CN116555102B - 一种产γ-氨基丁酸的植物乳杆菌、助睡眠益生菌组合物及其制备方法 - Google Patents
一种产γ-氨基丁酸的植物乳杆菌、助睡眠益生菌组合物及其制备方法 Download PDFInfo
- Publication number
- CN116555102B CN116555102B CN202310494077.8A CN202310494077A CN116555102B CN 116555102 B CN116555102 B CN 116555102B CN 202310494077 A CN202310494077 A CN 202310494077A CN 116555102 B CN116555102 B CN 116555102B
- Authority
- CN
- China
- Prior art keywords
- lactobacillus plantarum
- fphc0500
- culture
- aminobutyric acid
- sleep
- 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
- BTCSSZJGUNDROE-UHFFFAOYSA-N gamma-aminobutyric acid Chemical compound NCCCC(O)=O BTCSSZJGUNDROE-UHFFFAOYSA-N 0.000 title claims abstract description 88
- 240000006024 Lactobacillus plantarum Species 0.000 title claims abstract description 84
- 235000013965 Lactobacillus plantarum Nutrition 0.000 title claims abstract description 80
- 229940072205 lactobacillus plantarum Drugs 0.000 title claims abstract description 80
- OGNSCSPNOLGXSM-UHFFFAOYSA-N (+/-)-DABA Natural products NCCC(N)C(O)=O OGNSCSPNOLGXSM-UHFFFAOYSA-N 0.000 title claims abstract description 44
- 229960003692 gamma aminobutyric acid Drugs 0.000 title claims abstract description 44
- 239000000203 mixture Substances 0.000 title claims abstract description 28
- 239000006041 probiotic Substances 0.000 title claims abstract description 24
- 235000018291 probiotics Nutrition 0.000 title claims abstract description 24
- 230000000529 probiotic effect Effects 0.000 title claims abstract description 22
- 238000002360 preparation method Methods 0.000 title claims abstract description 16
- 238000004321 preservation Methods 0.000 claims abstract description 13
- 230000002906 microbiologic effect Effects 0.000 claims abstract description 4
- 238000009629 microbiological culture Methods 0.000 claims abstract description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 39
- 239000007788 liquid Substances 0.000 claims description 37
- 239000000284 extract Substances 0.000 claims description 29
- 238000000855 fermentation Methods 0.000 claims description 25
- 230000004151 fermentation Effects 0.000 claims description 25
- 238000001035 drying Methods 0.000 claims description 20
- 239000000047 product Substances 0.000 claims description 20
- 239000000843 powder Substances 0.000 claims description 18
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 16
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 16
- 239000001963 growth medium Substances 0.000 claims description 16
- LXNHXLLTXMVWPM-UHFFFAOYSA-N pyridoxine Chemical compound CC1=NC=C(CO)C(CO)=C1O LXNHXLLTXMVWPM-UHFFFAOYSA-N 0.000 claims description 16
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 15
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 15
- 239000003814 drug Substances 0.000 claims description 15
- 239000012046 mixed solvent Substances 0.000 claims description 15
- OKIXYCVPKXYQRH-UHFFFAOYSA-N ethanol;ethyl acetate;propan-2-one Chemical compound CCO.CC(C)=O.CCOC(C)=O OKIXYCVPKXYQRH-UHFFFAOYSA-N 0.000 claims description 14
- 239000000654 additive Substances 0.000 claims description 13
- 238000003756 stirring Methods 0.000 claims description 13
- NLXLAEXVIDQMFP-UHFFFAOYSA-N Ammonia chloride Chemical compound [NH4+].[Cl-] NLXLAEXVIDQMFP-UHFFFAOYSA-N 0.000 claims description 12
- 230000001580 bacterial effect Effects 0.000 claims description 12
- 238000010438 heat treatment Methods 0.000 claims description 12
- 239000002609 medium Substances 0.000 claims description 12
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 claims description 11
- 239000001110 calcium chloride Substances 0.000 claims description 11
- 229910001628 calcium chloride Inorganic materials 0.000 claims description 11
- 229960002713 calcium chloride Drugs 0.000 claims description 11
- 238000001914 filtration Methods 0.000 claims description 11
- 238000002156 mixing Methods 0.000 claims description 11
- 241000894006 Bacteria Species 0.000 claims description 10
- 241000234435 Lilium Species 0.000 claims description 10
- 244000197580 Poria cocos Species 0.000 claims description 10
- 235000008599 Poria cocos Nutrition 0.000 claims description 10
- 230000004913 activation Effects 0.000 claims description 10
- 230000000996 additive effect Effects 0.000 claims description 10
- 235000011148 calcium chloride Nutrition 0.000 claims description 10
- 238000004108 freeze drying Methods 0.000 claims description 10
- 229930003451 Vitamin B1 Natural products 0.000 claims description 8
- 238000012258 culturing Methods 0.000 claims description 8
- 238000000034 method Methods 0.000 claims description 8
- RADKZDMFGJYCBB-UHFFFAOYSA-N pyridoxal hydrochloride Natural products CC1=NC=C(CO)C(C=O)=C1O RADKZDMFGJYCBB-UHFFFAOYSA-N 0.000 claims description 8
- 239000011780 sodium chloride Substances 0.000 claims description 8
- 229960003495 thiamine Drugs 0.000 claims description 8
- DPJRMOMPQZCRJU-UHFFFAOYSA-M thiamine hydrochloride Chemical compound Cl.[Cl-].CC1=C(CCO)SC=[N+]1CC1=CN=C(C)N=C1N DPJRMOMPQZCRJU-UHFFFAOYSA-M 0.000 claims description 8
- 235000010374 vitamin B1 Nutrition 0.000 claims description 8
- 239000011691 vitamin B1 Substances 0.000 claims description 8
- 235000019158 vitamin B6 Nutrition 0.000 claims description 8
- 239000011726 vitamin B6 Substances 0.000 claims description 8
- 229940011671 vitamin b6 Drugs 0.000 claims description 8
- 229920001661 Chitosan Polymers 0.000 claims description 7
- 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 claims description 7
- 239000004372 Polyvinyl alcohol Substances 0.000 claims description 7
- 230000003213 activating effect Effects 0.000 claims description 7
- 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 claims description 7
- 229960001031 glucose Drugs 0.000 claims description 7
- 239000008103 glucose Substances 0.000 claims description 7
- 125000004029 hydroxymethyl group Chemical group [H]OC([H])([H])* 0.000 claims description 7
- 238000011081 inoculation Methods 0.000 claims description 7
- 229920002451 polyvinyl alcohol Polymers 0.000 claims description 7
- 229960002901 sodium glycerophosphate Drugs 0.000 claims description 7
- 239000008223 sterile water Substances 0.000 claims description 7
- LEHOTFFKMJEONL-UHFFFAOYSA-N Uric Acid Chemical compound N1C(=O)NC(=O)C2=C1NC(=O)N2 LEHOTFFKMJEONL-UHFFFAOYSA-N 0.000 claims description 6
- TVWHNULVHGKJHS-UHFFFAOYSA-N Uric acid Natural products N1C(=O)NC(=O)C2NC(=O)NC21 TVWHNULVHGKJHS-UHFFFAOYSA-N 0.000 claims description 6
- VSCWAEJMTAWNJL-UHFFFAOYSA-K aluminium trichloride Chemical compound Cl[Al](Cl)Cl VSCWAEJMTAWNJL-UHFFFAOYSA-K 0.000 claims description 6
- 229960001040 ammonium chloride Drugs 0.000 claims description 6
- 235000019270 ammonium chloride Nutrition 0.000 claims description 6
- 238000000605 extraction Methods 0.000 claims description 6
- 229910052751 metal Inorganic materials 0.000 claims description 6
- 239000002184 metal Substances 0.000 claims description 6
- 229960002668 sodium chloride Drugs 0.000 claims description 6
- 229940116269 uric acid Drugs 0.000 claims description 6
- 238000009835 boiling Methods 0.000 claims description 5
- 238000004140 cleaning Methods 0.000 claims description 5
- 239000000706 filtrate Substances 0.000 claims description 5
- 235000001727 glucose Nutrition 0.000 claims description 5
- 238000004519 manufacturing process Methods 0.000 claims description 5
- 230000000813 microbial effect Effects 0.000 claims description 5
- 230000002829 reductive effect Effects 0.000 claims description 5
- 150000003839 salts Chemical class 0.000 claims description 5
- 239000002904 solvent Substances 0.000 claims description 5
- 230000001954 sterilising effect Effects 0.000 claims description 5
- 238000005406 washing Methods 0.000 claims description 5
- 239000002994 raw material Substances 0.000 claims description 4
- 108020004465 16S ribosomal RNA Proteins 0.000 claims description 3
- 239000000049 pigment Substances 0.000 claims description 3
- 241000192125 Firmicutes Species 0.000 claims description 2
- 230000008569 process Effects 0.000 claims description 2
- 239000012266 salt solution Substances 0.000 claims description 2
- FMYHFHVAPZDDPJ-UHFFFAOYSA-N ethanol;ethyl acetate;hydrate Chemical compound O.CCO.CCOC(C)=O FMYHFHVAPZDDPJ-UHFFFAOYSA-N 0.000 claims 1
- 230000004083 survival effect Effects 0.000 abstract description 10
- 210000004211 gastric acid Anatomy 0.000 abstract description 6
- 229960001231 choline Drugs 0.000 abstract description 3
- OEYIOHPDSNJKLS-UHFFFAOYSA-N choline Chemical compound C[N+](C)(C)CCO OEYIOHPDSNJKLS-UHFFFAOYSA-N 0.000 abstract description 3
- 230000000052 comparative effect Effects 0.000 description 32
- QZAYGJVTTNCVMB-UHFFFAOYSA-N serotonin Chemical compound C1=C(O)C=C2C(CCN)=CNC2=C1 QZAYGJVTTNCVMB-UHFFFAOYSA-N 0.000 description 10
- 241000699670 Mus sp. Species 0.000 description 9
- 239000000243 solution Substances 0.000 description 9
- 240000001046 Lactobacillus acidophilus Species 0.000 description 6
- 235000013956 Lactobacillus acidophilus Nutrition 0.000 description 6
- 239000003833 bile salt Substances 0.000 description 6
- 229940039695 lactobacillus acidophilus Drugs 0.000 description 6
- 239000013641 positive control Substances 0.000 description 6
- 238000012216 screening Methods 0.000 description 6
- 230000000694 effects Effects 0.000 description 5
- 230000006872 improvement Effects 0.000 description 5
- 238000009630 liquid culture Methods 0.000 description 5
- 210000004556 brain Anatomy 0.000 description 4
- 210000004027 cell Anatomy 0.000 description 4
- 235000013305 food Nutrition 0.000 description 4
- 230000036541 health Effects 0.000 description 4
- 230000015784 hyperosmotic salinity response Effects 0.000 description 4
- 230000000147 hypnotic effect Effects 0.000 description 4
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 4
- 230000001737 promoting effect Effects 0.000 description 4
- 230000001105 regulatory effect Effects 0.000 description 4
- 239000007787 solid Substances 0.000 description 4
- 241000196324 Embryophyta Species 0.000 description 3
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 3
- 241001619461 Poria <basidiomycete fungus> Species 0.000 description 3
- 239000007864 aqueous solution Substances 0.000 description 3
- 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 3
- 238000011049 filling Methods 0.000 description 3
- 230000002496 gastric effect Effects 0.000 description 3
- 235000013923 monosodium glutamate Nutrition 0.000 description 3
- 238000005070 sampling Methods 0.000 description 3
- 229940073490 sodium glutamate Drugs 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- HOMSOWZTBJWNHP-UHFFFAOYSA-N 5-chlorothiadiazole Chemical compound ClC1=CN=NS1 HOMSOWZTBJWNHP-UHFFFAOYSA-N 0.000 description 2
- 229920001817 Agar Polymers 0.000 description 2
- 101000831205 Danio rerio Dynein axonemal assembly factor 11 Proteins 0.000 description 2
- 102100024282 Dynein axonemal assembly factor 11 Human genes 0.000 description 2
- 102000018997 Growth Hormone Human genes 0.000 description 2
- 108010051696 Growth Hormone Proteins 0.000 description 2
- 241001559542 Hippocampus hippocampus Species 0.000 description 2
- 101000831210 Homo sapiens Dynein axonemal assembly factor 11 Proteins 0.000 description 2
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 2
- 241000699666 Mus <mouse, genus> Species 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- 239000008272 agar Substances 0.000 description 2
- 229940093761 bile salts Drugs 0.000 description 2
- 238000010170 biological method Methods 0.000 description 2
- 230000036772 blood pressure Effects 0.000 description 2
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 description 2
- 235000015140 cultured milk Nutrition 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- VYFYYTLLBUKUHU-UHFFFAOYSA-N dopamine Chemical compound NCCC1=CC=C(O)C(O)=C1 VYFYYTLLBUKUHU-UHFFFAOYSA-N 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 210000004051 gastric juice Anatomy 0.000 description 2
- 239000000122 growth hormone Substances 0.000 description 2
- 238000000338 in vitro Methods 0.000 description 2
- 239000002054 inoculum Substances 0.000 description 2
- 230000006799 invasive growth in response to glucose limitation Effects 0.000 description 2
- 239000004310 lactic acid Substances 0.000 description 2
- 235000014655 lactic acid Nutrition 0.000 description 2
- 239000003960 organic solvent Substances 0.000 description 2
- VDYCLYGKCGVBHN-UHFFFAOYSA-N pachymaic acid Natural products CC12CCC(OC(C)=O)C(C)(C)C1CCC1=C2CCC2(C)C(C(CCC(=C)C(C)C)C(O)=O)C(O)CC21C VDYCLYGKCGVBHN-UHFFFAOYSA-N 0.000 description 2
- SRDNLMOBFKJOSD-UHFFFAOYSA-N pachymic acid Natural products CC12CCC(OC(C)=O)C(C)(C)C1CCC1=C2CCC2(C)C(C(CCC=C(C)C)C(O)=O)C(O)CC21C SRDNLMOBFKJOSD-UHFFFAOYSA-N 0.000 description 2
- VLTRZXGMWDSKGL-UHFFFAOYSA-N perchloric acid Chemical compound OCl(=O)(=O)=O VLTRZXGMWDSKGL-UHFFFAOYSA-N 0.000 description 2
- 239000002504 physiological saline solution Substances 0.000 description 2
- 230000028327 secretion Effects 0.000 description 2
- 238000011218 seed culture Methods 0.000 description 2
- 230000004622 sleep time Effects 0.000 description 2
- 210000002784 stomach Anatomy 0.000 description 2
- 239000000725 suspension Substances 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- -1 triterpene compound Chemical class 0.000 description 2
- 150000003648 triterpenes Chemical class 0.000 description 2
- AWFYPPSBLUWMFQ-UHFFFAOYSA-N 2-[5-[2-(2,3-dihydro-1H-inden-2-ylamino)pyrimidin-5-yl]-1,3,4-oxadiazol-2-yl]-1-(1,4,6,7-tetrahydropyrazolo[4,3-c]pyridin-5-yl)ethanone Chemical compound C1C(CC2=CC=CC=C12)NC1=NC=C(C=N1)C1=NN=C(O1)CC(=O)N1CC2=C(CC1)NN=C2 AWFYPPSBLUWMFQ-UHFFFAOYSA-N 0.000 description 1
- MGGVALXERJRIRO-UHFFFAOYSA-N 4-[2-(2,3-dihydro-1H-inden-2-ylamino)pyrimidin-5-yl]-2-[2-oxo-2-(2,4,6,7-tetrahydrotriazolo[4,5-c]pyridin-5-yl)ethyl]-1H-pyrazol-5-one Chemical compound C1C(CC2=CC=CC=C12)NC1=NC=C(C=N1)C=1C(=NN(C=1)CC(=O)N1CC2=C(CC1)NN=N2)O MGGVALXERJRIRO-UHFFFAOYSA-N 0.000 description 1
- JBYXPOFIGCOSSB-GOJKSUSPSA-N 9-cis,11-trans-octadecadienoic acid Chemical compound CCCCCC\C=C\C=C/CCCCCCCC(O)=O JBYXPOFIGCOSSB-GOJKSUSPSA-N 0.000 description 1
- 244000068988 Glycine max Species 0.000 description 1
- 235000010469 Glycine max Nutrition 0.000 description 1
- 206010020772 Hypertension Diseases 0.000 description 1
- 241000124008 Mammalia Species 0.000 description 1
- QGMRQYFBGABWDR-UHFFFAOYSA-M Pentobarbital sodium Chemical compound [Na+].CCCC(C)C1(CC)C(=O)NC(=O)[N-]C1=O QGMRQYFBGABWDR-UHFFFAOYSA-M 0.000 description 1
- 102000057297 Pepsin A Human genes 0.000 description 1
- 108090000284 Pepsin A Proteins 0.000 description 1
- 239000001888 Peptone Substances 0.000 description 1
- 108010080698 Peptones Proteins 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 1
- 210000000683 abdominal cavity Anatomy 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- 229940024606 amino acid Drugs 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 235000015278 beef Nutrition 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 102000006995 beta-Glucosidase Human genes 0.000 description 1
- 108010047754 beta-Glucosidase Proteins 0.000 description 1
- 230000007177 brain activity Effects 0.000 description 1
- 230000008344 brain blood flow Effects 0.000 description 1
- 125000002057 carboxymethyl group Chemical group [H]OC(=O)C([H])([H])[*] 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 235000012000 cholesterol Nutrition 0.000 description 1
- 229940108924 conjugated linoleic acid Drugs 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 235000005911 diet Nutrition 0.000 description 1
- 230000037213 diet Effects 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution 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
- 239000012153 distilled water Substances 0.000 description 1
- 229960003638 dopamine Drugs 0.000 description 1
- 235000021001 fermented dairy product Nutrition 0.000 description 1
- 235000013376 functional food Nutrition 0.000 description 1
- 229910001385 heavy metal Inorganic materials 0.000 description 1
- 210000001320 hippocampus Anatomy 0.000 description 1
- 239000000017 hydrogel Substances 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 238000009776 industrial production Methods 0.000 description 1
- 238000001802 infusion Methods 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 230000000968 intestinal effect Effects 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 210000003734 kidney Anatomy 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- OYHQOLUKZRVURQ-IXWMQOLASA-N linoleic acid Natural products CCCCC\C=C/C\C=C\CCCCCCCC(O)=O OYHQOLUKZRVURQ-IXWMQOLASA-N 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 1
- 235000019341 magnesium sulphate Nutrition 0.000 description 1
- 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
- 239000012528 membrane Substances 0.000 description 1
- 230000003340 mental effect Effects 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 229930014626 natural product Natural products 0.000 description 1
- 210000002569 neuron Anatomy 0.000 description 1
- 229960002275 pentobarbital sodium Drugs 0.000 description 1
- 229940111202 pepsin Drugs 0.000 description 1
- 235000019319 peptone Nutrition 0.000 description 1
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 description 1
- 229920000053 polysorbate 80 Polymers 0.000 description 1
- 230000002335 preservative effect Effects 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 239000000932 sedative agent Substances 0.000 description 1
- 230000001624 sedative effect Effects 0.000 description 1
- 230000004799 sedative–hypnotic effect Effects 0.000 description 1
- 210000000582 semen Anatomy 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 238000002864 sequence alignment Methods 0.000 description 1
- 238000004088 simulation Methods 0.000 description 1
- 230000036578 sleeping time Effects 0.000 description 1
- 239000001632 sodium acetate Substances 0.000 description 1
- 235000017281 sodium acetate Nutrition 0.000 description 1
- 238000000638 solvent extraction Methods 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 230000003068 static effect Effects 0.000 description 1
- 239000008174 sterile solution Substances 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 230000002195 synergetic effect Effects 0.000 description 1
- 230000002194 synthesizing effect Effects 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
- 239000001393 triammonium citrate Substances 0.000 description 1
- 235000011046 triammonium citrate Nutrition 0.000 description 1
- 238000003809 water extraction Methods 0.000 description 1
- 235000015099 wheat brans Nutrition 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
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
- A23L29/065—Microorganisms
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L31/00—Edible extracts or preparations of fungi; Preparation or treatment thereof
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
-
- 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
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
-
- 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
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/06—Fungi, e.g. yeasts
- A61K36/07—Basidiomycota, e.g. Cryptococcus
- A61K36/076—Poria
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/88—Liliopsida (monocotyledons)
- A61K36/896—Liliaceae (Lily family), e.g. daylily, plantain lily, Hyacinth or narcissus
- A61K36/8967—Lilium, e.g. tiger lily or Easter lily
-
- 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
-
- 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/10—Preparation or pretreatment of starting material
- A61K2236/19—Preparation or pretreatment of starting material involving fermentation using yeast, bacteria or both; enzymatic treatment
-
- 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
Abstract
本发明提出了一种产γ‑氨基丁酸的植物乳杆菌、助睡眠益生菌组合物及其制备方法,属于益生菌技术领域。所述植物乳杆菌FPHC0500分类命名为植物乳杆菌Lactobacillus plantarum,保藏编号为CGMCC NO.26761,保藏日期为2023年3月6日,保藏单位为中国微生物菌种保藏管理委员会普通微生物中心,地址:北京市朝阳区北辰西路1号院3号,中国科学院微生物研究所,邮编:100101。该植物乳杆菌FPHC0500具有高耐胃酸、耐胆碱存活率,具有发酵高产γ‑氨基丁酸的能力,该菌株易培养、繁殖速度快,在菌种混合培养时属于优势菌种。
Description
技术领域
本发明涉及益生菌技术领域,具体涉及一种产γ-氨基丁酸的植物乳杆菌、助睡眠益生菌组合物及其制备方法。
背景技术
γ-氨基丁酸(γ-aminobutyricacid,简称GABA)是哺乳动物神经中枢的一种抑制性递质,具有调节血压、促使精神安定、促进脑部血流、增进脑活力、营养神经细胞、增加生长激素分泌、健肝利肾、促进乙醇代谢(醒酒)等多种功效。2009年卫生部已正式批准GABA为新资源食品,富集GABA的原料可应用于生产功能性食品,通过饮食来降低和预防高血压。GABA的获取方法有化学合成法和生物法。化学合成法成本较高,产品中易残留化学物质,不属于天然产物,应用有条件限制。生物法主要从植物体和微生物发酵液中获取,相对于植物富集法的成本高、产率低和局限性大的缺点,微生物发酵法更有优越性,它不受空间、环境和资源的限制,具有成本低、无化学残留和产量高等显著优点,是富集γ-氨基丁酸的理想途径。
植物乳杆菌是一种重要的植物乳酸菌,其发酵过程中能产出特有的乳酸杆菌素,具有天然防腐作用,其产出的酸性物质能降解重金属,其β葡萄糖苷酶具有较高的活性,具有减毒作用,此外植物乳杆菌还具有合成共轭亚油酸的能力,2009年植物乳杆菌被批准为新资源食品。因此植物乳杆菌是一种优良γ-氨基丁酸生产菌种。目前已分离得到多株产γ-氨基丁酸的植物乳杆菌。中国农业大学梁岩等筛选到一株产γ-氨基丁酸的植物乳杆菌S35,产量为4.52g/L,范杰等筛选到一株产γ-氨基丁酸的植物乳杆菌,产量为4.98g/L;浙江师范大学蒋冬花分离到一株植物乳杆菌,γ-氨基丁酸产量为约4g/L;浙江工业大学丁兴等分离到一株植物乳杆菌,γ-氨基丁酸产量为0.527g/L,但其研究成果中产GABA能力都不高,也很少能应用于工业化生产。因此,从乳酸菌中筛选高产GABA的功能性植物乳酸菌对GABA产业的发展和人类健康均具有重要的意义。
发明内容
本发明的目的在于提出一种产γ-氨基丁酸的植物乳杆菌、助睡眠益生菌组合物及其制备方法,该植物乳杆菌FPHC0500具有高耐胃酸、耐胆碱存活率,具有发酵高产γ-氨基丁酸的能力,该菌株易培养、繁殖速度快,在菌种混合培养时属于优势菌种。
本发明的技术方案是这样实现的:
本发明提供一种产γ-氨基丁酸的植物乳杆菌FPHC0500,所述植物乳杆菌FPHC0500分类命名为植物乳杆菌Lactobacillus plantarum,保藏编号为CGMCCNO.26761,保藏日期为2023年3月6日,保藏单位为中国微生物菌种保藏管理委员会普通微生物中心,地址:北京市朝阳区北辰西路1号院3号,中国科学院微生物研究所,邮编:100101。
作为本发明的进一步改进,所述植物乳杆菌FPHC0500的16S rDNA序列如SEQ IDNo.1所示。
作为本发明的进一步改进,所述植物乳杆菌FPHC0500为革兰氏阳性菌,菌体呈短杆状,成单、成对或者成链,在MRS培养基上菌落呈圆形,边缘整齐,乳白色,表面湿润光滑,不产生色素。
本发明进一步保护含有上述植物乳杆菌FPHC0500的微生物制剂。
本发明进一步保护上述的植物乳杆菌FPHC0500在微生物发酵制备γ-氨基丁酸中的应用。
本发明进一步保护上述的植物乳杆菌FPHC0500在制备助睡眠产品中的应用。
本发明进一步保护一种助睡眠益生菌组合物的制备方法,包括以下步骤:
S1.将茯苓和百合分别洗净,干燥,粉碎,制得中药粉,将中药粉加入水中,加热煮沸提取,过滤,滤渣留用,干燥,得到水提取物;
S2.将步骤S1中的滤渣加入乙酸乙酯-乙醇-丙酮混合溶剂中,加热提取,过滤,滤渣洗涤,干燥,得到干滤渣,滤液减压除去溶剂,干燥,制得有机提取物;
S3.将上述产γ-氨基丁酸的植物乳杆菌FPHC0500接种至高氏培养基中,活化培养,制得菌种种子液;
S4.将步骤S2中的干滤渣加入无菌水中,加入添加剂,灭菌,接种步骤S3制得的植物乳杆菌FPHC0500菌种种子液,发酵培养,冷冻干燥,制得发酵产物;
S5.将聚乙烯醇和羟甲基壳聚糖溶于水中,加入步骤S4中的发酵产物,搅拌混合均匀,加入β-甘油磷酸钠和金属盐溶液,搅拌反应,冷冻干燥,粉碎,得到缓释体系,与步骤S1制得的水提取物、步骤S2制得的有机提取物混合均匀,制得助睡眠益生菌组合物。
作为本发明的进一步改进,步骤S1中所述茯苓和百合的质量比为10-15:7-12,所述中药粉和水的固液比为1:5-10g/mL,所述提取的时间为2-3h,步骤S2中所述乙酸乙酯-乙醇-丙酮混合溶剂中乙酸乙酯、乙醇和丙酮的体积比为3-5:5-7:1-2,所述滤渣和乙酸乙酯-乙醇-丙酮混合溶剂的固液比为1:3-5g/mL,所述加热提取的温度为50-70℃,时间为1-2h,步骤S3中所述活化培养的条件为缺氧条件下,37-39℃,50-70r/min,活化培养12-18h,所述菌种种子液的含菌量为108-109cfu/mL。
作为本发明的进一步改进,步骤S4中所述干滤渣、无菌水、添加剂的质量比为10-12:100:5-10,所述添加剂的组成为:氯化钠、氯化铵、尿酸、葡萄糖、维生素B1、维生素B6、氯化钙,质量比为1-2:2-4:1-2:3-5:0.1-0.2:0.2-0.3:0.1-0.2,所述植物乳杆菌FPHC0500菌种种子液的接种量为2-3%,所述发酵培养的条件为缺氧条件下,37-39℃,50-70r/min,活化培养38-48h,步骤S5中所述金属盐为氯化钙或氯化铝,所述聚乙烯醇、羟甲基壳聚糖、发酵产物、β-甘油磷酸钠和金属盐的质量比为7-10:5-7:6-8:2-3:0.5-1,所述缓释体系、水提取物、有机提取物的质量比为10-12:3-5:2-3。
本发明进一步保护一种上述的制备方法制得的助睡眠益生菌组合物。
本发明具有如下有益效果:本发明产γ-氨基丁酸的植物乳杆菌FPHC0500具有高耐胃酸、耐胆碱存活率,具有发酵高产γ-氨基丁酸的能力,该菌株易培养、繁殖速度快,在菌种混合培养时属于优势菌种。
本发明提供了一种助睡眠益生菌组合物,将茯苓和百合进行水提取、有机溶剂提取和利用植物乳杆菌FPHC0500发酵提取,得到的发酵产物经过缓释水凝胶包埋,不仅具有很好的缓释效果,能保护发酵产物在胃部不被分解释放,在肠道碱性环境下溶胀释药,且原料来源广,使用安全。
茯苓的主要有效成分为茯苓多糖和三萜类物质,茯苓酸是茯苓特有的一种四环三萜化合物。茯苓水煎液、茯苓中羧甲基茯苓多糖、茯苓总三萜及茯苓酸均有镇静催眠作用,茯苓酸的催眠效果与调节脑内γ-氨基丁酸A受体表达水平有关。百合具有较好的镇静、催眠作用,其提取物中的甾体皂苷类化合物具有很好的助眠作用。两者的提取物具有很好的协同作用,呈现明显的催眠作用,能够明显调节中枢系统中单胺类递质如多巴胺、5-羟色胺的含量,从而在睡眠调节中起着重要的作用。将两者经过提取后,经过植物乳杆菌FPHC0500发酵后,能产生大量的γ-氨基丁酸,GABA具有调节睡眠、降血压、促进生长激素分泌和降低胆固醇等多种保健功能,为开发新的富含γ-氨基丁酸的保健食品提供依据,将所述植物乳杆菌FPHC0500应用于食品、保健品的开发,具有广泛的应用前景。
附图说明
为了更清楚地说明本发明实施例或现有技术中的技术方案,下面将对实施例或现有技术描述中所需要使用的附图作简单地介绍,显而易见地,下面描述中的附图仅仅是本发明的一些实施例,对于本领域普通技术人员来讲,在不付出创造性劳动性的前提下,还可以根据这些附图获得其他的附图。
图1为植物乳杆菌FPHC0500的菌落形态图;
图2为植物乳杆菌FPHC0500的菌体形态图;
图3为各组菌耐胃酸能力的对比图;
图4为各组菌耐胆盐能力的对比图。
具体实施方式
下面将对本发明实施例中的技术方案进行清楚、完整地描述,显然,所描述的实施例仅仅是本发明一部分实施例,而不是全部的实施例。基于本发明中的实施例,本领域普通技术人员在没有做出创造性劳动前提下所获得的所有其他实施例,都属于本发明保护的范围。
本发明一种产γ-氨基丁酸的植物乳杆菌FPHC0500,所述植物乳杆菌FPHC0500分类命名为植物乳杆菌Lactobacillus plantarum,保藏编号为CGMCCNO.26761,保藏日期为2023年3月6日,保藏单位为中国微生物菌种保藏管理委员会普通微生物中心,地址:北京市朝阳区北辰西路1号院3号,中国科学院微生物研究所,邮编:100101。
实施例1植物乳杆菌FPHC0500的筛选及鉴定
1、植物乳杆菌FPHC0500的筛选
1.1样品来源
本发明所使用的菌株均收集自内蒙古传统的发酵乳制品,即本发明来自于发酵乳作为样品。
1.2培养基的配制
样品分离和菌株筛选所用的培养基为MRS液体培养基,成分如下:蛋白胨10g、牛肉粉5g、酵母粉4g、磷酸氢二钾2g、柠檬酸三铵2g、乙酸钠5g、葡萄糖20g、吐温80 1mL、硫酸镁0.2g、硫酸锰0.05g、琼脂15g、蒸馏水1000mL,pH=6.2±0.1。
1.3菌株的分离
将10g发酵乳样品加入含有0.7%的NaCl无菌溶液中,得到浑浊液,将浑浊液进行梯度稀释,取10-5、10-6、10-7三个稀释梯度的菌液100μL涂于含有无菌MRS固体培养基(购于北京路桥公司)的培养皿上,于有氧条件下37℃静置培养24h,至有明显单菌落形成后,从培养基中选择生长有65-90个单菌落的平板,挑取典型菌落,在MRS固体平板培养基上多次划线纯化,直至整个平板上的菌落形态一致,得培养,物做菌株鉴定。
1.4菌株的保藏
挑选单菌落到MRS液体培养基中,于37℃下,70r/min,培养36h,吸取500μL菌液加入1mL 60%(v/v)的甘油管中,于-80℃冷冻保藏。
2、植物乳杆菌FPHC0500的鉴定
2.1菌落特征
植物乳杆菌FPHC0500在MRS固体培养基中培养24h后,在MRS培养基上形成明显的菌落,直径在0.5-2.0mm之间,圆形,边缘整齐,乳白色,表面湿润光滑,不产生色素,见图1。
2.2显微镜下形态
植物乳杆菌FPHC0500菌落涂片:革兰氏染色阳性,菌体呈短杆状,成单、成对或者成链,见图2。
2.3 16S rDNA鉴定
鉴定单位:上海美吉生物医药科技有限公司。
鉴定序列:
GCTGGTTCCTAAAAGGTTACCCCACCGACTTTGGGTGTTACAAACTCTCATGGTGTGACGGGCGGTGTGTACAAGGCCCGGGAACGTATTCACCGCGGCATGCTGATCCGCGATTACTAGCGATTCCGACTTCATGTAGGCGAGTTGCAGC
CTACAATCCGAACTGAGAATGGCTTTAAGAGATTAGCTTACTCTCGCGAGTT
CGCAACTCGTTGTACCATCCATTGTAGCACGTGTGTAGCCCAGGTCATAAGG
GGCATGATGATTTGACGTCATCCCCACCTTCCTCCGGTTTGTCACCGGCAGT
CTCACCAGAGTGCCCAACTTAATGCTGGCAACTGATAATAAGGGTTGCGCT
CGTTGCGGGACTTAACCCAACATCTCACGACACGAGCTGACGACAACCATG
CACCACCTGTATCCATGTCCCCGAAGGGAACGTCTAATCTCTTAGATTTGCA
TAGTATGTCAAGACCTGGTAAGGTTCTTCGCGTAGCTTCGAATTAAACCACA
TGCTCCACCGCTTGTGCGGGCCCCCGTCAATTCCTTTGAGTTTCAGCCTTGC
GGCCGTACTCCCCAGGCGGAATGCTTAATGCGTTAGCTGCAGCACTGAAGG
GCGGAAACCCTCCAACACTTAGCATTCATCGTTTACGGTATGGACTACCAGG
GTATCTAATCCTGTTTGCTACCCATACTTTCGAGCCTCAGCGTCAGTTACAG
ACCAGACAGCCGCCTTCGCCACTGGTGTTCTTCCATATATCTACGCATTTCA
CCGCTACACATGGAGTTCCACTGTCCTCTTCTGCACTCAAGTTTCCCAGTTT
CCCGATGCACTTCTTCCGGTTTGAGCCGAAGGCTTTCACATCAGACTTAAA
AAACCGCCTGCGCTCGCTTTACGCCCAATAAATCCGGACAACGCTTGCCAC
CTACGTATTACCGCGGCTGCTGGCACGTAGTTAGCCGTGGCTTTCTGGTTAA
ATACCGTCAATACCTGAACAGTTACTCTCAGATATGTTCTTCTTTAACAACA
GAGTTTTACGAGCCGAAACCCTTCTTCACTCACGCGGCGTTGCTCCATCAG
ACTTTCGTCCATTGTGGAAGATTCCCTACTGCTGCCTCCCGTAGGAGTTTGG
GCCGTGTCTCAGTCCCAATGTGGCCGATTACCCTCTCAGGTCGGCTACGTAT
CATTGCCATGGTGAGCCGTTACCCCACCATCTAGCTAATACGCCGCGGGACC
ATCCAAAAGTGATAGCCGAAGCCATCTTTCAAGCTCGGACCATGCGGTCCA
AGTTGTTATGCGGTATTAGCATCTGTTTCCAGGTGTTATCCCCCGCTTCTGGG
CAGGTTTCCCACGTGTTACTCACCAGTTCGCCACTCACTCAGGATGTAAATCATGATGCAAGCACCAATCAATACCAGAGTCGTCGACTGCATTATA。
鉴定结果:根据菌株的序列比对结果和生理生化结果相结合,确定筛选的菌株为Lactobacillus plantarum(植物乳杆菌)。
实施例2植物乳杆菌FPHC0500菌株耐酸能力测试
人工胃液的配制:NaCl 0.2%、胃蛋白酶0.35%、用1mol/L的HCl调整pH值为3.0后,过滤除菌,现配现用。
37℃,MRS液体培养基培养24h,经5000r/min离心10min收集菌体后重悬于生理盐水,制成菌悬液,取1mL菌悬液与9mL pH3.0的人工胃液混合,摇匀,置于恒温振荡器中培养(37℃,300r/min),并分别在0h、1h、2h和3h取样,用MRS琼脂培养基倾注37℃培养48h。用平板计数法测定活菌数,计算其存活率(%):
存活率(%)=取样时的活菌数/0h的活菌数×100%
相同条件下以嗜酸乳杆菌La28(Lactobacillus acidophilus La28)、植物乳杆菌LP45(Lactobacillus plantarum LP45)(均由河北一然生物科技有限公司提供)、嗜酸乳杆菌LA16(由本公司提供)作为对照,菌株溶液、活菌量与植物乳杆菌FPHC0500相同。
模拟胃酸存活率的结果如图3。由图可知,植物乳杆菌FPHC0500在体外模拟胃酸实验时,在3h的存活率达到了86.9±3.06%,存活率远超过其他对照菌株。
实施例3植物乳杆菌FPHC0500菌株耐受胆盐的测定
将活化好的菌液按2%的接种量用移液枪接种于含0.3%牛胆盐的MRS培养基。在恒温振荡器中37℃培养6h后,以0h为对照,分别在0h、2h、4h和6h取样,测定上述溶液的活菌值,计算菌株对胆盐的耐受力(%)。
胆盐耐受力(%)=取样时溶液的活菌数/0h时溶液的活菌数×100%。
相同条件下以嗜酸乳杆菌La28(Lactobacillus acidophilus La28)、植物乳杆菌LP45(Lactobacillus plantarum LP45)(均由河北一然生物科技有限公司提供)、嗜酸乳杆菌LA16(由本公司提供)作为对照,菌株溶液、活菌量与植物乳杆菌FPHC0500相同。
胆盐耐受力的结果如图4。由图可知,植物乳杆菌FPHC0500在体外模拟胆盐耐受力实验时,在6h的存活率达到了70.2±5.08%,存活率远超过其他对照菌株。
实施例4产γ-氨基丁酸能力测定
(1)活化:将保藏的菌种接入MRS液体培养基,37℃静置活化12h;
(2)一级培养:将250mL三角瓶中装入100mL一级种子培养基(谷氨酸钠含量为3g/L的MRS液体培养基),取静置培养的活化培养液,按5%的接种量接种后,37℃静置培养20h作为一级种子液;
(3)二级培养:6L具塞三角瓶中装入5L二级种子培养基(谷氨酸钠含量为30g/L的MRS液体培养基),取一级种子液,按5%的接种量接种后,37℃静置培养40h作为二级种子液;
(4)700L发酵罐中装入500L量产发酵培养基(谷氨酸钠含量为50g/L的MRS液体培养基,另外添加了大豆粉10g/L,麦麸12g/L),按3%的接种量接种后,37℃静置培养56h,培养过程中培养过程中使用65%磷酸调节pH,控制发酵pH在6。经氨基酸酸自动分析仪检测为:47.8g/L。因此,本发明植物乳杆菌FPHC0500为一种高产γ-氨基丁酸的菌株。
实施例5
本实施例提供一种助睡眠益生菌组合物的制备方法,包括以下步骤:
S1.将10重量份茯苓和7重量份百合分别洗净,干燥,粉碎,制得中药粉,将中药粉加入水中,所述中药粉和水的固液比为1:5g/mL,加热煮沸提取2h,过滤,滤渣留用,干燥,得到水提取物;
S2.将步骤S1中的滤渣加入乙酸乙酯-乙醇-丙酮混合溶剂中,所述乙酸乙酯-乙醇-丙酮混合溶剂中乙酸乙酯、乙醇和丙酮的体积比为3:5:1,所述滤渣和乙酸乙酯-乙醇-丙酮混合溶剂的固液比为1:3g/mL,加热至50℃,提取1h,过滤,滤渣洗涤,干燥,得到干滤渣,滤液减压除去溶剂,干燥,制得有机提取物;
S3.将产γ-氨基丁酸的植物乳杆菌FPHC0500接种至高氏培养基中,缺氧条件下,39℃,50r/min,活化培养12h,制得菌种种子液,含菌量为108cfu/mL;
S4.将10重量份步骤S2中的干滤渣加入100重量份无菌水中,加入5重量份添加剂,紫外线灭菌,接种步骤S3制得的植物乳杆菌FPHC0500菌种种子液,所述植物乳杆菌FPHC0500菌种种子液的接种量为2%,缺氧条件下,39℃,50r/min,活化培养38h,冷冻干燥,制得发酵产物;
所述添加剂的组成为:氯化钠、氯化铵、尿酸、葡萄糖、维生素B1、维生素B6、氯化钙,质量比为1:2:1:3:0.1:0.2:0.1;
S5.将7重量份聚乙烯醇和5重量份羟甲基壳聚糖溶于100重量份水中,加入6重量份步骤S4中的发酵产物,搅拌混合均匀,加入20重量份含有2重量份β-甘油磷酸钠和0.5重量份氯化钙的水溶液,搅拌反应50min,冷冻干燥,粉碎,得到缓释体系,将10重量份缓释体系与3重量份步骤S1制得的水提取物、2重量份步骤S2制得的有机提取物搅拌混合15min,制得助睡眠益生菌组合物。
实施例6
本实施例提供一种助睡眠益生菌组合物的制备方法,包括以下步骤:
S1.将15重量份茯苓和12重量份百合分别洗净,干燥,粉碎,制得中药粉,将中药粉加入水中,所述中药粉和水的固液比为1:10g/mL,加热煮沸提取3h,过滤,滤渣留用,干燥,得到水提取物;
S2.将步骤S1中的滤渣加入乙酸乙酯-乙醇-丙酮混合溶剂中,所述乙酸乙酯-乙醇-丙酮混合溶剂中乙酸乙酯、乙醇和丙酮的体积比为5:7:2,所述滤渣和乙酸乙酯-乙醇-丙酮混合溶剂的固液比为1:5g/mL,加热至70℃,提取2h,过滤,滤渣洗涤,干燥,得到干滤渣,滤液减压除去溶剂,干燥,制得有机提取物;
S3.将产γ-氨基丁酸的植物乳杆菌FPHC0500接种至高氏培养基中,缺氧条件下,38℃,70r/min,活化培养18h,制得菌种种子液,含菌量为109cfu/mL;
S4.将12重量份步骤S2中的干滤渣加入100重量份无菌水中,加入10重量份添加剂,紫外线灭菌,接种步骤S3制得的植物乳杆菌FPHC0500菌种种子液,所述植物乳杆菌FPHC0500菌种种子液的接种量为3%,缺氧条件下,38℃,70r/min,活化培养48h,冷冻干燥,制得发酵产物;
所述添加剂的组成为:氯化钠、氯化铵、尿酸、葡萄糖、维生素B1、维生素B6、氯化钙,质量比为2:4:2:5:0.2:0.3:0.2;
S5.将10重量份聚乙烯醇和7重量份羟甲基壳聚糖溶于100重量份水中,加入8重量份步骤S4中的发酵产物,搅拌混合均匀,加入20重量份含有3重量份β-甘油磷酸钠和1重量份氯化铝的水溶液,搅拌反应50min,冷冻干燥,粉碎,得到缓释体系,将12重量份缓释体系与5重量份步骤S1制得的水提取物、3重量份步骤S2制得的有机提取物搅拌混合15min,制得助睡眠益生菌组合物。
实施例7
本实施例提供一种助睡眠益生菌组合物的制备方法,包括以下步骤:
S1.将12重量份茯苓和10重量份百合分别洗净,干燥,粉碎,制得中药粉,将中药粉加入水中,所述中药粉和水的固液比为1:7g/mL,加热煮沸提取2.5h,过滤,滤渣留用,干燥,得到水提取物;
S2.将步骤S1中的滤渣加入乙酸乙酯-乙醇-丙酮混合溶剂中,所述乙酸乙酯-乙醇-丙酮混合溶剂中乙酸乙酯、乙醇和丙酮的体积比为4:6:1.5,所述滤渣和乙酸乙酯-乙醇-丙酮混合溶剂的固液比为1:4g/mL,加热至60℃,提取1.5h,过滤,滤渣洗涤,干燥,得到干滤渣,滤液减压除去溶剂,干燥,制得有机提取物;
S3.将产γ-氨基丁酸的植物乳杆菌FPHC0500接种至高氏培养基中,缺氧条件下,37℃,60r/min,活化培养16h,制得菌种种子液,含菌量为109cfu/mL;
S4.将11重量份步骤S2中的干滤渣加入100重量份无菌水中,加入8重量份添加剂,紫外线灭菌,接种步骤S3制得的植物乳杆菌FPHC0500菌种种子液,所述植物乳杆菌FPHC0500菌种种子液的接种量为2.5%,缺氧条件下,37℃,60r/min,活化培养42h,冷冻干燥,制得发酵产物;
所述添加剂的组成为:氯化钠、氯化铵、尿酸、葡萄糖、维生素B1、维生素B6、氯化钙,质量比为1.5:3:1.5:4:0.15:0.25:0.15;
S5.将8.5重量份聚乙烯醇和6重量份羟甲基壳聚糖溶于100重量份水中,加入7重量份步骤S4中的发酵产物,搅拌混合均匀,加入20重量份含有2.5重量份β-甘油磷酸钠和0.7重量份氯化钙的水溶液,搅拌反应50min,冷冻干燥,粉碎,得到缓释体系,将11重量份缓释体系与4重量份步骤S1制得的水提取物、2.5重量份步骤S2制得的有机提取物搅拌混合15min,制得助睡眠益生菌组合物。
对比例1
与实施例7相比,不同之处在于,步骤S1中未添加茯苓。
对比例2
与实施例7相比,不同之处在于,步骤S1中未添加百合。
对比例3
与实施例7相比,不同之处在于,未进行步骤S1的水提取。
对比例4
与实施例7相比,不同之处在于,未进行步骤S2的有机溶剂提取。
对比例5
与实施例7相比,不同之处在于,步骤S4中添加剂未添加氯化钙。
对比例6
与实施例7相比,不同之处在于,步骤S4中添加剂维生素B1。
对比例7
与实施例7相比,不同之处在于,步骤S4中添加剂维生素B6。
对比例8
与实施例7相比,不同之处在于,步骤S4中添加剂维生素B1和维生素B6。
对比例9
与实施例7相比,不同之处在于,未进行步骤S4。
对比例10
与实施例7相比,不同之处在于,步骤S5中未进行包埋,直接将发酵产物、水提取物、有机提取物搅拌混合15min,制得助睡眠益生菌组合物。
测试例1助眠效果实验
取小鼠150只,体质量18-22g,随机分成15组,分别为实施例5-7组、对比例1-10组、空白对照组、阳性对照组,每组10只,雌雄各半,分别灌胃给药。灌胃剂量分别为:实施例5-7、对比例1-10组给与助睡眠益生菌组合物2g/kg;阳性对照组给酸枣仁提取物(购于广州合诚三先生物科技有限公司)2g/kg;空白对照组给等量生理盐水。每天1次,连续给药1周。1周后用监控摄像观察并记录:灌胃30min后各组小鼠的睡眠率;灌胃结束后12h内各组小鼠的睡眠时间。
结果见表1。
表1
组别 | 30min后睡眠小鼠数量(只) | 睡眠率(%) |
空白对照组 | 4 | 40 |
阳性对照组 | 8 | 80 |
实施例5 | 9 | 90 |
实施例6 | 10 | 100 |
实施例7 | 10 | 100 |
对比例1 | 6 | 60 |
对比例2 | 7 | 70 |
对比例3 | 8 | 80 |
对比例4 | 7 | 70 |
对比例5 | 8 | 80 |
对比例6 | 8 | 80 |
对比例7 | 8 | 80 |
对比例8 | 7 | 70 |
对比例9 | 6 | 60 |
对比例10 | 5 | 50 |
最后一次灌胃结束30min后,腹腔注射戊巴比妥钠70mg/kg,准确观察并记录小鼠的入睡时间和睡眠时间。
结果见表2。
表2
组别 | 睡眠时间(min) | 入睡时间(min) |
空白对照组 | 167.2±23.5 | 7.15±4.14 |
阳性对照组 | 277.3±30.1* | 4.09±2.11* |
实施例5 | 299.4±29.3# | 3.12±2.19# |
实施例6 | 302.1±32.1# | 3.08±2.01# |
实施例7 | 307.5±27.9# | 2.97±1.98# |
对比例1 | 260.1±31.2 | 4.24±2.57 |
对比例2 | 270.6±28.4 | 4.17±2.82 |
对比例3 | 272.3±29.4 | 4.10±2.78 |
对比例4 | 267.2±31.1 | 4.19±2.55 |
对比例5 | 282.2±27.9 | 4.04±2.62 |
对比例6 | 285.1±30.5 | 3.92±2.10 |
对比例7 | 284.9±28.5 | 3.97±2.04 |
对比例8 | 290.7±29.1 | 4.01±2.11 |
对比例9 | 262.2±26.8 | 4.19±2.35 |
对比例10 | 253.1±25.2 | 4.31±2.61 |
注释:*为与空白对照组相比,P<0.05,#为与阳性对照组相比,P<0.05。
使用异弗烷麻醉小鼠后进行经心灌注。将小鼠断头处死,迅速剥取全脑,置冰盘中,分离出小鼠左、右海马区,-80℃冰冻保存备用。称取海马区0.2g,加入0.1mol/L的高氯酸300μL后低温碾碎1min。以15000r/min离心5min,取上清液过0.22μm滤膜后,分析GABA、5-HT(5-羟色胺)的含量。
结果见表3。
表3
注释:*为与空白对照组相比,P<0.05,#为与阳性对照组相比,P<0.05。
由上表可知,本发明实施例5-7制得的助睡眠益生菌组合物能明显促进小鼠睡眠,其大脑中GABA、5-HT(5-羟色胺)的含量明显提高。
以上所述仅为本发明的较佳实施例而已,并不用以限制本发明,凡在本发明的精神和原则之内,所作的任何修改、等同替换、改进等,均应包含在本发明的保护范围之内。
Claims (10)
1.一种产γ-氨基丁酸的植物乳杆菌FPHC0500,其特征在于,所述植物乳杆菌FPHC0500分类命名为植物乳杆菌Lactobacillus plantarum,保藏编号为CGMCC NO.26761,保藏日期为2023年3月6日,保藏单位为中国微生物菌种保藏管理委员会普通微生物中心,地址:北京市朝阳区北辰西路1号院3号,中国科学院微生物研究所,邮编:100101。
2.根据权利要求1所述产γ-氨基丁酸的植物乳杆菌FPHC0500,其特征在于,所述植物乳杆菌FPHC0500的16S rDNA序列如SEQ ID No.1所示。
3.根据权利要求1所述产γ-氨基丁酸的植物乳杆菌FPHC0500,其特征在于,所述植物乳杆菌FPHC0500为革兰氏阳性菌,菌体呈短杆状,成单、成对或者成链,在MRS培养基上菌落呈圆形,边缘整齐,乳白色,表面湿润光滑,不产生色素。
4.含有权利要求1-3任一项所述植物乳杆菌FPHC0500的微生物制剂。
5.如权利要求1-3任一项所述的植物乳杆菌FPHC0500在微生物发酵制备γ-氨基丁酸中的应用。
6.如权利要求1-3任一项所述的植物乳杆菌FPHC0500在制备助睡眠产品中的应用。
7.一种助睡眠益生菌组合物的制备方法,其特征在于,包括以下步骤:
S1.将茯苓和百合分别洗净,干燥,粉碎,制得中药粉,将中药粉加入水中,加热煮沸提取,过滤,滤渣留用,干燥,得到水提取物;
S2.将步骤S1中的滤渣加入乙酸乙酯-乙醇-丙酮混合溶剂中,加热提取,过滤,滤渣洗涤,干燥,得到干滤渣,滤液减压除去溶剂,干燥,制得有机提取物;
S3.将权利要求1所述产γ-氨基丁酸的植物乳杆菌FPHC0500接种至高氏培养基中,活化培养,制得菌种种子液;
S4.将步骤S2中的干滤渣加入无菌水中,加入添加剂,灭菌,接种步骤S3制得的植物乳杆菌FPHC0500菌种种子液,发酵培养,冷冻干燥,制得发酵产物;
S5.将聚乙烯醇和羟甲基壳聚糖溶于水中,加入步骤S4中的发酵产物,搅拌混合均匀,加入β-甘油磷酸钠和金属盐溶液,搅拌反应,冷冻干燥,粉碎,得到缓释体系,与步骤S1制得的水提取物、步骤S2制得的有机提取物混合均匀,制得助睡眠益生菌组合物。
8.根据权利要求7所述的制备方法,其特征在于,步骤S1中所述茯苓和百合的质量比为10-15:7-12,所述中药粉和水的固液比为1:5-10g/mL,所述提取的时间为2-3h,步骤S2中所述乙酸乙酯-乙醇-丙酮混合溶剂中乙酸乙酯、乙醇和丙酮的体积比为3-5:5-7:1-2,所述滤渣和乙酸乙酯-乙醇-丙酮混合溶剂的固液比为1:3-5g/mL,所述加热提取的温度为50-70℃,时间为1-2h,步骤S3中所述活化培养的条件为缺氧条件下,37-39℃,50-70r/min,活化培养12-18h,所述菌种种子液的含菌量为108-109cfu/mL。
9.根据权利要求7所述的制备方法,其特征在于,步骤S4中所述干滤渣、无菌水、添加剂的质量比为10-12:100:5-10,所述添加剂的组成为:氯化钠、氯化铵、尿酸、葡萄糖、维生素B1、维生素B6、氯化钙,质量比为1-2:2-4:1-2:3-5:0.1-0.2:0.2-0.3:0.1-0.2,所述植物乳杆菌FPHC0500菌种种子液的接种量为2-3%,所述发酵培养的条件为缺氧条件下,37-39℃,50-70r/min,活化培养38-48h,步骤S5中所述金属盐为氯化钙或氯化铝,所述聚乙烯醇、羟甲基壳聚糖、发酵产物、β-甘油磷酸钠和金属盐的质量比为7-10:5-7:6-8:2-3:0.5-1,所述缓释体系、水提取物、有机提取物的质量比为10-12:3-5:2-3。
10.一种如权利要求7-9任一项所述的制备方法制得的助睡眠益生菌组合物。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202310494077.8A CN116555102B (zh) | 2023-05-05 | 2023-05-05 | 一种产γ-氨基丁酸的植物乳杆菌、助睡眠益生菌组合物及其制备方法 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202310494077.8A CN116555102B (zh) | 2023-05-05 | 2023-05-05 | 一种产γ-氨基丁酸的植物乳杆菌、助睡眠益生菌组合物及其制备方法 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN116555102A CN116555102A (zh) | 2023-08-08 |
CN116555102B true CN116555102B (zh) | 2023-11-21 |
Family
ID=87485543
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202310494077.8A Active CN116555102B (zh) | 2023-05-05 | 2023-05-05 | 一种产γ-氨基丁酸的植物乳杆菌、助睡眠益生菌组合物及其制备方法 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN116555102B (zh) |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN116622559B (zh) * | 2023-05-10 | 2023-12-01 | 江苏新申奥生物科技有限公司 | 一种产乙醛脱氢酶的干酪乳杆菌、解酒益生菌组合物及其制备方法 |
Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN115521889A (zh) * | 2022-10-30 | 2022-12-27 | 江南大学 | 一株产γ-氨基丁酸的植物乳杆菌WL02及其用途 |
-
2023
- 2023-05-05 CN CN202310494077.8A patent/CN116555102B/zh active Active
Patent Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN115521889A (zh) * | 2022-10-30 | 2022-12-27 | 江南大学 | 一株产γ-氨基丁酸的植物乳杆菌WL02及其用途 |
Also Published As
Publication number | Publication date |
---|---|
CN116555102A (zh) | 2023-08-08 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN108728382B (zh) | 一株具降胆固醇及促进肠道短链脂肪酸产生能力的植物乳杆菌及其应用 | |
CN104770575B (zh) | 一种黄芪微生态制剂及其制备方法和应用 | |
CN110272842A (zh) | 一株具有减肥降脂功能的植物乳杆菌lp104 | |
CN110106119B (zh) | 一株分离自母乳的鼠李糖乳杆菌m9及其应用 | |
CN102899276B (zh) | 一株具有降胆固醇能力的嗜热链球菌及其应用 | |
CN109628346B (zh) | 发酵乳杆菌cqpc04及其在制备发酵食品中的应用 | |
CN110218681A (zh) | 一株发酵乳杆菌kp101及其应用 | |
CN116024130B (zh) | 一株降血尿酸发酵乳杆菌a21215及其应用 | |
CN109497558A (zh) | 苦参药渣多糖合生元及其制备方法与应用 | |
CN110564638A (zh) | 一株具有益生特性的罗伊氏乳杆菌及其用途 | |
CN116555102B (zh) | 一种产γ-氨基丁酸的植物乳杆菌、助睡眠益生菌组合物及其制备方法 | |
CN116286551B (zh) | 长双歧杆菌婴儿亚种在调节体内脂肪代谢、塑形减脂改善肥胖方面的应用 | |
CN114574406B (zh) | 鼠李糖乳杆菌菌株wka55及其在制备防治酒精性肝损伤制品方面的用途与产品 | |
CN113604395A (zh) | 一株可发酵石斛且其发酵液可改善皮肤质量的植物乳杆菌 | |
CN115838676A (zh) | 利用新型植物乳植杆菌菌株中药发酵及应用 | |
CN110484459B (zh) | 一株植物乳杆菌及其应用 | |
CN111714572A (zh) | 一种基于植物乳杆菌的益生菌片剂及其制备方法 | |
CN113616715B (zh) | 治疗仔猪腹泻及改善其肠道菌群的发酵中药口服液 | |
CN114908020A (zh) | 一种抗幽门螺旋杆菌感染的植物乳杆菌及其在食用本草酵素产品中的应用 | |
CN114032190A (zh) | 一株可发酵石斛且其发酵液可有效修复日光皮炎的罗伊氏乳杆菌 | |
CN109207389B (zh) | 一种消栓降脂益生复合菌中药口服液及其制备方法 | |
CN109645490A (zh) | 植物乳杆菌cqpc02在制备预防糖尿病的食品或药品中的应用 | |
CN105567590A (zh) | 一种双歧杆菌乳酸菌混合发酵菌酶解法制备工艺及其运用 | |
CN115895966B (zh) | 一株辅助缓解痛风的两歧双歧杆菌bl002及其应用 | |
CN115927116A (zh) | 一种具有降甘油三酯、降胆固醇、减脂的发酵乳杆菌菌株及其应用 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
GR01 | Patent grant | ||
GR01 | Patent grant |