 3
- 229960004023 minocycline Drugs 0.000 description 3
- 230000000877 morphologic effect Effects 0.000 description 3
- 229960001180 norfloxacin Drugs 0.000 description 3
- OGJPXUAPXNRGGI-UHFFFAOYSA-N norfloxacin Chemical compound C1=C2N(CC)C=C(C(O)=O)C(=O)C2=CC(F)=C1N1CCNCC1 OGJPXUAPXNRGGI-UHFFFAOYSA-N 0.000 description 3
- 239000003960 organic solvent Substances 0.000 description 3
- 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 3
- 229940049954 penicillin Drugs 0.000 description 3
- 239000002504 physiological saline solution Substances 0.000 description 3
- NLKNQRATVPKPDG-UHFFFAOYSA-M potassium iodide Chemical compound [K+].[I-] NLKNQRATVPKPDG-UHFFFAOYSA-M 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 229960001225 rifampicin Drugs 0.000 description 3
- JQXXHWHPUNPDRT-WLSIYKJHSA-N rifampicin Chemical compound O([C@](C1=O)(C)O/C=C/[C@@H]([C@H]([C@@H](OC(C)=O)[C@H](C)[C@H](O)[C@H](C)[C@@H](O)[C@@H](C)\C=C\C=C(C)/C(=O)NC=2C(O)=C3C([O-])=C4C)C)OC)C4=C1C3=C(O)C=2\C=N\N1CC[NH+](C)CC1 JQXXHWHPUNPDRT-WLSIYKJHSA-N 0.000 description 3
- 230000001954 sterilising effect Effects 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- -1 teicholam Chemical compound 0.000 description 3
- 229960002180 tetracycline Drugs 0.000 description 3
- 229930101283 tetracycline Natural products 0.000 description 3
- 235000019364 tetracycline Nutrition 0.000 description 3
- 150000003522 tetracyclines Chemical class 0.000 description 3
- 238000003260 vortexing Methods 0.000 description 3
- GEYOCULIXLDCMW-UHFFFAOYSA-N 1,2-phenylenediamine Chemical compound NC1=CC=CC=C1N GEYOCULIXLDCMW-UHFFFAOYSA-N 0.000 description 2
- OLBNOBQOQZRLMP-UHFFFAOYSA-N 2,6-dimethoxy-p-benzoquinone Chemical compound COC1=CC(=O)C=C(OC)C1=O OLBNOBQOQZRLMP-UHFFFAOYSA-N 0.000 description 2
- 241000283690 Bos taurus 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
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- 241000192001 Pediococcus Species 0.000 description 2
- 229920002472 Starch Polymers 0.000 description 2
- 230000004913 activation Effects 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 229940093761 bile salts Drugs 0.000 description 2
- 238000004364 calculation method Methods 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 230000003247 decreasing effect Effects 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 239000012153 distilled water Substances 0.000 description 2
- 238000001914 filtration Methods 0.000 description 2
- 239000003205 fragrance Substances 0.000 description 2
- 239000008103 glucose Substances 0.000 description 2
- 238000000265 homogenisation Methods 0.000 description 2
- 238000000338 in vitro Methods 0.000 description 2
- 210000002490 intestinal epithelial cell Anatomy 0.000 description 2
- 239000008101 lactose Substances 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 239000008107 starch Substances 0.000 description 2
- 235000019698 starch Nutrition 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
- 238000004448 titration Methods 0.000 description 2
- YNJBWRMUSHSURL-UHFFFAOYSA-N trichloroacetic acid Chemical compound OC(=O)C(Cl)(Cl)Cl YNJBWRMUSHSURL-UHFFFAOYSA-N 0.000 description 2
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- GUBGYTABKSRVRQ-XLOQQCSPSA-N Alpha-Lactose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@H](O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-XLOQQCSPSA-N 0.000 description 1
- 241000193830 Bacillus <bacterium> Species 0.000 description 1
- 108010062877 Bacteriocins Proteins 0.000 description 1
- 241000186000 Bifidobacterium Species 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- GUBGYTABKSRVRQ-CUHNMECISA-N D-Cellobiose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-CUHNMECISA-N 0.000 description 1
- FBPFZTCFMRRESA-FSIIMWSLSA-N D-Glucitol Natural products OC[C@H](O)[C@H](O)[C@@H](O)[C@H](O)CO FBPFZTCFMRRESA-FSIIMWSLSA-N 0.000 description 1
- FBPFZTCFMRRESA-JGWLITMVSA-N D-glucitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-JGWLITMVSA-N 0.000 description 1
- SHZGCJCMOBCMKK-UHFFFAOYSA-N D-mannomethylose Natural products CC1OC(O)C(O)C(O)C1O SHZGCJCMOBCMKK-UHFFFAOYSA-N 0.000 description 1
- WQZGKKKJIJFFOK-QTVWNMPRSA-N D-mannopyranose Chemical compound OC[C@H]1OC(O)[C@@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-QTVWNMPRSA-N 0.000 description 1
- SRBFZHDQGSBBOR-IOVATXLUSA-N D-xylopyranose Chemical compound O[C@@H]1COC(O)[C@H](O)[C@H]1O SRBFZHDQGSBBOR-IOVATXLUSA-N 0.000 description 1
- 230000004544 DNA amplification Effects 0.000 description 1
- 241000252212 Danio rerio Species 0.000 description 1
- 229930091371 Fructose Natural products 0.000 description 1
- 239000005715 Fructose Substances 0.000 description 1
- RFSUNEUAIZKAJO-ARQDHWQXSA-N Fructose Chemical compound OC[C@H]1O[C@](O)(CO)[C@@H](O)[C@@H]1O RFSUNEUAIZKAJO-ARQDHWQXSA-N 0.000 description 1
- 230000005526 G1 to G0 transition Effects 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- 238000003794 Gram staining Methods 0.000 description 1
- 241000282412 Homo Species 0.000 description 1
- 238000002768 Kirby-Bauer method Methods 0.000 description 1
- XUJNEKJLAYXESH-REOHCLBHSA-N L-Cysteine Chemical compound SC[C@H](N)C(O)=O XUJNEKJLAYXESH-REOHCLBHSA-N 0.000 description 1
- PNNNRSAQSRJVSB-UHFFFAOYSA-N L-rhamnose Natural products CC(O)C(O)C(O)C(O)C=O PNNNRSAQSRJVSB-UHFFFAOYSA-N 0.000 description 1
- 235000013960 Lactobacillus bulgaricus Nutrition 0.000 description 1
- 241000186672 Lactobacillus delbrueckii subsp. bulgaricus Species 0.000 description 1
- 241000194036 Lactococcus Species 0.000 description 1
- 241000192132 Leuconostoc Species 0.000 description 1
- 229910002651 NO3 Inorganic materials 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- NHNBFGGVMKEFGY-UHFFFAOYSA-N Nitrate Chemical compound [O-][N+]([O-])=O NHNBFGGVMKEFGY-UHFFFAOYSA-N 0.000 description 1
- 102000057297 Pepsin A Human genes 0.000 description 1
- 108090000284 Pepsin A Proteins 0.000 description 1
- 108010040201 Polymyxins Proteins 0.000 description 1
- 241000186429 Propionibacterium Species 0.000 description 1
- MUPFEKGTMRGPLJ-RMMQSMQOSA-N Raffinose Natural products O(C[C@H]1[C@@H](O)[C@H](O)[C@@H](O)[C@@H](O[C@@]2(CO)[C@H](O)[C@@H](O)[C@@H](CO)O2)O1)[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 MUPFEKGTMRGPLJ-RMMQSMQOSA-N 0.000 description 1
- 238000012300 Sequence Analysis Methods 0.000 description 1
- 241000191940 Staphylococcus Species 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
- 241000209140 Triticum Species 0.000 description 1
- 235000021307 Triticum Nutrition 0.000 description 1
- 102000004142 Trypsin Human genes 0.000 description 1
- 108090000631 Trypsin Proteins 0.000 description 1
- MUPFEKGTMRGPLJ-UHFFFAOYSA-N UNPD196149 Natural products OC1C(O)C(CO)OC1(CO)OC1C(O)C(O)C(O)C(COC2C(C(O)C(O)C(CO)O2)O)O1 MUPFEKGTMRGPLJ-UHFFFAOYSA-N 0.000 description 1
- 108010059993 Vancomycin Proteins 0.000 description 1
- 241000607265 Vibrio vulnificus Species 0.000 description 1
- TVXBFESIOXBWNM-UHFFFAOYSA-N Xylitol Natural products OCCC(O)C(O)C(O)CCO TVXBFESIOXBWNM-UHFFFAOYSA-N 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 150000007513 acids Chemical class 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 238000000246 agarose gel electrophoresis Methods 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- PNNNRSAQSRJVSB-BXKVDMCESA-N aldehydo-L-rhamnose Chemical compound C[C@H](O)[C@H](O)[C@@H](O)[C@@H](O)C=O PNNNRSAQSRJVSB-BXKVDMCESA-N 0.000 description 1
- PYMYPHUHKUWMLA-WISUUJSJSA-N aldehydo-L-xylose Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)C=O PYMYPHUHKUWMLA-WISUUJSJSA-N 0.000 description 1
- 229930195726 aldehydo-L-xylose Natural products 0.000 description 1
- WQZGKKKJIJFFOK-PHYPRBDBSA-N alpha-D-galactose Chemical compound OC[C@H]1O[C@H](O)[C@H](O)[C@@H](O)[C@H]1O WQZGKKKJIJFFOK-PHYPRBDBSA-N 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 238000000137 annealing Methods 0.000 description 1
- 230000003042 antagnostic effect Effects 0.000 description 1
- PYMYPHUHKUWMLA-WDCZJNDASA-N arabinose Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)C=O PYMYPHUHKUWMLA-WDCZJNDASA-N 0.000 description 1
- PYMYPHUHKUWMLA-UHFFFAOYSA-N arabinose Natural products OCC(O)C(O)C(O)C=O PYMYPHUHKUWMLA-UHFFFAOYSA-N 0.000 description 1
- 239000011324 bead Substances 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 239000002775 capsule Substances 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 230000021164 cell adhesion Effects 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 238000005345 coagulation Methods 0.000 description 1
- 239000003086 colorant Substances 0.000 description 1
- 230000002596 correlated effect Effects 0.000 description 1
- 238000010227 cup method (microbiological evaluation) Methods 0.000 description 1
- 238000000354 decomposition reaction Methods 0.000 description 1
- 238000004925 denaturation Methods 0.000 description 1
- 230000036425 denaturation Effects 0.000 description 1
- 238000009792 diffusion process Methods 0.000 description 1
- 230000001079 digestive effect Effects 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 238000011049 filling Methods 0.000 description 1
- 210000003495 flagella Anatomy 0.000 description 1
- 238000009920 food preservation Methods 0.000 description 1
- 235000011389 fruit/vegetable juice Nutrition 0.000 description 1
- 229930182830 galactose Natural products 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 230000002008 hemorrhagic effect Effects 0.000 description 1
- 230000007366 host health Effects 0.000 description 1
- 230000002209 hydrophobic effect Effects 0.000 description 1
- 238000007654 immersion Methods 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 229940004208 lactobacillus bulgaricus Drugs 0.000 description 1
- 244000144972 livestock Species 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000035800 maturation Effects 0.000 description 1
- HEBKCHPVOIAQTA-UHFFFAOYSA-N meso ribitol Natural products OCC(O)C(O)C(O)CO HEBKCHPVOIAQTA-UHFFFAOYSA-N 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 238000001471 micro-filtration Methods 0.000 description 1
- 230000004899 motility Effects 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 229960001019 oxacillin Drugs 0.000 description 1
- UWYHMGVUTGAWSP-JKIFEVAISA-N oxacillin Chemical compound N([C@@H]1C(N2[C@H](C(C)(C)S[C@@H]21)C(O)=O)=O)C(=O)C1=C(C)ON=C1C1=CC=CC=C1 UWYHMGVUTGAWSP-JKIFEVAISA-N 0.000 description 1
- 230000020477 pH reduction Effects 0.000 description 1
- 238000009928 pasteurization Methods 0.000 description 1
- 229940111202 pepsin Drugs 0.000 description 1
- 230000035479 physiological effects, processes and functions Effects 0.000 description 1
- 238000006116 polymerization reaction Methods 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 238000012257 pre-denaturation Methods 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- MUPFEKGTMRGPLJ-ZQSKZDJDSA-N raffinose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO[C@@H]2[C@@H]([C@@H](O)[C@@H](O)[C@@H](CO)O2)O)O1 MUPFEKGTMRGPLJ-ZQSKZDJDSA-N 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 235000020122 reconstituted milk Nutrition 0.000 description 1
- 230000002040 relaxant effect Effects 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 238000004088 simulation Methods 0.000 description 1
- UIIMBOGNXHQVGW-UHFFFAOYSA-M sodium bicarbonate Substances [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 1
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 1
- 239000000600 sorbitol Substances 0.000 description 1
- 239000008223 sterile water Substances 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 229960005322 streptomycin Drugs 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 208000011580 syndromic disease Diseases 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- 239000012588 trypsin Substances 0.000 description 1
- 229960003165 vancomycin Drugs 0.000 description 1
- MYPYJXKWCTUITO-UHFFFAOYSA-N vancomycin Natural products O1C(C(=C2)Cl)=CC=C2C(O)C(C(NC(C2=CC(O)=CC(O)=C2C=2C(O)=CC=C3C=2)C(O)=O)=O)NC(=O)C3NC(=O)C2NC(=O)C(CC(N)=O)NC(=O)C(NC(=O)C(CC(C)C)NC)C(O)C(C=C3Cl)=CC=C3OC3=CC2=CC1=C3OC1OC(CO)C(O)C(O)C1OC1CC(C)(N)C(O)C(C)O1 MYPYJXKWCTUITO-UHFFFAOYSA-N 0.000 description 1
- MYPYJXKWCTUITO-LYRMYLQWSA-O vancomycin(1+) Chemical compound O([C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1=C2C=C3C=C1OC1=CC=C(C=C1Cl)[C@@H](O)[C@H](C(N[C@@H](CC(N)=O)C(=O)N[C@H]3C(=O)N[C@H]1C(=O)N[C@H](C(N[C@@H](C3=CC(O)=CC(O)=C3C=3C(O)=CC=C1C=3)C([O-])=O)=O)[C@H](O)C1=CC=C(C(=C1)Cl)O2)=O)NC(=O)[C@@H](CC(C)C)[NH2+]C)[C@H]1C[C@](C)([NH3+])[C@H](O)[C@H](C)O1 MYPYJXKWCTUITO-LYRMYLQWSA-O 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
- 239000000811 xylitol Substances 0.000 description 1
- HEBKCHPVOIAQTA-SCDXWVJYSA-N xylitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)CO HEBKCHPVOIAQTA-SCDXWVJYSA-N 0.000 description 1
- 229960002675 xylitol Drugs 0.000 description 1
- 235000010447 xylitol Nutrition 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
-
- 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
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
- A23C9/00—Milk preparations; Milk powder or milk powder preparations
- A23C9/12—Fermented milk preparations; Treatment using microorganisms or enzymes
- A23C9/123—Fermented milk preparations; Treatment using microorganisms or enzymes using only microorganisms of the genus lactobacteriaceae; Yoghurt
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- 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
-
- 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
-
- 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
-
- 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)
- General Health & Medical Sciences (AREA)
- Mycology (AREA)
- Genetics & Genomics (AREA)
- Medicinal Chemistry (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Wood Science & Technology (AREA)
- Biochemistry (AREA)
- Food Science & Technology (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- General Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Polymers & Plastics (AREA)
- Animal Behavior & Ethology (AREA)
- Pharmacology & Pharmacy (AREA)
- Virology (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Oncology (AREA)
- Communicable Diseases (AREA)
- Tropical Medicine & Parasitology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Epidemiology (AREA)
- Nutrition Science (AREA)
- Dairy Products (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention belongs to the technical field of microbial fermentation, and particularly relates to a pediococcus pentosaceus strain derived from donkey milk and application thereof. The Pediococcus pentosaceus (Pediococcus pentosaceus) HMTN.01 is preserved in China center for type culture Collection No. 5/25 in 2020, and the preservation number is CCTCC NO: m2020144. The strain has strong acid production, viscosity production and aroma production capabilities, has good hydrophobicity, self-aggregation capability and gastrointestinal tract tolerance capability, can produce good adhesion to the gastrointestinal tract, is beneficial to field planting in the gastrointestinal tract, can inhibit intestinal pathogenic bacteria and regulate intestinal flora, can be prepared into food or food additives, and the yogurt prepared by the strain has good flavor, taste and appearance, and can be used for large-scale preparation of yogurt.
Description
Technical Field
The invention belongs to the technical field of microbial fermentation, and particularly relates to donkey milk source pediococcus pentosaceus and application thereof.
Background
The yoghourt is a milk product which is prepared by taking fresh milk or reconstituted milk as a raw material, adding a leavening agent into the milk after pasteurization, fermenting at about 42 ℃, and then cooling and filling. At present, various yoghurts appear in the market, and are popular with consumers due to the unique flavor, comfortable mouthfeel, rich nutrition and good health care function. With the pursuit of the public to healthy life and the transformation of consumption concept, the consumption of various yoghurts is continuously increased, and the yoghurt industry becomes a new growth bright spot in the dairy industry. The quality and flavor of the yogurt depend on the quality, type and activity of the lactic acid bacteria starter. The production performance (acid production, viscosity production, aroma production, lactose decomposition and the like) and the functional characteristics (lactose intolerance relieving, bacteriostasis and the like) of the lactic acid bacteria are closely related to the performance of the fermented dairy product. The livestock resources in Xinjiang area are sufficient, and the milk source is rich. They contain abundant lactic acid bacteria resources, are a natural treasury for developing excellent lactic acid bacteria, and some strains with good passage and fermentation performance can be used as excellent starter strains.
Probiotics are defined as "taking up sufficient amounts of living microorganisms beneficial to the health of the host" (FAO/WHO), and yoghurt is considered to be the most suitable carrier for probiotics and can transport them to the human intestinal tract, exerting their probiotic function. The probiotics has the effects of improving immunity, maintaining the microecological balance of digestive intestinal tracts, resisting tumors and the like, and the yoghourt added with the probiotics is favorable for regulating the microecological balance of the intestinal tracts of a human body and relieving intestinal syndromes such as diarrhea. At present, most of probiotic strains which can be used for food are specified in China, and comprise bifidobacteria, lactobacilli, streptococcus, lactococcus, leuconostoc, propionibacterium, pediococcus, staphylococcus and bacillus, and a plurality of the strains are generally regarded as food-grade safe microorganisms. The newly isolated lactic acid bacteria can be considered as probiotics by utilizing various screening conditions, wherein one important condition is the tolerance capability to acid and bile salt, namely, the strain has to be capable of surviving under the severe environment of low acid, high bile salt and the like of the gastrointestinal tract; lactic acid bacteria can inhibit the growth of common pathogenic bacteria through antibacterial substances such as lactic acid, acetic acid, extracellular polysaccharide, bacteriocin and the like generated by metabolism, and the antibacterial substances are also a main condition for screening probiotics; the adhesion ability of the lactobacillus to intestinal cells is also an important condition for screening probiotics, the adhesion ability of the strain and the cells and the hydrophobicity and the self-aggregation rate of the strain and the cells to an organic solvent are in positive correlation, and the higher the hydrophobicity and the self-aggregation rate is, the better the adhesion ability of the strain is; in addition, part of antibiotic resistance genes carried by the lactic acid bacteria can be transferred to other bacteria and pathogenic bacteria in the intestinal tract of a host, so that the evaluation of the antibiotic resistance of the lactic acid bacteria is also extremely important for screening probiotics. However, due to the time and effort involved in vivo tests, the probiotic activity of lactic acid bacteria is generally studied by in vitro tests.
Most of the yogurt in the current market is prepared by fermenting streptococcus thermophilus, lactobacillus bulgaricus and auxiliary strains, the strains are conventional, the taste is single, and a single strain is difficult to have multiple functional characteristics of acid production, viscosity production, good aroma production, bacteriostatic activity, bacterial diarrhea relief and the like. The probiotic yogurt produced by the traditional bacterial strain has a certain inhibition effect on food-borne pathogenic bacteria, but has a general effect; traditional probiotic strains have poor colonization in the intestinal tract, and the probiotic effect is not fully and effectively exerted and is already discharged out of the body. At present, most of domestic dairy products are foreign patent strains, and probiotics with independent intellectual property rights only account for a small part and are applied less. Therefore, a lactic acid bacteria strain which has good antibacterial activity, strong adhesion capability to intestinal epithelial cells and high safety is to be screened, has good fermentation characteristics of acid production, viscosity production, aroma production and the like, and is combined with a common leavening agent to prepare the probiotic yogurt.
Pediococcus pentosaceus (Pediococcus pentosaceus) belongs to the genus Pediococcus of the family Streptococcus, is a gram-positive bacterium, and belongs to a group of lactic acid bacteria capable of producing lactic acid by fermenting glucose. Pediococcus pentosaceus has been used as a probiotic in the fields of food preservation, animal feed preparation, animal probiotics, and the like. For example, patent CN201510257748.4 discloses a pediococcus pentosaceus capable of producing 2, 6-dimethoxy-p-benzoquinone by fermenting wheat germ, but the pediococcus pentosaceus does not disclose its property of producing yogurt by fermentation; patent CN201811633701.3 discloses a pediococcus pentosaceus strain which has a remarkable effect of relaxing bowel, has a very remarkable inhibitory effect on the growth of pathogenic bacteria staphylococcus aureus, and can remarkably improve the survival rate of zebra fish after being attacked by vibrio vulnificus. However, the patent does not disclose the characteristics of yogurt prepared from pediococcus pentosaceus, and the yogurt prepared can relieve lactose intolerance and prevent and treat diarrhea and the like. Aiming at the technical problems, the pediococcus pentosaceus is extracted and separated from the donkey milk source unexpectedly, the pediococcus pentosaceus has good bacteriostatic effect, good gastrointestinal tract tolerance, strong adhesion capability to intestinal epithelial cells and high safety, the prepared single-strain fermented milk has good texture characteristics (hardness, viscosity, elasticity and the like), unique flavor and high viable bacteria content, can relieve lactose intolerance, prevent and treat diarrhea, regulate intestinal flora, keep intestinal microecological balance, improve the microenvironment of a human body, and has a good health care effect.
Disclosure of Invention
In order to solve the problems, the invention discloses Pediococcus pentosaceus (Pediococcus pentosaceus) HMTN.01 of donkey milk source, which is preserved in China center for type culture Collection No. 5/25 in 2020, with the preservation number of CCTCC NO: m2020144, survival status is survival, preservation address is Wuhan university in Wuhan city of Hubei province, postcode is 430072, and contact telephone is 027-68754833.
The Pediococcus pentosaceus (Pediococcus pentosaceus) HMTN.01 can be used for preparing medicines for inhibiting intestinal pathogenic bacteria.
Preferably, the enteropathogenic bacteria include pathogenic Escherichia coli, enterotoxigenic Escherichia coli, enterohemorrhagic Escherichia coli, serotype Salmonella enteritidis, Salmonella typhimurium, Listeria monocytogenes, and Staphylococcus aureus.
The Pediococcus pentosaceus (Pediococcus pentosaceus) HMTN.01 can be applied to the preparation of health care products for regulating intestinal flora, and the health care products can be prepared into probiotic bacteria agents or capsules.
The Pediococcus pentosaceus (Pediococcus pentosaceus) HMTN.01 can be used for preparing food or food additives.
Preferably, the food product is a fermented dairy product.
Preferably, the fermented dairy product is yogurt, cheese or fermented milk beverage.
The invention also provides a method for preparing yoghourt by fermenting pediococcus pentosaceus HMTN.01, and the process flow for preparing the yoghourt is as follows:
starter strain
↓
Skim milk → Heat treatment → Cooling → inoculation → culture → Cooling → storage
The specific steps of the operation are as follows:
(1) activating strains: inoculating strain HMTN.01 preserved at-80 deg.C into MRS liquid culture medium, anaerobically culturing at 37 deg.C, and continuously inoculating for 2-3 times to complete strain activation.
(2) Seed liquid: and (2) centrifuging the activated bacterium liquid obtained in the step (1), discarding the supernatant, repeatedly washing the bacterium mud twice with sterile normal saline, adding equivalent sterile normal saline for resuspension, and taking the obtained bacterium suspension as the seed liquid for subsequent fermentation.
(3) Working leavening agent: inoculating the seed liquid obtained in the step (2) into sterilized skim milk, culturing until the seed liquid is coagulated, transferring the seed liquid into another tank of sterilized skim milk, repeatedly inoculating for 2-3 times, obtaining a working starter after the seed liquid is coagulated, and placing the working starter into a refrigerator at 4 ℃ for later use.
(4) Preparation of fermented milk: and (4) inoculating the working leavening agent obtained in the step (3) into homogenized and sterilized skim milk containing 3% m/v of edible white granulated sugar, adding the mixture, fully stirring the mixture to uniformly mix the mixture, fermenting the mixture at 42 ℃ to curd, and then transferring the curd to a refrigerator at 4 ℃ for refrigeration and ripening for 24 hours to obtain the fermented milk.
Preferably, in the step (2), the centrifugation is carried out for 5min at 4000r/min at 20 ℃.
Preferably, in the step (3), the inoculation amount of the seed solution is 3% v/v, the inoculation amount transferred to the skim milk is 10% m/v, and the culture condition is anaerobic culture at 37 ℃.
Preferably, in the step (4), the homogenizing mode is primary homogenizing, and the homogenizing pressure is 23 MPa; the sterilization condition is that the constant temperature treatment is carried out for 15min at 95 ℃; the inoculation amount of the working leaven is 5-10% m/v.
The invention has the beneficial effects that:
(1) the pediococcus pentosaceus HMTN.01 provided by the invention is obtained by separating and purifying donkey milk in Hami area of Xinjiang, and is safe and reliable;
(2) the Pediococcus pentosaceus HMTN.01 provided by the invention has strong acid, viscosity and aroma producing capabilities, and the prepared fermented milk has good flavor, taste and appearance;
(3) the pediococcus pentosaceus HMTN.01 also has good hydrophobicity, self-aggregation capability and gastrointestinal tract tolerance capability, can generate good adhesion to the gastrointestinal tract, and is beneficial to field planting in the gastrointestinal tract;
(4) the pediococcus pentosaceus HMTN.01 also has broad-spectrum antibacterial activity, has a good inhibition effect on common food-borne pathogenic bacteria, and is beneficial to maintaining intestinal microecological balance and improving immunity;
(5) the pediococcus pentosaceus HMTN.01 provided by the invention is also sensitive or moderately sensitive to most common antibiotics, and is safe and reliable. Antibiotics include cefotaxime, minocycline, rifampin, kanamycin, chloramphenicol, erythromycin, clindamycin, penicillin, teicholam, cloxacillin, tetracycline, ciprofloxacin, norfloxacin, amoxicillin and gentamicin.
(6) The fermented milk prepared by the pediococcus pentosaceus HMTN.01 provided by the invention can also adjust intestinal flora, improve the intestinal micro-ecological environment, effectively relieve lactose intolerance and bacterial diarrhea, and has a good health care function.
Drawings
FIG. 1 is a morphological diagram of screened Pediococcus pentosaceus HMTN.01 under 100 times oil immersion
FIG. 2 is a graph showing the growth profile of Pediococcus pentosaceus HMTN.01
FIG. 3 shows that 16S rRNA gene sequence of Pediococcus pentosaceus HMTN.01 is subjected to homology comparison analysis, and MEGA v.5.0 software is used for establishing phylogenetic tree
FIG. 4 is a graph showing the effect of Pediococcus pentosaceus HMTN.01 in inhibiting food-borne pathogenic bacteria
FIG. 5 is a drug susceptibility test chart of Pediococcus pentosaceus HMTN.01
FIG. 6 is a picture of fermented milk prepared from Pediococcus pentosaceus HMTN.01
FIG. 7 is a graph showing sensory evaluation of fermented milk prepared from Pediococcus pentosaceus HMTN.01 at different storage times
FIG. 8 is a graph showing the determination of acid-producing ability of Pediococcus pentosaceus HMTN.01
FIG. 9 is a graph showing the post-acidification capacity determination of Pediococcus pentosaceus HMTN.01
FIG. 10 is a graph showing the standard curve of fermented lactondione
FIG. 11 is a graph showing the change of diacetyl and acetaldehyde content in fermented milk
FIG. 12 is a graph showing the change of viable cell count of fermented milk prepared from Pediococcus pentosaceus HMTN.01 under different storage times
Detailed Description
In order to more clearly illustrate the technical scheme of the embodiment of the invention, the invention is explained in detail by specific examples and experimental schemes below.
In the following examples of the invention, the experimental materials and reagent sources used are as follows:
pediococcus pentosaceus (Pediococcus pentosaceus) HMTN.01 is preserved in China center for type culture Collection No. 5/25 in 2020 with the preservation number of CCTCC NO: m2020144.
Pathogenic Escherichia coli (EC1), enterotoxigenic Escherichia coli (EC2), hemorrhagic Escherichia coli (EC3), serotype Salmonella enteritidis (SM1), Salmonella typhimurium (SM2), Listeria monocytogenes (LS1) and Staphylococcus Aureus (SA) are all purchased from China center for preservation and management of industrial microbial strains; streptococcus thermophilus D-2 is isolated from traditional dairy products in Yili area of Xinjiang by food biotechnology laboratory of food institute of Stone river university and is stored in the laboratory.
Example 1 screening and identification of strains
1.1 isolation and purification of the Strain
Firstly, preparing an improved MRS (containing 0.5g/L cysteine) culture medium, sterilizing, pouring a flat plate, solidifying, diluting a donkey milk sample collected from Hami area of Xinjiang by using sterile normal saline, and diluting with a dilution gradient of 10-2、10-3、10-4、10-5、10-6In the case of (2), 100. mu.L of each dilution was pipetted and applied to the solidified plate, the plate was numbered and recorded, and the applied plate was placed in an anaerobic chamber at 37 ℃ and cultured upside down for 36 hours.
Selecting a plate with appropriate dilution for each donkey milk sample, picking single colonies with different colors, sizes and shapes for microscopic examination, reserving the colonies with the cell morphology in a rod shape or a spherical shape, continuously transferring and carrying out streak culture for 3 times, picking pure single colonies on an improved culture medium, carrying out anaerobic inverted culture for 24 hours at 37 ℃, carrying out gram staining and catalase test on part of the culture, enriching gram positive and catalase negative strains, and then preserving to-80 ℃ for later use, thereby realizing the screening and separation of suspected lactobacillus.
56 pure cultures are screened and separated out, and 38 suspected lactic acid bacteria are obtained through preliminary sequencing identification.
And (3) performing probiotic characteristic and fermentation characteristic tests on the 38 suspected lactobacillus strains obtained by screening to screen out a suspected lactobacillus strain with better probiotic characteristic and fermentation characteristic.
1.2 identification of the Strain
The suspected lactobacillus is identified by a method combining morphological characteristics, growth characteristics, physiology and biochemistry and molecular identification.
(1) Morphological characteristics and growth characteristics of the strains:
the strain has spherical thallus, gram positive, no flagellum, no motion and no spore, and the thallus morphology is shown in figure 1. The colony on the MRS solid culture medium is milky white, and has regular and convex edges and smooth surface. The strain grows rapidly, the strain can enter a stationary phase within 14h, and the growth curve is shown in figure 2.
(2) Physiological and biochemical characteristics of the strain:
the physiological and biochemical characteristics of the strain are identified by referring to a 'Dongxu pearl, Chuia Miaoying' common bacteria system identification manual, and the specific identification results are as follows:
motility test, nitrate buffer test, H2The S experiment, the gelatin liquefaction experiment and the catalase experiment are all negative; in the aspect of carbon source utilization, glucose, lactose, galactose, sucrose, fructose, mannose, raffinose and L-rhamnose can be utilized, starch can be hydrolyzed, and arabinose, cellobiose, D-xylose, L-xylose, xylitol and sorbitol cannot be utilized.
(3) 16S rRNA gene sequence analysis of the strains:
the extraction of DNA from the strain was carried out by a phenol-chloroform-glass bead method modified by Matsukit or the like. 16S rRNA gene amplification was performed on the strains using 27F (5'-AGAGTTTGATCCTGGCTCAG-3') and 1492R (5'-TACGGTTACCTTGTTACGACTT-3') PCR universal primers.
The amplification conditions were: 35 cycles (pre-denaturation at 95 ℃ for 5min, denaturation at 94 ℃ for 30s, annealing at 55 ℃ for 40s, and extension at 72 ℃ for 1min), and final extension at 72 ℃ for 10 min. After 3. mu.L of the PCR product was aspirated and detected by 1.5% (w/v) agarose gel electrophoresis, the remaining PCR product was used as a sample to be sequenced by Meiji corporation, Shanghai. The 16S rRNA gene sequences thus determined were submitted to GenBank databases, subjected to sequence homology analysis using BLAST, and phylogenetic trees were established using MEGA v.7.0 software.
As shown in FIG. 3, the phylogenetic tree established revealed that the strain had the closest homology to 16Sr RNA of Pediococcus pentosaceus RKG 1-544 (ID: MT045930.1), Pediococcus pentosaceus RKG 1-479 (ID: MT045910.1), Pediococcus pentosaceus RKG 1-476 (ID: MT045907.1) in GenBank of NCBI, and the strain was identified as Pediococcus pentosaceus (Pediococcus pentosaceus) HMTN.01.
And Pediococcus pentosaceus (Pediococcus pentosaceus) HMTN.01 is preserved in China center for type culture Collection No. 5/25 in 2020 with the preservation number of CCTCC NO: m2020144, survival status is survival, preservation address is Wuhan university in Wuhan city of Hubei province, postcode is 430072, and contact telephone is 027-68754833.
Example 2 study of probiotic Properties of Pediococcus pentosaceus HMTN.01
2.1 study of acid tolerance and bile salt tolerance of Pediococcus pentosaceus HMTN.01
(1) Pediococcus pentosaceus HMTN.01 acid tolerance assay
After the Pediococcus pentosaceus HMTN.01 is activated, the Pediococcus pentosaceus is inoculated into a liquid MRS culture medium in an inoculation amount of 1 percent, and after standing culture for 18 hours at 37 ℃, the bacterium concentration is about 4.8 multiplied by 108The thalli is collected after CFU/mL and 4000r/min centrifugation for 10min, and is resuspended in liquid MRS culture medium with pH of 2.5 after repeated washing twice with sterile normal saline. And sucking 100 mu L of bacterial suspension at 0, 1, 2 and 3 hours respectively, performing gradient dilution by using sterile physiological saline, and selecting proper dilution to perform plate coating colony counting. The control strain was Streptococcus thermophilus D-2, and the acid tolerance test was performed 3 times in parallel as described above.
(2) Pediococcus pentosaceus HMTN.01 bile salt tolerance assay
Activating Pediococcus pentosaceus HMTN.01, inoculating in liquid MRS culture medium in 1% inoculation amount, and standing at 37 deg.C for 18 hr to obtain a strain concentration of about 5.0 × 108The thalli is collected after CFU/mL and 4000r/min centrifugation for 10min, and is repeatedly washed twice by sterile normal saline, and then is respectively resuspended in liquid MRS culture medium containing 0.3%, 0.5% and 1% of bovine bile salt. After standing and culturing for 3h, sucking 100 mu L of bacterial suspension, performing gradient dilution by using sterile normal saline, and selecting proper dilution to perform plate coating colony counting. The control strain is Streptococcus thermophilus D-2, and the bile salt tolerance experiment process is the same as above and is performed 3 times in parallel.
As shown in Table 1 below, the acid tolerance of Pediococcus pentosaceus HMTN.01 was good, and the viable cell count remained at 10 after 3 hours of treatment at pH2.56CFU/mL or more, the survival rate is 71.3%, while thermophilicThe survival rate of streptococcus D-2 is only 59%; the bile salt tolerance of Pediococcus pentosaceus HMTN.01 is also better than that of Streptococcus thermophilus D-2, and the viable count is still 10 after treatment for 3h under the conditions of 0.3% and 0.5% of bile salt concentration6CFU/mL or more, and after treatment for 3h under 1% condition, the survival rate of the strain is 43.1%, and the survival rate of the streptococcus thermophilus D-2 after treatment for 3h is only 27.4%. The result shows that the pediococcus pentosaceus HMTN.01 has better acid and bile salt tolerance and is more suitable for the intestinal environment.
TABLE 1 results of the resistance of Pediococcus pentosaceus HMTN.01 and Streptococcus thermophilus D-2 to acids and bile salts
2.2 Strain tolerance study in vitro simulation of gastrointestinal fluids
Preparing simulated gastric juice: 125mM NaCl, 7mM KCl, 45mM NaHCO3And 3g/L pepsin, adjusted to pH2.5 with concentrated hydrochloric acid and then sterilized through a 0.45 μm filter. Preparing simulated intestinal juice: 45mM NaCl, 1g/L trypsin, 3g/L bovine bile salts, pH adjusted to 8.0 with NaOH, and then sterilized through a 0.22 μm microfiltration membrane.
Inoculating activated strain Pediococcus pentosaceus HMTN.01 in a liquid MRS culture medium in an inoculation amount of 1%, standing and culturing at 37 ℃ for 18h, centrifuging at 4000r/min for 10min, collecting thalli, repeatedly washing twice with sterile normal saline, and then re-suspending the thalli. Transferring the resuspended thallus into simulated gastric juice at an inoculum size of 1% for culture, and sampling at 0min, 90min and 180min respectively for viable bacteria counting; the cells were inoculated in 1% of the culture medium in simulated intestinal fluid, and the cells were sampled at 120min and 240min for viable count. The control strain was S.thermophilus D-2. Each group of experiments was performed in triplicate and the results were counted.
The experimental results are shown in the following table 2, the Pediococcus pentosaceus HMTN.01 can survive well in simulated gastric juice, and after 180min treatment, the viable count is still 107The survival rate is 76 percent when the ratio of CFU/mL is higher than that of the streptococcus thermophilus D-2, and the survival rate of the streptococcus thermophilus D-2 is 67 percent; compared with simulated gastric fluid, the simulated intestinal fluid has more obvious effect on the bacterial strain, and the effect is over 240minAfter the treatment, the viable count of Pediococcus pentosaceus HMTN.01 was reduced to 107The survival rate is still as high as 69 percent under the condition of less than CFU/mL, and the viable count of the streptococcus thermophilus D-2 is reduced to 105Below CFU/mL, the survival rate is only 54%. The result shows that the pediococcus pentosaceus HMTN.01 has better gastrointestinal fluid tolerance.
TABLE 2 Pediococcus pentosaceus HMTN.01 results of mimicking the tolerance of gastrointestinal fluids
2.3 determination of ability of Pediococcus pentosaceus HMTN.01 to inhibit food-borne pathogenic bacteria
Preparation of pediococcus pentosaceus hmtn.01 cell-free fermentation supernatant (CFS): inoculating the activated strain into a liquid MRS culture medium with the inoculation amount of 1%, culturing at 37 ℃ for 18h, centrifuging the fermentation liquor to obtain supernatant, filtering and sterilizing by a 0.45-micrometer microporous filter membrane, and storing at 4 ℃ for later use.
7 common pathogenic bacteria: pathogenic Escherichia coli (EC1), enterotoxigenic Escherichia coli (EC2), enterohemorrhagic Escherichia coli (EC3), serotype salmonella enteritidis (SM1), salmonella typhimurium (SM2), Listeria monocytogenes (LS1) and Staphylococcus Aureus (SA) are used as indicator bacteria, and the antibacterial activity of the lactic acid bacteria is measured by an Oxford cup method, which comprises the following specific steps:
activating seven kinds of indicator bacteria with corresponding culture medium, diluting 100 μ L indicator bacteria culture solution to obtain concentration of about 105～106CFU/mL dilution. Pouring to the culture medium required by various indicator bacteria, after the indicator bacteria are solidified, respectively taking 100 mu L of bacterial suspension of each indicator bacteria, uniformly coating the bacterial suspension on the corresponding culture medium surface, then equally placing 4 Oxford cups on the culture medium surface, wherein 200 mu L of CFS is added into 3 Oxford cups, and adding a liquid MRS culture medium (blank control) without bacteria into the other 1 Oxford cup, and pre-diffusing at 4 DEG CAfter 6-8 h, culturing at 37 ℃ for 18h, measuring the diameter of the inhibition zone by using a vernier caliper, performing 3 parallels in each experiment, and calculating the average value.
The experimental results are shown in the following table 3, and the pediococcus pentosaceus HMTN.01 has a certain inhibition effect on seven common food-borne pathogenic bacteria, and is beneficial to the micro-ecological health of intestinal flora. The experimental effect is shown in figure 4, the pathogenic bacteria numbers of a-g are EC1, EC2, EC3, SM1, SM2, LS1 and SA in sequence, the size of the inhibition zone on the plate represents the antagonistic capacity of the strain to the pathogenic bacteria, and the pediococcus pentosaceus HMTN.01 has better inhibition capacity to 7 types of pathogenic bacteria.
TABLE 3 bacteriostatic test results of strain HMTN.01
2.4 determination of surface hydrophobicity
The surface hydrophobicity of the bacteria is an important index of nonspecific adhesion of the strain to the gastrointestinal tract, and can be judged by the affinity of the strain to an organic solvent, so that the surface hydrophobicity and the adhesion capability of the strain are positively correlated. Ethyl acetate investigated the electron accepting ability of the surface of the strain, while chloroform investigated the electron donating ability of the surface of the strain.
The experiment adopts a microbial adhesion organic solvent method, after the Pediococcus pentosaceus HMTN.01 is activated, the Pediococcus pentosaceus is centrifuged at 6000r/min for 8min, thallus is collected, is repeatedly washed twice by sterile physiological saline, and then is suspended in the sterile physiological saline, the concentration of the bacterial suspension is adjusted until the measured OD is reached600Value (A)0) 0.6 to 0.7. Dividing into three groups, namely a chloroform group, an ethyl acetate group and a sterile water group; the specific operation is as follows:
chloroform group: sucking 1mL chloroform, adding into 4mL bacterial suspension, mixing, vortexing for 1min, standing at room temperature for 30min, collecting water phase, and determining OD600Value (A)1)；
Ethyl acetate group: sucking 1mL ethyl acetate, adding into 4mL bacterial suspension, mixing, vortexing for 1min, standing at room temperature for 30min, collecting water phase, and determining OD600Value (A)1)；
Sterile water group: sucking 1mL sterile water, adding into 4mL bacterial suspension, mixing, vortexing for 1min, standing at room temperature for 30min, collecting water phase, and determining OD600Value (A)1) This group was referred to as a control group.
Each set of 3 replicates. The strain hydrophobicity (%) was represented by the following formula:
the experimental results are shown in the following table 4, the hydrophobicity of pediococcus pentosaceus HMTN.01 to chloroform is higher than that to ethyl acetate, and both are higher than that of the control group, which indicates that pediococcus pentosaceus HMTN.01 has stronger electron supply capability and hydrophobic capability and better adhesion to gastrointestinal tract.
TABLE 4 measurement results of surface hydrophobicity of Pediococcus pentosaceus HMTN.01
2.5 determination of self-polymerizing ability
The self-aggregation ability of the strain is one of indexes for evaluating the adhesion of the strain, the higher the self-aggregation rate of the strain is, the stronger the cell adhesion ability of the strain is, and meanwhile, the strain can be more firmly adhered to the gastrointestinal tract when being aggregated in a certain amount, thereby being beneficial to the colonization of the strain.
Activating Pediococcus pentosaceus HMTN.01, centrifuging at 6000r/min for 8min, collecting thallus, repeatedly washing with sterile normal saline twice, suspending in sterile normal saline, and adjusting the concentration of the bacterial suspension until the measured OD600Value (A)0) 0.6 to 0.7. Taking 5mL of bacterial suspension, whirling for 30s, standing at room temperature for 12h, taking the supernatant of the bacterial suspension every 2h, and determining the OD600Value (A)t). Each set of 3 replicates. The calculation formula of the self-polymerization capacity (%) of the strain is as follows:
the experimental results are shown in the following table 5, and the self-aggregation capability of pediococcus pentosaceus HMTN.01 is increased along with the increase of the standing time, which indicates that the strain has better self-aggregation rate and can be better adhered to the gastrointestinal tract.
TABLE 5 measurement results of self-aggregation ability of Pediococcus pentosaceus HMTN.01
Note: different letters indicate significant difference between groups (p <0.05)
EXAMPLE 3 determination of resistance to Strain
The tolerance and infection capacity of the probiotics to antibiotics have great influence on the safety of human bodies. When the probiotics antagonize pathogenic bacteria, if the drug resistance gene is transferred to the pathogenic bacteria, the pathogenic bacteria can generate drug resistance to corresponding antibiotics. Although lactic acid bacteria are widely recognized as safe, their safety needs to be evaluated.
The drug susceptibility test of 20 antibiotics was carried out on Pediococcus pentosaceus HMTN.01, as determined by the Kirby-Bauer method (K-B paper diffusion method) according to the drug susceptibility test method recommended by the WHO. The inoculated bacterial load was diluted to 10 as specified with reference to NCCLS7～108And CFU/mL, putting 120 mu L of bacterial suspension on the surface of an MRS agar plate, uniformly coating, after the surface of the plate is dried, sticking tablets on the surface of the plate by using a sterile forceps, sticking 4 tablets on the surface of each plate, performing inverted culture at 37 ℃ for 24h after sticking, observing an antibacterial result, measuring the diameter of an antibacterial ring by using a vernier caliper, and making 3 tablets in parallel. The results were evaluated according to the manual of methods of the American Clinical Laboratory Standards Institute (CLSI)2015 edition.
The results of the drug susceptibility tests are shown in Table 6 below, where Pediococcus pentosaceus HMTN.01 is sensitive to cefotaxime, minocycline, rifampin, kanamycin, chloramphenicol, erythromycin, clindamycin, penicillin, teicholine, and cloxacillin, is moderately sensitive to tetracycline, ciprofloxacin, norfloxacin, amoxicillin, and gentamicin, and is resistant to polymyxin, streptomycin, vancomycin, acamicin, and oxacillin. The experimental effect diagram is shown in figure 5, the drug resistance of the strain to antibiotics can be reflected by the size of the inhibition zone formed on the flat plate, and the inhibition zone is generated around most tablets, which shows that the bacteria is sensitive or moderately sensitive to most antibiotics, so that the pediococcus pentosaceus HMTN.01 has higher safety.
TABLE 6 antibiotic resistance of Pediococcus pentosaceus HMTN.01
Note: s represents sensitivity; i represents moderate sensitivity; r represents drug resistance
Example 4 preparation of fermented milk and sensory evaluation
4.1 preparation of fermented milks
Activating strains: inoculating Pediococcus pentosaceus HMTN.01 preserved at-80 ℃ to an MRS liquid culture medium, carrying out anaerobic culture at 37 ℃ for 18h, and continuously transferring for 2-3 times to complete strain activation.
Seed liquid: centrifuging the activated bacterium liquid for 5min at 20 ℃ and 4000r/min, discarding the supernatant, repeatedly washing the bacterium mud twice by using sterile normal saline, adding equivalent sterile normal saline for resuspending, and taking the obtained bacterium suspension as the seed liquid for subsequent fermentation.
Working leavening agent: inoculating the seed solution into sterilized skim milk at an inoculation amount of 3% v/v, culturing to obtain curd, inoculating into sterilized skim milk at an inoculation amount of 10% m/v, repeating the inoculation for 3 times, and placing into a refrigerator at 4 deg.C to obtain the working starter after curd.
Preparation of fermented milk: preheating the pre-treated skim milk to 55 ℃, and performing primary homogenization at a homogenization pressure of 23 MPa; homogenizing, treating at 95 deg.C for 15min, cooling to 45 deg.C, transferring to sterilized glass jar, adding 8% m/v working leaven, stirring, fermenting at 42 deg.C to curd, transferring to 4 deg.C refrigerator, refrigerating, and aging for 24 hr to obtain fermented milk, as shown in FIG. 6.
4.2 sensory evaluation of fermented milks
Fermented milks of different storage times were subjected to sensory evaluation by trained experimenters. The samples were evaluated for odor, appearance, texture, mouthfeel and overall acceptability at 20 points for each full score, the sum of the score values being the total sensory score, 100 full scores, and the evaluation criteria are given in table 7.
TABLE 7 sensory evaluation indices and criteria
The sensory evaluation results are shown in a radar chart of fig. 7, and within 14d of storage time, the sensory index evaluation of the fermented milk is high and is more than 15 points; after a storage time of more than 18 days, although part of the sensory index evaluations were reduced, of which the appearance and texture were more prominent, the overall evaluation was still 60 points or more. The results show that the sensory evaluation of the fermented milk prepared from pediococcus pentosaceus HMTN.01 reaches the standard within the shelf life of 21d and meets the requirements.
4.3 texture Properties of fermented milks
The measurement is carried out by a texture analyzer, the probe is A/BE with the diameter of 35mm, and the measurement conditions are as follows: the speed before measurement is 1.00mm/s, the speed after measurement is 2.00mm/s, the measurement distance is 30mm, and the surface trigger force is 5 g. The fermented milk after maturation was taken out and placed in a 500ml glass container 64mm in diameter and 70mm in height for testing.
The test results are shown in the following table 8, and the hardness, viscosity, elasticity and the like of the fermented milk prepared from pediococcus pentosaceus HMTN.01 are moderate, which indicates that the texture characteristics of the fermented milk are good.
TABLE 8 measurement results of texture Properties of fermented milks
Example 5 acidification Capacity assay of Pediococcus pentosaceus HMTN.01
5.1 determination of acid-producing ability of Strain
Inoculating the activated bacterial liquid into sterilized skim milk at the inoculation amount of 3%, fully stirring to uniformly mix, and then fermenting and culturing at 42 ℃. Sampling every 1h, determining the total acid content, and making an acid production curve of the total acid content along with the change of time.
The acid production curve is shown in FIG. 8, the acid production rate of Pediococcus pentosaceus HMTN.01 is slow at the early stage, the acid production amount is general, but after 6 hours, the acid production rate is rapidly increased, and the final acid production amount reaches 1.52g/100mL, which indicates that the acid production capability of the strain is good.
5.2 determination of post-acidification Capacity of strains
After-ripening, the fermented milk samples were refrigerated at 4 ℃ and sampled at 1d, 4d, 8d, 12d, 16d and 20d to determine the total acid content, and the total acid content was plotted against time.
FIG. 9 shows the change curve of total acid content, and the fermented milk prepared from Pediococcus pentosaceus HMTN.01 only has 0.06g/100mL of total acid content after being stored at 4 ℃ for 20 days, has little influence on the acidity of the fermented milk, has a final acidity of 82 DEG T, and is within the range specified by the national standard (70-110 DEG T), which indicates that the strain has weak post-acidification capability.
Example 6 measurement of fragrance-producing ability of Pediococcus pentosaceus HMTN.01
The acetaldehyde and diacetyl content of the fermented milk are important indexes for evaluating the aroma-producing capability of the strain, and the fermented milk has good flavor only when the ratio of the acetaldehyde to the diacetyl content is more than 3: 1. And (3) refrigerating a plurality of batches of fermented milk in a refrigerator at 4 ℃, and sampling for 12 th, 24 th, 36 th, 48 th, 60 th and 72 th hours to measure the content of diacetyl and acetaldehyde.
6.1 measurement of diacetyl content in fermented milk
(1) Drawing of diacetyl standard curve
Dissolving 30 μ L butanedione in distilled water, and adding waterTo 1L. Respectively measuring 0mL, 2mL, 4mL, 6mL, 8mL and 10mL of butanedione solution in 12 test tubes arranged in two parallel rows, and adding water to constant volume to 10 mL. 5mL of the above butanedione solution is added into 5mL of 16% trichloroacetic acid solution to obtain butanedione standard solutions with concentrations of 0mg/L, 3mg/L, 6mg/L, 9mg/L, 12mg/L and 15mg/L, respectively. 0.5mL of 1% o-phenylenediamine solution is added into 6 test tubes in the front row, and the test tubes in the rear row are not added, fully and uniformly mixed, and then placed in a dark place for reaction for 0.5 h. After the reaction was completed, 2mL and 2.5mL of 4.0mol/L hydrochloric acid were added to the front and rear test tubes, respectively, to terminate the reaction, and after mixing, OD was measured335The value is obtained. The back row of test tubes is used as a blank control, and a standard curve is drawn by taking the concentration of the butanedione as an abscissa and the absorbance value as an ordinate. The standard curve is shown in FIG. 10.
(2) Determination of diacetyl content
Adding equal volume of 16% trichloroacetic acid solution into 30mL of fermented milk, mixing, standing for 10min, centrifuging at 6000r/min for 8min, and filtering the supernatant with filter paper to ensure that the supernatant is clear and has no impurities. And simultaneously adding 10mL of filtered supernatant into 2 empty test tubes, numbering 1 and 2, adding 0.5mL of 1% o-phenylenediamine solution into No. 1, not adding No. 2, fully mixing, and placing in a dark place for reaction for 0.5 h. After completion of the reaction, 2mL and 2.5mL of 4.0mol/L hydrochloric acid were added to the test tubes of rows 1 and 2, respectively, to terminate the reaction, and after mixing, OD was measured335The value is obtained. And (5) calculating the diacetyl content of the fermented milk by contrasting the standard curve.
6.2 determination of acetaldehyde content in fermented milk
(1) Preparation of reagents
0.1mol/L iodine standard solution: weighing 13g of iodine into a beaker, adding 35g of potassium iodide, adding distilled water for dissolving, transferring to a 1L brown volumetric flask, and shaking up to a constant volume.
0.01moL/L iodine standard solution: accurately measuring 10mL of 0.1mol/L iodine standard solution in a 100mL brown volumetric flask, adding water to a constant volume, and shaking up. It is prepared as before use.
(2) Determination of acetaldehyde content
The pretreatment of the fermented milk is the same as in step 5.1. 5mL of 1% NaHSO was taken3Dissolving in 25 deg.CAnd adding 25mL of the pretreated supernatant into a 0mL conical flask, uniformly mixing, standing for 1h, adding 1mL of 1% starch solution, uniformly mixing, titrating to be close to light blue purple by using 0.1mol/L iodine solution, titrating to be light blue purple by using 0.01mol/L iodine solution, and keeping the color unchanged within 30 s. Then 20mL of 1mol/L NaHCO was added3And (3) fully and uniformly mixing the solution to eliminate the bluish purple color of the solution, finally titrating the solution to a bluish purple end point by using 0.01mol/L iodine solution, recording the volume of the consumed iodine solution, wherein each group contains 3 parallel iodine solutions, and fresh milk is used as a blank control. The calculation formula is as follows:
acetaldehyde content (mg/L) ═ V1-V2)×0.022×C×106/25
In the formula: v1Titration of sample consumes volume of iodine standard solution (mL); v2Titration of blank depletion iodine standard solution volume (mL); concentration of C-iodine standard solution (mol/L); 25-sample size (mL); 0.022-acetaldehyde reaction chemical basic unit (g).
As shown in the following FIG. 11, after 24 hours of after-ripening, the acetaldehyde and diacetyl content reached the maximum, respectively at 18.48mg/L and 4.80 mg/L; after the after-ripening time exceeded 24 hours, although the contents of the two slightly decreased, the ratio of the two was still greater than 3:1, indicating that the fermented milk prepared from pediococcus pentosaceus hmtn.01 had a better flavor.
Example 7 determination of viable cell count during Cold storage of fermented milk
The national relevant standard stipulates that the viable count of the fermented milk is higher than 106CFU/mL can produce beneficial effects in humans. A plurality of batches of fermented milk prepared from after-ripening Pediococcus pentosaceus HMTN.01 are refrigerated in a refrigerator at 4 ℃, and the viable count of the fermented milk is measured by sampling at 1 st, 4 th, 8 th, 12 th, 16 th and 20 th days respectively.
The plate counting method is adopted for determination, and the specific steps are as follows, 1mL of the fermented milk sample taken at each time point is diluted by 9mL of sterile normal saline, and then the diluted fermented milk sample is sequentially diluted to 108And taking 0.1mL of each dilution sample, uniformly coating the sample on an MRS solid culture medium, and counting after culturing for 24h at 37 ℃. Each set of 3 replicates.
As shown in FIG. 12, the viable cell count of the fermented milk changed, but the viable cell count of the fermented milk decreased after 20 days of storage, but it was still larger than the predetermined valueConstant viable bacteria content 106CFU/mL indicates that the pediococcus pentosaceus HMTN.01 can keep better survival ability in the refrigeration process and meets the requirements.
In conclusion, the pediococcus pentosaceus HMTN.01 separated and purified from donkey milk in Hami area of Xinjiang has strong acid, viscosity and fragrance producing capabilities, and the prepared fermented milk has good flavor, taste and appearance; the product also has good hydrophobicity, self-coagulation ability and gastrointestinal tract tolerance ability, can generate good adhesion to the gastrointestinal tract, and is beneficial to permanent planting in the gastrointestinal tract; the product also has broad-spectrum antibacterial activity, has good inhibition effect on common food-borne pathogenic bacteria, and is beneficial to maintaining intestinal microecological balance and improving immunity; but also shows sensitivity or moderate sensitivity to most common antibiotics, and has reliable safety. Antibiotics include cefotaxime, minocycline, rifampin, kanamycin, chloramphenicol, erythromycin, clindamycin, penicillin, teicholam, cloxacillin, tetracycline, ciprofloxacin, norfloxacin, amoxicillin and gentamicin; the prepared fermented milk can also adjust intestinal flora, improve intestinal micro-ecological environment, effectively relieve lactose intolerance and bacterial diarrhea, and has good health care function.
Claims (10)
1. The Pediococcus pentosaceus (Pediococcus pentosaceus) HMTN.01 of donkey milk source is deposited in China center for type culture Collection No. 5/25 in 2020 with the deposition number of CCTCC NO: m2020144.
2. Use of Pediococcus pentosaceus (Pediococcus pentosaceus) hmtn.01 according to claim 1 for the preparation of a medicament for the inhibition of enteropathogenic bacteria.
3. The use of claim 2, wherein the enteropathogenic bacteria comprise pathogenic escherichia coli, enterotoxigenic escherichia coli, enterohemorrhagic escherichia coli, serotype salmonella enteritidis, salmonella typhimurium, listeria monocytogenes, and staphylococcus aureus.
4. Use of Pediococcus pentosaceus (Pediococcus pentosaceus) hmtn.01 according to claim 1 for the preparation of a health product for regulating intestinal flora.
5. Use of Pediococcus pentosaceus (Pediococcus pentosaceus) hmtn.01 according to claim 1 for the preparation of a food product or food additive.
6. Use according to claim 5, wherein the food product is a fermented dairy product.
7. The use according to claim 6, wherein the fermented milk product is yogurt, cheese, fermented milk beverage.
8. A method for preparing yogurt by using Pediococcus pentosaceus (HMTN.01), wherein the method comprises the following steps:
(1) activating strains: inoculating Pediococcus pentosaceus (HMTN.01) preserved at-80 deg.C into MRS liquid culture medium, performing anaerobic culture at 37 deg.C, and continuously transferring for 2-3 times;
(2) seed liquid: centrifuging the activated bacterium liquid obtained in the step (1), discarding supernatant, repeatedly washing the bacterium mud twice with sterile normal saline, adding equivalent sterile normal saline for resuspension, and taking the obtained bacterium suspension as seed liquid for subsequent fermentation;
(3) working leavening agent: inoculating the seed liquid obtained in the step (2) into sterilized skim milk, culturing until the seed liquid is coagulated, transferring the skim milk in a coagulated state into another pot of sterilized skim milk, and repeatedly inoculating for 2-3 times to obtain a working starter after the seed liquid is coagulated;
(4) preparation of fermented milk: and (4) inoculating the working leavening agent obtained in the step (3) into homogenized and sterilized skim milk containing 3% m/v of edible white granulated sugar, adding the mixture, fully stirring the mixture to uniformly mix the mixture, fermenting the mixture at 42 ℃ to curd, and then transferring the curd to a refrigerator at 4 ℃ for refrigeration and ripening for 24 hours to obtain the fermented milk.
9. The method of claim 8, wherein the inoculum size of the seed solution in step (3) is 3% v/v, the inoculum size transferred to skim milk is 10% m/v, and the culture conditions are anaerobic culture at 37 ℃.
10. The method of claim 8, wherein the working starter culture in step (4) is inoculated in an amount of 5% to 10% m/v.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010593449.9A CN112375696B (en) | 2020-06-27 | 2020-06-27 | Donkey milk source pediococcus pentosaceus and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010593449.9A CN112375696B (en) | 2020-06-27 | 2020-06-27 | Donkey milk source pediococcus pentosaceus and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN112375696A true CN112375696A (en) | 2021-02-19 |
| CN112375696B CN112375696B (en) | 2022-04-01 |
Family
ID=74586447
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010593449.9A Active CN112375696B (en) | 2020-06-27 | 2020-06-27 | Donkey milk source pediococcus pentosaceus and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN112375696B (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113151064A (en) * | 2021-03-25 | 2021-07-23 | 中国科学院合肥物质科学研究院 | Pediococcus pentosaceus and application thereof |
| CN113528381A (en) * | 2021-06-30 | 2021-10-22 | 广东省科学院微生物研究所(广东省微生物分析检测中心) | An antagonistic escherichia coli O157: h7 pediococcus pentosaceus and application thereof |
| CN117264850A (en) * | 2023-11-09 | 2023-12-22 | 潍坊君薇生物科技有限责任公司 | Pediococcus pentosaceus SW006 with auxiliary treatment of colpitis and immunity enhancing functions and application thereof |
| CN117264850B (en) * | 2023-11-09 | 2024-05-14 | 潍坊君薇生物科技有限责任公司 | Pediococcus pentosaceus SW006 with auxiliary treatment of colpitis and immunity enhancing functions and application thereof |
Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101033456A (en) * | 2007-02-06 | 2007-09-12 | 中国农业科学院北京畜牧兽医研究所 | Special-purpose bacterial strain for pediococcus pentosaceus used for low-acid fermentation meat products processing and preparing method for leaven thereof |
| US20110189132A1 (en) * | 2010-02-01 | 2011-08-04 | Microbios, Inc. | Microbial product containing multiple microorganisms |
| CN102599348A (en) * | 2012-03-26 | 2012-07-25 | 山东宝来利来生物工程股份有限公司 | Lactic acid bacteria semi-solid fermentation product for increasing growth performance of animals and preparation process thereof |
| WO2013019801A1 (en) * | 2011-08-01 | 2013-02-07 | Nutrition Physiology Company, Llc | Use of lactic acid bacteria to reduce pathogens and as a bio-sanitizer |
| CN106047759A (en) * | 2016-07-12 | 2016-10-26 | 西北农林科技大学 | Pediococcus pentosaceus and application thereof |
| WO2017048098A1 (en) * | 2015-09-14 | 2017-03-23 | 일동제약(주) | Functional hydrated hyaluronic acid and method for producing coated lactic acid bacteria having excellent intestinal mucoadhesive ability and selective antagonistic action using same |
| CN108660182A (en) * | 2018-05-22 | 2018-10-16 | 江南大学 | The culture medium of lactobacillus fermenti and its application in a kind of separation screening human body intestinal canal |
| WO2019093611A1 (en) * | 2017-11-10 | 2019-05-16 | (주)앰틱스바이오 | Novel pediococcus pentosaceus ab 160011 strain and composition containing same |
| EP3649866A2 (en) * | 2018-10-18 | 2020-05-13 | Universidade Católica Portuguesa - UCP | New lactic acid bacteria for preventing infections with listeria species, compositions comprising said strains and uses thereof |
-
2020
- 2020-06-27 CN CN202010593449.9A patent/CN112375696B/en active Active
Patent Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101033456A (en) * | 2007-02-06 | 2007-09-12 | 中国农业科学院北京畜牧兽医研究所 | Special-purpose bacterial strain for pediococcus pentosaceus used for low-acid fermentation meat products processing and preparing method for leaven thereof |
| US20110189132A1 (en) * | 2010-02-01 | 2011-08-04 | Microbios, Inc. | Microbial product containing multiple microorganisms |
| WO2013019801A1 (en) * | 2011-08-01 | 2013-02-07 | Nutrition Physiology Company, Llc | Use of lactic acid bacteria to reduce pathogens and as a bio-sanitizer |
| CN102599348A (en) * | 2012-03-26 | 2012-07-25 | 山东宝来利来生物工程股份有限公司 | Lactic acid bacteria semi-solid fermentation product for increasing growth performance of animals and preparation process thereof |
| WO2017048098A1 (en) * | 2015-09-14 | 2017-03-23 | 일동제약(주) | Functional hydrated hyaluronic acid and method for producing coated lactic acid bacteria having excellent intestinal mucoadhesive ability and selective antagonistic action using same |
| CN106047759A (en) * | 2016-07-12 | 2016-10-26 | 西北农林科技大学 | Pediococcus pentosaceus and application thereof |
| WO2019093611A1 (en) * | 2017-11-10 | 2019-05-16 | (주)앰틱스바이오 | Novel pediococcus pentosaceus ab 160011 strain and composition containing same |
| CN108660182A (en) * | 2018-05-22 | 2018-10-16 | 江南大学 | The culture medium of lactobacillus fermenti and its application in a kind of separation screening human body intestinal canal |
| EP3649866A2 (en) * | 2018-10-18 | 2020-05-13 | Universidade Católica Portuguesa - UCP | New lactic acid bacteria for preventing infections with listeria species, compositions comprising said strains and uses thereof |
Non-Patent Citations (6)
| Title |
|---|
| AHMAD NASROLLAHZADEH等: "Identification and Evaluation of the Antimicrobial Potential of Strains Derived from Traditional Fermented Dairy Products of Iran as A Biological Preservative Against Listeria monocytogenes,Staphylococcus aureus, Salmonella enterica and Escherichia coli", 《IRANIAN JOURNAL OF MEDICAL MICROBIOLOGY》 * |
| VENKATASUBRAMANIAN VIDHYASAGAR等: "Evaluation of Pediococcus pentosaceus strains isolated from Idly batter for probiotic properties in vitro", 《JOURNAL OF FUNCTIONAL FOODS》 * |
| 曾雪峰等: "湘西传统酸鱼中乳酸菌的分离及特性研究", 《食品与发酵工业》 * |
| 杜金城等: "具有抑制大肠杆菌作用的乳酸菌的初步筛选及其益生潜能的研究", 《食品工业科技》 * |
| 靳亚梅等: "评估中国新疆牧区原马奶中潜在益生乳酸菌在发酵乳制品中的应用潜力", 《中国食品科学技术学会第十七届年会摘要集》 * |
| 高云云等: "新疆传统乳品中产胞外多糖乳酸菌的筛选及益生特性的研究", 《中国酿造》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113151064A (en) * | 2021-03-25 | 2021-07-23 | 中国科学院合肥物质科学研究院 | Pediococcus pentosaceus and application thereof |
| CN113528381A (en) * | 2021-06-30 | 2021-10-22 | 广东省科学院微生物研究所(广东省微生物分析检测中心) | An antagonistic escherichia coli O157: h7 pediococcus pentosaceus and application thereof |
| CN117264850A (en) * | 2023-11-09 | 2023-12-22 | 潍坊君薇生物科技有限责任公司 | Pediococcus pentosaceus SW006 with auxiliary treatment of colpitis and immunity enhancing functions and application thereof |
| CN117264850B (en) * | 2023-11-09 | 2024-05-14 | 潍坊君薇生物科技有限责任公司 | Pediococcus pentosaceus SW006 with auxiliary treatment of colpitis and immunity enhancing functions and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN112375696B (en) | 2022-04-01 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Yerlikaya | Probiotic potential and biochemical and technological properties of Lactococcus lactis ssp. lactis strains isolated from raw milk and kefir grains | |
| CN110577912B (en) | Lactobacillus gasseri and application thereof in preparing fermented milk | |
| CN105255752B (en) | High acid plant lactobacillus ZDY2013 | |
| CN112143680B (en) | Lactobacillus paracasei ZJUIDS05 with antioxidant effect and application thereof | |
| CN113699063A (en) | Lactobacillus paracasei for reducing cholesterol and application thereof | |
| CN112375696B (en) | Donkey milk source pediococcus pentosaceus and application thereof | |
| CN112812999B (en) | Lactobacillus plantarum SLB01 with inhibition effect on enterobacter cloacae and derivative product and application thereof | |
| CN111925961A (en) | Lactobacillus plantarum Lp2 and application thereof | |
| CN111534447A (en) | Lactobacillus johnsonii and application thereof | |
| CN114774327B (en) | Lactobacillus plantarum HB13-2 and application thereof | |
| CN110129220B (en) | Lactobacillus bulgaricus BSTS6-4 and application thereof | |
| CN107828703B (en) | Lactobacillus reuteri Fullarton-9-35 and application thereof | |
| Ar et al. | Assessment of potential probiotic properties lactic acid bacteria from shrimp paste or belacan | |
| CN117143767A (en) | Breast milk-derived fermented lactobacillus mucilaginosus MSJK0025 capable of regulating intestinal flora and application thereof | |
| CN107937316A (en) | Space lactobacillus reuteri Fullarton 9 71 and application | |
| JP4457364B2 (en) | New lactic acid bacteria and various products processed using the lactic acid bacteria | |
| CN114717150B (en) | Lactobacillus plantarum CRS33 and application thereof | |
| Arıcı et al. | Some technological and functional properties of lactic acid bacteria isolated from hardaliye | |
| CN113881592B (en) | Lactobacillus reuteri and application thereof | |
| CN107974425A (en) | Space lactobacillus reuteri Fullarton-9-79 and application | |
| KR100720025B1 (en) | Probiotic lactic acid bacteria and composition comprising the same | |
| CN108094527B (en) | Lactobacillus reuteri Fullarton-9-87 and application thereof | |
| JP2017012163A (en) | Lactobacillus plantarum llp5193 with high acid- and bile-resistance and high cell attaching ability and products containing it as the effective ingredient | |
| CN110141583A (en) | A kind of application of Kefir grains lactobacillus M3 in active constituent antibacterial and as treatment type II diabetes medicament | |
| Jikang et al. | Characterization and in vitro properties of Lactobacillus plantarum and Leuconostoc mesenteroides for probiotic potential and nitrite degradation. |
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 | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20230508 Address after: Room 4-27, 429, No. 19-3 Beisan East Road, Development Zone, Shihezi City, Xinjiang Uygur Autonomous Region, 832061 Patentee after: Xinjiang Lingshui Biotechnology Co.,Ltd. Address before: 832003 North four road, Shihezi, the Xinjiang Uygur Autonomous Region 221 Patentee before: SHIHEZI University |
|
| TR01 | Transfer of patent right | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20231222 Address after: 401120 plant 4 (No. 01), No. 100, Baohuan Third Road, Huixing street, Yubei District, Chongqing Patentee after: Chongqing Lingshui Biotechnology Co.,Ltd. Address before: 832061 Xinjiang Uygur Autonomous Region Provincial and County level Administrative Division, Shihezi City Development Zone, No. 19-3 Beisan East Road, Room 4-27, 429 Patentee before: Xinjiang Lingshui Biotechnology Co.,Ltd. |
|
| TR01 | Transfer of patent right |