CN116676226B - Lactobacillus plantarum LP-28 for relieving anxiety, yoghourt and application - Google Patents
Lactobacillus plantarum LP-28 for relieving anxiety, yoghourt and application Download PDFInfo
- Publication number
- CN116676226B CN116676226B CN202310645941.XA CN202310645941A CN116676226B CN 116676226 B CN116676226 B CN 116676226B CN 202310645941 A CN202310645941 A CN 202310645941A CN 116676226 B CN116676226 B CN 116676226B
- Authority
- CN
- China
- Prior art keywords
- strain
- lactobacillus plantarum
- fermentation
- lactobacillus
- culture
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 240000006024 Lactobacillus plantarum Species 0.000 title claims abstract description 88
- 235000013965 Lactobacillus plantarum Nutrition 0.000 title claims abstract description 88
- 229940072205 lactobacillus plantarum Drugs 0.000 title claims abstract description 88
- 235000013618 yogurt Nutrition 0.000 title claims abstract description 45
- 208000019901 Anxiety disease Diseases 0.000 title abstract description 15
- 230000036506 anxiety Effects 0.000 title abstract description 15
- BTCSSZJGUNDROE-UHFFFAOYSA-N gamma-aminobutyric acid Chemical compound NCCCC(O)=O BTCSSZJGUNDROE-UHFFFAOYSA-N 0.000 claims abstract description 74
- OGNSCSPNOLGXSM-UHFFFAOYSA-N (+/-)-DABA Natural products NCCC(N)C(O)=O OGNSCSPNOLGXSM-UHFFFAOYSA-N 0.000 claims abstract description 37
- 229960003692 gamma aminobutyric acid Drugs 0.000 claims abstract description 37
- 230000000968 intestinal effect Effects 0.000 claims abstract description 26
- 241000218588 Lactobacillus rhamnosus Species 0.000 claims abstract description 25
- 238000004321 preservation Methods 0.000 claims abstract description 17
- 230000003860 sleep quality Effects 0.000 claims abstract description 14
- 101100342977 Neurospora crassa (strain ATCC 24698 / 74-OR23-1A / CBS 708.71 / DSM 1257 / FGSC 987) leu-1 gene Proteins 0.000 claims abstract description 8
- 238000009629 microbiological culture Methods 0.000 claims abstract description 8
- 238000000855 fermentation Methods 0.000 claims description 74
- 230000004151 fermentation Effects 0.000 claims description 70
- 239000001963 growth medium Substances 0.000 claims description 23
- 238000012258 culturing Methods 0.000 claims description 13
- 238000004519 manufacturing process Methods 0.000 claims description 13
- 238000009630 liquid culture Methods 0.000 claims description 11
- 238000000034 method Methods 0.000 claims description 11
- 238000011081 inoculation Methods 0.000 claims description 8
- 239000000843 powder Substances 0.000 claims description 8
- 235000013336 milk Nutrition 0.000 claims description 6
- 239000008267 milk Substances 0.000 claims description 6
- 210000004080 milk Anatomy 0.000 claims description 6
- 239000002994 raw material Substances 0.000 claims description 5
- 235000013305 food Nutrition 0.000 claims description 4
- 230000036541 health Effects 0.000 claims description 3
- 238000002360 preparation method Methods 0.000 claims description 3
- 238000011218 seed culture Methods 0.000 claims description 3
- 235000003599 food sweetener Nutrition 0.000 claims description 2
- 238000001694 spray drying Methods 0.000 claims description 2
- 239000003381 stabilizer Substances 0.000 claims description 2
- 239000003765 sweetening agent Substances 0.000 claims description 2
- 230000002906 microbiologic effect Effects 0.000 claims 1
- 239000006041 probiotic Substances 0.000 abstract description 12
- 235000018291 probiotics Nutrition 0.000 abstract description 12
- 230000000529 probiotic effect Effects 0.000 abstract description 11
- 239000000796 flavoring agent Substances 0.000 abstract description 5
- 235000019634 flavors Nutrition 0.000 abstract description 5
- 230000036039 immunity Effects 0.000 abstract description 4
- 244000005700 microbiome Species 0.000 abstract description 3
- 241000894006 Bacteria Species 0.000 description 14
- 239000007788 liquid Substances 0.000 description 13
- 230000001580 bacterial effect Effects 0.000 description 12
- 239000000047 product Substances 0.000 description 12
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 11
- 230000000694 effects Effects 0.000 description 10
- 210000001035 gastrointestinal tract Anatomy 0.000 description 10
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 10
- 239000002054 inoculum Substances 0.000 description 9
- 238000002156 mixing Methods 0.000 description 9
- 239000006228 supernatant Substances 0.000 description 9
- 229920002472 Starch Polymers 0.000 description 8
- 238000010790 dilution Methods 0.000 description 8
- 239000012895 dilution Substances 0.000 description 8
- PXEDJBXQKAGXNJ-QTNFYWBSSA-L disodium L-glutamate Chemical compound [Na+].[Na+].[O-]C(=O)[C@@H](N)CCC([O-])=O PXEDJBXQKAGXNJ-QTNFYWBSSA-L 0.000 description 8
- 235000013923 monosodium glutamate Nutrition 0.000 description 8
- 229940073490 sodium glutamate Drugs 0.000 description 8
- 239000000243 solution Substances 0.000 description 8
- 239000008107 starch Substances 0.000 description 8
- 235000019698 starch Nutrition 0.000 description 8
- 239000002253 acid Substances 0.000 description 7
- 230000002550 fecal effect Effects 0.000 description 7
- 230000006872 improvement Effects 0.000 description 7
- 238000000338 in vitro Methods 0.000 description 7
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 6
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 6
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 6
- 238000001816 cooling Methods 0.000 description 6
- 239000006071 cream Substances 0.000 description 6
- 238000012216 screening Methods 0.000 description 6
- 238000012360 testing method Methods 0.000 description 6
- 206010002869 Anxiety symptoms Diseases 0.000 description 5
- 102000008214 Glutamate decarboxylase Human genes 0.000 description 5
- 108091022930 Glutamate decarboxylase Proteins 0.000 description 5
- 239000003833 bile salt Substances 0.000 description 5
- 239000012530 fluid Substances 0.000 description 5
- 239000004310 lactic acid Substances 0.000 description 5
- 235000014655 lactic acid Nutrition 0.000 description 5
- 230000004060 metabolic process Effects 0.000 description 5
- 230000000813 microbial effect Effects 0.000 description 5
- 238000004088 simulation Methods 0.000 description 5
- 108020004465 16S ribosomal RNA Proteins 0.000 description 4
- WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 description 4
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 description 4
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 description 4
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 4
- 210000004027 cell Anatomy 0.000 description 4
- 230000002496 gastric effect Effects 0.000 description 4
- 235000021552 granulated sugar Nutrition 0.000 description 4
- 239000000463 material Substances 0.000 description 4
- 239000002609 medium Substances 0.000 description 4
- 230000008569 process Effects 0.000 description 4
- 239000007787 solid Substances 0.000 description 4
- 230000002195 synergetic effect Effects 0.000 description 4
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 3
- 239000001888 Peptone Substances 0.000 description 3
- 108010080698 Peptones Proteins 0.000 description 3
- 241000607142 Salmonella Species 0.000 description 3
- 208000013738 Sleep Initiation and Maintenance disease Diseases 0.000 description 3
- UIIMBOGNXHQVGW-DEQYMQKBSA-M Sodium bicarbonate-14C Chemical compound [Na+].O[14C]([O-])=O UIIMBOGNXHQVGW-DEQYMQKBSA-M 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 241000191967 Staphylococcus aureus Species 0.000 description 3
- 230000000844 anti-bacterial effect Effects 0.000 description 3
- 230000008901 benefit Effects 0.000 description 3
- 229940093761 bile salts Drugs 0.000 description 3
- 210000001072 colon Anatomy 0.000 description 3
- 238000011049 filling Methods 0.000 description 3
- 206010022437 insomnia Diseases 0.000 description 3
- 230000014759 maintenance of location Effects 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 235000019319 peptone Nutrition 0.000 description 3
- 229920000136 polysorbate Polymers 0.000 description 3
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 description 3
- 230000000630 rising effect Effects 0.000 description 3
- 238000007789 sealing Methods 0.000 description 3
- 235000017557 sodium bicarbonate Nutrition 0.000 description 3
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 3
- 230000001954 sterilising effect Effects 0.000 description 3
- 238000003756 stirring Methods 0.000 description 3
- 230000007306 turnover Effects 0.000 description 3
- SATHPVQTSSUFFW-UHFFFAOYSA-N 4-[6-[(3,5-dihydroxy-4-methoxyoxan-2-yl)oxymethyl]-3,5-dihydroxy-4-methoxyoxan-2-yl]oxy-2-(hydroxymethyl)-6-methyloxane-3,5-diol Chemical compound OC1C(OC)C(O)COC1OCC1C(O)C(OC)C(O)C(OC2C(C(CO)OC(C)C2O)O)O1 SATHPVQTSSUFFW-UHFFFAOYSA-N 0.000 description 2
- 239000001904 Arabinogalactan Substances 0.000 description 2
- 229920000189 Arabinogalactan Polymers 0.000 description 2
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 2
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 description 2
- 206010013954 Dysphoria Diseases 0.000 description 2
- 241000588724 Escherichia coli Species 0.000 description 2
- 229920002907 Guar gum Polymers 0.000 description 2
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 2
- SIKJAQJRHWYJAI-UHFFFAOYSA-N Indole Chemical compound C1=CC=C2NC=CC2=C1 SIKJAQJRHWYJAI-UHFFFAOYSA-N 0.000 description 2
- 229920001202 Inulin Polymers 0.000 description 2
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 2
- 108010019160 Pancreatin Proteins 0.000 description 2
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 2
- 229930006000 Sucrose Natural products 0.000 description 2
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 2
- 235000011054 acetic acid Nutrition 0.000 description 2
- 230000003213 activating effect Effects 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 235000019312 arabinogalactan Nutrition 0.000 description 2
- 230000003385 bacteriostatic effect Effects 0.000 description 2
- 239000001110 calcium chloride Substances 0.000 description 2
- 229910001628 calcium chloride Inorganic materials 0.000 description 2
- 229940041514 candida albicans extract Drugs 0.000 description 2
- 239000005018 casein Substances 0.000 description 2
- BECPQYXYKAMYBN-UHFFFAOYSA-N casein, tech. Chemical compound NCCCCC(C(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(CC(C)C)N=C(O)C(CCC(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(C(C)O)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(COP(O)(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(N)CC1=CC=CC=C1 BECPQYXYKAMYBN-UHFFFAOYSA-N 0.000 description 2
- 235000021240 caseins Nutrition 0.000 description 2
- 239000011248 coating agent Substances 0.000 description 2
- 238000000576 coating method Methods 0.000 description 2
- 229960002433 cysteine Drugs 0.000 description 2
- 235000018417 cysteine Nutrition 0.000 description 2
- XUJNEKJLAYXESH-UHFFFAOYSA-N cysteine Natural products SCC(N)C(O)=O XUJNEKJLAYXESH-UHFFFAOYSA-N 0.000 description 2
- 230000029087 digestion Effects 0.000 description 2
- XBDQKXXYIPTUBI-UHFFFAOYSA-N dimethylselenoniopropionate Natural products CCC(O)=O XBDQKXXYIPTUBI-UHFFFAOYSA-N 0.000 description 2
- 210000004921 distal colon Anatomy 0.000 description 2
- 230000008451 emotion Effects 0.000 description 2
- 229910052564 epsomite Inorganic materials 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 210000003608 fece Anatomy 0.000 description 2
- 238000001914 filtration Methods 0.000 description 2
- 235000011389 fruit/vegetable juice Nutrition 0.000 description 2
- 210000004211 gastric acid Anatomy 0.000 description 2
- 210000004051 gastric juice Anatomy 0.000 description 2
- 235000013922 glutamic acid Nutrition 0.000 description 2
- 239000004220 glutamic acid Substances 0.000 description 2
- 229960002989 glutamic acid Drugs 0.000 description 2
- 230000012010 growth Effects 0.000 description 2
- 239000000665 guar gum Substances 0.000 description 2
- 229960002154 guar gum Drugs 0.000 description 2
- 235000010417 guar gum Nutrition 0.000 description 2
- 150000003278 haem Chemical class 0.000 description 2
- 238000004128 high performance liquid chromatography Methods 0.000 description 2
- 229910052739 hydrogen Inorganic materials 0.000 description 2
- 230000002401 inhibitory effect Effects 0.000 description 2
- 239000007924 injection Substances 0.000 description 2
- 238000002347 injection Methods 0.000 description 2
- JYJIGFIDKWBXDU-MNNPPOADSA-N inulin Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)OC[C@]1(OC[C@]2(OC[C@]3(OC[C@]4(OC[C@]5(OC[C@]6(OC[C@]7(OC[C@]8(OC[C@]9(OC[C@]%10(OC[C@]%11(OC[C@]%12(OC[C@]%13(OC[C@]%14(OC[C@]%15(OC[C@]%16(OC[C@]%17(OC[C@]%18(OC[C@]%19(OC[C@]%20(OC[C@]%21(OC[C@]%22(OC[C@]%23(OC[C@]%24(OC[C@]%25(OC[C@]%26(OC[C@]%27(OC[C@]%28(OC[C@]%29(OC[C@]%30(OC[C@]%31(OC[C@]%32(OC[C@]%33(OC[C@]%34(OC[C@]%35(OC[C@]%36(O[C@@H]%37[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O%37)O)[C@H]([C@H](O)[C@@H](CO)O%36)O)[C@H]([C@H](O)[C@@H](CO)O%35)O)[C@H]([C@H](O)[C@@H](CO)O%34)O)[C@H]([C@H](O)[C@@H](CO)O%33)O)[C@H]([C@H](O)[C@@H](CO)O%32)O)[C@H]([C@H](O)[C@@H](CO)O%31)O)[C@H]([C@H](O)[C@@H](CO)O%30)O)[C@H]([C@H](O)[C@@H](CO)O%29)O)[C@H]([C@H](O)[C@@H](CO)O%28)O)[C@H]([C@H](O)[C@@H](CO)O%27)O)[C@H]([C@H](O)[C@@H](CO)O%26)O)[C@H]([C@H](O)[C@@H](CO)O%25)O)[C@H]([C@H](O)[C@@H](CO)O%24)O)[C@H]([C@H](O)[C@@H](CO)O%23)O)[C@H]([C@H](O)[C@@H](CO)O%22)O)[C@H]([C@H](O)[C@@H](CO)O%21)O)[C@H]([C@H](O)[C@@H](CO)O%20)O)[C@H]([C@H](O)[C@@H](CO)O%19)O)[C@H]([C@H](O)[C@@H](CO)O%18)O)[C@H]([C@H](O)[C@@H](CO)O%17)O)[C@H]([C@H](O)[C@@H](CO)O%16)O)[C@H]([C@H](O)[C@@H](CO)O%15)O)[C@H]([C@H](O)[C@@H](CO)O%14)O)[C@H]([C@H](O)[C@@H](CO)O%13)O)[C@H]([C@H](O)[C@@H](CO)O%12)O)[C@H]([C@H](O)[C@@H](CO)O%11)O)[C@H]([C@H](O)[C@@H](CO)O%10)O)[C@H]([C@H](O)[C@@H](CO)O9)O)[C@H]([C@H](O)[C@@H](CO)O8)O)[C@H]([C@H](O)[C@@H](CO)O7)O)[C@H]([C@H](O)[C@@H](CO)O6)O)[C@H]([C@H](O)[C@@H](CO)O5)O)[C@H]([C@H](O)[C@@H](CO)O4)O)[C@H]([C@H](O)[C@@H](CO)O3)O)[C@H]([C@H](O)[C@@H](CO)O2)O)[C@@H](O)[C@H](O)[C@@H](CO)O1 JYJIGFIDKWBXDU-MNNPPOADSA-N 0.000 description 2
- 229940029339 inulin Drugs 0.000 description 2
- 230000006799 invasive growth in response to glucose limitation Effects 0.000 description 2
- 238000011835 investigation Methods 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 2
- 235000019796 monopotassium phosphate Nutrition 0.000 description 2
- 210000002569 neuron Anatomy 0.000 description 2
- 229940055695 pancreatin Drugs 0.000 description 2
- 239000001814 pectin Substances 0.000 description 2
- 229920001277 pectin Polymers 0.000 description 2
- 235000010987 pectin Nutrition 0.000 description 2
- 230000002572 peristaltic effect Effects 0.000 description 2
- 239000001103 potassium chloride Substances 0.000 description 2
- 235000011164 potassium chloride Nutrition 0.000 description 2
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 description 2
- 235000020185 raw untreated milk Nutrition 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 235000020183 skimmed milk Nutrition 0.000 description 2
- 208000019116 sleep disease Diseases 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- 235000019614 sour taste Nutrition 0.000 description 2
- 239000011550 stock solution Substances 0.000 description 2
- 210000002784 stomach Anatomy 0.000 description 2
- 239000012137 tryptone Substances 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- 235000008939 whole milk Nutrition 0.000 description 2
- 229920001221 xylan Polymers 0.000 description 2
- 150000004823 xylans Chemical class 0.000 description 2
- 239000012138 yeast extract Substances 0.000 description 2
- 230000002407 ATP formation Effects 0.000 description 1
- 241000304886 Bacilli Species 0.000 description 1
- 108010062877 Bacteriocins Proteins 0.000 description 1
- 208000024172 Cardiovascular disease Diseases 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 102000005915 GABA Receptors Human genes 0.000 description 1
- 108010005551 GABA Receptors Proteins 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 238000003794 Gram staining Methods 0.000 description 1
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 1
- 108090000604 Hydrolases Proteins 0.000 description 1
- 102000004157 Hydrolases Human genes 0.000 description 1
- 206010020772 Hypertension Diseases 0.000 description 1
- 241000186660 Lactobacillus Species 0.000 description 1
- 244000199885 Lactobacillus bulgaricus Species 0.000 description 1
- 235000013960 Lactobacillus bulgaricus Nutrition 0.000 description 1
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 description 1
- 108091005804 Peptidases Proteins 0.000 description 1
- 206010062519 Poor quality sleep Diseases 0.000 description 1
- 239000004365 Protease Substances 0.000 description 1
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 238000012300 Sequence Analysis Methods 0.000 description 1
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 1
- 241000194020 Streptococcus thermophilus Species 0.000 description 1
- VLSOAXRVHARBEQ-UHFFFAOYSA-N [4-fluoro-2-(hydroxymethyl)phenyl]methanol Chemical compound OCC1=CC=C(F)C=C1CO VLSOAXRVHARBEQ-UHFFFAOYSA-N 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 150000007513 acids Chemical class 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 150000001412 amines Chemical class 0.000 description 1
- 229940024606 amino acid Drugs 0.000 description 1
- 235000001014 amino acid Nutrition 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 230000004888 barrier function Effects 0.000 description 1
- 235000015278 beef Nutrition 0.000 description 1
- 235000013361 beverage Nutrition 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 229910000019 calcium carbonate Inorganic materials 0.000 description 1
- 230000021164 cell adhesion Effects 0.000 description 1
- 210000003169 central nervous system Anatomy 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 208000029078 coronary artery disease Diseases 0.000 description 1
- 235000013365 dairy product Nutrition 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 230000000994 depressogenic effect Effects 0.000 description 1
- 206010012601 diabetes mellitus Diseases 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 description 1
- 229910000396 dipotassium phosphate Inorganic materials 0.000 description 1
- 235000019797 dipotassium phosphate Nutrition 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 229940088598 enzyme Drugs 0.000 description 1
- 230000005284 excitation Effects 0.000 description 1
- 230000029142 excretion Effects 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 210000003736 gastrointestinal content Anatomy 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 230000005802 health problem Effects 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- PZOUSPYUWWUPPK-UHFFFAOYSA-N indole Natural products CC1=CC=CC2=C1C=CN2 PZOUSPYUWWUPPK-UHFFFAOYSA-N 0.000 description 1
- RKJUIXBNRJVNHR-UHFFFAOYSA-N indolenine Natural products C1=CC=C2CC=NC2=C1 RKJUIXBNRJVNHR-UHFFFAOYSA-N 0.000 description 1
- 210000004347 intestinal mucosa Anatomy 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 229940039696 lactobacillus Drugs 0.000 description 1
- 229940004208 lactobacillus bulgaricus Drugs 0.000 description 1
- 239000008101 lactose Substances 0.000 description 1
- 235000015073 liquid stocks Nutrition 0.000 description 1
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 1
- 235000019341 magnesium sulphate Nutrition 0.000 description 1
- 229940099596 manganese sulfate Drugs 0.000 description 1
- 239000011702 manganese sulphate Substances 0.000 description 1
- 235000007079 manganese sulphate Nutrition 0.000 description 1
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- 230000007269 microbial metabolism Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 210000005036 nerve Anatomy 0.000 description 1
- 230000000926 neurological effect Effects 0.000 description 1
- 239000002858 neurotransmitter agent Substances 0.000 description 1
- FEMOMIGRRWSMCU-UHFFFAOYSA-N ninhydrin Chemical compound C1=CC=C2C(=O)C(O)(O)C(=O)C2=C1 FEMOMIGRRWSMCU-UHFFFAOYSA-N 0.000 description 1
- 230000008520 organization Effects 0.000 description 1
- 230000001766 physiological effect Effects 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 235000019260 propionic acid Nutrition 0.000 description 1
- 235000018102 proteins Nutrition 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 230000009323 psychological health Effects 0.000 description 1
- IUVKMZGDUIUOCP-BTNSXGMBSA-N quinbolone Chemical compound O([C@H]1CC[C@H]2[C@H]3[C@@H]([C@]4(C=CC(=O)C=C4CC3)C)CC[C@@]21C)C1=CCCC1 IUVKMZGDUIUOCP-BTNSXGMBSA-N 0.000 description 1
- 229910052704 radon Inorganic materials 0.000 description 1
- SYUHGPGVQRZVTB-UHFFFAOYSA-N radon atom Chemical compound [Rn] SYUHGPGVQRZVTB-UHFFFAOYSA-N 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000035806 respiratory chain Effects 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 235000021391 short chain fatty acids Nutrition 0.000 description 1
- 150000004666 short chain fatty acids Chemical class 0.000 description 1
- 239000002356 single layer Substances 0.000 description 1
- 230000004622 sleep time Effects 0.000 description 1
- 239000001632 sodium acetate Substances 0.000 description 1
- 235000017281 sodium acetate Nutrition 0.000 description 1
- 238000005507 spraying Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- YWYZEGXAUVWDED-UHFFFAOYSA-N triammonium citrate Chemical compound [NH4+].[NH4+].[NH4+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O YWYZEGXAUVWDED-UHFFFAOYSA-N 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
- A23C9/00—Milk preparations; Milk powder or milk powder preparations
- A23C9/12—Fermented milk preparations; Treatment using microorganisms or enzymes
- A23C9/123—Fermented milk preparations; Treatment using microorganisms or enzymes using only microorganisms of the genus lactobacteriaceae; Yoghurt
- A23C9/1238—Fermented milk preparations; Treatment using microorganisms or enzymes using only microorganisms of the genus lactobacteriaceae; Yoghurt using specific L. bulgaricus or S. thermophilus microorganisms; using entrapped or encapsulated yoghurt bacteria; Physical or chemical treatment of L. bulgaricus or S. thermophilus cultures; Fermentation only with L. bulgaricus or only with S. thermophilus
-
- 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
- A23C9/16—Agglomerating or granulating milk powder; Making instant milk powder; Products obtained thereby
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
- A61K35/741—Probiotics
- A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
- A61K35/747—Lactobacilli, e.g. L. acidophilus or L. brevis
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
- A61P25/20—Hypnotics; Sedatives
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
- A61P25/22—Anxiolytics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
- A61P25/24—Antidepressants
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P13/00—Preparation of nitrogen-containing organic compounds
- C12P13/005—Amino acids other than alpha- or beta amino acids, e.g. gamma amino acids
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/225—Lactobacillus
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/225—Lactobacillus
- C12R2001/25—Lactobacillus plantarum
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Microbiology (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Biotechnology (AREA)
- Biomedical Technology (AREA)
- Genetics & Genomics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Pharmacology & Pharmacy (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- General Chemical & Material Sciences (AREA)
- Mycology (AREA)
- Neurosurgery (AREA)
- General Engineering & Computer Science (AREA)
- Polymers & Plastics (AREA)
- Food Science & Technology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Biochemistry (AREA)
- Neurology (AREA)
- Virology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Molecular Biology (AREA)
- Pain & Pain Management (AREA)
- Psychiatry (AREA)
- Anesthesiology (AREA)
- Nutrition Science (AREA)
- Epidemiology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention belongs to the technical field of microorganisms, and relates to a Lactobacillus plantarum LP-28 for relieving anxiety, yoghourt and application, wherein the LP-28 strain is preserved in China general microbiological culture collection center (CGMCC) with the preservation number of 7.463 in the year 6 and 25 of 2022; the preservation address is the Beijing Chaoyang district North Star West Leu 1, 3. The lactobacillus plantarum LP-28 strain can be directly metabolized in the intestinal environment of a human body to produce gamma-aminobutyric acid, can be applied to products for improving sleep quality, improves the balance of intestinal flora, and enhances the immunity of organisms. Meanwhile, the yogurt prepared by fermenting the probiotic EM28 formed by combining the lactobacillus plantarum LP-28 strain and lactobacillus rhamnosus LR-28 strain is proper in sweetness and sourness and good in flavor.
Description
Technical Field
The invention belongs to the technical field of microorganisms, and particularly relates to lactobacillus plantarum LP-28 for relieving anxiety, yoghourt and application thereof.
Background
Investigation by the world health organization data shows that 27% of people worldwide have sleep disorders, with china accounting for 38.2%. From 2013 to 2018, the average sleep time of Chinese is shortened from 8.8 hours to 6.5 hours. China becomes a country with high sleep problems, and insomnia becomes a health problem of particular concern to modern people.
Poor sleep quality is a risk factor of various diseases, has close relation with hypertension, coronary heart disease, diabetes and the like, and can influence the psychological health condition of people, so that people are tired, anxious and depressed, negative emotion is increased, and the attention degree and satisfaction degree to the external matters are reduced. Anxiety and depression symptoms increase the prevalence and incidence of cardiovascular diseases.
Gamma-aminobutyric acid (Gamma-aminobutyric acid, GABA) is an important central nervous system inhibitory neurotransmitter, and GABA as a small molecular weight non-protein amino acid has edible safety, and can be used for the production of foods such as beverages. Research shows that the intake of a certain amount of GABA has the physiological effects of improving anxiety emotion of organisms and improving sleep quality.
Currently, GABA is synthesized by utilizing microbial metabolism to convert sodium glutamate or glutamic acid into GABA, and sodium glutamate or glutamic acid substrate is additionally provided in the process. In the prior art, strains for directly converting and producing GABA are cultured by microbial fermentation, but the fermentation process for producing GABA is carried out in an in-vitro fermentation tank, the culture medium used under the fermentation condition and the culture environment have great difference from the complex human intestinal environment, and the strains cannot be directly metabolized in the human intestinal environment to produce GABA. Therefore, the advantage of GABA production by the above strain needs to be exerted by means of in vitro fermentation, but is not necessarily applicable as a product of direct metabolism in human intestinal environment for GABA production.
On the other hand, yogurt is a common dairy product obtained by fermentation with lactic acid bacteria, and the sour taste of the yogurt is derived from acid substances generated in the fermentation process of lactic acid bacteria. The acidogenesis of lactobacillus plantarum and lactobacillus rhamnosus in the metabolic process is too much, so that the acidity of the yoghurt after reaching the fermentation end point is too high (the acidity is 170-250 DEG T), and the mouthfeel is seriously influenced. Therefore, the yogurt sold in the market at present does not adopt the two strains for fermentation, and the streptococcus thermophilus and the lactobacillus bulgaricus are adopted for synergistic fermentation to control the acidity, but generally only have fermentation effect, and are not probiotics.
Disclosure of Invention
Aiming at the technical problems, the invention provides lactobacillus plantarum LP-28, yoghourt and application thereof, and the lactobacillus plantarum LP-28 can regulate the balance of intestinal microbial flora, enhance the immunity of organisms and directly metabolize and produce GABA in the intestinal environment of human bodies. Meanwhile, the yoghurt prepared by the combined fermentation of the lactobacillus plantarum LP-28 strain and the lactobacillus rhamnosus LR-28 strain is improved by a novel process, and has proper sour taste and good flavor.
In order to achieve the aim of the invention, the invention adopts the following technical scheme:
A lactobacillus plantarum LP-28 strain, which is classified as lactobacillus plantarum (Lactobacillus plantarum) and preserved in China general microbiological culture Collection center (CGMCC) with the preservation number of CGMCC No.7.463 in the year 6 and 25 of 2022; the preservation address is the Beijing Chaoyang district North Star West Leu 1, 3.
Compared with the prior art, the lactobacillus plantarum LP-28 strain with the preservation number of CGMCC No.7.463 provided by the invention has the following advantages:
(1) The lactobacillus plantarum LP-28 strain provided by the invention belongs to lactobacillus and can effectively resist gastric acid and bile salts, and is subjected to field planting in intestinal tracts so as to improve intestinal microbial flora, regulate the balance of the intestinal microbial flora and strengthen the immunity of organisms.
(2) The living bacterium of the lactobacillus plantarum LP-28 provided by the invention can produce high-activity glutamate decarboxylase (GAD) in an intestinal environment, and Glu is decarboxylated to form GABA by utilizing the GAD activity. GABA absorbs and enters the human body, enters nerve cells along with blood, and combines with GABA receptors among neurons, thereby playing a role in inhibiting nerve excitation, thereby playing roles in relieving dysphoria and improving sleep quality.
The 16S rRNA sequence is shown below:
AGGACGTGCCGCGTGCCTATGATGCAAGTCGAACGAACTCTGGTATTGATTGGTGCTTGCATCATGATTTACATTTGAGTGAGTGGCGAACTGGTGAGTAACACGTGGGAAACCTGCCCAGAAGCGGGGGATAACACCTGGAAACAGATGCTAATACCGCATAACAACTTGGACCGCATGGTCCGAGCTTGAAAGATGGCTTCGGCTATCACTTTTGGATGGTCCCGCGGCGTATTAGCTAGATGGTGGGGTAACGGCTCACCATGGCAATGATACGTAGCCGACCTGAGAGGGTAATCGGCCACATTGGGACTGAGACACGGCCCAAACTCCTACGGGAGGCAGCAGTAGGGAATCTTCCACAATGGACGAAAGTCTGATGGAGCAACGCCGCGTGAGTGAAGAAGGGTTTCGGCTCGTAAAACTCTGTTGTTAAAGAAGAACATATCTGAGAGTAACTGTTCAGGTATTGACGGTATTTAACCAGAAAGCCACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTGTCCGGATTTATTGGGCGTAAAGCGAGCGCAGGCGGTTTTTTAAGTCTGATGTGAAAGCCTTCGGCTCAACCGAAGAAGTGCATCGGAAACTGGGAAACTTGAGTGCAGAAGAGGACAGTGGAACTCCATGTGTAGCGGTGAAATGCGTAGATATATGGAAGAACACCAGTGGCGAAGGCGGCTGTCTGGTCTGTAACTGACGCTGAGGCTCGAAAGTATGGGTAGCAAACAGGATTAGATACCCTGGTAGTCCATACCGTAAACGATGAATGCTAAGTGTTGGAGGGTTTCCGCCCTTCAGTGCTGCAGCTAACGCATTAAGCATTCCGCCTGGGGAGTACGGCCGCAAGGCTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCTACGCGAAGAACCTTACCAGGTCTTGACATACTATGCAAATCTAAGAGATTAGACGTTCCCTTCGGGGACATGGATACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTATTATCAGTTGCCAGCATTAAGTTGGGCACTCTGGTGAGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAATCATCATGCCCCTTATGACCTGGGCTACACACGTGCTACAATGGATGGTACAACGAGTTGCGAACTCGCGAGAGTAAGCTAATCTCTTAAAGCCATTCTCAGTTCGGATTGTAGGCTGCAACTCGCCTACATGAAGTCGGAATCGCTAGTAATCGCGGATCAGCATGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCATGAGAGTTTGTAACACCCAAAGTCGGTGGGGTAACCTTTTAGGAACCAGCCGCCTAAGGGGGCCCATTGC
in a second aspect, the invention also provides the use of the lactobacillus plantarum LP-28 strain described above for the production of gamma-aminobutyric acid.
In a third aspect, the invention also provides application of the lactobacillus plantarum LP-28 strain in preparing health care products for improving sleep quality.
The lactobacillus plantarum LP-28 strain provided by the invention can produce high-activity glutamate decarboxylase, and can directly utilize intestinal contents to produce GABA through a metabolic process in a complex intestinal environment of a human body, so that the lactobacillus plantarum LP-28 strain can be prepared into a sleep-aiding product, and the GABA produced by in vivo metabolism of the lactobacillus plantarum LP-28 strain is utilized to achieve the aim of improving sleep quality.
In a fourth aspect, the invention also provides a fermentation product of the above strain of Lactobacillus plantarum LP-28, said fermentation product being obtained by fermentation of a strain of Lactobacillus plantarum LP-28. The fermented product can be further processed to improve sleep quality.
The lactobacillus plantarum LP-28 strain can also be cultured by a culture medium, and Glu is decarboxylated to form GABA by utilizing the high activity of GAD. Therefore, the product for improving sleep quality can also be prepared by adopting an in-vitro fermentation culture mode and utilizing a fermentation product or a fermentation extract thereof.
In a fifth aspect, the present invention also provides a method for producing gamma-aminobutyric acid by using the lactobacillus plantarum LP-28 strain, which specifically comprises the following steps: seed culture is carried out on lactobacillus plantarum LP-28 strain in an improved MRS liquid culture medium, and anaerobic fermentation culture is carried out on the cultured fermentation broth on an intestinal simulated culture medium.
Further, the anaerobic fermentation culture conditions are as follows: the fermentation temperature is 35-37 ℃, the pH is 5-7, and the culture is carried out for more than 48 hours.
Alternatively, the method may specifically employ the following procedure:
Seed culture stage: inoculating lactobacillus plantarum LP-28 seed solution into an improved MRS liquid culture medium according to an inoculum size of 3%, shaking and mixing uniformly, culturing for 24 hours at 37 ℃, sequentially and continuously activating for three generations, taking 100ml of the cultured fermentation liquid, and obtaining a fermentation liquid stock solution sample, and refrigerating at 4 ℃ for later use.
CMGM composition (g/L): starch, 5.0; pectin, 2.0; guar gum, 1.0; mucin (porcine stomach type III), 4.0; xylan, 2.0; arabinogalactan, 2.0; inulin, 1.0; casein, 3.0; peptone water, 5.0; tryptone, 5.0; bile salts, 0.4; yeast extract, 4.5; feSO 4·7H2 O,0.005; sodium chloride, 4.5; potassium chloride ,4.5;KH2PO4,0.5;MgSO4·7H2O,1.25;CaCl2·6H2O,0.15;NaHCO3,1.5; cysteine, 0.8; heme, 0.05; tween 80,1.0.
The intestinal tract simulation culture medium is placed in an intestinal tract in-vitro fermentation simulation system fermentation tank and is used for carrying out fermentation culture on the fermentation liquid: the human colon three-stage continuous culture intestinal model system consists of three fermentors, a simulated proximal fermentor (V1, 280 mL), a transverse fermentor (V2, 300 mL) and a distal colon fermentor (V3, 320, mL), respectively. Three fermenters in series were maintained at 37 ℃ and pH values were maintained at 5.5 (V1), 6.2 (V2) and 6.8 (V3), respectively, and anaerobic conditions were created by continuous N 2 introduction. V1, V2 and V3 are all introduced CMGM by peristaltic pumps. Fresh healthy human fecal samples were collected, stored in an anaerobic cabinet (10% H 2、10% CO2、80% N2), and 1:5 (w/w) fecal dilutions in anaerobic PBS (0.1 mol/L PBS, pH 7.4) were prepared within 15 minutes after collection. Each fermenter of the intestinal model was inoculated with 100 mL faeces dilutions. The total system transit time was set to 48 hours based on the average retention time of healthy individuals. After inoculation with fecal dilutions, the intestinal model was run for 24 hours to stabilize the bacterial population prior to injection into the culture medium. After 24 hours, the system was run for 8 full capacity cycles to achieve steady state, and then the above fermentation broth was added to V1 in an amount of 1% (w/V) of the volume of the contents in the tank, followed by eight volume cycles to reach steady state, and further culture for 24 hours.
In a sixth aspect, the invention provides a probiotic EM28 comprising the above strain of Lactobacillus plantarum LP-28 and Lactobacillus rhamnosus LR-28.
In a seventh aspect, the present invention provides a yoghurt product prepared from the probiotic group EM28 as described above after fermentation of milk.
Lactobacillus rhamnosus LR-28 is preserved in China general microbiological culture Collection center (CGMCC) with a preservation number of 7.464 in the 6 th month of 2022; the preservation address is the Beijing Chaoyang district North Star West Leu 1, 3.
Microorganisms increase the expression of the proton respiratory chain complex while producing GABA, promote ATP synthesis, and up-regulate the activity of F1F0-ATP hydrolase, promote the ATP-dependent H + excretion process under acidic conditions, thereby changing the pH of the extracellular environment. More importantly, GABA is zwitterionic in the physiological environment and therefore plays a role in acid-base regulation.
The probiotic EM28 formed by combining lactobacillus rhamnosus LR-28 and lactobacillus plantarum LP-28 can produce synergistic effect in the process of fermenting milk, and the GABA yield is far higher than that of the pure superposition of the two bacteria. Therefore, the acidity of the yoghurt provided by the invention is finally between 90 and 100 ℃ and the pH value is between 4.25 and 4.35, and compared with the unflavoured yoghurt obtained by fermentation of other lactic acid bacteria, the yoghurt provided by the invention has proper acidity and does not need to be additionally added with additives to improve the flavor.
Further, the ratio of the viable count of the lactobacillus plantarum LP-28 strain to the lactobacillus rhamnosus LR-28 strain is: 1-3:4-7.
In different periods of yoghurt production, due to different pH values in the yoghurt, the metabolic states of the yoghurt and the yoghurt are different, and only if the two strains keep a good symbiotic relationship and a synergistic effect, the yoghurt can obtain good flavor. The lactobacillus plantarum LP-28 strain and lactobacillus rhamnosus LR-28 strain can maintain a good symbiotic relationship within the above proportion range, so that a synergistic effect is generated, and the yoghurt has good flavor.
Further, the total inoculation amount of the lactobacillus plantarum LP-28 strain and the lactobacillus rhamnosus LR-28 strain is 0.002-0.006%; the fermentation conditions are as follows: culturing at 38-43deg.C for 10-20 hr.
In the production of the yoghourt, the primary fermentation is mainly acid production, and the strain can utilize lactose metabolism to produce lactic acid, acetic acid, propionic acid and other acids to endow the yoghourt with certain acidity and cool mouthfeel. Therefore, the inoculation amount and fermentation conditions need to be strictly controlled to achieve the optimal acidity.
Further, one or more of food materials, stabilizers or sweeteners may be added to the yogurt material. The food material comprises fruits, grains, etc.
In an eighth aspect, the invention provides a yoghurt powder, which is prepared by spray drying the yoghurt product.
Drawings
FIG. 1 shows the bacterial morphology of Lactobacillus plantarum LP-28 strain in example 1 of the present invention;
FIG. 2 (a) shows the bacteriostatic ability of Lactobacillus plantarum LP-28 strain against E.coli;
FIG. 2 (b) shows the bacteriostatic ability of Lactobacillus plantarum LP-28 strain against Staphylococcus aureus;
FIG. 2 (c) shows the antibacterial ability of Lactobacillus plantarum LP-28 strain against Salmonella;
FIG. 3 is a sleep survey of group A people in example 7 of the present invention;
FIG. 4 is a sleep survey of group B people in example 7 of the present invention;
FIG. 5 is a sleep survey of group C people in example 7 of the present invention;
FIG. 6 shows the improvement of Hamiltonian anxiety among groups of people in example 7 of the present invention.
Detailed Description
The present invention will be described in further detail with reference to specific embodiments in order to make the objects, technical solutions and advantages of the present invention more apparent. It should be understood that the specific embodiments described herein are for purposes of illustration only and are not intended to limit the scope of the invention.
The sources of the strains used in the following examples of the invention are as follows:
Lactobacillus plantarum LP-28, which is classified under the name Lactobacillus plantarum (Lactobacillus plantarum), is preserved in China general microbiological culture Collection center (CGMCC) with the preservation number of CGMCC No.7.463 at the date of 25 and 6 in 2022; the preservation address is the Beijing Chaoyang district North Star West Leu 1, 3.
Lactobacillus rhamnosus LR-28, which is classified as lactobacillus rhamnosus (Lactobacillus rhamnosus) and is preserved in the China general microbiological culture Collection center (CGMCC) with the preservation number of 7.464 in the 6 th month of 2022; the preservation address is the Beijing Chaoyang district North Star West Leu 1, 3.
Modified MRS liquid Medium (g/L): peptone 8.0-10.0; 9.0 to 10.0 portions of beef powder; 3.0 to 5.0 percent of yeast powder; glucose 15.0-20.0; magnesium sulfate, 0.08-0.1; sodium acetate 1.8-2.0; ammonium citrate, 4.0-5.0; 1.6 to 2.0 portions of radon dipotassium phosphate; manganese sulfate, 0.03-0.05; tween 80,0.8-1.0; l-cysteine hydrochloride, 0.05-0.07 (pH 6.2.+ -. 0.2).
CMGM composition (g/L): starch, 5.0; pectin, 2.0; guar gum, 1.0; mucin (porcine stomach type III), 4.0; xylan, 2.0; arabinogalactan, 2.0; inulin, 1.0; casein, 3.0; peptone water, 5.0; tryptone, 5.0; bile salts, 0.4; yeast extract, 4.5; feSO 4·7H2 O,0.005; sodium chloride, 4.5; potassium chloride ,4.5;KH2PO4,0.5;MgSO4·7H2O,1.25;CaCl2·6H2O,0.15;NaHCO3,1.5; cysteine, 0.8; heme, 0.05; tween 80,1.0.
Intestinal tract in vitro fermentation simulation system: the human colon three-stage continuous culture intestinal model system consists of three fermentors, a simulated proximal fermentor (V1, 280 mL), a transverse fermentor (V2, 300 mL) and a distal colon fermentor (V3, 320, mL), respectively. Three fermenters in series were maintained at 37 ℃, pH was maintained at 5.5 (V1), 6.2 (V2) and 6.8 (V3), and anaerobic conditions were created by continuous N 2 introduction. V1, V2 and V3 are all introduced CMGM by peristaltic pumps. Fresh healthy human fecal samples were collected, stored in an anaerobic cabinet (10% H 2、10% CO2、80% N2), and 1:5 (w/w) fecal dilutions in anaerobic PBS (0.1 mol/L PBS pH 7.4) were prepared within 15 minutes after collection. Each fermenter of the intestinal model was inoculated with 100 mL faeces dilutions. The total system transit time was set to 48 hours based on the average retention time of healthy individuals. After inoculation with fecal dilutions, the intestinal model was run for 24 hours to stabilize the bacterial population prior to injection into the culture medium. After 24 hours, the system was run for 8 full capacity cycles to achieve steady state, and then the above fermentation broth was added to V1 in an amount of 1% (w/V) of the volume of the contents in the tank, followed by eight volume cycles to reach steady state, and further culture for 24 hours.
Other experimental materials and instruments were commercially available unless otherwise specified.
Example 1
The present example provides a strain of Lactobacillus plantarum LP-28 with reduced anxiety, which strain was selected by the following procedure:
1. isolation and screening of Lactobacillus plantarum LP-28
1.1 Bacterial source:
9 months 2019, tashikuer, tajike county, traditional yogurt in herlands.
1.2 Separation
Taking 10g of traditional yoghurt in the herdsman's home, adding the traditional yoghurt into 100ml of physiological saline, fully and uniformly mixing, and diluting the mixture, wherein the dilution gradient is 10 -1,10-2,10-3,10-4,10-5; each gradient was applied in 100. Mu.L to modified MRS solid medium supplemented with calcium carbonate, and each gradient was repeated. Placing in a constant temperature incubator at 37 ℃ for culturing for 48 hours.
Selecting a proper gradient (10 -5) from the above culture concentration gradients, selecting 15-25 single colonies with clear boundaries and transparent circles, subculturing the selected colonies on a newly prepared MRS solid culture medium by a streaking method for 24-48h, and repeating the steps three times again to obtain the monoclonal strain. After the strain is cultured for 24 hours by an MRS liquid culture medium, bacterial liquid and 50% glycerol are uniformly mixed according to a ratio of 1:1 (the final concentration of the glycerol is 25%), and the strain is preserved at-75 ℃, and simultaneously inoculated to an MRS solid culture medium test tube inclined plane for temporary preservation.
1.3 Preliminary screening of Strain
The glycerol-preserved bacterial liquid was inoculated into 1.5mL of MRS liquid medium at an inoculum size of 2% and placed in a constant temperature incubator at 37℃for 10 hours as seed bacterial liquid for the next experiment.
Inoculating the seed bacterial liquid into 2mL EP tube filled with MRS liquid culture medium according to the inoculation amount of 2%, and culturing for 48h at 37 ℃; picking single colony on MRS liquid culture medium, and culturing at 37deg.C for 16 hr; transferring to GYP liquid culture medium, standing at 37deg.C for 48 hr; taking 1 mu L of supernatant fluid of the fermentation liquor, spotting the supernatant fluid on filter paper (each 10 samples of +1 standard/piece of filter paper), centrifuging the supernatant fluid for 10min at 2000rpm, drying the filter paper after 4h of chromatography in a developing agent, spraying 8g/L ninhydrin on the filter paper, developing the color in a baking oven at 70 ℃ for 5min, and primarily screening 22 strains with gamma-aminobutyric acid (GABA) production.
1.4 Strain double Screen
Inoculating the seed bacterial liquid of 22 strains obtained by primary screening into 2mL EP tube filled with MRS liquid culture medium according to the inoculum size of 2%, and culturing for 48h at 37 ℃; picking single colony on MRS liquid culture medium, and culturing at 37deg.C for 16 hr; then transferring to GYP liquid culture medium, standing and culturing for 48h at 37 ℃. Centrifuging to obtain supernatant, and quantitatively determining GABA production capacity of 18 strains by adopting high performance liquid chromatography. The 22 strains obtained by separation have GABA producing capacity, wherein the strain numbered LP-28 has the strongest GABA producing capacity, and GABA yield is up to 3.653g/L.
1.5 Identification of LP-28 Strain
1.5.1 Colony morphology and fungus morphology characteristics of LP-28 Strain
Gram staining: positive; colony morphology observation: the surface is smooth and glossy; cell morphology:
The polymorphic bacilli are shown in FIG. 1.
1.5.2 Sequencing analysis of 16S rRNA of LP-28 Strain
TABLE 1 LP-28 similarity of 16S rRNA sequences of strains with reference strains
In summary, the identification of LP-28 as a Lactobacillus plantarum (Lactobacillus plantarum) strain was performed in combination with the results of 16S rRNA sequence analysis and physiological and biochemical identification.
The lactobacillus plantarum LP-28 strain is preserved in China general microbiological culture Collection center (CGMCC) at 25-6-2022, and the preservation number is CGMCC No.7.463; the preservation address is the Beijing Chaoyang district North Star West Leu 1, 3.
2. Gastrointestinal digestion resistance of Lactobacillus plantarum LP-28 Strain
Gastric juice is simulated manually: taking 16.4mL of dilute hydrochloric acid, adding 800mL of water and 10g of protease, shaking uniformly, adding water to dilute into 1000mL, mixing uniformly, and filtering by a sterile filter membrane with the thickness of 0.22 mu m.
Manually simulating intestinal juice: taking 6.8g of monopotassium phosphate, adding 500mL of water to dissolve the monopotassium phosphate, and adjusting the pH to 6.8 by using 0.1mol/L sodium hydroxide solution; and (3) taking 10g of pancreatin, adding a proper amount of water to dissolve, mixing the two solutions, adding water to dilute to 1000mL, uniformly mixing, and filtering with a 0.22 mu m sterile filter membrane to obtain the pancreatic enzyme.
The lactobacillus plantarum LP-28 screened in the example 1 is cultured and activated with common commercial lactobacillus plantarum to obtain an inoculation liquid with the bacterial liquid concentration of 1 multiplied by 10 9 CFU/mL.
Gastric acid tolerance test: 1mL of Lactobacillus plantarum LP-28 strain inoculum and common commercial Lactobacillus plantarum inoculum (strain No. ATCC 8014) were inoculated and respectively preheated to a test tube filled with 9mL of simulated gastric fluid, cultured in an incubator at 37℃and 80r/min, and viable counts of 0, 2 and 4 hours were determined (plate count method).
Bile salt resistance test: 1mL of Lactobacillus plantarum LP-28 strain inoculum and common commercial Lactobacillus plantarum inoculum (strain No. ATCC 8014) were inoculated and respectively preheated to a test tube filled with 9mL of simulated intestinal fluid, and cultured in an incubator at 37℃and 80r/min, and viable counts of 0, 2 and 4 hours were measured (plate count method).
TABLE 2 survival rate of strains by simulation of gastrointestinal digests (100 mL bacteria count)
Compared with common commercial strains, the lactobacillus plantarum LP-28 has very high acid resistance and bile salt resistance, can resist artificial gastric juice and artificial intestinal juice, and ensures that enough viable bacteria are planted in intestinal tracts after the gastric and intestinal digestions.
3. High adhesion capability of lactobacillus plantarum LP-28 to Caco-2 cells
Inoculating Caco-2 cells with the concentration of 1X 10/mL at one time to a 12-well plate, culturing to form single-layer cells under the condition of 37 ℃ and 5% CO, discarding culture solution, lightly washing three times by using sterile PBS buffer solution, adding 1mL of DMEM stock solution, respectively adding lactobacillus plantarum LP-28 strain inoculating solution and 100 mu L of common commercial lactobacillus plantarum (strain number ATCC 8014) bacterial suspension (suspended by DMEM, the concentration is about 10 0 cfu/mL), making three holes for each strain, after culturing for 2 hours, discarding supernatant, washing three times by using sterile PBS, adding 300 mu L of 0.25% pancreatin into each hole to digest for 5-8min, adding 300 mu L of incomplete DMEM culture medium (containing serum without double antibodies), and then performing plate dilution coating to calculate the number of adhered strains.
Adhesion (%) = number of adhesion strains/initial inoculation number x 100%.
TABLE 3 cell adhesion ability of strains
Compared with common commercial lactobacillus plantarum, the lactobacillus plantarum LP-28 can be more effectively adhered to intestinal mucosa to form a biological barrier, protect intestinal tracts and provide important preconditions for colonisation in human intestinal tracts.
4. Antibacterial ability of Lactobacillus plantarum LP-28 Strain
The activated Lactobacillus plantarum LP-28 was inoculated at an inoculum size of 2% into 2mL EP tubes containing MRS liquid medium and incubated in a 37℃incubator for 18h. Taking 500g of lactobacillus plantarum LP-28 culture solution, and centrifuging for 10min to obtain a supernatant for later use.
Coli (ATCC 25922), staphylococcus aureus (ATCC 43300) and salmonella (ATCC 50760) were used as indicator bacteria. Taking 100 mu L of activated indicator bacteria, adjusting the concentration of the indicator bacteria to 1.0X10 5 CFU/mL, coating a plate on a corresponding culture medium, placing the coated culture dish on a semi-open working table for 10min, adding 200 mu L of lactobacillus plantarum LP-28 supernatant into oxford cups placed on a solid culture medium, culturing at 37 ℃ for 16h steadily, observing and measuring the diameter of a bacteriostasis ring of each bacterial liquid, and performing three groups of experiments in parallel.
The results show that the lactobacillus plantarum LP-28 can inhibit the growth of escherichia coli, staphylococcus aureus and salmonella, and has antibacterial effect. The lactobacillus plantarum LP-28 can be effectively adhered to human intestinal tracts, short-chain fatty acids (such as acetic acid and lactic acid), hydrogen peroxide, bacteriocin and the like generated by the lactobacillus plantarum LP-28 can effectively inhibit the growth of putrefying bacteria and the generation of harmful metabolites such as amine, phenol, indole and the like, and reduce the pH value in the intestinal tracts.
Example 2
The embodiment provides a fermentation product of lactobacillus plantarum LP-28 strain, which is prepared by the following steps: inoculating seed solution of lactobacillus plantarum LP-28 strain into an improved MRS liquid culture medium according to an inoculum size of 3%, shaking and mixing uniformly, culturing for 24 hours at 37 ℃, and sequentially and continuously activating for three generations to obtain fermentation liquor.
Taking the cultured fermentation liquor, adding the fermentation liquor into a fermentation tank (V1) of an intestinal in-vitro fermentation simulation system which is connected with CMGM and fecal diluent and completes 8 full-capacity turnover by adding the fermentation liquor into the tank in an adding amount of 1% (w/V) of the volume of the content, performing fermentation culture, performing volume turnover for eight times, reaching a steady state, centrifuging, and taking supernatant.
Considering the operating volume (900 mL) and retention time (48 h) of the colon model system, fermentation broth was added to V1 at 1% (w/V) per day, followed by eight additional volume turnovers to steady state, centrifugation, and supernatant removal.
The GABA content in the fermentation broth obtained in example 2 was quantitatively determined by high performance liquid chromatography, and the GABA content was 10.+ -. 4.2mg/100ml. Lactobacillus plantarum LP-28 was shown to be capable of producing GABA under in vitro simulated fermentation conditions in the intestinal tract.
Example 3
The present example provides a probiotic EM28 comprising Lactobacillus plantarum LP-28 strain obtained in example 1 in combination with Lactobacillus rhamnosus LR-28 strain. The mass ratio of the lactobacillus plantarum LP-28 strain to the lactobacillus rhamnosus LR-28 strain is 1:2.5.
Example 4
This example provides a yoghurt prepared from milk fermented with the probiotic EM28 of example 3.
Is prepared from the following raw materials: raw milk: 865.5g; white granulated sugar: 110g; cream: 5g; starch: 16g; sodium glutamate: 1g; sodium bicarbonate: 1.3g; probiotic EM28:0.035g (Lactobacillus plantarum LP-28 strain: 0.01g; lactobacillus rhamnosus LR-28 strain: 0.025 g).
Preparing yoghourt: mixing raw milk, white granulated sugar, cream and starch, sterilizing at 95deg.C for 5min, cooling to 40deg.C, adding sodium glutamate and sodium bicarbonate, inoculating lactobacillus plantarum LP-28 strain and lactobacillus rhamnosus LR-28 strain, and sealing for fermentation. The fermentation is carried out for 7h for the first time, the fermentation termination time is predicted according to the acid rising condition, the total fermentation time is about 10-11h, and the fermentation is terminated when the pH value is 4.30+/-0.05 (the pH value is used as the standard for judging the fermentation end point). Demulsification stirring after fermentation, cooling and filling. The final viable bacteria content of the obtained yoghourt is as follows: 1.0X10 8 CFU/ml; the acidity is: 98 deg.t.
Example 5
This example provides a yoghurt prepared from milk fermented with the probiotic EM28 of example 3.
The preparation method comprises the following steps: whole milk powder: 103g; water: 762.5g; white granulated sugar: 110g; cream: 5g; starch: 16g; sodium glutamate: 1g; sodium bicarbonate: 1.3g; probiotic EM28:0.035g (Lactobacillus plantarum LP-28 strain: 0.01g; lactobacillus rhamnosus LR-28 strain: 0.025 g).
Preparing yoghourt: mixing whole milk powder, water, white sugar, cream and starch, sterilizing at 95deg.C for 5min, cooling to 40deg.C, adding sodium glutamate and sodium bicarbonate, inoculating lactobacillus plantarum strain LP-28 and lactobacillus rhamnosus strain LR-28, and sealing for fermentation. The fermentation is carried out for 7h for the first time, the fermentation termination time is predicted according to the acid rising condition, the total fermentation time is about 10-11h, and the fermentation is terminated when the pH value is 4.30+/-0.05 (the pH value is used as the standard for judging the fermentation end point). Demulsification stirring after fermentation, cooling and filling. The final viable bacteria content of the obtained yoghourt is as follows: 1.0X10 8 CFU/ml; the acidity is: 97T.
Example 6
This example provides a yoghurt prepared from milk fermented with the probiotic EM28 of example 3.
Is prepared from the following raw materials: skimmed milk powder: 98g; water: 767.5g; white granulated sugar: 110g; cream: 5g; starch: 16g; sodium glutamate: 1g; sodium bicarbonate: 1.3g; probiotic EM28:0.035g (Lactobacillus plantarum LP-28 strain: 0.01g; lactobacillus rhamnosus LR-28 strain: 0.025 g).
Preparing yoghourt: mixing skimmed milk powder, water, white sugar, cream and starch, sterilizing at 95deg.C for 5min, cooling to 40deg.C, adding sodium glutamate and sodium bicarbonate, inoculating lactobacillus plantarum strain LP-28 and lactobacillus rhamnosus strain LR-28, and sealing for fermentation. The fermentation is carried out for 7h for the first time, the fermentation termination time is predicted according to the acid rising condition, the total fermentation time is about 10-11h, and the fermentation is terminated when the pH value is 4.30+/-0.05 (the pH value is used as the standard for judging the fermentation end point). Demulsification stirring after fermentation, cooling and filling. The final viable bacteria content of the obtained yoghourt is as follows: 1.0X10 8 CFU/ml; the acidity is: 95T.
Example 7
The present example provides the use of lactobacillus plantarum LP-28 strain as a raw material for the preparation of a product for improving sleep quality.
Hamilton anxiety scale (Hamilton Anxiey Scale, HAMA), which was one of the scales commonly used in psychiatric clinics, was compiled by Hamilton in 1959 and was used primarily to assess the severity of anxiety in neurological and other patients.
1. Test procedure
Insomnia patients enrolled 195 for anxiety symptoms (screening criteria: HAMA (Hamiltonian anxiety) > 7 population screening into groups), group A (20-35 years) were grouped by age: 60 people; group B (35-59 years): 70 people; group C (over 60 years): 65 persons. The yogurt obtained in example 5 was administered to patients suffering from anxiety and insomnia, 250ml per day, with a period of 2 weeks. The anxiety symptoms of the three groups are investigated, and the hamiltonian anxiety scale is investigated.
2. Test results and analysis
2.1 Sleep findings for 2 weeks:
group A has 60 anxiety symptoms, 38 people have obviously improved sleep quality, 22 people have no obvious effect, and the effective rate is 63.30 percent (shown in figure 1);
group B has 70 anxiety symptoms, 32 people have obviously improved sleep quality, 38 people have no obvious effect, and the effective rate is 45.70 percent (shown in figure 2);
Group C has 65 anxiety symptoms, 36 people have obviously improved sleep quality, 29 people have no obvious effect, and the effective rate is 55.30 percent (shown in figure 3);
2.2A, B, C results of three group of symptomatic improvement studies
Table 4A group results of symptomatic improvement study
Table 5B group results of symptomatic improvement study
Table 6C group results of symptomatic improvement study
2.3 Improvement amount was determined using hamiltonian anxiety scale (HAMA) investigation
The results are shown in Table 7 and FIG. 4.
Tables 7A and B, C three groups were examined using hamiltonian anxiety scale (HAMA) to determine improvement
From the above tables 5 to 7, it can be seen that the yogurt provided by the invention has good improving effects in relieving dysphoria and improving sleep quality.
In conclusion, the lactobacillus plantarum LP-28 strain provided by the invention has the capability of high-yielding GABA in the human intestinal environment, can improve and regulate the balance of intestinal microbial flora, and can enhance the immunity of organisms.
The foregoing description of the preferred embodiments of the invention is not intended to be limiting, but rather is intended to cover all modifications, equivalents, or alternatives falling within the spirit and principles of the invention.
Claims (7)
1. The application of the lactobacillus plantarum LP-28 strain in preparing health care products for improving sleep quality is characterized in that the lactobacillus plantarum LP-28 strain is classified as lactobacillus plantarum (Lactobacillus plantarum) which is preserved in China general microbiological culture Collection center (CGMCC) No.7.463 in the 6 th month 25 of 2022; the preservation address is the Beijing Chaoyang district North Star West Leu 1, 3.
2. Use of the lactobacillus plantarum LP-28 strain as claimed in claim 1, for the production of gamma-aminobutyric acid.
3. A method for producing gamma-aminobutyric acid using lactobacillus plantarum LP-28 strain as claimed in claim 1, characterized in that the method comprises in particular the following operations: seed culture is carried out on lactobacillus plantarum LP-28 strain in an improved MRS liquid culture medium, and anaerobic fermentation culture is carried out on the cultured fermentation broth on an intestinal simulated culture medium.
4. A method for producing gamma-aminobutyric acid by lactobacillus plantarum LP-28 strain as claimed in claim 3, wherein the anaerobic fermentation culture conditions are: the fermentation temperature is 35-40 ℃, the pH is 4.0-4.7, and the culture is carried out for more than 18 hours.
5. A yoghurt product, which is prepared by fermenting a milk product with the lactobacillus plantarum LP-28 strain and the lactobacillus rhamnosus (Lactobacillus rhamnosus) LR-28 strain of claim 1, wherein the lactobacillus rhamnosus LR-28 strain is preserved in the general microbiological center of the China general microbiological culture collection center (cmv) at the month 26 of 2022, and the preservation number is CGMCC No.7.464; the preservation address is the Beijing Chaoyang district North Star West Leu 1, 3.
6. Yoghurt preparation in accordance with claim 5, characterized in that the ratio of viable count of lactobacillus plantarum LP-28 strain to lactobacillus rhamnosus LR-28 strain is: 1-3:4-7; and/or
The total inoculation amount of the lactobacillus plantarum LP-28 strain and lactobacillus rhamnosus LR-28 strain is 0.002-0.006%; the fermentation conditions are as follows: culturing at 38-43deg.C for 10-20 hr; and/or
The raw materials for preparing the yoghurt product also comprise one or more than two of food raw materials, stabilizers or sweeteners.
7. Yoghurt powder, characterized in that it is obtained from a yoghurt product as claimed in claim 5 by spray drying.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310645941.XA CN116676226B (en) | 2023-05-23 | 2023-05-23 | Lactobacillus plantarum LP-28 for relieving anxiety, yoghourt and application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310645941.XA CN116676226B (en) | 2023-05-23 | 2023-05-23 | Lactobacillus plantarum LP-28 for relieving anxiety, yoghourt and application |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN116676226A CN116676226A (en) | 2023-09-01 |
| CN116676226B true CN116676226B (en) | 2024-04-26 |
Family
ID=87790252
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202310645941.XA Active CN116676226B (en) | 2023-05-23 | 2023-05-23 | Lactobacillus plantarum LP-28 for relieving anxiety, yoghourt and application |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN116676226B (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117887608B (en) * | 2023-06-19 | 2024-08-23 | 河北源民生物科技有限公司 | Lactic acid bacteria YM-3 with blood lipid reducing and body fat reducing effects, and preparation method and application thereof |
| CN117327631B (en) * | 2023-11-29 | 2024-01-26 | 山东环亿生物科技有限公司 | Lactobacillus plantarum strain 10A-2 for improving anxiety and depression and application thereof |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109136130A (en) * | 2018-08-27 | 2019-01-04 | 南昌大学 | One plant of Lactobacillus rhamnosus NCU2217 |
| CN111728111A (en) * | 2020-06-28 | 2020-10-02 | 武汉康复得生物科技股份有限公司 | Probiotic composition for relieving anxiety or depression and application thereof |
| CN112501063A (en) * | 2020-12-03 | 2021-03-16 | 兰州大学 | Lactobacillus plantarum LZU-J-TSL6 capable of highly producing GABA and application thereof |
| CN115161250A (en) * | 2022-09-06 | 2022-10-11 | 广东益可维生物技术有限公司 | Lactobacillus plantarum with functions of improving sleep, strengthening brain and improving intelligence and application thereof |
| CN115521889A (en) * | 2022-10-30 | 2022-12-27 | 江南大学 | Lactobacillus plantarum WL02 capable of producing gamma-aminobutyric acid and application thereof |
| CN116676225A (en) * | 2023-05-23 | 2023-09-01 | 河北源民生物科技有限公司 | Lactobacillus rhamnosus LR-28 strain with nerve soothing and sleep aiding effects, fermentation product, hypnotic fungus group mixture and application |
-
2023
- 2023-05-23 CN CN202310645941.XA patent/CN116676226B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109136130A (en) * | 2018-08-27 | 2019-01-04 | 南昌大学 | One plant of Lactobacillus rhamnosus NCU2217 |
| CN111728111A (en) * | 2020-06-28 | 2020-10-02 | 武汉康复得生物科技股份有限公司 | Probiotic composition for relieving anxiety or depression and application thereof |
| CN112501063A (en) * | 2020-12-03 | 2021-03-16 | 兰州大学 | Lactobacillus plantarum LZU-J-TSL6 capable of highly producing GABA and application thereof |
| CN115161250A (en) * | 2022-09-06 | 2022-10-11 | 广东益可维生物技术有限公司 | Lactobacillus plantarum with functions of improving sleep, strengthening brain and improving intelligence and application thereof |
| CN115521889A (en) * | 2022-10-30 | 2022-12-27 | 江南大学 | Lactobacillus plantarum WL02 capable of producing gamma-aminobutyric acid and application thereof |
| CN116676225A (en) * | 2023-05-23 | 2023-09-01 | 河北源民生物科技有限公司 | Lactobacillus rhamnosus LR-28 strain with nerve soothing and sleep aiding effects, fermentation product, hypnotic fungus group mixture and application |
Non-Patent Citations (1)
| Title |
|---|
| 石慧等.《食品分子微生物学》.中国农业大学出版社,2019,(2019年5月第1版),第79页. * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN116676226A (en) | 2023-09-01 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN111575207B (en) | Lactobacillus paracasei separated from yak yogurt and application thereof | |
| CN116676226B (en) | Lactobacillus plantarum LP-28 for relieving anxiety, yoghourt and application | |
| US20230413837A1 (en) | Streptococcus thermophilus producing gamma-aminobutyric acid and application thereof | |
| CN110106119B (en) | Lactobacillus rhamnosus M9 separated from breast milk and application thereof | |
| CN116676225B (en) | Lactobacillus rhamnosus LR-28 strain with nerve soothing and sleep aiding effects, fermentation product, hypnotic fungus group mixture and application | |
| WO2020140319A1 (en) | Lactobacillus paracasei and use thereof | |
| CN101331900A (en) | Four kinds of lactobacillus for generating bilesalt hydrolase and extracellular polysaccharide and functionality yoghourt production technique thereof | |
| CN110157650B (en) | Bifidobacterium lactis M8 separated from breast milk and application thereof | |
| CN115261264A (en) | Lactobacillus paracasei PC804 and application thereof | |
| CN116814481B (en) | Probiotic Lactobacillus paracasei PC646 derived from fermented mare milk and artificial intelligent screening method thereof | |
| CN115340965A (en) | Lactobacillus paracasei PC724 and application thereof | |
| CN113444664B (en) | Lactobacillus brevis for producing gamma-aminobutyric acid and application thereof | |
| CN110129220A (en) | A kind of lactobacillus bulgaricus BSTS6-4 and its application | |
| CN113355375B (en) | Method for synthesizing B vitamins by fermenting lactobacillus paracasei K56 and application | |
| CN109810917A (en) | Lactobacillus salivarius and its application | |
| CN113755370A (en) | Application of lactobacillus acidophilus LA85 in preparation of blood fat reducing medicines or health-care foods | |
| CN116769654B (en) | Bifidobacterium animalis subspecies lactis and application thereof | |
| CN104774783A (en) | Lactobacillus casei capable of reducing cholesterol and method for producing probiotic yogurt from same | |
| CN111631262A (en) | Functional polysaccharide yoghourt starter and preparation method of functional polysaccharide yoghourt | |
| CN114947027B (en) | Preparation method of fermented fruit and vegetable juice rich in gamma-aminobutyric acid, acetic acid and lactic acid | |
| CN113817632B (en) | Streptococcus thermophilus, sleep-aiding fermented milk base material rich in GABA (gamma-amino acid residues), lactobacillus beverage and preparation method | |
| CN113355374B (en) | Method for synthesizing B vitamins by fermenting lactobacillus paracasei ET-22 and application | |
| CN115895974A (en) | Lactobacillus plantarum rich in selenium and capable of producing gamma-aminobutyric acid at high yield and application of lactobacillus plantarum | |
| CN107788119A (en) | A kind of application for the method and the Paenibacillus polymyxa that acidified milk is prepared using Paenibacillus polymyxa | |
| CN109706098B (en) | Lactobacillus paracasei and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |