KR20220162284A - A composition for increased activity of alcohol and acetaldehyde degrading enzymes of the comprising heat-killed lactobacillus plantarum v135 as an active ingredient - Google Patents
A composition for increased activity of alcohol and acetaldehyde degrading enzymes of the comprising heat-killed lactobacillus plantarum v135 as an active ingredient Download PDFInfo
- Publication number
- KR20220162284A KR20220162284A KR1020210070567A KR20210070567A KR20220162284A KR 20220162284 A KR20220162284 A KR 20220162284A KR 1020210070567 A KR1020210070567 A KR 1020210070567A KR 20210070567 A KR20210070567 A KR 20210070567A KR 20220162284 A KR20220162284 A KR 20220162284A
- Authority
- KR
- South Korea
- Prior art keywords
- lactobacillus plantarum
- alcohol
- strain
- activity
- acetaldehyde
- Prior art date
Links
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 title claims abstract description 133
- IKHGUXGNUITLKF-UHFFFAOYSA-N Acetaldehyde Chemical compound CC=O IKHGUXGNUITLKF-UHFFFAOYSA-N 0.000 title claims abstract description 126
- 240000006024 Lactobacillus plantarum Species 0.000 title claims abstract description 98
- 235000013965 Lactobacillus plantarum Nutrition 0.000 title claims abstract description 98
- 229940072205 lactobacillus plantarum Drugs 0.000 title claims abstract description 98
- 230000000694 effects Effects 0.000 title claims abstract description 51
- 230000001965 increasing effect Effects 0.000 title claims abstract description 41
- 239000000203 mixture Substances 0.000 title claims abstract description 40
- 102000004190 Enzymes Human genes 0.000 title claims abstract description 38
- 108090000790 Enzymes Proteins 0.000 title claims abstract description 38
- 230000000593 degrading effect Effects 0.000 title claims abstract description 32
- 239000004480 active ingredient Substances 0.000 title claims abstract description 19
- 235000013373 food additive Nutrition 0.000 claims abstract description 26
- 239000002778 food additive Substances 0.000 claims abstract description 26
- 239000008194 pharmaceutical composition Substances 0.000 claims abstract description 26
- 238000000034 method Methods 0.000 claims description 24
- 238000010438 heat treatment Methods 0.000 claims description 14
- 230000008569 process Effects 0.000 claims description 14
- 238000001816 cooling Methods 0.000 claims description 6
- 210000004027 cell Anatomy 0.000 description 72
- 241000894006 Bacteria Species 0.000 description 44
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 44
- 229940088598 enzyme Drugs 0.000 description 27
- 210000004369 blood Anatomy 0.000 description 25
- 239000008280 blood Substances 0.000 description 25
- 235000014655 lactic acid Nutrition 0.000 description 22
- 239000004310 lactic acid Substances 0.000 description 22
- 239000002609 medium Substances 0.000 description 22
- 239000000047 product Substances 0.000 description 15
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 13
- 235000013305 food Nutrition 0.000 description 12
- 230000000844 anti-bacterial effect Effects 0.000 description 11
- 239000012153 distilled water Substances 0.000 description 9
- 239000006041 probiotic Substances 0.000 description 9
- 235000018291 probiotics Nutrition 0.000 description 9
- 239000000523 sample Substances 0.000 description 9
- 230000003078 antioxidant effect Effects 0.000 description 8
- 238000004364 calculation method Methods 0.000 description 8
- 239000001963 growth medium Substances 0.000 description 8
- 239000007788 liquid Substances 0.000 description 8
- 239000013641 positive control Substances 0.000 description 8
- 239000000126 substance Substances 0.000 description 8
- 241001465754 Metazoa Species 0.000 description 7
- 230000002292 Radical scavenging effect Effects 0.000 description 7
- 239000002253 acid Substances 0.000 description 7
- MGJZITXUQXWAKY-UHFFFAOYSA-N diphenyl-(2,4,6-trinitrophenyl)iminoazanium Chemical compound [O-][N+](=O)C1=CC([N+](=O)[O-])=CC([N+]([O-])=O)=C1N=[N+](C=1C=CC=CC=1)C1=CC=CC=C1 MGJZITXUQXWAKY-UHFFFAOYSA-N 0.000 description 7
- 238000002474 experimental method Methods 0.000 description 7
- 210000000936 intestine Anatomy 0.000 description 7
- 238000002835 absorbance Methods 0.000 description 6
- 238000010171 animal model Methods 0.000 description 6
- 230000008901 benefit Effects 0.000 description 6
- 239000000463 material Substances 0.000 description 6
- 235000015097 nutrients Nutrition 0.000 description 6
- 239000000243 solution Substances 0.000 description 6
- 238000012360 testing method Methods 0.000 description 6
- 241000293869 Salmonella enterica subsp. enterica serovar Typhimurium Species 0.000 description 5
- 210000000941 bile Anatomy 0.000 description 5
- 235000021109 kimchi Nutrition 0.000 description 5
- 244000005700 microbiome Species 0.000 description 5
- 239000002953 phosphate buffered saline Substances 0.000 description 5
- 239000000725 suspension Substances 0.000 description 5
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 4
- 241000588724 Escherichia coli Species 0.000 description 4
- 241001333951 Escherichia coli O157 Species 0.000 description 4
- 241000699670 Mus sp. Species 0.000 description 4
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 4
- 201000010099 disease Diseases 0.000 description 4
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 4
- 238000000855 fermentation Methods 0.000 description 4
- 230000004151 fermentation Effects 0.000 description 4
- 230000002147 killing effect Effects 0.000 description 4
- 238000004519 manufacturing process Methods 0.000 description 4
- 239000006228 supernatant Substances 0.000 description 4
- 241000193755 Bacillus cereus Species 0.000 description 3
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 3
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 3
- 238000004220 aggregation Methods 0.000 description 3
- 230000009286 beneficial effect Effects 0.000 description 3
- BLFLLBZGZJTVJG-UHFFFAOYSA-N benzocaine Chemical compound CCOC(=O)C1=CC=C(N)C=C1 BLFLLBZGZJTVJG-UHFFFAOYSA-N 0.000 description 3
- 235000013361 beverage Nutrition 0.000 description 3
- 239000003833 bile salt Substances 0.000 description 3
- 229940093761 bile salts Drugs 0.000 description 3
- 230000037396 body weight Effects 0.000 description 3
- 210000005056 cell body Anatomy 0.000 description 3
- 210000003850 cellular structure Anatomy 0.000 description 3
- 238000005119 centrifugation Methods 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- HHEAADYXPMHMCT-UHFFFAOYSA-N dpph Chemical compound [O-][N+](=O)C1=CC([N+](=O)[O-])=CC([N+]([O-])=O)=C1[N]N(C=1C=CC=CC=1)C1=CC=CC=C1 HHEAADYXPMHMCT-UHFFFAOYSA-N 0.000 description 3
- 239000003814 drug Substances 0.000 description 3
- 238000011156 evaluation Methods 0.000 description 3
- 235000013376 functional food Nutrition 0.000 description 3
- 210000001035 gastrointestinal tract Anatomy 0.000 description 3
- 230000002503 metabolic effect Effects 0.000 description 3
- 150000007524 organic acids Chemical class 0.000 description 3
- 230000001766 physiological effect Effects 0.000 description 3
- 239000000843 powder Substances 0.000 description 3
- 238000002360 preparation method Methods 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 239000007787 solid Substances 0.000 description 3
- 230000001954 sterilising effect Effects 0.000 description 3
- 238000004659 sterilization and disinfection Methods 0.000 description 3
- 238000003860 storage Methods 0.000 description 3
- 108010067770 Endopeptidase K Proteins 0.000 description 2
- 102000004317 Lyases Human genes 0.000 description 2
- 108090000856 Lyases Proteins 0.000 description 2
- 241000320380 Silybum Species 0.000 description 2
- 235000010841 Silybum marianum Nutrition 0.000 description 2
- DBMJMQXJHONAFJ-UHFFFAOYSA-M Sodium laurylsulphate Chemical compound [Na+].CCCCCCCCCCCCOS([O-])(=O)=O DBMJMQXJHONAFJ-UHFFFAOYSA-M 0.000 description 2
- 241000191967 Staphylococcus aureus Species 0.000 description 2
- 230000002776 aggregation Effects 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 239000003242 anti bacterial agent Substances 0.000 description 2
- 229940088710 antibiotic agent Drugs 0.000 description 2
- 210000001367 artery Anatomy 0.000 description 2
- 235000010323 ascorbic acid Nutrition 0.000 description 2
- 229960005070 ascorbic acid Drugs 0.000 description 2
- 239000011668 ascorbic acid Substances 0.000 description 2
- 230000001580 bacterial effect Effects 0.000 description 2
- 238000001574 biopsy Methods 0.000 description 2
- 238000009395 breeding Methods 0.000 description 2
- 230000001488 breeding effect Effects 0.000 description 2
- 239000002775 capsule Substances 0.000 description 2
- 150000001720 carbohydrates Chemical class 0.000 description 2
- 235000014633 carbohydrates Nutrition 0.000 description 2
- 238000012258 culturing Methods 0.000 description 2
- 230000003247 decreasing effect Effects 0.000 description 2
- 235000015872 dietary supplement Nutrition 0.000 description 2
- 238000007865 diluting Methods 0.000 description 2
- 238000009826 distribution Methods 0.000 description 2
- 230000002708 enhancing effect Effects 0.000 description 2
- 239000000284 extract Substances 0.000 description 2
- 239000003925 fat Substances 0.000 description 2
- 235000020510 functional beverage Nutrition 0.000 description 2
- 230000002068 genetic effect Effects 0.000 description 2
- 239000008187 granular material Substances 0.000 description 2
- 229940093915 gynecological organic acid Drugs 0.000 description 2
- 230000036541 health Effects 0.000 description 2
- 230000005764 inhibitory process Effects 0.000 description 2
- 230000000968 intestinal effect Effects 0.000 description 2
- 239000011259 mixed solution Substances 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 235000005985 organic acids Nutrition 0.000 description 2
- 239000000546 pharmaceutical excipient Substances 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 230000002829 reductive effect Effects 0.000 description 2
- 230000002441 reversible effect Effects 0.000 description 2
- 210000002966 serum Anatomy 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- 239000008223 sterile water Substances 0.000 description 2
- 235000013311 vegetables Nutrition 0.000 description 2
- MZOFCQQQCNRIBI-VMXHOPILSA-N (3s)-4-[[(2s)-1-[[(2s)-1-[[(1s)-1-carboxy-2-hydroxyethyl]amino]-4-methyl-1-oxopentan-2-yl]amino]-5-(diaminomethylideneamino)-1-oxopentan-2-yl]amino]-3-[[2-[[(2s)-2,6-diaminohexanoyl]amino]acetyl]amino]-4-oxobutanoic acid Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CC(O)=O)NC(=O)CNC(=O)[C@@H](N)CCCCN MZOFCQQQCNRIBI-VMXHOPILSA-N 0.000 description 1
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 1
- 108020004465 16S ribosomal RNA Proteins 0.000 description 1
- FWMNVWWHGCHHJJ-SKKKGAJSSA-N 4-amino-1-[(2r)-6-amino-2-[[(2r)-2-[[(2r)-2-[[(2r)-2-amino-3-phenylpropanoyl]amino]-3-phenylpropanoyl]amino]-4-methylpentanoyl]amino]hexanoyl]piperidine-4-carboxylic acid Chemical compound C([C@H](C(=O)N[C@H](CC(C)C)C(=O)N[C@H](CCCCN)C(=O)N1CCC(N)(CC1)C(O)=O)NC(=O)[C@H](N)CC=1C=CC=CC=1)C1=CC=CC=C1 FWMNVWWHGCHHJJ-SKKKGAJSSA-N 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- 108020002663 Aldehyde Dehydrogenase Proteins 0.000 description 1
- 208000035285 Allergic Seasonal Rhinitis Diseases 0.000 description 1
- USFZMSVCRYTOJT-UHFFFAOYSA-N Ammonium acetate Chemical compound N.CC(O)=O USFZMSVCRYTOJT-UHFFFAOYSA-N 0.000 description 1
- 239000005695 Ammonium acetate Substances 0.000 description 1
- 239000004375 Dextrin Substances 0.000 description 1
- 229920001353 Dextrin Polymers 0.000 description 1
- 230000005526 G1 to G0 transition Effects 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 108090001005 Interleukin-6 Proteins 0.000 description 1
- 241000186660 Lactobacillus Species 0.000 description 1
- 229920002774 Maltodextrin Polymers 0.000 description 1
- 239000005913 Maltodextrin Substances 0.000 description 1
- 102000016943 Muramidase Human genes 0.000 description 1
- 108010014251 Muramidase Proteins 0.000 description 1
- MSFSPUZXLOGKHJ-UHFFFAOYSA-N Muraminsaeure Natural products OC(=O)C(C)OC1C(N)C(O)OC(CO)C1O MSFSPUZXLOGKHJ-UHFFFAOYSA-N 0.000 description 1
- 241000699666 Mus <mouse, genus> Species 0.000 description 1
- 108010062010 N-Acetylmuramoyl-L-alanine Amidase Proteins 0.000 description 1
- BAWFJGJZGIEFAR-NNYOXOHSSA-O NAD(+) Chemical compound NC(=O)C1=CC=C[N+]([C@H]2[C@@H]([C@H](O)[C@@H](COP(O)(=O)OP(O)(=O)OC[C@@H]3[C@H]([C@@H](O)[C@@H](O3)N3C4=NC=NC(N)=C4N=C3)O)O2)O)=C1 BAWFJGJZGIEFAR-NNYOXOHSSA-O 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 108010013639 Peptidoglycan Proteins 0.000 description 1
- 102000006382 Ribonucleases Human genes 0.000 description 1
- 108010083644 Ribonucleases Proteins 0.000 description 1
- 241000283984 Rodentia Species 0.000 description 1
- 206010039491 Sarcoma Diseases 0.000 description 1
- 206010048908 Seasonal allergy Diseases 0.000 description 1
- 244000269722 Thea sinensis Species 0.000 description 1
- 239000007984 Tris EDTA buffer Substances 0.000 description 1
- 108060008682 Tumor Necrosis Factor Proteins 0.000 description 1
- 102000000852 Tumor Necrosis Factor-alpha Human genes 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 230000006978 adaptation Effects 0.000 description 1
- 239000000443 aerosol Substances 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 239000011543 agarose gel Substances 0.000 description 1
- 235000019257 ammonium acetate Nutrition 0.000 description 1
- 229940043376 ammonium acetate Drugs 0.000 description 1
- 239000003674 animal food additive Substances 0.000 description 1
- 230000003110 anti-inflammatory effect Effects 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 235000006708 antioxidants Nutrition 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 238000003149 assay kit Methods 0.000 description 1
- 244000052616 bacterial pathogen Species 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 239000000969 carrier Substances 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 210000002421 cell wall Anatomy 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000004182 chemical digestion Methods 0.000 description 1
- 239000007795 chemical reaction product Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 235000015218 chewing gum Nutrition 0.000 description 1
- 229940112822 chewing gum Drugs 0.000 description 1
- ZYWFEOZQIUMEGL-UHFFFAOYSA-N chloroform;3-methylbutan-1-ol;phenol Chemical compound ClC(Cl)Cl.CC(C)CCO.OC1=CC=CC=C1 ZYWFEOZQIUMEGL-UHFFFAOYSA-N 0.000 description 1
- 238000005345 coagulation Methods 0.000 description 1
- 230000015271 coagulation Effects 0.000 description 1
- 206010009887 colitis Diseases 0.000 description 1
- 230000000295 complement effect Effects 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 239000002537 cosmetic Substances 0.000 description 1
- 210000000805 cytoplasm Anatomy 0.000 description 1
- 230000009849 deactivation Effects 0.000 description 1
- 238000000354 decomposition reaction Methods 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 235000019425 dextrin Nutrition 0.000 description 1
- 235000005911 diet Nutrition 0.000 description 1
- 230000037213 diet Effects 0.000 description 1
- 238000009792 diffusion process Methods 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 230000001079 digestive effect Effects 0.000 description 1
- 102000038379 digestive enzymes Human genes 0.000 description 1
- 108091007734 digestive enzymes Proteins 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 238000003113 dilution method Methods 0.000 description 1
- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical compound P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 239000000839 emulsion Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- XJRPTMORGOIMMI-UHFFFAOYSA-N ethyl 2-amino-4-(trifluoromethyl)-1,3-thiazole-5-carboxylate Chemical compound CCOC(=O)C=1SC(N)=NC=1C(F)(F)F XJRPTMORGOIMMI-UHFFFAOYSA-N 0.000 description 1
- 235000021121 fermented vegetables Nutrition 0.000 description 1
- 230000037406 food intake Effects 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 235000011389 fruit/vegetable juice Nutrition 0.000 description 1
- 210000004211 gastric acid Anatomy 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 210000002865 immune cell Anatomy 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 230000008676 import Effects 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 238000001802 infusion Methods 0.000 description 1
- -1 infusions Substances 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 230000003871 intestinal function Effects 0.000 description 1
- 229940039696 lactobacillus Drugs 0.000 description 1
- 235000010335 lysozyme Nutrition 0.000 description 1
- 239000004325 lysozyme Substances 0.000 description 1
- 229960000274 lysozyme Drugs 0.000 description 1
- 210000002540 macrophage Anatomy 0.000 description 1
- 239000002075 main ingredient Substances 0.000 description 1
- 229940035034 maltodextrin Drugs 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 238000000691 measurement method Methods 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 239000006872 mrs medium Substances 0.000 description 1
- 210000000822 natural killer cell Anatomy 0.000 description 1
- 229930014626 natural product Natural products 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 239000002773 nucleotide Substances 0.000 description 1
- 125000003729 nucleotide group Chemical group 0.000 description 1
- 239000006916 nutrient agar Substances 0.000 description 1
- 238000009931 pascalization Methods 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 239000008188 pellet Substances 0.000 description 1
- 239000008363 phosphate buffer Substances 0.000 description 1
- 238000000053 physical method Methods 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 239000006187 pill Substances 0.000 description 1
- 201000004338 pollen allergy Diseases 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 239000003223 protective agent Substances 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 230000002000 scavenging effect Effects 0.000 description 1
- 239000013049 sediment Substances 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 235000021391 short chain fatty acids Nutrition 0.000 description 1
- 150000004666 short chain fatty acids Chemical class 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 238000000527 sonication Methods 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
- 238000013222 sprague-dawley male rat Methods 0.000 description 1
- 238000001694 spray drying Methods 0.000 description 1
- 230000007480 spreading Effects 0.000 description 1
- 238000003892 spreading Methods 0.000 description 1
- 210000002784 stomach Anatomy 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000013595 supernatant sample Substances 0.000 description 1
- 239000000829 suppository Substances 0.000 description 1
- 239000006188 syrup Substances 0.000 description 1
- 235000020357 syrup Nutrition 0.000 description 1
- 239000003826 tablet Substances 0.000 description 1
- 235000013616 tea Nutrition 0.000 description 1
- 238000009281 ultraviolet germicidal irradiation Methods 0.000 description 1
- 230000035899 viability Effects 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 150000003722 vitamin derivatives Chemical class 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Images
Classifications
-
- 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
- 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
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Mycology (AREA)
- General Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Veterinary Medicine (AREA)
- Animal Behavior & Ethology (AREA)
- Microbiology (AREA)
- Public Health (AREA)
- Epidemiology (AREA)
- Molecular Biology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Nutrition Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
Description
본 발명은 락토바실러스 플란타룸 V135 균주의 사균체를 유효성분으로 함유하는 알코올 및 아세트알데히드 분해효소의 활성 증가용 조성물에 관한 것으로, 보다 구체적으로는 락토바실러스 플란타룸 V135(Lactobacillus plantarum V135) KCTC18796P 균주의 사균체를 유효성분으로 함유하는 알코올 및 아세트알데히드 분해효소의 활성 증가용 약학적 조성물과 식품첨가제 조성물에 관한 것이다.The present invention relates to a composition for increasing the activity of alcohol and acetaldehyde degrading enzymes containing dead cells of the Lactobacillus plantarum V135 strain as an active ingredient, and more specifically, to a composition for increasing the activity of Lactobacillus plantarum V135 ( Lactobacillus plantarum V135) KCTC18796P It relates to a pharmaceutical composition and a food additive composition for increasing the activity of alcohol and acetaldehyde degrading enzyme containing dead cells of a strain as an active ingredient.
식물성 유산균은 동물성 유산균에 비하여 영양분이 부족하고, 낮은 pH, 높은 염 농도 등의 척박한 환경에서도 강한 생존력을 보이기 때문에 장내에 안정적으로 도달할 수 있게 한다는 장점이 있다. 또한, 동물성 유산균은 배지 영양분으로 탄수화물, 단백질, 지방 등을 사용하여 장시간 발효된 이후 단백질과 지방에 의해 부패되는 균주의 수가 증가하지만, 식물성 유산균은 배지 영양분으로 탄수화물만을 사용하므로 발효 시 부패하지 않는다. 이로 인해, 국내에서는 김치 등의 식물성 발효 식품에서 분리한 유산균의 우수성을 평가한 연구결과를 지속적으로 발표하고 있다.Compared to animal lactic acid bacteria, vegetable lactic acid bacteria lack nutrients and show strong viability even in harsh environments such as low pH and high salt concentration, so they have the advantage of being able to stably reach the intestines. In addition, animal lactic acid bacteria are fermented for a long time using carbohydrates, proteins, fats, etc. as medium nutrients, and then the number of strains corrupted by proteins and fats increases, but vegetable lactic acid bacteria use only carbohydrates as medium nutrients, so they do not decay during fermentation. For this reason, in Korea, research results evaluating the excellence of lactic acid bacteria isolated from fermented vegetable foods such as kimchi are continuously being published.
여러 가지 유익한 작용을 하는 미생물을 프로바이오틱스(probiotics)라고 하는데, 프로바이오틱스의 정의는 점차 확대되어 1989년 Fuller는 프로바이오티스를 “장내 균총을 개선시켜 줌으로써 숙주에게 유익한 영향을 주는 생균제제”라고 정의하였고 살아있는 미생물로서의 정립이 이루어졌지만 1999년 Salminen 등은 “숙주에 유익한 작용을 갖는 미생물 또는 미생물의 성분”으로 정의함으로써 프로바이오틱스의 범위를 사균 및 그 균체 성분으로까지 확대시켰다. 최근 들어서는 사균 및 그 균체 성분으로까지 확대되어 정의되던 프로바이오틱스 개념에서 새로운 포스트바이오틱스(postbiotics)라는 새로운 개념이 나타났다. 포스트바이오틱스는 생균과 대사(발효)산물들을 열처리 등에 의해 균의 성장이 일어나지 못하도록한 형태로 균체성분과 대사(발효)산물은 직접 또는 간접적으로 장 내 면역을 조절한다고 알려져 있다(Aguilar-Toal￡ JE et al., Trends Food Sci Technol, 2018). 균체성분으로 세포질(cytoplasm), 세포벽(cell wall), 리포테이코산(lipoteichoic acid), 테이코산(eichoic acid), 펩티도글리칸(peptidoglycan), DNA 등이 있으며, 대사(발효)산물은 유기산(organic acid), 단쇄지방산(short chain fatty acid), 다당류(polysaccharides) 등을 포함하고 있다. 대표적인 부산물인 유기산은 면역세포가 집결되어 있는 파이엘판을 자극하여 면역물질이 나오게 하며 주변의 유해물질을 억제하는 작용을 한다. 포스트바이오틱스는 여러 가지 사균화 방법이 보고되고 있는데 물리적 방법으로는 기계적 파괴(mechanical disruption), 열처리(heat treatment), 감마 또는 UV 조사(gamma- or UV irradiation), 고수압(High hydrostatic pressure), 동결건조(Freeze-drying), 음파처리(Sonication)가 있으며, 화학적 방법으로는 산성 탈수활성(Acid deactivation), 효소처리(enzymatic treatment), 용매추출(solvent extract)법이 있다. 포스트바이오틱스는 heat-treated probiotics, ghost probiotics, inactivated probiotics, paraprobiotics, 유산균 사균체, 열처리 유산균, 유산균 추출물 등 다양한 용어로 불리고 있다.Microorganisms with various beneficial actions are called probiotics. Although it was established as a microorganism, Salminen et al. in 1999 expanded the scope of probiotics to dead bacteria and their cell components by defining them as "microorganisms or components of microorganisms that have beneficial effects on the host". Recently, a new concept called postbiotics has emerged from the concept of probiotics, which was defined by extending to dead cells and their cell components. Postbiotics is a form in which live bacteria and metabolic (fermentation) products are prevented from growing by heat treatment, etc., and it is known that bacterial cell components and metabolic (fermentation) products directly or indirectly regulate immunity in the intestine (Aguilar-Toal￡ JE et al., Trends Food Sci Technol, 2018). Cell body components include cytoplasm, cell wall, lipoteichoic acid, eichoic acid, peptidoglycan, and DNA, and metabolic (fermentation) products are organic acids ( organic acids), short chain fatty acids, and polysaccharides. Organic acid, a typical by-product, stimulates Piel plate, where immune cells are concentrated, to produce immune substances and suppresses surrounding harmful substances. Several methods of killing postbiotics have been reported. Physical methods include mechanical disruption, heat treatment, gamma or UV irradiation, high hydrostatic pressure, There are freeze-drying and sonication, and chemical methods include acid deactivation, enzymatic treatment, and solvent extract. Postbiotics are called various terms such as heat-treated probiotics, ghost probiotics, inactivated probiotics, paraprobiotics, dead lactic acid bacteria, heat-treated lactic acid bacteria, and lactic acid bacteria extracts.
포스트바이오틱스(사균)는 프로바이오틱스(생균) 보다 다양한 장점을 가지고 있어 이미 일본, 미국 및 유럽에서는 다양한 형태로 상용화되고 있다. 프로바이오틱스는 위장관 내 물리, 화학적(위산, 담즙, 소화효소) 소화과정에 의해 대부분 사멸하여 외부환경에 의한 안정성이 떨어지고 열에 불안정하여 유통과정의 저온보관이 필요하고 이로 인한 저온보관 및 저온배송이 필요함에 따라 생산비용이 증가하게 된다. 또한 열에 불안정하여 살균처리 공정이 포함된 제품 등의 적용이 불가능하다는 단점이 발생한다. 포스트바이오틱스의 경우, 이미 균의 성장이 일어나지 못하도록 한 형태로 위장관 내 물리, 화학적 소화과정에 영향을 받지 않으며, 열에 안정하여 보관기간별 일정한 기능성을 나타내고, 저온보관 및 저온배송이 불필요함에 따라 생산비용을 절감할 수 있다. 또한 열 안정성이 있어 살균처리 공정이 포함된 제품에 적용이 가능하여 적용 제품의 다양성을 가진다. 이러한 특징으로 포스트바이오틱스는 프로바이오틱스 보다 소재 안정성이 있어 산업적 적용의 범위가 넓고, 유통과정에서 다루기가 쉽고, 일반식품에도 첨가되어 식품의 기능성을 강화하면서 면역조절 기능에서 프로바이오틱스와 동일한 효과로 건강기능식품, 식품첨가제, 의약품, 동물사료 등과 같은 기존의 유산균이 적용되어온 분야 외에도 화장품 원료로도 적용되어 그 시장이 확대되고 있다. 또한 항생제 사용에 대한 규제가 강화되고 있기 때문에 대체제로서 활용성과 아직 사균체 제품 생산에 본격적으로 뛰어든 업체가 손에 꼽을 정도이기 때문에 시장성과 성장 가능성은 크다고 할 수 있다. 특히 일본에서는 이미 사균체 제품이 많이 출시되어 꽃가루 알러지 등 면역 관련 효능을 기반으로 홍보하고 있다. 하지만 국내는 아직 개별인정소재로 인정받은 소재가 없을 뿐 아니라 관련 제품 또한 수입에 의존하고 있다.Postbiotics (dead bacteria) have various advantages over probiotics (live bacteria) and are already commercialized in various forms in Japan, the United States and Europe. Probiotics are mostly killed by the physical and chemical (gastric acid, bile, digestive enzymes) digestion process in the gastrointestinal tract, so they are less stable in the external environment and unstable to heat, so they require low-temperature storage in the distribution process, which requires low-temperature storage and low-temperature delivery. As a result, production costs increase. In addition, since it is unstable to heat, there is a disadvantage that it is impossible to apply products including a sterilization process. In the case of postbiotics, the growth of bacteria has already been prevented, so they are not affected by physical and chemical digestion processes in the gastrointestinal tract, are stable to heat, and show constant functionality for each storage period. can save In addition, since it has thermal stability, it can be applied to products that include a sterilization process, and thus has a variety of applied products. Due to these characteristics, postbiotics have a wider range of industrial applications because they are more stable than probiotics, and are easy to handle in the distribution process. In addition to fields where lactic acid bacteria have been applied, such as food additives, pharmaceuticals, and animal feed, the market is expanding as it is also applied as a raw material for cosmetics. In addition, since regulations on the use of antibiotics are being strengthened, it can be used as an alternative and there are only a handful of companies that have yet entered into the production of dead cell products in earnest, so it can be said that the marketability and growth potential are great. In particular, many dead cell products have already been released in Japan, and they are promoted based on their immune-related efficacy, such as pollen allergy. However, in Korea, there is no material recognized as an individual recognized material yet, and related products are also dependent on imports.
유산균 사균체의 생리활성에 대해 보고된 연구들에 따르면 DSS(Dextran sulfate sodium)로 유도된 대장염을 앓고 있는 마우스의 경우 유산균 사균체를 섭취한 마우스에서 DSS 완화 효과를 보였으며 이식한 육종암세포(Sarcoma-180)의 증식이 감소되고 NK세포가 활성화됨이 보고되었다(Tadano et al., J. Japan Mibyou System association,2011). 또한 유해균의 억제 및 정장 작용에 관해서, 항생제를 투여한 마우스에서 장내 대조군에 비해 유익균은 빠르게 증식하고 유해균은 억제시키는 효능이 개시되어 있다(Simohashi et al., Medicine and biology, 2002). 아울러, 유산균 사균체의 다양한 생리활성은 사균체의 특성상 열과 pH에 영향을 받지 않아 다양한 형태의 제제로 가공이 가능한 장점이 있다(Kan, Food industry, 2001).According to studies reported on the physiological activity of killed lactic acid bacteria, in the case of mice suffering from colitis induced by DSS (Dextran sulfate sodium), mice ingested lactobacillus killed cells showed a DSS alleviating effect, and transplanted sarcoma cancer cells -180) and activation of NK cells were reported (Tadano et al., J. Japan Mibyou System association, 2011). In addition, regarding the inhibition of harmful bacteria and the intestinal function, beneficial bacteria proliferate rapidly and harmful bacteria are suppressed in mice administered with antibiotics compared to the intestinal control group (Simohashi et al., Medicine and Biology, 2002). In addition, the various physiological activities of dead lactic acid bacteria are not affected by heat and pH due to the nature of dead cells, so there is an advantage that it can be processed into various types of preparations (Kan, Food industry, 2001).
한편, 대한민국 등록특허 제10-2028744호에는 면역증강 활성, 항산화 활성 및 소화액 내성을 갖는 락토바실러스 플란타룸(Lactobacillus plantarum) HY7717 균주 및 이를 포함하는 약학 조성물 및 식품 조성물에 관련된 내용이 개시되어 있으며, 대한민국 등록특허 제10-1919938호에는 대식세포 및 마우스 동물 모델에서 면역 지표인 TNF-α 및 IL-6의 생산을 유도함으로써 면역 증강 활성을 가지는, 신규한 락토바실러스 플란타룸(Lactobacillus plantarum) LM1004 및 이를 유효성분으로 함유하는 면역 증강용 조성물, 약학적 조성물, 건강기능식품, 사료첨가제, 기능성 음료, 및 발효제품에 관련된 내용이 개시되어 있으나, 락토바실러스 플란타룸 V135(Lactobacillus plantarum V135) KCTC18796P 균주의 사균체를 유효성분으로 함유하는 알코올 및 아세트알데히드 분해효소의 활성 증가용 약학적 조성물과 식품첨가제 조성물에 대해서는 밝혀진 바가 없다.On the other hand, Korean Patent Registration No. 10-2028744 discloses a Lactobacillus plantarum HY7717 strain having immune enhancing activity, antioxidant activity and digestive juice resistance, and pharmaceutical compositions and food compositions containing the same. Disclosed, Korean Patent Registration No. 10-1919938 discloses novel Lactobacillus plantarum LM1004, which has immune enhancing activity by inducing the production of TNF-α and IL-6, which are immune indicators, in macrophages and mouse animal models, and Immunity-enhancing compositions, pharmaceutical compositions, health functional foods, feed additives, functional beverages, and fermented products containing it as an active ingredient are disclosed, but the Lactobacillus plantarum V135 ( Lactobacillus plantarum V135) KCTC18796P strain A pharmaceutical composition and a food additive composition for increasing the activity of alcohol and acetaldehyde degrading enzyme containing dead cells as an active ingredient have not been identified.
본 발명은 상술한 것과 같은 문제점을 해결하고 필요한 기술을 제공하기 위하여 안출된 것으로서,The present invention has been made to solve the problems described above and provide necessary technology,
본 발명은 락토바실러스 플란타룸 V135(Lactobacillus plantarum V135) KCTC18796P 균주의 사균체를 유효성분으로 함유하는 알코올 및 아세트알데히드 분해효소의 활성 증가용 약학적 조성물을 제공함에 그 목적이 있다.An object of the present invention is to provide a pharmaceutical composition for increasing the activity of alcohol and acetaldehyde degrading enzymes containing dead cells of Lactobacillus plantarum V135 KCTC18796P strain as an active ingredient.
또한, 본 발명은 락토바실러스 플란타룸 V135(Lactobacillus plantarum V135) KCTC18796P 균주의 사균체를 유효성분으로 함유하는 알코올 및 아세트알데히드 분해효소의 활성 증가용 식품첨가제 조성물을 제공함에 다른 목적이 있다.In addition, another object of the present invention is to provide a food additive composition for increasing the activity of alcohol and acetaldehyde degrading enzyme containing dead cells of Lactobacillus plantarum V135 KCTC18796P strain as an active ingredient.
상기와 같은 목적을 달성하기 위한 본 발명의 일 실시형태로서,As one embodiment of the present invention for achieving the above object,
본 발명은 락토바실러스 플란타룸 V135(Lactobacillus plantarum V135) KCTC18796P 균주의 사균체를 유효성분으로 함유하는 알코올 및 아세트알데히드 분해효소의 활성 증가용 약학적 조성물을 제공한다.The present invention provides a pharmaceutical composition for increasing the activity of alcohol and acetaldehyde degrading enzymes containing dead cells of Lactobacillus plantarum V135 KCTC18796P strain as an active ingredient.
이때, 상기 알코올 및 아세트알데히드 분해효소의 활성 증가용 약학적 조성물에 함유된 균주의 사균체는 락토바실러스 플란타룸 V135(Lactobacillus plantarum V135) KCTC18796P 균주를 식용배지에 접종하여 배양액을 제조하고, 배양액을 80℃ 이상의 온도로 가열한 뒤 60℃ 이하의 온도가 되도록 급속 냉각시키는 공정을 1회 이상 반복 실시하여 열처리한 것임을 특징으로 한다.At this time, the dead cells of the strain contained in the pharmaceutical composition for increasing the activity of alcohol and acetaldehyde degrading enzyme are inoculated with Lactobacillus plantarum V135 KCTC18796P strain in an edible medium to prepare a culture medium, It is characterized in that the heat treatment is performed by repeatedly performing a process of heating to a temperature of 80 ° C or higher and then rapidly cooling to a temperature of 60 ° C or lower.
또한, 상기 알코올 및 아세트알데히드 분해효소의 활성 증가용 약학적 조성물은 락토바실러스 플란타룸 V135(Lactobacillus plantarum V135) KCTC18796P 균주의 사균체가 조성물 1g당 1×105cfu 내지 1×1012cfu로 포함되는 것을 특징으로 한다.In addition, the pharmaceutical composition for increasing the activity of alcohol and acetaldehyde degrading enzyme includes dead cells of Lactobacillus plantarum V135 KCTC18796P strain at 1×10 5 cfu to 1×10 12 cfu per 1 g of the composition. characterized by being
본 발명은 락토바실러스 플란타룸 V135(Lactobacillus plantarum V135) KCTC18796P 균주의 사균체를 유효성분으로 함유하는 알코올 및 아세트알데히드 분해효소의 활성 증가용 식품첨가제 조성물을 제공한다.The present invention provides a food additive composition for increasing the activity of alcohol and acetaldehyde degrading enzymes containing dead cells of Lactobacillus plantarum V135 KCTC18796P strain as an active ingredient.
이때, 상기 알코올 및 아세트알데히드 분해효소의 활성 증가용 식품첨가제 조성물에 함유된 균주의 사균체는 락토바실러스 플란타룸 V135(Lactobacillus plantarum V135) KCTC18796P 균주를 식용배지에 접종하여 배양액을 제조하고, 배양액을 80℃ 이상의 온도로 가열한 뒤 60℃ 이하의 온도가 되도록 급속 냉각시키는 공정을 1회 이상 반복 실시하여 열처리한 것임을 특징으로 한다.At this time, the dead cells of the strain contained in the food additive composition for increasing the activity of alcohol and acetaldehyde decomposing enzyme are inoculated with Lactobacillus plantarum V135 KCTC18796P strain in an edible medium to prepare a culture solution, It is characterized in that the heat treatment is performed by repeatedly performing a process of heating to a temperature of 80 ° C or higher and then rapidly cooling to a temperature of 60 ° C or lower.
또한, 상기 알코올 및 아세트알데히드 분해효소의 활성 증가용 식품첨가제 조성물은 락토바실러스 플란타룸 V135(Lactobacillus plantarum V135) KCTC18796P 균주의 사균체가 조성물 1g당 1×105cfu 내지 1×1012cfu로 포함되는 것을 특징으로 한다.In addition, the food additive composition for increasing the activity of alcohol and acetaldehyde degrading enzyme includes dead cells of Lactobacillus plantarum V135 KCTC18796P strain at 1×10 5 cfu to 1×10 12 cfu per 1 g of the composition. characterized by being
본 발명의 일 실시형태에 따른 락토바실러스 플란타룸 V135(Lactobacillus plantarum V135) KCTC18796P 균주의 사균체를 유효성분으로 함유하는 알코올 및 아세트알데히드 분해효소의 활성 증가용 조성물은 열처리 공정을 수행함에 따라 세포 독성이 없고, 알코올 및 아세트알데히드 분해효소의 활성을 증가시킴으로써 혈중 알코올 및 아세트알데히드 농도를 감소시키는 효과가 있어 약학적 조성물 또는 식품첨가제 조성물로 사용될 수 있다.A composition for increasing the activity of alcohol and acetaldehyde degrading enzyme containing dead cells of Lactobacillus plantarum V135 KCTC18796P strain according to an embodiment of the present invention as an active ingredient is subjected to a heat treatment process to cell cells. It is non-toxic and has the effect of reducing the concentration of alcohol and acetaldehyde in the blood by increasing the activity of alcohol and acetaldehyde degrading enzymes, so it can be used as a pharmaceutical composition or a food additive composition.
즉, 본 발명의 락토바실러스 플란타룸 V135(Lactobacillus plantarum V135) KCTC18796P 균주의 열처리를 통한 사균화는 생균의 단점(안정성, 제품 적용성 등)을 개선하여 안정하고 다양한 제품으로의 적용이 가능하면서 생균과의 동등 이상의 기능성을 보유한 소재를 제공할 수 있는 장점이 있기 때문에 알코올 및 아세트알데히드 분해효소의 활성 증가용 약학적 조성물 또는 식품첨가제 조성물로 사용될 수 있다.That is, killing through heat treatment of the Lactobacillus plantarum V135 KCTC18796P strain of the present invention improves the disadvantages (stability, product applicability, etc.) of live bacteria, enabling stable and application to various products. Since it has the advantage of providing a material having functionality equal to or greater than that of live bacteria, it can be used as a pharmaceutical composition or food additive composition for increasing the activity of alcohol and acetaldehyde degrading enzyme.
도 1은 본 발명에서 분리한 락토바실러스 플란타룸 V135 균주의 플레이트 배양 사진이다.
도 2는 락토바실러스 플란타룸 V135 사균체의 Escherichia coli(A), Salmonella typhimurium(B), Bacillus cereus(C)에 대한 항균활성을 평가한 사진이다.
도 3은 락토바실러스 플란타룸 V135 사균체의 항산화 활성을 평가한 그래프이다.
도 4는 알코올 투여 30분 전 락토바실러스 플란타룸 V135 균주의 사균체를 투여한 실험동물의 혈중 알코올 및 아세트알데히드의 농도 감소 효과를 관찰한 결과를 나타내는 그래프이다.
도 5는 알코올 투여 30분 후 락토바실러스 플란타룸 V135 균주의 사균체를 투여한 실험동물의 혈중 알코올 및 아세트알데히드의 농도 감소 효과를 관찰한 결과를 나타내는 그래프이다.1 is a plate culture photograph of the Lactobacillus plantarum V135 strain isolated in the present invention.
2 is a photograph evaluating the antibacterial activity of Lactobacillus plantarum V135 dead cells against Escherichia coli (A), Salmonella typhimurium (B), and Bacillus cereus (C).
Figure 3 is a graph evaluating the antioxidant activity of Lactobacillus plantarum V135 dead cells.
Figure 4 is a graph showing the results of observing the effect of reducing the concentration of alcohol and acetaldehyde in laboratory animals administered with dead cells of Lactobacillus plantarum V135 strain 30 minutes before alcohol administration.
Figure 5 is a graph showing the results of observing the effect of reducing the concentration of alcohol and acetaldehyde in laboratory animals administered with dead cells of Lactobacillus plantarum V135 strain 30 minutes after alcohol administration.
이하, 본 발명이 속하는 기술분야에서 통상의 지식을 가진 자가 용이하게 실시할 수 있도록 본 발명의 실시형태를 들어 상세히 설명한다. 본 발명의 실시형태는 당업계에서 평균적인 지식을 가진 자에게 본 발명을 더욱 완전하게 설명하기 위해서 제공되는 것이다. 따라서, 본 발명의 실시형태는 여러 가지 다른 형태로 변형될 수 있으며, 본 발명의 범위가 이하 설명하는 실시형태로 한정되는 것은 아니다.Hereinafter, embodiments of the present invention will be described in detail so that those skilled in the art can easily implement the present invention. Embodiments of the present invention are provided to more completely explain the present invention to those skilled in the art. Accordingly, the embodiments of the present invention can be modified in many different forms, and the scope of the present invention is not limited to the embodiments described below.
본 발명의 명세서 전체에서, 어떤 부분이 어떤 구성요소를 “포함”한다고 할 때, 이는 특별히 반대되는 기재가 없는 한 다른 구성요소를 제외하는 것이 아니라 다른 구성요소를 더 포함하는 것을 의미한다.Throughout the specification of the present invention, when a part "includes" a certain component, it means that it further includes other components, not excluding other components unless otherwise stated.
본 발명의 명세서 전체에서 사용되는 정도의 용어 “약”, “실질적으로” 등은 언급된 의미에 고유한 제조 및 물질 허용 오차가 제시될 때 그 수치에서 또는 그 수치에 근접한 의미로 사용되고, 본 발명의 이해를 돕기 위해 정확하거나 절대적인 수치가 언급된 개시 내용을 비양심적인 침해자가 부당하게 이용하는 것을 방지하기 위해 사용된다.The terms "about," "substantially," and the like used throughout the specification of the present invention are used at or close to that value when manufacturing and material tolerances inherent in the stated meaning are given, and the present invention Accurate or absolute figures are used to prevent unfair use by unscrupulous infringers of the disclosed disclosures mentioned for the sake of understanding.
본 발명은 락토바실러스 플란타룸 V135(Lactobacillus plantarum V135) KCTC18796P 균주의 사균체를 유효성분으로 함유하는 알코올 및 아세트알데히드 분해효소의 활성 증가용 약학적 조성물(이하, ‘약학적 조성물’이라고도 함)을 제공한다.The present invention is a pharmaceutical composition for increasing the activity of alcohol and acetaldehyde degrading enzyme containing dead cells of Lactobacillus plantarum V135 KCTC18796P strain as an active ingredient (hereinafter also referred to as 'pharmaceutical composition') provides
또한, 본 발명은 락토바실러스 플란타룸 V135(Lactobacillus plantarum V135) KCTC18796P 균주의 사균체를 유효성분으로 함유하는 알코올 및 아세트알데히드 분해효소의 활성 증가용 식품첨가제 조성물(이하, ‘식품첨가제 조성물’이라고도 함)을 제공한다.In addition, the present invention is a food additive composition for increasing the activity of alcohol and acetaldehyde degrading enzyme containing dead cells of Lactobacillus plantarum V135 KCTC18796P strain as an active ingredient (hereinafter also referred to as 'food additive composition'). ) is provided.
본 발명에 따른 약학적 조성물 및 식품첨가제 조성물은 락토바실러스 플란타룸 V135(Lactobacillus plantarum V135) KCTC18796P 균주의 사균체를 유효성분으로 함유하는 것으로, 알코올 및 아세트알데히드 분해효소의 활성을 증가시킴으로써 혈중 알코올 및 아세트알데히드 농도를 감소시키는 효과가 있다.The pharmaceutical composition and food additive composition according to the present invention contain dead cells of Lactobacillus plantarum V1 35 KCTC18796P strain as an active ingredient, and increase the activity of alcohol and acetaldehyde degrading enzyme in the blood. It has the effect of reducing the concentration of alcohol and acetaldehyde.
본 발명의 일 실시형태에 따르면, 상기 알코올 및 아세트알데히드 분해효소의 활성 증가용 약학적 조성물 및 식품첨가제 조성물에 함유된 균주의 사균체는 락토바실러스 플란타룸 V135(Lactobacillus plantarum V135) KCTC18796P 균주를 식용배지에 접종하여 배양액을 제조하고, 배양액을 미생물의 사멸을 유도 할 수 있는 80℃ 이상의 온도로 가열한 뒤 60℃ 이하의 온도가 되도록 급속 냉각시키는 공정을 1회 이상 반복 실시하여 열처리한 것임을 특징으로 할 수 있다.According to one embodiment of the present invention, the dead cells of the strain contained in the pharmaceutical composition and food additive composition for increasing the activity of alcohol and acetaldehyde degrading enzyme are Lactobacillus plantarum V135 (L actobacillus plantarum V135) KCTC18796P strain It is characterized by heat treatment by inoculating an edible medium to prepare a culture medium, heating the culture medium to a temperature of 80 ° C or higher that can induce the death of microorganisms, and then rapidly cooling to a temperature of 60 ° C or lower by repeating the process one or more times. can be done with
또한, 본 발명의 일 실시형태에 따르면, 상기 알코올 및 아세트알데히드 분해효소의 활성 증가용 약학적 조성물 및 식품첨가제 조성물은 락토바실러스 플란타룸 V135(Lactobacillus plantarum V135) KCTC18796P 균주의 사균체가 조성물 1g당 1×105cfu 내지 1×1012cfu로 포함되는 것을 특징으로 할 수 있다.In addition, according to one embodiment of the present invention, the pharmaceutical composition and food additive composition for increasing the activity of alcohol and acetaldehyde degrading enzyme include dead cells of Lactobacillus plantarum V135 KCTC18796P
이는, 락토바실러스 플란타룸 V135(Lactobacillus plantarum V135) KCTC18796P 균주의 사균체가 조성물 1g당 1×105cfu 미만의 농도로 포함될 경우 알코올 섭취로 인한 혈중 알코올 및 아세트알데히드 농도 감소 효과를 기대하지 못할 우려가 있기 때문이며, 균 수 범위(농도)는 전임상 섭취농도를 인체적용시험을 위한 체표면적 대비 환산계와 성인체중(60㎏)을 계산하여 1×1012cfu 까지 범위로 설정하였다.This is, when the dead cells of the Lactobacillus plantarum V135 KCTC18796P strain are included at a concentration of less than 1×10 5 cfu per 1 g of the composition, the effect of reducing blood alcohol and acetaldehyde concentrations due to alcohol consumption cannot be expected. This is because there is a concern, and the range (concentration) of the number of bacteria was set in the range up to 1 × 10 12 cfu by calculating the body surface area conversion system and adult weight (60 kg) for the preclinical intake concentration for the human application test.
본 발명의 락토바실러스 플란타룸 V135(Lactobacillus plantarum V135) KCTC18796P 균주의 사균체를 유효성분으로 함유하는 알코올 및 아세트알데히드 분해효소의 활성 증가용 약학적 조성물은 혈중 알코올 및 아세트알데히드 농도를 감소시키기 위한 약학적 조성물로 사용될 수 있다. 본 발명의 약학적 조성물을 사용할 수 있는 질환 또는 질병은 이에 제한되는 것은 아니며, 알코올 섭취로 인해 혈중 알코올 및 아세트알데히드 농도 증가가 야기된 관련 질환 또는 질병은 모두 포함될 수 있다.A pharmaceutical composition for increasing the activity of alcohol and acetaldehyde degrading enzymes containing dead cells of Lactobacillus plantarum V135 KCTC18796P strain of the present invention as an active ingredient is for reducing blood alcohol and acetaldehyde concentrations It can be used as a pharmaceutical composition. Diseases or diseases for which the pharmaceutical composition of the present invention can be used are not limited thereto, and all related diseases or diseases caused by increased blood alcohol and acetaldehyde levels due to alcohol intake may be included.
본 발명의 약학적 조성물을 사용하는 경우 약학적 조성물의 제조에 통상적으로 사용하는 적절한 담체, 부형제 및 희석제를 더 포함할 수 있다.When using the pharmaceutical composition of the present invention, suitable carriers, excipients and diluents commonly used in the preparation of pharmaceutical compositions may be further included.
또한, 본 발명에 따른 약학적 조성물은 통상의 방법에 따라 산제, 과립제, 정제, 캡슐제, 현탁액, 에멀젼, 시럽, 에어로졸 등의 경구형 제형, 외용제, 좌제 및 멸균 주사용액의 형태로 제형화하여 사용될 수 있다.In addition, the pharmaceutical composition according to the present invention is formulated in the form of oral formulations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, external preparations, suppositories and sterile injection solutions according to conventional methods. can be used
본 발명의 락토바실러스 플란타룸 V135(Lactobacillus plantarum V135) KCTC18796P 균주의 사균체를 유효성분으로 함유하는 알코올 및 아세트알데히드 분해효소의 활성 증가용 식품첨가제 조성물은 혈중 알코올 및 아세트알데히드 농도 감소 효과를 가지는 것으로, 이를 목적으로 하는 식품에 첨가될 수 있다. 이에 제한되는 것은 아니나, 본 발명에 따른 식품첨가제 조성물은 식품의 주원료, 부원료, 식품 첨가제, 기능성 식품 또는 음료에 사용될 수 있다.The food additive composition for increasing the activity of alcohol and acetaldehyde degrading enzymes containing dead cells of Lactobacillus plantarum V135 KCTC18796P strain of the present invention as an active ingredient has an effect of reducing blood alcohol and acetaldehyde concentrations As such, it can be added to foods intended for this purpose. Although not limited thereto, the food additive composition according to the present invention may be used for main ingredients, supplementary ingredients, food additives, functional foods or beverages of food.
본 발명에서 ‘식품’이라 함은 영양소를 한 가지 또는 그 이상 함유하고 있는 천연물 또는 가공품을 의미하며, 바람직하게는 어느 정도의 가공 공정을 거쳐 직접 먹을 수 있는 상태가 된 것을 의미하고, 통상적인 의미로서, 식품, 식품 첨가제, 건강 기능성 식품, 건강 보조식품 및 음료를 모두 포함한다.In the present invention, 'food' means a natural product or processed product containing one or more nutrients, preferably means a state that can be eaten directly through a certain degree of processing process, and usually means As such, it includes all foods, food additives, health functional foods, health supplements and beverages.
본 발명의 식품첨가제 조성물을 첨가할 수 있는 식품으로는 각종 식품류, 예를 들어, 음료, 껌, 차, 비타민 복합제, 건강보조 식품류 등이 있으며, 환제, 분말, 과립, 침제, 정제, 캡슐 또는 음료 형태로 제형화될 수 있다.Foods to which the food additive composition of the present invention can be added include various foods, such as beverages, chewing gum, tea, vitamin complexes, health supplements, pills, powders, granules, infusions, tablets, capsules or beverages. It can be formulated into a form.
이하, 본 발명을 구체적인 실시예에 따라 상세히 설명한다. 단, 하기의 실시예는 본 발명을 예시로 하는 것일 뿐, 본 발명의 내용이 하기 실시예에 한정되는 것은 아니다.Hereinafter, the present invention will be described in detail according to specific examples. However, the following examples are merely illustrative of the present invention, and the content of the present invention is not limited to the following examples.
락토바실러스 플란타룸 V135(LLactobacillus Plantarum V135(L actobacillus plantarum actobacillus plantarum V135) KCTC18796P 균주V135) KCTC18796P strain
1. 항균활성이 높은 유산균의 확보1. Securing lactic acid bacteria with high antibacterial activity
유산균 분리를 위해 김치 시료는 가정에서 담근 김치를 사용하였으며, 각종 김치를 무균적으로 잘게 다진 다음, 이 중 1g을 취하여 멸균 생리식염수(0.85% NaCl) 9㎖에 현탁하고 단계적으로 희석하여 혼합용액 1㎖를 취하여 십진희석법으로 희석한 후 MRS 고체배지(Difco, MI, USA)에 도말하여 37℃, 48시간 배양하였다. 각 시료별 배지에서 형성된 단일 콜로니는 bromocresol purple이 첨가된 MRS 고체배지에 도말하여 노란색을 보이는 콜로니를 최종 선별하였다. 선별된 균주의 플레이트 배양 사진은 도 1과 같다.For the separation of lactic acid bacteria, kimchi samples made at home were used as kimchi samples. Aseptically, 1 g of the kimchi was finely chopped, suspended in 9 ml of sterilized physiological saline (0.85% NaCl), and diluted step by step to obtain
선별된 균주는 항균활성을 평가하여 항균활성이 가장 뛰어난 1종의 유산균을 선발하였으며, 보관은 20% glycerol stock으로 -70℃에서 보관하였다.The selected strains were evaluated for antibacterial activity, and one type of lactic acid bacteria with the most excellent antibacterial activity was selected, and stored at -70 ° C as a 20% glycerol stock.
2. 김치로부터 분리한 유산균의 유전학적 동정2. Genetic identification of lactic acid bacteria isolated from kimchi
분리 균주의 유전학적 동정을 위하여 MRS 액체배지에 접종한 다음 37℃에서 24시간 2차 계대 배양하여 배양액 2㎖를 4,000rpm에서 10분 동안 원심분리하였다. 원심분리하여 얻은 세포 침전물을 0.85% NaCl로 3회 세척하였다. 세척 후, 리소자임(lysozyme, 10㎎/㎖) 0.5㎖를 첨가하여 37℃, 1시간 동안 처리하였다. 단백질 분해효소인 프로테아제 K(Protease K) 20㎕와 10% SDS(sodium dodecyl sulfate) 25㎕를 첨가한 후, 60℃ 항온수조(water bath)에서 30분 처리한 후 RNA 분해효소인 RNase 1㎕를 첨가하여 37℃, 1시간 처리하였다. 동량의 페놀(Phenol)-클로로포름(Chloroform)-이소아밀 알콜(Isoamyl alcohol)을 25:24:1의 비율로 첨가하여 현탁한 후, 14,000rpm으로 5분 동안 4℃에서 원심분리하여 상층액만 취한 다음, 이와 같은 과정을 2회 반복하였다. 상층액 양의 1/2배 양의 3M 암모늄 아세테이트(ammonium acetate, pH 4.8)와 2배 양의 100% alcohol을 첨가하고 -20℃에서 1시간 정치하였다. 3,000rpm으로 5분 동안 4℃에서 원심분리 하여 세포 침전물을 확인하고, 상등액을 완전히 제거한 후에 70% ethanol 1㎖/ℓ를 넣고 다시 3,000rpm으로 5분 동안 4℃에서 원심분리 하였다. 상층액 제거 후에 건조시키고 TE 버퍼나 증류수로 현탁하여 DNA를 분리하였다. 미생물의 16S rRNA를 증폭하기 위해 정방향 프라이머(27f) : (5'- AGA GTT TGA TCM TGG CTC AG -3' ; 서열번호 2)와 역방향 프라이머(1492r) : (5'- GGT TAC CTT TGT TAC GAC TT -3' ; 서열번호 3)를 사용하였다. PCR 프리믹스(Bioneer, Cat No. K-2012)에 증류수 17㎕, 정방향 프라이머 1㎕, 역방향 프라이머 1㎕, DNA 1㎕를 첨가하여 혼합한 후 PCR을 수행하였다. PCR 조건은 94℃에서 5분간 처리 후, 94℃에서 1분, 62℃에서 40초, 72℃에서 40초로 30회를 반복하였으며, 72℃에서 7분으로 반응을 종료하였다. PCR 반응 산물을 0.8% 아가로스 겔(agarose gel)로 전기영동을 실시하여 확인하였다. 최종 염기서열을 분석하고 이를 데이터베이스와 비교하여 유산균주를 동정한 결과, 본 발명에 의해 분리된 균주는 락토바실러스 플란타룸으로 나타났으며, 상기 유산균주는 한국생명공학연구원 생물자원센터에 2019년 12월 3일자로 기탁하였다(KCTC18796P).For genetic identification of the isolated strain, it was inoculated into MRS liquid medium and subcultured for 24 hours at 37 ° C., and 2 ml of the culture medium was centrifuged at 4,000 rpm for 10 minutes. Cell pellets obtained by centrifugation were washed three times with 0.85% NaCl. After washing, 0.5 ml of lysozyme (10 mg/ml) was added and treated at 37° C. for 1 hour. After adding 20 μl of protease K (Protease K) and 25 μl of 10% SDS (sodium dodecyl sulfate), treated in a water bath at 60 ° C for 30 minutes, 1 μl of RNase, an RNA degrading enzyme, was added. was added and treated at 37°C for 1 hour. After suspension by adding the same amount of phenol-chloroform-isoamyl alcohol at a ratio of 25:24:1, centrifuged at 14,000 rpm for 5 minutes at 4°C to take only the supernatant. Next, this process was repeated twice. After adding 1/2 the amount of 3M ammonium acetate (pH 4.8) and 2 times the amount of 100% alcohol to the amount of the supernatant, it was allowed to stand at -20 ° C for 1 hour. Cell sediment was confirmed by centrifugation at 3,000 rpm at 4° C. for 5 minutes, and after completely removing the supernatant, 1 ml/L of 70% ethanol was added thereto, followed by centrifugation at 3,000 rpm for 5 minutes at 4° C. After removing the supernatant, it was dried and suspended in TE buffer or distilled water to separate DNA. To amplify microbial 16S rRNA, forward primer (27f): (5'- AGA GTT TGA TCM TGG CTC AG -3'; SEQ ID NO: 2) and reverse primer (1492r): (5'- GGT TAC CTT TGT TAC GAC TT-3′; SEQ ID NO: 3) was used. 17 μl of distilled water, 1 μl of forward primer, 1 μl of reverse primer, and 1 μl of DNA were added to a PCR premix (Bioneer, Cat No. K-2012), and PCR was performed. PCR conditions were treated at 94 ° C for 5 minutes, followed by 30 cycles of 94 ° C for 1 minute, 62 ° C for 40 seconds, and 72 ° C for 40 seconds, and the reaction was terminated at 72 ° C for 7 minutes. PCR reaction products were confirmed by performing electrophoresis on a 0.8% agarose gel. As a result of analyzing the final nucleotide sequence and comparing it with the database to identify the lactic acid strain, the strain isolated by the present invention was found to be Lactobacillus plantarum. It was deposited on the 3rd of the month (KCTC18796P).
락토바실러스 플란타룸 V135(LLactobacillus Plantarum V135(L actobacillus plantarum V1actobacillus plantarum V1 35) KCTC18796P 균주에 대한 활용 특성 분석35) Utilization characteristic analysis for KCTC18796P strain
1. 락토바실러스 플란타룸 V135 균주의 내산성 평가1. Acid resistance evaluation of Lactobacillus plantarum V135 strain
유산균이 장내 도착하기 위해서는 섭취 후 pH 2인 위를 통과하여야 한다. 이러한 극한의 조건을 견뎌내는 내산성 조사하기 위하여 락토바실러스 플란타룸 V135 균주를 MRS 액체배지(MRS broth medium; Difco, USA)에 접종한 후 37℃에서 24시간 동안 배양하였다. 원심분리(4,000×g, 4℃, 5min)하여 Phosphate-buffered saline(PBS, pH7)에 2번 세척하였고, PBS로 균을 OD600 = 1.0(108~109cfu/㎖)으로 조정하여 사용하였다. 유산균 희석액 1㎖를 9㎖ PBS(pH 2와 7)에 첨가 후, 진탕한 뒤 37℃에서 배양하였다. 배양 초기부터 시간별(0hour, 30min, 1hour, 3hour)로 샘플링한 후 MRS 액체배지로 희석하고 MRS 고체배지에 도말한 후 37℃에서 48시간 배양 후 평판배지 위의 집락 수를 계수하여 생균수를 측정하였으며 그 결과는 하기 표 1과 같다.In order for lactic acid bacteria to reach the intestine, they must pass through the stomach, which has a pH of 2 after ingestion. In order to investigate acid resistance that endures these extreme conditions, the Lactobacillus plantarum V135 strain was inoculated into MRS broth medium (Difco, USA) and then cultured at 37° C. for 24 hours. Centrifuged (4,000×g, 4℃, 5min), washed twice in Phosphate-buffered saline (PBS, pH7), and used by adjusting the bacteria to OD 600 = 1.0 (10 8 ~10 9 cfu/ml) with PBS did After adding 1 ml of the lactic acid bacteria dilution to 9 ml of PBS (pH 2 and 7), shaken and incubated at 37 ° C. After sampling hourly (0hour, 30min, 1hour, 3hour) from the beginning of culture, diluting with MRS liquid medium, spreading on MRS solid medium, and culturing at 37℃ for 48 hours, counting the number of colonies on the plate medium to measure the number of viable cells. And the results are shown in Table 1 below.
상기 표 1에 나타나 바와 같이, 락토바실러스 플란타룸 V135 균주는 생리적 활성보다 낮은 pH 2에서도 생존 균수의 감소가 적어 내산성이 우수함을 확인하였다.As shown in Table 1, it was confirmed that the Lactobacillus plantarum V135 strain had excellent acid resistance with a small decrease in the number of viable bacteria even at a pH lower than physiological activity of 2.
2. 락토바실러스 플란타룸 V135 균주의 내담즙성 평가2. Evaluation of bile resistance of Lactobacillus plantarum V135 strain
본 발명에 따른 락토바실러스 플란타룸 V135 균주의 내담즙성을 평가하였다. 장관 내 담즙산염 농도가 0.1% 내외임을 감안하여 담즙산염(Porcine bile salts)농도가 각각 0, 1, 3%가 되도록 MRS 액체배지를 제조하였다. 본 발명에 따른 락토바실러스 플란타룸 V135 균주를 멸균된 MRS 액체배지에 접종하여 37℃에서 24시간 동안 배양한 후 담즙산염이 함유된 MRS 액체배지에 0.1%(v/v)씩 접종하였다. 접종 후 37℃에서 배양하면서 0, 0.5, 1, 3시간 뒤 시료를 채취한 후에 멸균된 MRS 액체배지에 희석하고 MRS 평판배지에 도말한 다음 37℃에서 48시간동안 배양한 후, 평판배지 위의 집락 수를 계수하여 생균수를 측정하였다. 그 결과는 하기 표 2에 나타내었으며, 락토바실러스 플란타룸 V135 균주는 장내 실제 농도와 유사한 1% 뿐만 아니라 더 높은 3%에서도 적정 균수를 유지하였기 때문에 락토바실러스 플란타룸 V135 균주는 담즙에 의한 사멸이 없이 인체나 동물의 장 내에서도 충분히 생존할 수 있음을 확인 할 수 있는 근거가 될 수 있다.The bile resistance of the Lactobacillus plantarum V135 strain according to the present invention was evaluated. Considering that the concentration of bile salts in the intestinal tract was around 0.1%, MRS liquid medium was prepared so that the concentrations of porcine bile salts were 0, 1, and 3%, respectively. The Lactobacillus plantarum V135 strain according to the present invention was inoculated into a sterilized MRS liquid medium, cultured at 37 ° C. for 24 hours, and then inoculated into MRS liquid medium containing bile salts at 0.1% (v / v). After inoculation, samples were taken after 0, 0.5, 1, and 3 hours while incubating at 37°C, diluted in sterilized MRS liquid medium, spread on MRS plate medium, and then incubated at 37°C for 48 hours. The number of viable cells was measured by counting the number of colonies. The results are shown in Table 2 below. Since the Lactobacillus plantarum V135 strain maintained an appropriate number of bacteria at a higher 3% as well as 1% similar to the actual concentration in the intestine, the Lactobacillus plantarum V135 strain was killed by bile Without this, it can be the basis for confirming that it can survive sufficiently in the intestines of humans or animals.
3. 락토바실러스 플란타룸 V135 균주의 공응집능 평가3. Evaluation of coagulation ability of Lactobacillus plantarum V135 strain
유산균의 항염 작용에 있어 유산균이 장내에 점착하면서 장내 병원성 유해세균과의 응집력이 매우 중요한 요인이다. 본 발명에서는 지시균으로 대장균(Escherichia coli), 대장균(Escherichia coli O157), 황색포도상구균(Staphylococcus aureus), 장티푸스(Salmonella typhimurium) 균주를 사용하였다. 배양 배지로 락토바실러스 플란타룸 V135 균주와 대장균을 각각 MRS 액체배지와 Nutrient 액체배지(Difco; USA)를 사용하였다. 락토바실러스 플란타룸 V135 균주를 전 배양한 배양액을 OD600 = 1.0(108~109cfu/㎖)으로 조정하여 새로운 5㎖ MRS 배지에 1% 접종한 뒤, 24시간 배양하고 원심분리(4,000×g, 4℃, 5min)하여 세포를 회수하였다. 병원성 균은 배양배지로 전배양한 후 OD600 = 0.1으로 조정하여 접종하고 37℃에서 24시간 동안 배양하여 사용하였다. 이를 PBS 완충용액에 다시 현탁하여 생균수를 1.0×108cfu/㎖로 조정하였다. 락토바실러스 플란타룸 V135 균주와 유해균 현탁액 각각 2㎖씩 혼합하고 10초간 교반한 후 37℃에서 5시간 동안 정치 보관하면서 각 시간별(0hour, 5hour)로 공응집능을 조사하였다. 공응집능은 Kos 등의 방법(2003)에 따라 하기 [계산식 1]로 계산하였으며, 그 결과는 하기 표 3과 같다.In the anti-inflammatory action of lactic acid bacteria, cohesion with pathogenic harmful bacteria in the intestine is a very important factor while lactic acid bacteria adhere to the intestine. In the present invention, strains of Escherichia coli , Escherichia coli O157, Staphylococcus aureus , and Salmonella typhimurium were used as indicator bacteria. As the culture medium, MRS liquid medium and Nutrient liquid medium (Difco; USA) were used for the Lactobacillus plantarum V135 strain and Escherichia coli, respectively. After adjusting the OD 600 = 1.0 (10 8 ~ 10 9 cfu / ml) of the pre-cultured culture medium of the Lactobacillus plantarum V135 strain and inoculating 1% in a new 5 ml MRS medium, incubating for 24 hours and centrifuging (4,000 ×g, 4°C, 5 min) to recover the cells. Pathogenic bacteria were inoculated by adjusting the OD 600 = 0.1 after pre-cultivation with a culture medium, and incubated at 37 ° C. for 24 hours before use. This was resuspended in PBS buffer solution to adjust the viable cell count to 1.0×10 8 cfu/ml. Lactobacillus plantarum V135 strain and harmful bacteria suspension were mixed by 2 ml each, stirred for 10 seconds, and then stored at 37 ° C. for 5 hours, and co-aggregation ability was investigated at each time (0 hour, 5 hour). The coagulability was calculated according to [Calculation 1] according to the method of Kos et al. (2003), and the results are shown in Table 3 below.
[계산식 1][Calculation 1]
- Ax : 락토바실러스 플란타룸 V135 현탁액의 OD600 - A x : OD 600 of Lactobacillus plantarum V135 suspension
- Ay : 유해균 현탁액의 OD600 - A y : OD 600 of harmful bacteria suspension
- Ax+y : 락토바실러스 플란타룸 V135와 유해균 혼합액의 OD600 - A x+y : OD 600 of a mixture of Lactobacillus plantarum V135 and harmful bacteria
colicoli
coli O157 Escherichia
coli O157
typhimuriumtyphimurium
aureusaureus
상기 표 3에 나타난 바와 같이 실험에 사용한 유해균 모두 락토바실러스 플란타룸 V135 균주와의 응집률이 높았다. 특히, 락토바실러스 플란타룸 V135 유산균과 대장균의 응집률이 74%로 장관 흡착능의 우수성을 보였다.As shown in Table 3, all of the harmful bacteria used in the experiment had a high rate of aggregation with the Lactobacillus plantarum V135 strain. In particular, the aggregation rate of Lactobacillus plantarum V135 lactic acid bacteria and Escherichia coli was 74%, showing excellent intestinal adsorption capacity.
락토바실러스 플란타룸 V135(LLactobacillus Plantarum V135(L actobacillus plantarum V1actobacillus plantarum V1 35) KCTC18796P 균주의 사균체35) Dead cells of KCTC18796P strain
락토바실러스 플란타룸 V135 균주 사균체의 제조방법은 하기와 같다.The method for preparing dead cells of the Lactobacillus plantarum V135 strain is as follows.
락토바실러스 플란타룸 V135 균주를 식용배지에 접종하고 37℃에서 24시간 배양하여 배양액을 제조하였다. 정지기(Stationary phase)로 세포 증식이 최고로 도달하고 다량의 대사산물을 포함하고 있는 시기에 상기 배양액을 100℃로 1차 가열(살균)하고 50℃ 이하로 급속냉각하였다. 완전한 사균화를 위하여 다시 100℃로 2차 가열(살균)하고 50℃ 이하로 급속냉각하여 열처리 유산균(이하, ‘사균체’이라고 함)을 제조하였다. 이후 상기 사균체에 부형제로 덱스트린, 말토덱스트린, 포도당 등의 부형제(또는 보호제)를 혼합하여 분무건조하여 분말화 하였다. 이때, 분무건조의 입구 온도는 180℃, 출구온도는 100℃ 이하로 하여 분무건조하였다.The Lactobacillus plantarum V135 strain was inoculated into an edible medium and cultured at 37° C. for 24 hours to prepare a culture solution. In the stationary phase, when cell proliferation reached its peak and contained a large amount of metabolites, the culture medium was first heated (sterilized) at 100 ° C and rapidly cooled below 50 ° C. For complete killing, heat-treated lactic acid bacteria (hereinafter referred to as 'killed cells') were prepared by secondary heating (sterilization) at 100 ° C and rapid cooling at 50 ° C or less. Thereafter, excipients (or protective agents) such as dextrin, maltodextrin, and glucose were mixed with the dead cells, and spray-dried to powder. At this time, the inlet temperature of the spray drying was 180 ℃, the outlet temperature was spray-dried to 100 ℃ or less.
락토바실러스 플란타룸 V135(LLactobacillus Plantarum V135(L actobacillus plantarum V1actobacillus plantarum V1 35) KCTC18796P 균주의 사균체에 대한 활용 특성 분석35) Analysis of utilization characteristics of dead cells of KCTC18796P strain
1. 락토바실러스 플란타룸 V135 사균체의 항균활성1. Antibacterial activity of dead cells of Lactobacillus plantarum V135
신규의 락토바실러스 플란타룸 V135 균주의 사균체에 대한 항균 활성은 agar well diffusion assay를 이용하여 확인하였다. 식품 유해균인 Escherichia coli O157, Salmonella typhimurium, Bacillus cereus를 대상으로 NB(Nutrient broth) 배지에서 24시간 이상 배양한 후 균액을 영양한천배지(Nutrient argar)에 1㎖를 분주하여 건조 하였다. 이후 멸균 상태의 biopsy punch(Stiefel Biopsy Punch; Stiefel Laboratories, Research Triangle Park, NC, USA)를 이용하여 well을 만들었다. 각 well에 대조구(a)와 락토바실러스 플란타룸 V135 사균체(b)를 각 well당 100㎕씩 분주하고 37℃에서 24시간 배양하였다. 배양 후, well 주위에 형성된 저해환을 확인하여 항균활성의 유무를 확인하였으며, 그 결과는 하기 도 2와 같다.The antibacterial activity of the novel Lactobacillus plantarum V135 strain against dead cells was confirmed using an agar well diffusion assay. Escherichia coli O157, Salmonella typhimurium , and Bacillus cereus , which are food harmful bacteria, were cultured in NB (Nutrient broth) medium for more than 24 hours, and then 1 ml of the bacterial solution was dispensed onto Nutrient agar and dried. Then, wells were made using a sterile biopsy punch (Stiefel Biopsy Punch; Stiefel Laboratories, Research Triangle Park, NC, USA). 100 μl of control (a) and dead cells (b) of Lactobacillus plantarum V135 were dispensed into each well and incubated at 37° C. for 24 hours. After culturing, the presence or absence of antibacterial activity was confirmed by checking the ring of inhibition formed around the well, and the results are shown in FIG. 2 below.
하기 도 2에 도시된 바와 같이, 락토바실러스 플란타룸 V135 균주는 Escherichia coli O157, Salmonella typhimurium 및 Bacillus cereus에 대한 높은 항균활성을 보였으며 락토바실러스 플란타룸 V135 균주의 사균체 또한 3종의 균주에 대한 항균활성이 높은 것으로 확인되었다.As shown in Figure 2, the Lactobacillus plantarum V135 strain showed high antibacterial activity against Escherichia coli O157, Salmonella typhimurium and Bacillus cereus , and the dead cells of the Lactobacillus plantarum V135 strain were also found in the three strains. It was found to have high antibacterial activity against
2. 락토바실러스 플란타룸 V135 사균체의 항산화 활성2. Antioxidant activity of dead cells of Lactobacillus plantarum V135
신규의 락토바실러스 플란타룸 V135 균주의 사균체에 대한 항균 활성은 DPPH radical 소거능 측정법을 이용하여 확인하였다. 락토바실러스 플란타룸 V135 균주의 사균체는 0.5, 1, 5, 10%의 농도가 되도록 멸균수에 녹여 실험에 사용하였다. DPPH 용액은 DMSO에 녹여 0.4mM의 농도로 제조하였다. DPPH radical 소거능은 시료 50㎕와 DPPH 용액 50㎕ 혼합하여 상온에서 10분간 반응한 후 분광광도계를 이용하여 517㎚에서 흡광도를 측정하였다. 대조구는 시료 대신 50㎕의 멸균수를 첨가하여 반응 후 얻은 흡광도 값을 사용하였다. 양성 대조구로는 항산화제로 잘 알려진 ascorbic acid를 사용하여 락토바실러스 플란타룸 V135 균주 사균체의 항산화 활성을 비교하였다. DPPH 라디칼 소거활성(%)은 하기의 [계산식 2]로 계산하였으며, 그 결과는 하기 도 3과 같다. The antibacterial activity of the novel Lactobacillus plantarum V135 strain against dead cells was confirmed using the DPPH radical scavenging activity measurement method. The dead cells of the Lactobacillus plantarum V135 strain were dissolved in sterile water to a concentration of 0.5, 1, 5, and 10% and used for the experiment. DPPH solution was prepared by dissolving in DMSO at a concentration of 0.4 mM. DPPH radical scavenging ability was measured by mixing 50 μl of sample and 50 μl of DPPH solution, reacting at room temperature for 10 minutes, and then measuring absorbance at 517 nm using a spectrophotometer. As a control, the absorbance value obtained after the reaction by adding 50 μl of sterile water instead of the sample was used. As a positive control, ascorbic acid, which is well known as an antioxidant, was used to compare the antioxidant activity of dead cells of Lactobacillus plantarum V135 strain. DPPH radical scavenging activity (%) was calculated by the following [Calculation 2], and the results are shown in FIG. 3 below.
[계산식 2][Calculation 2]
DPPH 라디칼 소거활성(%) = (B-A)/B × 100DPPH radical scavenging activity (%) = (B-A) / B × 100
(A: 시료의 흡광도, B: 대조구의 흡광도)(A: absorbance of sample, B: absorbance of control)
DPPH 라디칼 소거활성(%)을 측정한 결과, 하기 도 3에 도시된 바와 같이 락토바실러스 플란타룸 V135 사균체는 0.5, 1, 5, 10, 20%에서 각각 14.5, 27.6, 89.2, 94.3, 94.5%의 DPPH 라디칼 소거능(항산화능)을 나타내는 것으로 확인되었다. 양성 대조구로 사용된 ascorbic acid에서는 0.001, 0.01%에서 각각 2.14, 38.4%의 DPPH 라디칼 소거능을 나타내었다. 따라서 락토바실러스 플란타룸 V135 사균체는 뛰어난 DPPH 소거능을 가져 항산화 활성을 보유한 것으로 확인되었다.As a result of measuring the DPPH radical scavenging activity (%), as shown in FIG. 3, the Lactobacillus plantarum V135 dead cells were 14.5, 27.6, 89.2, 94.3, and 94.5 at 0.5, 1, 5, 10, and 20%, respectively. % of DPPH radical scavenging activity (antioxidant activity). Ascorbic acid used as a positive control showed DPPH radical scavenging activity of 2.14 and 38.4% at 0.001 and 0.01%, respectively. Therefore, it was confirmed that the dead cells of Lactobacillus plantarum V135 had an excellent DPPH scavenging ability and possessed antioxidant activity.
이러한 결과는 락토바실러스 플란타룸 V135 사균체는 높은 항균활성 및 항산화 효과를 나타내면서 균주가 가지는 단점인 소재의 안전성, 외부환경(내산성, 내담즙성) 및 열에 대한 안정성 부분을 보완한 사균체의 약학적 조성물 및 식품첨가제 조성물로서의 활용가능성을 보이는 결과이다.These results show that the dead cells of Lactobacillus plantarum V135 show high antibacterial activity and antioxidative effect, and the pharmaceuticals of dead cells complement the safety of the material, external environment (acid resistance, resistance to bile) and heat, which are the disadvantages of the strain. It is a result showing the applicability as a red composition and a food additive composition.
3. 락토바실러스 플란타룸 V135 사균체 투여로 인한 혈중 알코올 및 아세트알데히드 농도 측정 실험3. Blood alcohol and acetaldehyde concentration test due to administration of dead cells of Lactobacillus plantarum V135
락토바실러스 플란타룸 V135 사균체를 실험동물에 경구 투여하여 혈중 알코올 농도 및 아세트알데히드 농도를 측정하는 실험을 실시하였다.Lactobacillus plantarum V135 dead cells were orally administered to experimental animals to measure blood alcohol concentration and acetaldehyde concentration.
가. 실험군 설정 및 시료 투어go. Experiment group setup and sample tour
본 발명은 (주)오리엔트바이오(Seongnam, Korea)로부터 공급 받은 7주령의 수 컷 Sprague-Dawley rat를 일주일 동안 설치류 사육실에서 적응시킨 후, 적응기간 중 일반 상태를 관찰하여 건강한 개체만 실험에 사용하였다. 사육환경은 온도 22±2℃, 습도 55±5% 및 명/암 주기는 12시간으로 유지하고, 식이와 물을 자유롭게 공급하여 일주일간 기본식이로 적응시켰다. 순화기간 중 건강하다고 판정된 마우스에 한하여 무작위법을 이용하여 군을 분리하였다.In the present invention, 7-week-old male Sprague-Dawley rats supplied by Orient Bio Co., Ltd. (Seongnam, Korea) were acclimatized in a rodent breeding room for a week, and then, during the adaptation period, the general condition was observed and only healthy individuals were used in the experiment. . The breeding environment was maintained at a temperature of 22 ± 2 ° C, humidity of 55 ± 5%, and a light / dark cycle of 12 hours, and food and water were supplied freely to adapt to the basic diet for a week. Groups were separated using a random method only for mice determined to be healthy during the acclimatization period.
락토바실러스 플란타룸 V135 사균체의 알코올 및 아세트알데히드 분해효소의 활성 증가 효과를 알아보기 위해 하기 표 4에서와 같이, 정상대조군(NC), 에탄올군(EtOH), 양성대조군(milk thistle 200㎎/㎏/day for 10days, p.o.)(PC), 락토바실러스 플란타룸 V135 균주의 사균체(2×1010cfu/g)(V135) 200㎎/㎏/day 투여군 총 3군으로 군당 동일한 체중을 지닌 개체 8마리로 구성하여 실험을 진행하였다.In order to examine the effect of increasing the activity of alcohol and acetaldehyde degrading enzymes of Lactobacillus plantarum V135 dead cells, as shown in Table 4 below, normal control group (NC), ethanol group (EtOH), and positive control group (milk thistle 200 mg / kg/day for 10days, po) (PC), dead cell of Lactobacillus plantarum V135 strain (2×10 10 cfu/g) (V135) 200mg/kg/day administration group, 3 groups, each group having the same body weight The experiment was conducted with 8 individuals.
나. 실험 방법me. Experiment method
락토바실러스 플란타룸 V135 사균체의 알코올 및 아세트알데히드 분해효소의 활성 증가 효과를 평가하기 위해서, 알코올 섭취 30분 전 및 30분 후에 락토바실러스 플란타룸 V135 사균체를 포함한 배양물을 섭취하게 한 뒤 혈중 알코올 및 아세트알데히드 농도 측정하는 실험을 실시하였다.In order to evaluate the effect of increasing the activity of alcohol and acetaldehyde degrading enzymes of dead cells of Lactobacillus plantarum V135, cultures containing dead cells of Lactobacillus plantarum V135 were ingested 30 minutes before and 30 minutes after alcohol consumption An experiment was conducted to measure blood alcohol and acetaldehyde concentrations.
알코올 섭취 30분 전 락토바실러스 플란타룸 V135 사균체 섭취 시 락토바실러스 플란타룸 V135 사균체의 알코올 및 아세트알데히드 분해효소의 활성 증가 효과를 평가하기 위해서 NC와 EtOH군은 멸균한 증류수를 경구 투여 하였고, 양성대조군(PC), 락토바실러스 플란타룸 V135 사균체(V135)군은 멸균한 증류수에 각각의 시험물질(200㎎/㎏ body weight/d)을 준비하여 경구 투여하였다. 멸균 증류수와 각각의 시험물질을 경구 투여하고 30분이 경과한 후, 정상대조군(NC)은 멸균 증류수를 경구 투여하였고, 에탄올군(EtOH), 양성대조군(PC), 락토바실러스 플란타룸 V135 균주 사균체(V135)군은 25% 에탄올을 10㎖/㎏으로 경구 투여하였다. 에탄올을 경구 투여 한 후 1시간, 3시간, 5시간 경과 후, 각각 미동맥에서 채혈 하여 혈중 알코올 및 아세트알데히드 농도 측정하였다.In order to evaluate the effect of increasing the activity of alcohol and acetaldehyde decomposing enzyme of Lactobacillus plantarum V135 dead cells when ingesting dead Lactobacillus plantarum V135 cells 30 minutes before alcohol intake, sterilized distilled water was orally administered to the NC and EtOH groups. , The positive control group (PC), and the Lactobacillus plantarum V135 dead cells (V135) group prepared each test substance (200 mg/kg body weight/d) in sterilized distilled water and administered orally. After 30 minutes of oral administration of sterile distilled water and each test substance, the normal control group (NC) was orally administered sterile distilled water, and the ethanol group (EtOH), positive control group (PC), and Lactobacillus plantarum V135 strain were used. The cell (V135) group was orally administered with 10 ml/kg of 25% ethanol. After 1 hour, 3 hours, and 5 hours after oral administration of ethanol, blood was collected from the tail artery, respectively, and blood alcohol and acetaldehyde concentrations were measured.
또한, 알코올 섭취 30분 후 락토바실러스 플란타룸 V135 사균체 섭취 시 락토바실러스 플란타룸 V135 사균체의 알코올 및 아세트알데히드 분해효소의 활성 증가 효과를 평가하기 위해서 정상대조군(NC)은 멸균 증류수를 경구 투여하고, 에탄올군(EtOH), 양성대조군(PC), 락토바실러스 플란타룸 V135 균주 사균체(V135)군은 25% 에탄올을 10㎖/㎏으로 경구 투여하고 30분이 경과한 후, NC와 EtOH군은 멸균한 증류수를 경구 투여 하였고, 양성대조군(PC), 락토바실러스 플란타룸 V135 사균체(V135)군은 멸균한 증류수에 각각의 시험물질(200㎎/㎏ body weight/d)을 준비하여 경구 투여하였다. 에탄올을 경구 투여 한 후 1시간, 3시간, 5시간 경과 후, 각각 미동맥에서 채혈 하여 혈중 알코올 및 아세트알데히드 농도 측정하였다.In addition, in order to evaluate the effect of increasing the activity of alcohol and acetaldehyde lyase of Lactobacillus plantarum V135 dead cells when ingesting dead Lactobacillus plantarum V135 cells 30 minutes after alcohol consumption, the normal control group (NC) was given sterile distilled water orally. administration, and the ethanol group (EtOH), positive control group (PC), and Lactobacillus plantarum V135 strain killed cells (V135) group were orally administered with 25% ethanol at 10 ml/kg and after 30 minutes had elapsed, NC and EtOH The group was orally administered with sterilized distilled water, and the positive control group (PC) and Lactobacillus plantarum V135 dead cell (V135) group prepared each test substance (200 mg / kg body weight / d) in sterilized distilled water. Orally administered. After 1 hour, 3 hours, and 5 hours after oral administration of ethanol, blood was collected from the tail artery, respectively, and blood alcohol and acetaldehyde concentrations were measured.
다. 혈중 알코올 및 아세트알데히드 농도 측정 방법All. How to measure blood alcohol and acetaldehyde concentrations
에탄올 투여 후 시간별(1시간, 3시간 및 5시간)로 채취 한 혈액은 4℃, 3000rpm에서 10분간 원심분리하여 혈청을 얻었다. 채취한 혈청의 에탄올 함량을 측정하기 위해서 제조된 에탄올 및 아세트알데히드 측정용 assay kit(Germany, Roche Co., Darmstadt)를 이용하였다. Photassium phosphate buffer(pH 9)와 NAD+를 혼합한 후 혈액 상등액 샘플을 첨가 하여 20℃에서 5분간 반응 시킨 후 micro reader를 이용 하여 340㎚에서 흡광도를 측정하였다(A1). 또한 혼합액의 ADH와 ALDH 50㎕를 첨가하여 20℃에서 5분간 반응시킨 후 340㎚에서 흡광도를 측정하였다(A2). 이를 바탕으로 시간별 혈중 알코올의 농도는 하기의 [계산식 3]에 대입하여 정량하였으며, 혈중 아세트알데히드의 농도는 하기의 [계산식 4]에 대입하여 정량하였다.After ethanol administration, blood collected hourly (1 hour, 3 hours, and 5 hours) was centrifuged at 4°C and 3000 rpm for 10 minutes to obtain serum. In order to measure the ethanol content of the collected serum, an assay kit (Germany, Roche Co., Darmstadt) for measuring ethanol and acetaldehyde was used. After mixing Photassium phosphate buffer (pH 9) and NAD+, a blood supernatant sample was added and reacted at 20 ° C for 5 minutes, and then the absorbance was measured at 340 nm using a micro reader (A1). In addition, 50 μl of ADH and ALDH of the mixed solution was added and reacted at 20° C. for 5 minutes, and then absorbance was measured at 340 nm (A2). Based on this, the hourly blood alcohol concentration was quantified by substituting into [Calculation 3] below, and the concentration of acetaldehyde in blood was quantified by substituting into [Calculation 4] below.
[계산식 3][Calculation 3]
Concentration = 0.7259/3.6×△AConcentration = 0.7259/3.6×ΔA
△A = sample(A2-A1)-blank(A2-A1)ΔA = sample(A2-A1)-blank(A2-A1)
[계산식 4][Calculation 4]
Concentration = 0.7158/3.6×△AConcentration = 0.7158/3.6×ΔA
△A = sample(A2-A1)-blank(A2-A1)ΔA = sample(A2-A1)-blank(A2-A1)
라. 혈중 알코올 및 아세트알데히드 농도 측정 결과la. Measurement of blood alcohol and acetaldehyde concentrations
실험동물에 에탄올 투여 30분 전 각각의 시료를 투여하여 혈액의 알코올(에탄올) 및 아세트알데히드 함량 변화를 관찰한 실험 결과는 도 4와 같다. 하기 도 4에 도시된 바와 같이, 에탄올 투여 후 1시간까지는 알코올 및 아세트알데히드 함량이 급격히 증가 하였다가 그 이후 감소하였으며, 락토바실러스 플란타룸 V135 사균체(V135) 투여군은 5시간에서 유의적으로 혈중 알코올 및 아세트알데히드 함량이 감소되는 것으로 확인되었다.Experimental results obtained by administering each sample 30 minutes before ethanol administration to experimental animals and observing changes in blood alcohol (ethanol) and acetaldehyde contents are shown in FIG. 4 . As shown in FIG. 4 below, the alcohol and acetaldehyde contents increased rapidly up to 1 hour after ethanol administration and then decreased thereafter, and the Lactobacillus plantarum V135 dead cell body (V135) administration group was significantly blood at 5 hours It was found that the alcohol and acetaldehyde contents were reduced.
실험동물에 에탄올 투여 30분 후 각각의 시료를 투여하여 혈액의 알코올(에탄올) 및 아세트알데히드 함량 변화를 관찰한 실험 결과는 도 5와 같다. 하기 도 5에 도시된 바와 같이, 에탄올 투여 후 1시간까지는 알코올 및 아세트알데히드 함량이 급격히 증가하였지만 그 이후 감소하였으며, 양성대조군(PC)과 락토바실러스 플란타룸 V135 사균체(V135) 투여군은 5시간에서 모두 유의적으로 혈중 알코올 및 아세트알데히드 함량이 감소되는 것으로 확인되었다.Experimental results obtained by administering each sample 30 minutes after ethanol administration to experimental animals and observing changes in blood alcohol (ethanol) and acetaldehyde content are shown in FIG. 5 . As shown in FIG. 5 below, the alcohol and acetaldehyde contents increased rapidly up to 1 hour after ethanol administration, but decreased thereafter, and the positive control group (PC) and the Lactobacillus plantarum V135 dead cell body (V135) administration group were 5 hours It was confirmed that the blood alcohol and acetaldehyde content were significantly reduced in all of them.
이러한 결과를 종합할 때 락토바실러스 플란타룸 V135 사균체가 알코올 및 아세트알데히드 분해효소의 활성을 증가시킴으로써 혈중 알코올 및 아세트알데히드 농도를 감소시키는 효과가 있음을 설명할 수 있다.Taking these results together, it can be explained that dead cells of Lactobacillus plantarum V135 increase the activity of alcohol and acetaldehyde degrading enzymes, thereby reducing blood alcohol and acetaldehyde concentrations.
결론적으로, 상기 실시예 1 내지 4의 실험 결과를 통해, 락토바실러스 플란타룸 V135 균주의 사균체는 알코올 및 아세트알데히드 분해효소의 활성을 증가시킴으로써 혈중 알코올 및 아세트알데히드 농도를 감소시키는 효과를 나타내므로 알코올 및 아세트알데히드 분해효소의 활성 증가용 약학적 조성물 또는 식품첨가제 조성물로 사용될 수 있는 장점이 있으며, 락토바실러스 플란타룸 V135(Lactobacillus plantarum V135) KCTC 균주의 열처리를 통한 사균화는 생균의 단점(안정성, 제품 적용성 등)을 개선하여 안정하고 다양한 제품으로의 적용이 가능하면서 생균과의 동등 이상의 기능성을 보유한 소재를 제공할 수 있는 장점이 있음을 확인하였다.In conclusion, through the experimental results of Examples 1 to 4, the dead cells of the Lactobacillus plantarum V135 strain show the effect of reducing blood alcohol and acetaldehyde concentrations by increasing the activity of alcohol and acetaldehyde decomposition enzymes. It has the advantage of being able to be used as a pharmaceutical composition or food additive composition for increasing the activity of alcohol and acetaldehyde lyase, and killing Lactobacillus plantarum V135 KCTC strain through heat treatment has the disadvantage of live bacteria (stability , product applicability, etc.), it was confirmed that there is an advantage in providing a material that is stable and can be applied to various products and has functionality equal to or higher than that of live bacteria.
이상, 실시예를 들어 본 발명을 상세하게 설명하였으나, 본 발명은 상기 실시예에 한정되지 않으며, 여러 가지 다양한 형태로 변형될 수 있고, 본 발명의 기술적 사상 내에서 당 분야에서 통상의 지식을 가진 자에 의하여 여러 가지 많은 변형이 가능함이 명백하다. 또한, 청구범위의 기재된 본 발명의 기술적 사상을 벗어나지 않는 범위 내에서 당 기술분야의 통상의 지식을 가진 자에 의해 다양한 형태의 치환, 변형 및 변경이 가능할 것이며, 이 또한 본 발명의 범위에 속한다고 할 것이다.Above, the present invention has been described in detail with examples, but the present invention is not limited to the above embodiments, and can be modified in various forms, and within the technical spirit of the present invention, those skilled in the art It is clear that many variations are possible by the ruler. In addition, various forms of substitution, modification and change will be possible by those skilled in the art within the scope of the technical spirit of the present invention described in the claims, which also falls within the scope of the present invention. something to do.
Claims (6)
상기 균주의 사균체는,
락토바실러스 플란타룸 V135(Lactobacillus plantarum V135) KCTC18796P 균주를 식용배지에 접종하여 배양액을 제조하고, 배양액을 80℃ 이상의 온도로 가열한 뒤 60℃ 이하의 온도가 되도록 급속 냉각시키는 공정을 1회 이상 반복 실시하여 열처리한 것임을 특징으로 하는 알코올 및 아세트알데히드 분해효소의 활성 증가용 약학적 조성물.The method of claim 1,
Dead cells of the strain,
Lactobacillus plantarum V135 ( Lactobacillus plantarum V135) KCTC18796P strain is inoculated into an edible medium to prepare a culture solution, heating the culture solution to a temperature of 80 ° C or higher and then rapidly cooling to a temperature of 60 ° C or less, repeating the process one or more times. A pharmaceutical composition for increasing the activity of alcohol and acetaldehyde decomposing enzyme, characterized in that it is heat-treated by carrying out.
상기 알코올 및 아세트알데히드 분해효소의 활성 증가용 약학적 조성물은,
락토바실러스 플란타룸 V135(Lactobacillus plantarum V135) KCTC18796P 균주의 사균체가 조성물 1g당 1×105cfu 내지 1×1012cfu로 포함되는 것을 특징으로 하는 알코올 및 아세트알데히드 분해효소의 활성 증가용 약학적 조성물.The method of claim 1,
The pharmaceutical composition for increasing the activity of alcohol and acetaldehyde degrading enzyme,
Lactobacillus plantarum V135 ( Lactobacillus plantarum V135 ) Pharmaceutical for increasing the activity of alcohol and acetaldehyde degrading enzyme, characterized in that dead cells of KCTC18796P strain are included in 1 × 10 5 cfu to 1 × 10 12 cfu per 1 g of the composition composition.
상기 균주의 사균체는,
락토바실러스 플란타룸 V135(Lactobacillus plantarum V135) KCTC18796P 균주를 식용배지에 접종하여 배양액을 제조하고, 배양액을 80℃ 이상의 온도로 가열한 뒤 60℃ 이하의 온도가 되도록 급속 냉각시키는 공정을 1회 이상 반복 실시하여 열처리한 것임을 특징으로 하는 알코올 및 아세트알데히드 분해효소의 활성 증가용 식품첨가제 조성물.The method of claim 4,
Dead cells of the strain,
Lactobacillus plantarum V135 ( Lactobacillus plantarum V135) KCTC18796P strain is inoculated into an edible medium to prepare a culture solution, heating the culture solution to a temperature of 80 ° C or higher and then rapidly cooling to a temperature of 60 ° C or less, repeating the process one or more times. A food additive composition for increasing the activity of alcohol and acetaldehyde decomposing enzyme, characterized in that it is heat-treated by carrying out.
상기 알코올 및 아세트알데히드 분해효소의 활성 증가용 식품첨가제 조성물은,
락토바실러스 플란타룸 V135(Lactobacillus plantarum V135) KCTC18796P 균주의 사균체가 조성물 1g당 1×105cfu 내지 1×1012cfu로 포함되는 것을 특징으로 하는 알코올 및 아세트알데히드 분해효소의 활성 증가용 식품첨가제 조성물.The method of claim 4,
The food additive composition for increasing the activity of the alcohol and acetaldehyde degrading enzyme,
Lactobacillus plantarum V135 ( Lactobacillus plantarum V135 ) Food additive for increasing activity of alcohol and acetaldehyde degrading enzyme, characterized in that dead cells of KCTC18796P strain are contained in 1 × 10 5 cfu to 1 × 10 12 cfu per 1 g of composition composition.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| KR1020210070567A KR20220162284A (en) | 2021-06-01 | 2021-06-01 | A composition for increased activity of alcohol and acetaldehyde degrading enzymes of the comprising heat-killed lactobacillus plantarum v135 as an active ingredient |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| KR1020210070567A KR20220162284A (en) | 2021-06-01 | 2021-06-01 | A composition for increased activity of alcohol and acetaldehyde degrading enzymes of the comprising heat-killed lactobacillus plantarum v135 as an active ingredient |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| KR20220162284A true KR20220162284A (en) | 2022-12-08 |
Family
ID=84436872
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| KR1020210070567A KR20220162284A (en) | 2021-06-01 | 2021-06-01 | A composition for increased activity of alcohol and acetaldehyde degrading enzymes of the comprising heat-killed lactobacillus plantarum v135 as an active ingredient |
Country Status (1)
| Country | Link |
|---|---|
| KR (1) | KR20220162284A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20230134888A (en) * | 2022-03-15 | 2023-09-22 | 강원대학교산학협력단 | Mutant strain Lactobacillus fermentum JY563 with increased exopolysaccharide yield and increased alcohol dehydrogenase activity and increased antioxidant activity |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR101919938B1 (en) | 2017-06-16 | 2019-02-08 | 주식회사 락토메이슨 | Composition comprising heat-killed lactobacillus plantarum lm1004 having immunostimulating activity |
| KR102028744B1 (en) | 2018-07-23 | 2019-10-07 | 주식회사한국야쿠르트 | Lactobacillus plantarum HY7717 strain having immune-enhancing activity, antioxidative activity and digestive fluid resistance and use thereof |
-
2021
- 2021-06-01 KR KR1020210070567A patent/KR20220162284A/en not_active Application Discontinuation
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR101919938B1 (en) | 2017-06-16 | 2019-02-08 | 주식회사 락토메이슨 | Composition comprising heat-killed lactobacillus plantarum lm1004 having immunostimulating activity |
| KR102028744B1 (en) | 2018-07-23 | 2019-10-07 | 주식회사한국야쿠르트 | Lactobacillus plantarum HY7717 strain having immune-enhancing activity, antioxidative activity and digestive fluid resistance and use thereof |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20230134888A (en) * | 2022-03-15 | 2023-09-22 | 강원대학교산학협력단 | Mutant strain Lactobacillus fermentum JY563 with increased exopolysaccharide yield and increased alcohol dehydrogenase activity and increased antioxidant activity |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN110964655B (en) | Bifidobacterium lactis BL-99 and application thereof | |
| KR100367553B1 (en) | A Method for Preparing Functional Cultures | |
| Falah et al. | Optimization of γ-aminobutyric acid (GABA) production by Lactobacillus spp. from agro-food waste | |
| KR102251295B1 (en) | A composition for preventing or relieving hangover of the comprising heat-killed lactobacillus salivarius v133 as an active ingredient | |
| KR101868517B1 (en) | Lactobacillus fermentum PL9119 with biofunctional activities and high heat stability as a probiotic without antibiotic resistance | |
| CN101812414A (en) | Lactobacillus plantarum and bacteriocins produced by lactobacillus plantarum and capable of inhibiting Gram negative bacteria | |
| Rezaei et al. | Isolation of lactic acid probiotic strains from Iranian camel milk: technological and antioxidant properties | |
| KR102251294B1 (en) | A composition for preventing, improving or treating alcoholic gastritis of the comprising heat-killed lactobacillus salivarius v133 as an active ingredient | |
| CN114468306B (en) | Application of bacillus coagulans BC99 in preparation of colitis relieving product or immunoregulation product | |
| Pawar | Lactobacillus rhamnosus ARJD as a functional food with potential antioxidant and antibacterial abilities | |
| Soares et al. | Sporeforming probiotic bacteria: Characteristics, health benefits, and technological aspects for their applications in foods and beverages | |
| CN110607256A (en) | Lactobacillus reuteri JYLB-131 for improving activity of alcohol dehydrogenase after drinking and application thereof in food and medicine | |
| Guan et al. | Characterization and in vitro assessment of probiotic potential of Lactiplantibacillus plantarum BXM2 from fermented honey passion fruit beverage | |
| CN113088468B (en) | Lactobacillus casei Ma. GLRGJ1 and application thereof | |
| KR102262646B1 (en) | A composition for preventing or relieving hangover of the comprising heat-killed lactobacillus plantarum v135 as an active ingredient | |
| JP3520012B2 (en) | Composition suitable as a nutrient supplement and for treating intestinal disorders and modifying the bacterial flora | |
| KR101836365B1 (en) | Kimchi seasoning containing Leuconostoc mesenteroides WiKim32 and kimchi prepared by using the same | |
| JP2013116089A (en) | Lactobacillus strain having anti-helicobacter pylori activity, anti-helicobacter pylori medicine containing the lactobacillus strain, and food and drink containing the same | |
| KR102290381B1 (en) | A composition for preventing, improving or treating alcoholic hepatitis of the comprising lactobacillus plantarum v135 or heat-killed lactobacillus plantarum v135 as an active ingredient | |
| Zhang et al. | Safety evaluation of Lactobacillus paracasei subsp. paracasei LC-01, a probiotic bacterium | |
| Sherwani | Probiotics in processed dairy products and their role in gut microbiota health | |
| KR20220162284A (en) | A composition for increased activity of alcohol and acetaldehyde degrading enzymes of the comprising heat-killed lactobacillus plantarum v135 as an active ingredient | |
| KR100720025B1 (en) | Probiotic lactic acid bacteria and composition comprising the same | |
| KR101670955B1 (en) | Feed for farming-fish and Farming-fish farmed using that | |
| Mahrous et al. | The role of some probiotic lactic acid bacteria in the reduction of cholesterol on mice |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| E902 | Notification of reason for refusal | ||
| E601 | Decision to refuse application |