CN116814464B - Fermented lactobacillus mucilaginosus JF5 and application thereof in preparation of lipid-reducing and digestion-aiding foods and medicines - Google Patents
Fermented lactobacillus mucilaginosus JF5 and application thereof in preparation of lipid-reducing and digestion-aiding foods and medicines Download PDFInfo
- Publication number
- CN116814464B CN116814464B CN202310235233.9A CN202310235233A CN116814464B CN 116814464 B CN116814464 B CN 116814464B CN 202310235233 A CN202310235233 A CN 202310235233A CN 116814464 B CN116814464 B CN 116814464B
- Authority
- CN
- China
- Prior art keywords
- lactobacillus
- mucilaginosus
- strain
- fermentum
- limosilactobacillus
- 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
- 241000186660 Lactobacillus Species 0.000 title claims abstract description 80
- 229940039696 lactobacillus Drugs 0.000 title claims abstract description 80
- 238000002360 preparation method Methods 0.000 title claims abstract description 10
- 235000013305 food Nutrition 0.000 title abstract description 13
- 239000003814 drug Substances 0.000 title abstract description 6
- 229940079593 drug Drugs 0.000 title abstract description 5
- BTCSSZJGUNDROE-UHFFFAOYSA-N gamma-aminobutyric acid Chemical compound NCCCC(O)=O BTCSSZJGUNDROE-UHFFFAOYSA-N 0.000 claims abstract description 40
- 239000006041 probiotic Substances 0.000 claims abstract description 35
- 235000018291 probiotics Nutrition 0.000 claims abstract description 35
- KIUKXJAPPMFGSW-DNGZLQJQSA-N (2S,3S,4S,5R,6R)-6-[(2S,3R,4R,5S,6R)-3-Acetamido-2-[(2S,3S,4R,5R,6R)-6-[(2R,3R,4R,5S,6R)-3-acetamido-2,5-dihydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-2-carboxy-4,5-dihydroxyoxan-3-yl]oxy-5-hydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-3,4,5-trihydroxyoxane-2-carboxylic acid Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@H](O3)C(O)=O)O)[C@H](O)[C@@H](CO)O2)NC(C)=O)[C@@H](C(O)=O)O1 KIUKXJAPPMFGSW-DNGZLQJQSA-N 0.000 claims abstract description 27
- 229920002674 hyaluronan Polymers 0.000 claims abstract description 27
- 229960003160 hyaluronic acid Drugs 0.000 claims abstract description 27
- OGNSCSPNOLGXSM-UHFFFAOYSA-N (+/-)-DABA Natural products NCCC(N)C(O)=O OGNSCSPNOLGXSM-UHFFFAOYSA-N 0.000 claims abstract description 20
- 229960003692 gamma aminobutyric acid Drugs 0.000 claims abstract description 20
- 108091005804 Peptidases Proteins 0.000 claims abstract description 10
- 239000004365 Protease Substances 0.000 claims abstract description 10
- 230000000529 probiotic effect Effects 0.000 claims description 20
- 238000004519 manufacturing process Methods 0.000 claims description 12
- 230000001580 bacterial effect Effects 0.000 claims description 11
- 239000003795 chemical substances by application Substances 0.000 claims description 9
- 230000015556 catabolic process Effects 0.000 claims description 6
- 239000000544 cholinesterase inhibitor Substances 0.000 claims description 6
- 108020004465 16S ribosomal RNA Proteins 0.000 claims description 5
- 229940100578 Acetylcholinesterase inhibitor Drugs 0.000 claims description 5
- 229940077274 Alpha glucosidase inhibitor Drugs 0.000 claims description 5
- 239000003888 alpha glucosidase inhibitor Substances 0.000 claims description 5
- 238000006731 degradation reaction Methods 0.000 claims description 5
- 230000000813 microbial effect Effects 0.000 claims description 5
- 108010084185 Cellulases Proteins 0.000 claims description 3
- 102000005575 Cellulases Human genes 0.000 claims description 3
- 102000014171 Milk Proteins Human genes 0.000 claims description 2
- 108010011756 Milk Proteins Proteins 0.000 claims description 2
- 235000021239 milk protein Nutrition 0.000 claims description 2
- 239000000203 mixture Substances 0.000 claims description 2
- 102000035195 Peptidases Human genes 0.000 claims 1
- 230000000694 effects Effects 0.000 abstract description 37
- 102100024295 Maltase-glucoamylase Human genes 0.000 abstract description 16
- 108010028144 alpha-Glucosidases Proteins 0.000 abstract description 16
- 238000010521 absorption reaction Methods 0.000 abstract description 14
- 102000012440 Acetylcholinesterase Human genes 0.000 abstract description 12
- 108010022752 Acetylcholinesterase Proteins 0.000 abstract description 12
- 229940022698 acetylcholinesterase Drugs 0.000 abstract description 12
- 239000001913 cellulose Substances 0.000 abstract description 12
- 229920002678 cellulose Polymers 0.000 abstract description 12
- 230000001737 promoting effect Effects 0.000 abstract description 12
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 abstract description 9
- 230000002401 inhibitory effect Effects 0.000 abstract description 9
- 108010059892 Cellulase Proteins 0.000 abstract description 8
- 239000008280 blood Substances 0.000 abstract description 8
- 210000004369 blood Anatomy 0.000 abstract description 8
- 229940106157 cellulase Drugs 0.000 abstract description 8
- 230000006870 function Effects 0.000 abstract description 8
- 102000004169 proteins and genes Human genes 0.000 abstract description 8
- 108090000623 proteins and genes Proteins 0.000 abstract description 8
- 230000029087 digestion Effects 0.000 abstract description 7
- 206010010774 Constipation Diseases 0.000 abstract description 5
- 230000008855 peristalsis Effects 0.000 abstract description 5
- 235000021110 pickles Nutrition 0.000 abstract description 5
- 206010016946 Food allergy Diseases 0.000 abstract description 3
- 230000009286 beneficial effect Effects 0.000 abstract description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 abstract description 2
- 230000031891 intestinal absorption Effects 0.000 abstract description 2
- 230000032683 aging Effects 0.000 abstract 1
- 241000186840 Lactobacillus fermentum Species 0.000 description 29
- 229940012969 lactobacillus fermentum Drugs 0.000 description 27
- 238000000855 fermentation Methods 0.000 description 18
- 230000004151 fermentation Effects 0.000 description 18
- 239000002609 medium Substances 0.000 description 16
- 230000005526 G1 to G0 transition Effects 0.000 description 10
- 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 9
- 239000008103 glucose Substances 0.000 description 9
- 239000007788 liquid Substances 0.000 description 9
- 238000011160 research Methods 0.000 description 9
- 239000006228 supernatant Substances 0.000 description 9
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 8
- 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 8
- 210000001035 gastrointestinal tract Anatomy 0.000 description 8
- 239000001963 growth medium Substances 0.000 description 8
- 230000005764 inhibitory process Effects 0.000 description 8
- 235000018102 proteins Nutrition 0.000 description 7
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 6
- 230000000968 intestinal effect Effects 0.000 description 6
- 241001465754 Metazoa Species 0.000 description 5
- 238000010790 dilution Methods 0.000 description 5
- 239000012895 dilution Substances 0.000 description 5
- 229940099596 manganese sulfate Drugs 0.000 description 5
- 239000011702 manganese sulphate Substances 0.000 description 5
- 235000007079 manganese sulphate Nutrition 0.000 description 5
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 5
- 229920001817 Agar Polymers 0.000 description 4
- 241000894006 Bacteria Species 0.000 description 4
- 241000282326 Felis catus Species 0.000 description 4
- 239000001888 Peptone Substances 0.000 description 4
- 108010080698 Peptones Proteins 0.000 description 4
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 4
- 239000008272 agar Substances 0.000 description 4
- 230000001153 anti-wrinkle effect Effects 0.000 description 4
- 235000015278 beef Nutrition 0.000 description 4
- 229940041514 candida albicans extract Drugs 0.000 description 4
- 235000012000 cholesterol Nutrition 0.000 description 4
- 230000001276 controlling effect Effects 0.000 description 4
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 description 4
- 239000000284 extract Substances 0.000 description 4
- 230000002496 gastric effect Effects 0.000 description 4
- 230000036541 health Effects 0.000 description 4
- 238000009630 liquid culture Methods 0.000 description 4
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 4
- 235000019341 magnesium sulphate Nutrition 0.000 description 4
- 238000000034 method Methods 0.000 description 4
- 235000019319 peptone Nutrition 0.000 description 4
- 235000021574 pickled cabbage Nutrition 0.000 description 4
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 description 4
- 229920000053 polysorbate 80 Polymers 0.000 description 4
- 238000012216 screening Methods 0.000 description 4
- 230000028327 secretion Effects 0.000 description 4
- 239000001632 sodium acetate Substances 0.000 description 4
- 235000017281 sodium acetate Nutrition 0.000 description 4
- 239000000725 suspension Substances 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- 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 4
- 235000011046 triammonium citrate Nutrition 0.000 description 4
- 239000001393 triammonium citrate Substances 0.000 description 4
- 239000012138 yeast extract Substances 0.000 description 4
- 208000019901 Anxiety disease Diseases 0.000 description 3
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 3
- 206010061218 Inflammation Diseases 0.000 description 3
- 208000008589 Obesity Diseases 0.000 description 3
- 150000001413 amino acids Chemical class 0.000 description 3
- 230000003712 anti-aging effect Effects 0.000 description 3
- 230000036506 anxiety Effects 0.000 description 3
- 210000004027 cell Anatomy 0.000 description 3
- 238000005119 centrifugation Methods 0.000 description 3
- 238000012258 culturing Methods 0.000 description 3
- 230000000593 degrading effect Effects 0.000 description 3
- 239000002552 dosage form Substances 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 238000000227 grinding Methods 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 235000020824 obesity Nutrition 0.000 description 3
- 239000000546 pharmaceutical excipient Substances 0.000 description 3
- 238000004321 preservation Methods 0.000 description 3
- 238000000926 separation method Methods 0.000 description 3
- 239000011780 sodium chloride Substances 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 229920002134 Carboxymethyl cellulose Polymers 0.000 description 2
- 238000008157 ELISA kit Methods 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- 206010020751 Hypersensitivity Diseases 0.000 description 2
- 206010052428 Wound Diseases 0.000 description 2
- 208000027418 Wounds and injury Diseases 0.000 description 2
- OIPILFWXSMYKGL-UHFFFAOYSA-N acetylcholine Chemical compound CC(=O)OCC[N+](C)(C)C OIPILFWXSMYKGL-UHFFFAOYSA-N 0.000 description 2
- 229960004373 acetylcholine Drugs 0.000 description 2
- 230000009471 action Effects 0.000 description 2
- 208000026935 allergic disease Diseases 0.000 description 2
- 230000007815 allergy Effects 0.000 description 2
- 230000003527 anti-angiogenesis Effects 0.000 description 2
- 230000000844 anti-bacterial effect Effects 0.000 description 2
- 230000003110 anti-inflammatory effect Effects 0.000 description 2
- 230000036772 blood pressure Effects 0.000 description 2
- 210000000481 breast Anatomy 0.000 description 2
- 239000000872 buffer Substances 0.000 description 2
- 235000010948 carboxy methyl cellulose Nutrition 0.000 description 2
- 210000002421 cell wall Anatomy 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 239000002361 compost Substances 0.000 description 2
- 230000007547 defect Effects 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 206010012601 diabetes mellitus Diseases 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 239000012153 distilled water Substances 0.000 description 2
- 230000002708 enhancing effect Effects 0.000 description 2
- 229940088598 enzyme Drugs 0.000 description 2
- 210000003754 fetus Anatomy 0.000 description 2
- 239000012530 fluid Substances 0.000 description 2
- 244000053095 fungal pathogen Species 0.000 description 2
- 230000002068 genetic effect Effects 0.000 description 2
- 230000035876 healing Effects 0.000 description 2
- 201000001421 hyperglycemia Diseases 0.000 description 2
- 230000036039 immunity Effects 0.000 description 2
- NOESYZHRGYRDHS-UHFFFAOYSA-N insulin Chemical compound N1C(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(NC(=O)CN)C(C)CC)CSSCC(C(NC(CO)C(=O)NC(CC(C)C)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CCC(N)=O)C(=O)NC(CC(C)C)C(=O)NC(CCC(O)=O)C(=O)NC(CC(N)=O)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CSSCC(NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2C=CC(O)=CC=2)NC(=O)C(CC(C)C)NC(=O)C(C)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2NC=NC=2)NC(=O)C(CO)NC(=O)CNC2=O)C(=O)NCC(=O)NC(CCC(O)=O)C(=O)NC(CCCNC(N)=N)C(=O)NCC(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC(O)=CC=3)C(=O)NC(C(C)O)C(=O)N3C(CCC3)C(=O)NC(CCCCN)C(=O)NC(C)C(O)=O)C(=O)NC(CC(N)=O)C(O)=O)=O)NC(=O)C(C(C)CC)NC(=O)C(CO)NC(=O)C(C(C)O)NC(=O)C1CSSCC2NC(=O)C(CC(C)C)NC(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CC(N)=O)NC(=O)C(NC(=O)C(N)CC=1C=CC=CC=1)C(C)C)CC1=CN=CN1 NOESYZHRGYRDHS-UHFFFAOYSA-N 0.000 description 2
- 230000007413 intestinal health Effects 0.000 description 2
- 235000021109 kimchi Nutrition 0.000 description 2
- 230000037356 lipid metabolism Effects 0.000 description 2
- 244000144972 livestock Species 0.000 description 2
- 208000004396 mastitis Diseases 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 230000004060 metabolic process Effects 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000003020 moisturizing effect Effects 0.000 description 2
- 239000004626 polylactic acid Substances 0.000 description 2
- 230000002980 postoperative effect Effects 0.000 description 2
- 244000144977 poultry Species 0.000 description 2
- 235000013594 poultry meat Nutrition 0.000 description 2
- 108090000765 processed proteins & peptides Proteins 0.000 description 2
- 210000002027 skeletal muscle Anatomy 0.000 description 2
- 235000020183 skimmed milk Nutrition 0.000 description 2
- 210000002460 smooth muscle Anatomy 0.000 description 2
- 239000008223 sterile water Substances 0.000 description 2
- 208000001072 type 2 diabetes mellitus Diseases 0.000 description 2
- 206010000060 Abdominal distension Diseases 0.000 description 1
- 208000024827 Alzheimer disease Diseases 0.000 description 1
- 241000304886 Bacilli Species 0.000 description 1
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 description 1
- 229920001661 Chitosan Polymers 0.000 description 1
- 108010009685 Cholinergic Receptors Proteins 0.000 description 1
- 206010011224 Cough Diseases 0.000 description 1
- 229920000858 Cyclodextrin Polymers 0.000 description 1
- 208000002249 Diabetes Complications Diseases 0.000 description 1
- 206010012655 Diabetic complications Diseases 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 208000004262 Food Hypersensitivity Diseases 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 241000287828 Gallus gallus Species 0.000 description 1
- 206010019133 Hangover Diseases 0.000 description 1
- 241000282412 Homo Species 0.000 description 1
- 208000035150 Hypercholesterolemia Diseases 0.000 description 1
- 208000031226 Hyperlipidaemia Diseases 0.000 description 1
- 102000004877 Insulin Human genes 0.000 description 1
- 108090001061 Insulin Proteins 0.000 description 1
- 241000233805 Phoenix Species 0.000 description 1
- 206010057190 Respiratory tract infections Diseases 0.000 description 1
- 241000282887 Suidae Species 0.000 description 1
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 1
- 206010046555 Urinary retention Diseases 0.000 description 1
- 206010046914 Vaginal infection Diseases 0.000 description 1
- DPXJVFZANSGRMM-UHFFFAOYSA-N acetic acid;2,3,4,5,6-pentahydroxyhexanal;sodium Chemical compound [Na].CC(O)=O.OCC(O)C(O)C(O)C(O)C=O DPXJVFZANSGRMM-UHFFFAOYSA-N 0.000 description 1
- 102000034337 acetylcholine receptors Human genes 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 210000001789 adipocyte Anatomy 0.000 description 1
- 238000013272 agar well diffusion method Methods 0.000 description 1
- 230000002009 allergenic effect Effects 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 1
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 1
- 235000011130 ammonium sulphate Nutrition 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 208000011775 arteriosclerosis disease Diseases 0.000 description 1
- 206010003246 arthritis Diseases 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 230000001363 autoimmune Effects 0.000 description 1
- 239000003833 bile salt Substances 0.000 description 1
- 239000011230 binding agent Substances 0.000 description 1
- 230000008827 biological function Effects 0.000 description 1
- 210000004204 blood vessel Anatomy 0.000 description 1
- 210000004556 brain Anatomy 0.000 description 1
- 230000007177 brain activity Effects 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 239000001110 calcium chloride Substances 0.000 description 1
- 229910001628 calcium chloride Inorganic materials 0.000 description 1
- 239000002775 capsule Substances 0.000 description 1
- 239000004202 carbamide Substances 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 235000014633 carbohydrates Nutrition 0.000 description 1
- 239000001768 carboxy methyl cellulose Substances 0.000 description 1
- 230000003915 cell function Effects 0.000 description 1
- 239000013592 cell lysate Substances 0.000 description 1
- 210000003169 central nervous system Anatomy 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 235000013330 chicken meat Nutrition 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 230000003930 cognitive ability Effects 0.000 description 1
- 208000010877 cognitive disease Diseases 0.000 description 1
- 235000009508 confectionery Nutrition 0.000 description 1
- IQFVPQOLBLOTPF-HKXUKFGYSA-L congo red Chemical compound [Na+].[Na+].C1=CC=CC2=C(N)C(/N=N/C3=CC=C(C=C3)C3=CC=C(C=C3)/N=N/C3=C(C4=CC=CC=C4C(=C3)S([O-])(=O)=O)N)=CC(S([O-])(=O)=O)=C21 IQFVPQOLBLOTPF-HKXUKFGYSA-L 0.000 description 1
- 208000029078 coronary artery disease Diseases 0.000 description 1
- 239000002537 cosmetic Substances 0.000 description 1
- 230000001186 cumulative effect Effects 0.000 description 1
- 238000000354 decomposition reaction Methods 0.000 description 1
- 238000009792 diffusion process Methods 0.000 description 1
- 230000001079 digestive effect Effects 0.000 description 1
- 210000002249 digestive system Anatomy 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- 239000007884 disintegrant Substances 0.000 description 1
- 239000003937 drug carrier Substances 0.000 description 1
- 201000006549 dyspepsia Diseases 0.000 description 1
- 239000003995 emulsifying agent Substances 0.000 description 1
- 239000002662 enteric coated tablet Substances 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 230000004149 ethanol metabolism Effects 0.000 description 1
- 210000003608 fece Anatomy 0.000 description 1
- 239000000835 fiber Substances 0.000 description 1
- 235000013373 food additive Nutrition 0.000 description 1
- 239000002778 food additive Substances 0.000 description 1
- 235000020932 food allergy Nutrition 0.000 description 1
- 235000012041 food component Nutrition 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 230000008014 freezing Effects 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 238000009499 grossing Methods 0.000 description 1
- 230000007236 host immunity Effects 0.000 description 1
- 230000002218 hypoglycaemic effect Effects 0.000 description 1
- 239000007943 implant Substances 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 230000015788 innate immune response Effects 0.000 description 1
- 229940125396 insulin Drugs 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 210000003734 kidney Anatomy 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 239000000314 lubricant Substances 0.000 description 1
- 230000001050 lubricating effect Effects 0.000 description 1
- 230000002934 lysing effect Effects 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 230000003446 memory effect Effects 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- 239000002068 microbial inoculum Substances 0.000 description 1
- 230000005012 migration Effects 0.000 description 1
- 238000013508 migration Methods 0.000 description 1
- 235000013336 milk Nutrition 0.000 description 1
- 239000008267 milk Substances 0.000 description 1
- 210000004080 milk Anatomy 0.000 description 1
- 239000004570 mortar (masonry) Substances 0.000 description 1
- 239000006873 mp medium Substances 0.000 description 1
- 210000003097 mucus Anatomy 0.000 description 1
- 210000000663 muscle cell Anatomy 0.000 description 1
- 201000009240 nasopharyngitis Diseases 0.000 description 1
- 210000004126 nerve fiber Anatomy 0.000 description 1
- 239000002858 neurotransmitter agent Substances 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 239000008194 pharmaceutical composition Substances 0.000 description 1
- 229920003168 pharmaceutical polymer Polymers 0.000 description 1
- 230000001766 physiological effect Effects 0.000 description 1
- 239000006187 pill Substances 0.000 description 1
- 229920000747 poly(lactic acid) Polymers 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 239000002861 polymer material Substances 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 230000002797 proteolythic effect Effects 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 230000006965 reversible inhibition Effects 0.000 description 1
- HFHDHCJBZVLPGP-UHFFFAOYSA-N schardinger α-dextrin Chemical compound O1C(C(C2O)O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC(C(O)C2O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC2C(O)C(O)C1OC2CO HFHDHCJBZVLPGP-UHFFFAOYSA-N 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 235000019812 sodium carboxymethyl cellulose Nutrition 0.000 description 1
- 229920001027 sodium carboxymethylcellulose Polymers 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 230000007480 spreading Effects 0.000 description 1
- 238000003892 spreading Methods 0.000 description 1
- 239000003381 stabilizer Substances 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 238000005728 strengthening Methods 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000004094 surface-active agent Substances 0.000 description 1
- 238000001356 surgical procedure Methods 0.000 description 1
- 239000000375 suspending agent Substances 0.000 description 1
- 238000013268 sustained release Methods 0.000 description 1
- 239000012730 sustained-release form Substances 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 210000000225 synapse Anatomy 0.000 description 1
- 208000011580 syndromic disease Diseases 0.000 description 1
- 239000003826 tablet Substances 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 235000013311 vegetables Nutrition 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 238000012070 whole genome sequencing analysis Methods 0.000 description 1
- 230000029663 wound healing Effects 0.000 description 1
- 230000037303 wrinkles Effects 0.000 description 1
Classifications
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Abstract
The invention belongs to the technical field of probiotics and application thereof, and particularly relates to a lactobacillus mucilaginosus JF5 strain and application thereof in preparation of lipid-reducing and digestion-aiding foods and medicines. In order to further develop and utilize the probiotics function of the fermented lactobacillus, the invention separates a fermented lactobacillus JF5 strain from pickle samples, and the strain has various functions of producing protease, cellulase, gamma-aminobutyric acid, hyaluronic acid, inhibiting alpha-glucosidase activity, inhibiting acetylcholinesterase activity and the like. Therefore, the strain has the beneficial effects of promoting the digestion and absorption of protein food to improve protein allergy, promoting the absorption of cellulose to improve constipation, resisting depression and alcohol, resisting aging and crease, reducing blood sugar, promoting intestinal absorption and peristalsis and the like, and has important application value and economic value.
Description
Technical Field
The invention belongs to the technical field of probiotics and application thereof, and particularly relates to a lactobacillus mucilaginosus JF5 strain and application thereof in preparation of lipid-reducing and digestion-aiding foods and medicines.
Background
Lactobacillus fermentum (limosilactis), also known as lactobacillus fermentum, gram positive bacilli, facultative anaerobism. The lactobacillus fermentum has higher tolerance and can adapt to the environment with low acid and high bile salt in human intestinal tracts. In addition, the lactobacillus fermentum also has higher safety and genetic stability, is one of the earliest planted probiotics in human intestinal tracts, can continuously change along with the growth of human ages and the change of human bodies, and can influence the health of human bodies when the number of the probiotics is lower than a normal value.
Lactobacillus fermentum is widely present in the intestinal tract of humans and animals and is excreted with faeces. Lactobacillus mucilaginosus is a part of the normal flora of the human body, which is one of the important components of the normal microbial system of the intestinal tract and accompanies the host for life, and has important significance for maintaining the microecological balance of the intestinal tract. Meanwhile, as probiotics in intestinal flora, the fermented mucus lactobacillus is also edible lactobacillus, has no toxic or side effect, can play a role in helping intestinal peristalsis, and can also enhance the digestion capability of human bodies, improve immunity and the like. At present, fermented lactobacillus mucilaginosus has become a hot spot of research as a probiotic lactobacillus mucilaginosus with great potential and is being continuously used to make probiotic preparations suitable for human and animals.
Currently, different strains of lactobacillus fermentum have been reported to have different probiotic functions: (1) enhancing host immunity. For example, the lactobacillus mucilaginosus CECT5716 can effectively prevent, relieve and treat common uncomfortable symptoms of pregnant women and lactating women, help the fetus to pre-establish an intestinal immune protection system before birth, and remarkably improve the immunity and the resistance of infants. (2) antibacterial: preventing respiratory tract infection, reducing incidence of common cold, cough, etc., and preventing vaginal infection. (3) improving intestinal health: promoting health of gastrointestinal digestive system, reducing harmful substances in intestinal tract, and relieving constipation. (4) Helping the fetus build an autoimmune system, and laying a foundation for the innate immunity of the baby. (5) Effectively preventing and treating mastitis, relieving mammitis, relieving breast soreness, relieving pain, promoting breast health, and relieving tension and anxiety.
It can be seen that the diversity of sources of lactobacillus mucilaginosus results in genetic and functional diversity. However, the current studies on the separation and identification, the probiotics characteristics and the metabolic mechanism of the lactobacillus fermentum are still relatively few, which also affects the further development and utilization of the lactobacillus fermentum to a certain extent. Therefore, it is necessary to make lactobacillus mucilaginosus function better according to different sources of the lactobacillus mucilaginosus, such as determining the efficacy according to the functions of the strain or the probiotics metabolite, and defining the application prospect. In conclusion, the research and application of the probiotic fermented lactobacillus mucilaginosus have wider development space.
Disclosure of Invention
In order to overcome the defects in the prior art, the invention separates a lactobacillus mucilaginosus (JF 5) strain from pickle samples, and the strain has the functions of cellulase activity, hyaluronic acid production, alpha-glucosidase activity inhibition and the like, and has great potential application value.
In order to achieve the above purpose, the technical scheme adopted by the invention is as follows:
the first aspect of the present invention provides a lactobacillus fermentum JF5 strain, which is deposited with the chinese collection of typical cultures at 10 months 4 of 2022, with accession number: cctccc No. M20221511; the 16S rDNA complete sequence of the lactobacillus mucilaginosus JF5 strain is shown in SEQ ID No: 1.
In a second aspect the invention provides the use of a lactobacillus fermentum JF5 strain according to the first aspect for the production of cellulases.
The research shows that the probiotics fermented lactobacillus mucilaginosus JF5 can produce protease, which suggests that the fermented lactobacillus mucilaginosus JF5 is hopeful to be used for degrading cell walls of pathogenic fungi and controlling diseases; decomposing cellulose in the compost; preparing livestock and poultry breeding feed and the like.
In a third aspect the invention provides the use of a lactobacillus fermentum JF5 strain according to the first aspect for the production of hyaluronic acid.
The research shows that the probiotics lactobacillus mucilaginosus JF5 can produce Hyaluronic Acid (HA), which suggests that the probiotics lactobacillus mucilaginosus JF5 is expected to be used in the fields of anti-inflammation, anti-angiogenesis, anti-aging, wound inflammation promotion, healing and the like.
In a fourth aspect the invention provides the use of a lactobacillus fermentum JF5 strain according to the first aspect for the preparation of an alpha-glucosidase inhibitor.
Through researches, the probiotic fermented lactobacillus mucilaginosus JF5 can effectively inhibit the activity of alpha-glucosidase, and the alpha-glucosidase is related to type 2 diabetes, and the inhibition of the alpha-glucosidase is one of methods for controlling postprandial hyperglycemia, so that the fermented lactobacillus mucilaginosus JF5 is expected to be applied to the fields of reducing blood sugar, inhibiting obesity and the like.
In a fifth aspect the invention provides the use of a lactobacillus fermentum JF5 strain according to the first aspect for the production of a protease.
The research shows that the probiotics fermented lactobacillus mucilaginosus JF5 can produce cellulase, which suggests that the fermented lactobacillus mucilaginosus JF5 is expected to be used in the fields of promoting the digestion and absorption of protein in food, improving the absorption of small peptide and amino acid, resisting allergy and the like.
In a sixth aspect the invention provides the use of a lactobacillus fermentum JF5 strain according to the first aspect for the production of gamma-aminobutyric acid.
The research shows that the probiotics fermented lactobacillus mucilaginosus JF5 can produce gamma-aminobutyric acid (GABA), which suggests that the fermented lactobacillus mucilaginosus JF5 is expected to be used in the fields of improving the sleeping quality of organisms, resisting depression, resisting anxiety, reducing blood pressure, improving lipid metabolism and the like.
In a seventh aspect, the invention provides the use of a lactobacillus fermentum JF5 strain according to the first aspect for the preparation of an acetylcholinesterase inhibitor.
The research shows that the probiotics lactobacillus fermentum JF5 can effectively inhibit the activity of acetylcholinesterase, and suggests that the probiotics lactobacillus fermentum JF5 is expected to be used in the fields of promoting gastrointestinal absorption, peristalsis and the like.
According to an eighth aspect of the present invention there is provided a bacterial agent for use in a probiotic apparatus, said bacterial agent comprising a lactobacillus mucilaginosus (JF 5) strain according to the first aspect.
Preferably, the lactobacillus mucilaginosus (JF 5) strain is a fermented cell mixture. The culture medium used for fermentation is MRS culture medium.
Preferably, in the field of pharmaceutical application, the microbial agent further comprises a pharmaceutically acceptable carrier and/or excipient.
More preferably, the above excipients refer to diluents, binders, lubricants, disintegrants, co-solvents, stabilizers, etc. and some pharmaceutically acceptable bases which are useful in the pharmaceutical field. The carrier is a functional pharmaceutical adjuvant available in the pharmaceutical field and comprises a surfactant, a suspending agent, an emulsifying agent and a plurality of novel pharmaceutical polymer materials, such as cyclodextrin, chitosan, polylactic acid (PLA), polyglycolic acid-polylactic acid copolymer (PLGA), hyaluronic acid and the like.
Preferably, in the field of medical application, the dosage forms of the microbial inoculum comprise tablets, granules, capsules, dripping pills, sustained release agents, oral liquid preparations and injections.
More preferably, the above-mentioned dosage forms refer to clinically usual dosage forms. Pharmaceutical formulations may be administered orally or parenterally (e.g., intravenously, subcutaneously, intraperitoneally, or topically), and if some drugs are unstable under gastric conditions, they may be formulated as enteric coated tablets.
Compared with the prior art, the invention has the beneficial effects that:
the invention separates a strain of fermented lactobacillus JF5 from pickle samples, and the fermented lactobacillus JF5 has various probiotics effects, including excellent protease activity, excellent cellulase activity, gamma-aminobutyric acid production and secretion, hyaluronic acid production and secretion, alpha-glucosidase activity inhibition, acetylcholinesterase activity inhibition and the like. Thus, lactobacillus mucilaginosus JF5 has the effects of promoting digestion and absorption of protein foods and improving protein allergy; promoting absorption of cellulose and improving constipation; can be used for resisting depression and relieving hangover; anti-aging and anti-wrinkle; reducing blood glucose; promoting intestinal absorption and peristalsis. Therefore, the newly separated fermented lactobacillus mucilaginosus strain JF5 has various probiotics effects, can be used in the fields of improving intestinal tolerance, improving intestinal health, reducing blood fat, helping digestion and the like, for example, can be prepared into food and medicine for reducing blood fat and helping digestion, and has important application value and economic value.
Drawings
FIG. 1 is a phylogenetic tree of Lactobacillus mucilaginosus JF5 and other homologous strains;
FIG. 2 shows the degradation of milk plates by Lactobacillus mucilaginosus JF5 (left, blank; right, experimental group);
FIG. 3 shows the degradation experiment of Lactobacillus mucilaginosus JF5 on cellulose plates (left, blank; right, experimental group);
FIG. 4 shows that Lactobacillus mucilaginosus JF5 produces gamma-aminobutyric acid;
FIG. 5 shows that Lactobacillus mucilaginosus JF5 produces and secretes hyaluronic acid;
FIG. 6 shows that the secretion of substances by Lactobacillus mucilaginous fermentation broth JF5 significantly inhibits alpha-glucosidase activity;
fig. 7 shows that the secretion of substances by lactobacillus mucilaginosus JF5 fermentation broth significantly inhibited acetylcholinesterase activity.
Detailed Description
The following describes the invention in more detail. The description of these embodiments is provided to assist understanding of the present invention, but is not intended to limit the present invention. In addition, the technical features of the embodiments of the present invention described below may be combined with each other as long as they do not collide with each other.
The experimental methods in the following examples, unless otherwise specified, are conventional, and the experimental materials used in the following examples, unless otherwise specified, are commercially available.
The following examples relate to the following experimental materials:
(1) Strains: lactobacillus mucilaginosus (JF 5) strain isolated from kimchi in Guangdong area of China by the national institute of university of middle mountain student's intestinal microbiome study moisturizing laboratory and stored in a glycerol tube at-80 ℃ by freezing. In general, it is inoculated on the surface of a plate of MRS solid medium and cultured upside down in a constant temperature anaerobic incubator at 37℃for 24 hours to obtain colonies, or cultured in a constant temperature anaerobic incubator at 37℃for 24-48 hours with shaking in MRS liquid medium to obtain fermentation broth.
(2) The kit comprises: gamma-aminobutyric acid (GABA) detection kit (Cloud-Clone corp., cat: CEA900 Ge), hyaluronic acid (aka hyaluronic acid, HA) detection kit (Cloud-Clone corp., cat: CEA182 Ge), alpha-glucosidase inhibitor screening kit (abcam, cat: ab 284520), acetylcholinesterase inhibitor screening kit (abnova, cat: KA 6219).
(3) MRS plate: 10g of beef extract, 10g of peptone, 5g of yeast extract, 2g of triammonium citrate, 5g of sodium acetate, 20g of glucose, 2g of dipotassium hydrogen phosphate, 1mL of Tween 80, 0.58g of magnesium sulfate, 0.25g of manganese sulfate, 15g of agar and ddH 2 Filling O to 1L, adjusting pH to 6.2-6.6, and autoclaving at 121deg.C for 20min to obtain MRS plate.
(4) MRS liquid medium: 10g of beef extract, 10g of peptone, 5g of yeast extract, 2g of triammonium citrate, 5g of sodium acetate, 20g of glucose, 2g of dipotassium hydrogen phosphate, 1mL of Tween 80, 0.58g of magnesium sulfate, 0.25g of manganese sulfate and ddH 2 Adding O to 1L, adjusting pH to 6.2-6.6, and autoclaving at 121deg.C for 20min to obtain MRS liquid culture medium.
(5) MP plate: 10g of skimmed milk powder, 1g of sodium chloride, 10g of beef extract, 10g of peptone, 5g of yeast extract, 20g of glucose, 2g of tri-ammonium citrate, 5g of sodium acetate, 2g of dipotassium hydrogen phosphate, 0.5mL of Tween 80, 0.58g of magnesium sulfate, 0.25g of manganese sulfate, 15g of agar and ddH 2 Filling O to 1L, adjusting pH to 6.2-6.6, autoclaving at 121deg.C for 20min, and making into MP plate.
(6) CMC plate: 10g of sodium carboxymethyl cellulose, 1.5g of ammonium sulfate, 0.3g of manganese sulfate, 0.2g of calcium chloride, 5g of sodium chloride, 0.3g of urea, 10g of beef extract, 10g of peptone, 5g of yeast extract, 20g of glucose, 2g of tri-ammonium citrate, 5g of sodium acetate, 2g of dipotassium hydrogen phosphate, 0.5mL of tween 80, 0.58g of magnesium sulfate, 0.25g of manganese sulfate, 15g of agar and ddH 2 Filling O to 1L, adjusting pH to 6.2-6.6, autoclaving at 121deg.C for 20min, and preparing into CMC plate.
Example 1 isolation and characterization of Lactobacillus fermentum JF5
The lactobacillus mucilaginosus Limosilactobacillusfermentum JF strain is separated from pickle samples in Guangdong province of China, and is specifically as follows:
repeatedly cleaning pickled cabbage samples for one month purchased from the new phoenix comprehensive meat and vegetable market in Guangzhou sea pearl area with sterile water for 3 times, placing the pickled cabbage samples in a mortar, adding sterile water (v/m=1:10), thoroughly grinding the pickled cabbage samples into homogenate, sucking a proper amount of grinding fluid by a pipette, coating the grinding fluid on an MRS flat plate, and culturing the pickled cabbage pickle samples at room temperature for 3 days. The colonies to be streaked and purified in the separation experiment plates were numbered with a marker and strain numbers were marked on the plates accordingly. Colonies were picked and inoculated onto MRS plates and the strains were purified by plate streaking. If the strain cannot be separated by the method, colonies need to be picked from the enrichment plate, and the colonies are coated on the MRS plate after being subjected to gradient dilution by the MRS liquid culture medium. Reference is made to the "Berger's Manual of identification of bacteria" (eighth edition) and the "manual of identification of fungi" which identify strains belonging to bacteria first, and then the growth of colonies is observed. The purified strain is obtained through preliminary separation, the strain number is JT1, and after 24 hours of culture, bacterial colonies of the strain are observed to be round, milky white, smooth in surface, raised and neat in edge.
Next, after molecular identification by 16S rDNA universal primer (27F: AGAGTTTGATCCTGGCTCAG,1492R: TACGGCTACCTTGTTACGACTT), the isolated L.fermentum JF5 strain was subjected to whole genome sequencing by Beijing Baimeike biosciences Corp. The resulting 16S rDNA sequence (SEQ ID No: 1) was subjected to BLAST alignment at NCBI' S Genome database. The results show that the JF5 strain has >99% homology with the 16S rDNA sequence of the known lactobacillus fermentum (l.fertum); and performing evolution analysis with homologous strains (FIG. 1), confirming that strain JF5 is a different strain of Lactobacillus mucilaginosus.
Finally, strain JF5 is preserved, and the preservation information is as follows: preservation time: 2022, 10 and 4; preservation unit name: china Center for Type Culture Collection (CCTCC); deposit number: cctccc No. M20221511; deposit unit address: chinese university of Wuhan; classification naming: limosilactobacillus referenced.
Lactobacillus fermentum is a probiotic bacterial strain approved by the country for use in food, has wide probiotic effects such as antibacterial, anti-inflammatory and improving health of mother and infant, but different sources of bacterial strains have different effects, which means that the novel lactobacillus fermentum JF5 isolated from kimchi samples according to the present invention can be used as a probiotic and may have novel effects and functions.
L.fermentum JF516S rDNA sequence(1450bp，SEQ ID No：1)：
TACTGCAGTCGACGCGTTGGCCCAATTGATTGATGGTGCTTGCACCTGATTGATTTTGGTCGCCAACGAGTGGCGGACGGGTGAGTAACACGTAGGTAACCTGCCCAGAAGCGGGGGACAACATTTGGAAACAGATGCTAATACCGCATAACAACGTTGTTCGCATGAACAACGCTTAAAAGATGGCTTCTCGCTATCACTTCTGGATGGACCTGCGGTGCATTAGCTTGTTGGTGGGGTAAnGGCCTACCAAGGCGATGATGCATAGCCGAGTTGAGAGACTGATCGGCCACAATGGGACTGAGACACGGCCCATACTCCTACGGGAGGCAGCAGTAGGGAATCTTCCACAATGGGCGCAAGCCTGATGGAGCAACACCGCGTGAGTGAAGAAGGGTTTCGGCTCGTAAAGCTCTGTTGTTAAAGAAGAACACGTATGAGAGTAACTGTTCATACGTTGACGGTATTTAACCAGAAAGTCACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTATCCGGATTTATTGGGCGTAAAGAGAGTGCAGGCGGTTTTCTAAGTCTGATGTGAAAGCCTTCGGCTTAACCGGAGAAGTGCATCGGAAACTGGATAACTTGAGTGCAGAAGAGGGTAGTGGAACTCCATGTGTAGCGGTGGAATGCGTAGATATATGGAAGAACACCAGTGGCGAAGGCGGCTACCTGGTCTGCAACTGACGCTGAGACTCGAAAGCATGGGTAGCGAACAGGATTAGATACCCTGGTAGTCCATGCCGTAAACGATGAGTGCTAGGTGTTGGAGGGTTTCCGCCCTTCAGTGCCGGAGCTAACGCATTAAGCACTCCGCCTGGGGAGTACGACCGCAAGGTTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCTACGCGAAGAACCTTACCAGGTCTTGACATCTTGCGCCAACCCTAGAGATAGGGCGTTTCCTTCGGGAACGCAATGACAGGTGGTGCATGGTCGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTGTTACTAGTTGCCAGCATTAAGTTGGGCACTCTAGTGAGACTGCCGGTGACAAACCGGAGGAAGGTGGGGACGACGTCAGATCATCATGCCCCTTATGACCTGGGCTACACACGTGCTACAATGGACGGTACAACGAGTCGCGAACTCGCGAGGGCAAGCAAATCTCTTAAAACCGTTCTCAGTTCGGACTGCAGGCTGCAACTCGCCTGCACGAAGTCGGAATCGCTAGTAATCGCGGATCAGCATGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCATGAGAGTTTGTAACACCCAAAGTCGGTGGGGTAACCTTTAGGAGCCAGCCGCCTAAGT。
Example 2 function of Lactobacillus mucilaginosus (Lactobacillus fermentum) JF5 and use thereof
(1) Protease capable of producing degradable milk protein by fermenting lactobacillus mucilaginosus JF5 strain
The identification and measurement of the ability of lactobacillus mucilaginosus JF5 to secrete proteolytic proteins was performed according to the agar well diffusion assay using a skim milk plate medium (MP plate). In the test, 3uL of lactobacillus mucilaginosus JF5 bacteria solution with the concentration of 10Abs is dripped into the test group, and 3uL of blank MRS culture medium is dripped into the control group. The cells were cultured in an anaerobic incubator at 37℃for 3 days in an inverted manner. The results showed that JF5 significantly degraded protein and formed a distinct degradation circle (fig. 2) compared to the control with the blank medium, indicating that lactobacillus mucilaginosus JF5 produced protease.
Therefore, the lactobacillus mucilaginosus JF5 can promote the digestion and absorption of protein in food by human body and improve the absorption of small peptide and amino acid when used as a probiotic bacterial strain. And can be used for resisting allergy (improving food allergy caused by protein dyspepsia or non-absorption). In addition, the method can also be used for extracting protease and is applied to the production in the protease fields of food industry, washing industry and the like; can also be used in microbial feed to help animals digest and absorb nutrition, and improve the utilization rate of the feed.
(2) Cellulase produced by fermenting lactobacillus mucilaginosus JF5 strain
The identification and measurement of the cellulose degrading ability of lactobacillus mucilaginosus JF5 were carried out according to the agar well diffusion method, with slight modification, and the cellulose plate medium used was MP medium. In the test, 3uL of lactobacillus mucilaginosus JF5 bacteria solution with the concentration of 10Abs is dripped into the test group, and 3uL of blank MRS culture medium is dripped into the control group. Congo red staining was performed after 3 days of inversion culture in an anaerobic incubator at 37 ℃. The results show that JF5 can significantly degrade cellulose and form a distinct degradation loop compared to the control with the blank medium (fig. 3), indicating that lactobacillus mucilaginosus JF5 can produce cellulase.
It can be seen that probiotic fermented lactobacillus mucilaginosus JF5 strains can serve several purposes by producing the efficacy of cellulases: (1) to facilitate the digestive absorption of dietary components; (2) improving constipation: the cellulose is decomposed in the intestinal canal to generate certain moisture, and the effect of softening the stool is achieved; (3) Cholesterol reduction is the most important effect of cellulose, because the soluble fiber can inhibit the absorption of cholesterol by human body after being absorbed by human body, can be combined with cholesterol in intestinal tract, quickens the metabolism of cholesterol in human body, prevents human body from inducing hyperlipidemia due to hypercholesteremia, and can also reduce the incidence rate of arteriosclerosis and coronary heart disease; (4) The non-sweet 'sugar' of cellulose can slow down the absorption of glucose by blood, balance the concentration of blood sugar and promote the sensitivity of muscle and fat cells to insulin, thereby preventing and assisting in treating diabetes.
In addition, the lactobacillus mucilaginosus JF5 can also be used for fermenting and extracting cellulase, and has wide application in the food industry and the environmental industry; can be applied to degrading cell walls of pathogenic fungi and controlling diseases; can be applied to the decomposition of cellulose in compost; can also be used for preparing livestock and poultry raising feed, such as monogastric animal feed for pigs, chickens and the like, so as to overcome the defect that cellulose cannot be utilized.
(3) Fermentation of Lactobacillus mucilaginosus JF5 strain produces gamma-aminobutyric acid (GABA)
Culturing Lactobacillus fermentum JF5 in MRS liquid culture medium to stationary phase, spreading in new MRS liquid culture medium at dilution ratio of 1:30, collecting bacterial suspension when culturing to stationary phase for 24 hr, centrifuging at 10,000Xg and 4deg.C for 10min, collecting cultured thallus, and collecting thallus with buffer PBS (8 g NaCl, 0.2g KCl, 1.44g Na are weighed 2 HPO 4 、0.24gKH 2 PO 4 Dissolving in 800mL distilled water, regulating the solution to 7.2 with HCl, adding distilled water to a volume of 1L to obtain PBS buffer solution, and determining GABA concentration of cultured thallus by using GABA specific ELISA kit (CEA 900 Ge). The results showed that the concentration of GABA in the cell lysate of JF5 was significantly increased compared to buffer PBS used for lysing cells, and that the cumulative amount was 18.16pg/mL, indicating that Lactobacillus fermentum JF5 can produce gamma-aminobutyric acid in the stationary phase (FIG. 4).
Gamma-aminobutyric acid is an important central nervous system inhibitory neurotransmitter, and is widely present in animals, plants and microorganisms. It has been demonstrated that GABA, a small molecular weight non-protein amino acid, is food safe and can be used as a food additive. Research shows that intake of a certain amount of GABA has the physiological effects of improving sleeping quality of organisms, resisting depression, resisting anxiety, reducing blood pressure, improving lipid metabolism, enhancing memory and brain activity, accelerating brain metabolism, strengthening liver and kidney, promoting ethanol metabolism (dispelling alcohol effect), improving climacteric syndrome and the like.
Thus, the probiotic fermented lactobacillus mucilaginosus JF5 strain can exert the above multiple uses by the effect of producing gamma-aminobutyric acid.
(4) The lactobacillus mucilaginosus JF5 strain can produce and secrete Hyaluronic Acid (HA)
Lactobacillus fermentum JF5 cultured with MRS liquid medium to stationary phase was expanded into new MRS liquid medium at a dilution factor of 1:30, bacterial suspension was collected at 24h of stationary phase, and after centrifugation at 10,000×g at 4 ℃ for 10min, the supernatant of the fermentation broth was collected, and the HA concentration of the supernatant of the fermentation broth was determined by means of hyaluronic acid (also known as hyaluronic acid, HA) specific ELISA kit (CEA 182 Ge). The results showed that the concentration of HA in the fermentation supernatant of JF5 was significantly increased compared to the low concentration of HA in the blank medium MRS, with an accumulated amount of 9.96ng/mL, indicating that lactobacillus mucilaginosus JF5 can produce and secrete hyaluronic acid in the stationary phase (fig. 5).
Hyaluronic acid, also known as hyaluronic acid, is a biodegradable, biocompatible, non-toxic, non-allergenic polymer with a variety of biological functions. Has anti-inflammatory and anti-angiogenesis effects, and has strong anti-aging, moisturizing and wrinkle smoothing abilities. The anti-wrinkle agent is beneficial to skin anti-wrinkle, promotes wound anti-inflammation and healing, can be used as an anti-wrinkle agent, and has the potential of developing skin cosmetics. In addition, HA HAs high lubricating, water absorbing and retaining ability, and can affect various cell functions such as migration, adhesion and proliferation, so that HA is also widely used in biomedical fields such as ophthalmic surgery, arthritis treatment, wound healing scaffolds, tissue engineering, implant materials, and the like.
Thus, the probiotic fermented lactobacillus mucilaginosus JF5 strain can exert the above multiple uses by the efficacy of hyaluronic acid production.
(5) The fermentation liquor of the lactobacillus mucilaginosus JF5 strain can effectively inhibit the activity of alpha-glucosidase
Lactobacillus fermentum JF5 cultured with MRS liquid medium to stationary phase was expanded into new MRS liquid medium at a dilution ratio of 1:30, bacterial suspension was collected at 24h of stationary phase, and after centrifugation at 10,000×g at 4 ℃ for 10min, the supernatant of the fermentation broth was collected, and the effect of the supernatant of the fermentation broth on the enzyme activity ability of α -glucosidase to hydrolyze glucose was measured by an α -glucosidase inhibitor screening kit (ab 284520). The results showed that the fermentation supernatant of JF5 significantly inhibited the ability of α -glucosidase to hydrolyze glucose compared to the non-inhibitory effect of the blank medium MRS, with an inhibition rate of about 58.82%, indicating that the fermentation broth of lactobacillus mucilaginosus JF5 can effectively inhibit the activity of α -glucosidase (fig. 6).
Alpha-glucosidase, an enzyme that plays a role in carbohydrate breakdown, is associated with type 2 diabetes, and inhibition of alpha-glucosidase is one of the methods of controlling postprandial hyperglycemia, thereby contributing to the treatment of diabetes. Thus, α -glucosidase inhibitors help to maintain blood glucose levels and can improve diabetic complications. Furthermore, inhibition of α -glucosidase activity may control obesity.
Thus, the probiotic fermented lactobacillus mucilaginosus JF5 strain has an alpha-glucosidase inhibitory activity, which makes it a potential hypoglycemic and obesity inhibiting probiotic.
(6) Fermented lactobacillus mucilaginosus JF5 strain fermentation liquor can effectively inhibit acetylcholinesterase activity
Lactobacillus fermentum JF5 cultured with MRS liquid medium to stationary phase was expanded into new MRS liquid medium at a dilution ratio of 1:30, bacterial suspension was collected at 24h of stationary phase, and after centrifugation at 10,000×g at 4 ℃ for 10min, the supernatant of the fermentation broth was collected, and the inhibitory ability of the supernatant of the fermentation broth against acetylcholinesterase (AchE) activity was measured by means of acetylcholinesterase inhibitor screening kit (KA 6219). The results showed that the fermentation supernatant of JF5 significantly inhibited the activity of acetylcholinesterase compared to the non-inhibitory effect of the blank medium MRS, with an inhibition rate of about 3.47%, indicating that the fermentation broth of lactobacillus mucilaginosus JF5 can effectively inhibit the activity of acetylcholinesterase (fig. 7).
The acetylcholinesterase inhibitor can continuously accumulate at synapses under the action of acetylcholine released by nerve fiber tips through reversible inhibition of acetylcholinesterase, excite cholinergic receptors, and prolong and increase the action of acetylcholine. Acetylcholinesterase inhibitors can enhance cognitive ability and memory (improve Alzheimer's disease, improve cognitive dysfunction), dilate blood vessels (improve heart rate overspeed), excite skeletal and smooth muscles (improve constipation, postoperative abdominal distension, postoperative urinary retention).
Thus, the probiotic fermented lactobacillus mucilaginosus JF5 strain can inhibit acetylcholinesterase activity, and excite skeletal muscle and smooth muscle of intestinal tract, which makes it a potential probiotic for promoting gastrointestinal absorption and peristalsis.
Taken together, the novel isolated lactobacillus mucilaginosus strain JF5 of the present invention has a variety of probiotic effects: (1) has excellent protease activity; (2) has excellent cellulase activity; (3) gamma-aminobutyric acid can be produced and secreted; (4) can produce and secrete hyaluronic acid; (5) can inhibit alpha-glucosidase activity; (6) can inhibit acetylcholinesterase activity. Therefore, the lactobacillus mucilaginosus strain JF5 has important application value and economic value.
The embodiments of the present invention have been described in detail above, but the present invention is not limited to the described embodiments. It will be apparent to those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the invention, and yet fall within the scope of the invention.
Claims (9)
1. A lactobacillus mucilaginosus (Limosilactobacillus fermentum) JF5 strain, wherein the lactobacillus mucilaginosus JF5 strain was deposited with the chinese collection of typical cultures at 2022, 9 and 27 days, under the accession number: cctccc No. M20221511; the 16S rDNA complete sequence of the lactobacillus mucilaginosus JF5 strain is shown in SEQ ID No: 1.
2. Use of a lactobacillus mucilaginosus (Limosilactobacillus fermentum) JF5 strain according to claim 1 for producing cellulases.
3. Use of a lactobacillus mucilaginosus (Limosilactobacillus fermentum) JF5 strain according to claim 1 for the production of hyaluronic acid.
4. Use of a lactobacillus mucilaginosus (Limosilactobacillus fermentum) JF5 strain according to claim 1 for the preparation of an acetylcholinesterase inhibitor.
5. Use of a lactobacillus mucilaginosus (Limosilactobacillus fermentum) JF5 strain according to claim 1 for the production of proteases for the degradation of milk proteins.
6. Use of a lactobacillus mucilaginosus (Limosilactobacillus fermentum) JF5 strain according to claim 1 for the production of gamma-aminobutyric acid.
7. Use of a lactobacillus mucilaginosus (Limosilactobacillus fermentum) JF5 strain according to claim 1 for the preparation of an α -glucosidase inhibitor.
8. A probiotic functional bacterial agent, characterized in that it comprises the lactobacillus mucilaginosus (Limosilactobacillus fermentum) JF5 strain of claim 1.
9. The probiotic functional microbial agent according to claim 8, wherein the microbial agent is a fermented cell mixture of lactobacillus mucilaginosus (Limosilactobacillus fermentum) JF5 strain.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310235233.9A CN116814464B (en) | 2023-03-13 | 2023-03-13 | Fermented lactobacillus mucilaginosus JF5 and application thereof in preparation of lipid-reducing and digestion-aiding foods and medicines |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310235233.9A CN116814464B (en) | 2023-03-13 | 2023-03-13 | Fermented lactobacillus mucilaginosus JF5 and application thereof in preparation of lipid-reducing and digestion-aiding foods and medicines |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN116814464A CN116814464A (en) | 2023-09-29 |
| CN116814464B true CN116814464B (en) | 2024-02-02 |
Family
ID=88124749
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202310235233.9A Active CN116814464B (en) | 2023-03-13 | 2023-03-13 | Fermented lactobacillus mucilaginosus JF5 and application thereof in preparation of lipid-reducing and digestion-aiding foods and medicines |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN116814464B (en) |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107502575A (en) * | 2017-09-20 | 2017-12-22 | 中国农业科学院农产品加工研究所 | One plant of Lactobacillus plantarum with the high inhibitory activity of α glucuroides |
| KR101825836B1 (en) * | 2016-10-13 | 2018-02-07 | 건국대학교 산학협력단 | Novel Lactobacillus plantarum Lb41 strain and compositions for the prevention and treatment of diabetes or insulin resistance syndrome containing the same |
| CN110741073A (en) * | 2017-06-09 | 2020-01-31 | 曾根农场股份公司 | Novel lactic acid bacteria and use thereof |
| CN114381403A (en) * | 2022-01-21 | 2022-04-22 | 中山大学 | Bacillus belgii LOH112 and application thereof |
-
2023
- 2023-03-13 CN CN202310235233.9A patent/CN116814464B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR101825836B1 (en) * | 2016-10-13 | 2018-02-07 | 건국대학교 산학협력단 | Novel Lactobacillus plantarum Lb41 strain and compositions for the prevention and treatment of diabetes or insulin resistance syndrome containing the same |
| CN110741073A (en) * | 2017-06-09 | 2020-01-31 | 曾根农场股份公司 | Novel lactic acid bacteria and use thereof |
| CN107502575A (en) * | 2017-09-20 | 2017-12-22 | 中国农业科学院农产品加工研究所 | One plant of Lactobacillus plantarum with the high inhibitory activity of α glucuroides |
| CN114381403A (en) * | 2022-01-21 | 2022-04-22 | 中山大学 | Bacillus belgii LOH112 and application thereof |
Non-Patent Citations (1)
| Title |
|---|
| 响应面法优化发酵乳杆菌产γ-氨基丁酸的培养条件;上官文菲等;中国酿造;第38卷(第12期);87-91 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN116814464A (en) | 2023-09-29 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103333847B (en) | Lactobacillus fermenti grx08 with function of assisting blood fat reduction and application of lactobacillus fermenti grx08 | |
| CN110106119B (en) | Lactobacillus rhamnosus M9 separated from breast milk and application thereof | |
| CN116555076B (en) | Bifidobacterium longum subspecies longum MY1 and application thereof in preparation of food and medicine for relaxing bowels and protecting intestines | |
| CN116555074B (en) | Lactobacillus brevis JT1 and application thereof in preparation of hypoglycemic drugs | |
| CN117004503B (en) | Saliva combined lactobacillus MB1 and application thereof in preparation of food and medicine for assisting sleep and regulating intestines and stomach | |
| CN116555075B (en) | Lactobacillus plantarum JF1 and application thereof in preparation of anti-aging food and drug | |
| CN115895966B (en) | Bifidobacterium bifidum BL002 for assisting in relieving gout and application thereof | |
| CN114686405B (en) | Bifidobacterium bifidum with functions of reducing fat, relieving hyperglycemia and regulating intestinal immunity and application thereof | |
| CN116814464B (en) | Fermented lactobacillus mucilaginosus JF5 and application thereof in preparation of lipid-reducing and digestion-aiding foods and medicines | |
| CN114836349A (en) | Lactobacillus acidophilus LA16 for antagonizing helicobacter pylori and application thereof | |
| CN116286519B (en) | Lactobacillus paracasei KS3 and application thereof in preparation of anti-aging and digestion-aiding foods and medicines | |
| CN116656526B (en) | Lactobacillus plantarum JF4 and application thereof in preparation of blood sugar and cholesterol reducing foods and medicines | |
| CN116218733B (en) | Lactobacillus rhamnosus XY5 and application thereof in preparing antiallergic and digestion-promoting food and drug | |
| CN117286045B (en) | Bifidobacterium longum subspecies longum KS2 and application thereof in preparation of anti-aging medicines | |
| CN116574634B (en) | Streptococcus salivarius thermophilus subspecies JF2 and application thereof in preparation of anti-inflammatory and lipid-relieving food and drug | |
| KR102495246B1 (en) | Lactobacillus helveticus BCC-LH-04 having body fat-reducing activity and compositions comprising the same | |
| CN116478890B (en) | Lactobacillus casei for regulating high blood sugar level, preparation and application thereof | |
| CN117402768B (en) | Lactobacillus reuteri KA1 and application thereof in preparation of anti-inflammatory and anti-tumor medicines | |
| CN116970530B (en) | Bifidobacterium breve FPHC4024 for preventing and treating infantile diarrhea and application thereof | |
| CN117384790A (en) | Pediococcus pentosaceus KS5 and application thereof in preparation of anti-inflammatory and sleep-aiding foods and medicines | |
| CN117343875A (en) | Lactobacillus gasseri MY5 and application thereof in preparation of anti-inflammatory, laxative and intestine-protecting food and medicine | |
| CN117431179A (en) | Lactobacillus gasseri MY4 and application thereof in preparing food and medicine for regulating intestines and stomach and reducing uric acid | |
| CN117721033A (en) | Lactobacillus mucilaginosus KS6 and application thereof in preparation of anti-inflammatory and sleep-aiding foods and medicines | |
| CN117946913A (en) | Enterococcus faecalis XY3 and application thereof in preparation of hypoglycemic and anti-aging foods and medicines | |
| CN117946936A (en) | Lactobacillus plantarum XY4 and application thereof in preparation of digestion-aiding and anti-inflammatory food and drug |
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 |