JP2006271220A - Cultured microorganism obtained from medium consisting mainly of extracts of green tea, black tea and mugwort - Google Patents
Cultured microorganism obtained from medium consisting mainly of extracts of green tea, black tea and mugwort Download PDFInfo
- Publication number
- JP2006271220A JP2006271220A JP2005091624A JP2005091624A JP2006271220A JP 2006271220 A JP2006271220 A JP 2006271220A JP 2005091624 A JP2005091624 A JP 2005091624A JP 2005091624 A JP2005091624 A JP 2005091624A JP 2006271220 A JP2006271220 A JP 2006271220A
- Authority
- JP
- Japan
- Prior art keywords
- microorganism
- green tea
- culturing
- black tea
- tea
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 244000005700 microbiome Species 0.000 title claims abstract description 97
- 244000269722 Thea sinensis Species 0.000 title claims abstract description 54
- 239000000284 extract Substances 0.000 title claims abstract description 33
- 235000003261 Artemisia vulgaris Nutrition 0.000 title claims abstract description 28
- 240000006891 Artemisia vulgaris Species 0.000 title claims abstract description 28
- 235000009569 green tea Nutrition 0.000 title claims abstract description 27
- 235000006468 Thea sinensis Nutrition 0.000 title claims abstract description 26
- 235000020279 black tea Nutrition 0.000 title claims abstract description 26
- 235000015141 kefir Nutrition 0.000 claims abstract description 30
- 239000002537 cosmetic Substances 0.000 claims abstract description 23
- 238000012258 culturing Methods 0.000 claims abstract description 21
- 230000002401 inhibitory effect Effects 0.000 claims abstract description 21
- 235000013305 food Nutrition 0.000 claims abstract description 20
- 235000013361 beverage Nutrition 0.000 claims abstract description 18
- 239000003963 antioxidant agent Substances 0.000 claims description 43
- 230000003078 antioxidant effect Effects 0.000 claims description 42
- 241001468155 Lactobacillaceae Species 0.000 claims description 21
- 230000008099 melanin synthesis Effects 0.000 claims description 19
- 238000009629 microbiological culture Methods 0.000 claims description 15
- 241000894007 species Species 0.000 claims description 10
- 238000000034 method Methods 0.000 claims 1
- XUMBMVFBXHLACL-UHFFFAOYSA-N Melanin Chemical compound O=C1C(=O)C(C2=CNC3=C(C(C(=O)C4=C32)=O)C)=C2C4=CNC2=C1C XUMBMVFBXHLACL-UHFFFAOYSA-N 0.000 abstract description 26
- 230000000694 effects Effects 0.000 abstract description 13
- 230000003647 oxidation Effects 0.000 abstract description 11
- 238000007254 oxidation reaction Methods 0.000 abstract description 11
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 abstract description 9
- 239000001301 oxygen Substances 0.000 abstract description 9
- 229910052760 oxygen Inorganic materials 0.000 abstract description 9
- 239000002994 raw material Substances 0.000 abstract description 9
- 241001465754 Metazoa Species 0.000 abstract description 6
- 230000006866 deterioration Effects 0.000 abstract description 3
- 239000000654 additive Substances 0.000 abstract description 2
- 230000000996 additive effect Effects 0.000 abstract description 2
- 230000003064 anti-oxidating effect Effects 0.000 abstract description 2
- 239000002245 particle Substances 0.000 abstract 1
- 235000006708 antioxidants Nutrition 0.000 description 42
- 239000002609 medium Substances 0.000 description 23
- 239000000243 solution Substances 0.000 description 13
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 12
- 210000003491 skin Anatomy 0.000 description 10
- 239000000047 product Substances 0.000 description 9
- 210000004027 cell Anatomy 0.000 description 8
- 239000000126 substance Substances 0.000 description 8
- 241000186610 Lactobacillus sp. Species 0.000 description 6
- 201000010099 disease Diseases 0.000 description 5
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 5
- 238000001727 in vivo Methods 0.000 description 5
- WZNJWVWKTVETCG-UHFFFAOYSA-N kojic acid Natural products OC(=O)C(N)CN1C=CC(=O)C(O)=C1 WZNJWVWKTVETCG-UHFFFAOYSA-N 0.000 description 5
- BEJNERDRQOWKJM-UHFFFAOYSA-N kojic acid Chemical compound OCC1=CC(=O)C(O)=CO1 BEJNERDRQOWKJM-UHFFFAOYSA-N 0.000 description 5
- 229960004705 kojic acid Drugs 0.000 description 5
- QIGBRXMKCJKVMJ-UHFFFAOYSA-N Hydroquinone Chemical compound OC1=CC=C(O)C=C1 QIGBRXMKCJKVMJ-UHFFFAOYSA-N 0.000 description 4
- 206010028980 Neoplasm Diseases 0.000 description 4
- 230000002292 Radical scavenging effect Effects 0.000 description 4
- MBMBGCFOFBJSGT-KUBAVDMBSA-N all-cis-docosa-4,7,10,13,16,19-hexaenoic acid Chemical compound CC\C=C/C\C=C/C\C=C/C\C=C/C\C=C/C\C=C/CCC(O)=O MBMBGCFOFBJSGT-KUBAVDMBSA-N 0.000 description 4
- 229960005070 ascorbic acid Drugs 0.000 description 4
- 235000010354 butylated hydroxytoluene Nutrition 0.000 description 4
- 201000011510 cancer Diseases 0.000 description 4
- GVJHHUAWPYXKBD-UHFFFAOYSA-N d-alpha-tocopherol Natural products OC1=C(C)C(C)=C2OC(CCCC(C)CCCC(C)CCCC(C)C)(C)CCC2=C1C GVJHHUAWPYXKBD-UHFFFAOYSA-N 0.000 description 4
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 4
- 229930014626 natural product Natural products 0.000 description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 4
- 241000196324 Embryophyta Species 0.000 description 3
- 206010014970 Ephelides Diseases 0.000 description 3
- 208000003351 Melanosis Diseases 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 238000002835 absorbance Methods 0.000 description 3
- 230000032683 aging Effects 0.000 description 3
- CZBZUDVBLSSABA-UHFFFAOYSA-N butylated hydroxyanisole Chemical compound COC1=CC=C(O)C(C(C)(C)C)=C1.COC1=CC=C(O)C=C1C(C)(C)C CZBZUDVBLSSABA-UHFFFAOYSA-N 0.000 description 3
- 239000012153 distilled water Substances 0.000 description 3
- 239000003925 fat Substances 0.000 description 3
- 235000019197 fats Nutrition 0.000 description 3
- 239000003921 oil Substances 0.000 description 3
- 239000000049 pigment Substances 0.000 description 3
- 230000002087 whitening effect Effects 0.000 description 3
- SPSPIUSUWPLVKD-UHFFFAOYSA-N 2,3-dibutyl-6-methylphenol Chemical compound CCCCC1=CC=C(C)C(O)=C1CCCC SPSPIUSUWPLVKD-UHFFFAOYSA-N 0.000 description 2
- 206010003210 Arteriosclerosis Diseases 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 2
- CIWBSHSKHKDKBQ-DUZGATOHSA-N D-araboascorbic acid Natural products OC[C@@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-DUZGATOHSA-N 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 2
- 239000002211 L-ascorbic acid Substances 0.000 description 2
- 235000000069 L-ascorbic acid Nutrition 0.000 description 2
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 2
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 description 2
- 206010042496 Sunburn Diseases 0.000 description 2
- -1 VC) Chemical compound 0.000 description 2
- JAZBEHYOTPTENJ-JLNKQSITSA-N all-cis-5,8,11,14,17-icosapentaenoic acid Chemical compound CC\C=C/C\C=C/C\C=C/C\C=C/C\C=C/CCCC(O)=O JAZBEHYOTPTENJ-JLNKQSITSA-N 0.000 description 2
- 208000011775 arteriosclerosis disease Diseases 0.000 description 2
- 235000010323 ascorbic acid Nutrition 0.000 description 2
- 239000011668 ascorbic acid Substances 0.000 description 2
- 230000015556 catabolic process Effects 0.000 description 2
- ADRVNXBAWSRFAJ-UHFFFAOYSA-N catechin Natural products OC1Cc2cc(O)cc(O)c2OC1c3ccc(O)c(O)c3 ADRVNXBAWSRFAJ-UHFFFAOYSA-N 0.000 description 2
- 235000005487 catechin Nutrition 0.000 description 2
- 150000001765 catechin Chemical class 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 238000006731 degradation reaction Methods 0.000 description 2
- 206010012601 diabetes mellitus Diseases 0.000 description 2
- 235000020669 docosahexaenoic acid Nutrition 0.000 description 2
- 229940090949 docosahexaenoic acid Drugs 0.000 description 2
- 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 2
- JAZBEHYOTPTENJ-UHFFFAOYSA-N eicosapentaenoic acid Natural products CCC=CCC=CCC=CCC=CCC=CCCCC(O)=O JAZBEHYOTPTENJ-UHFFFAOYSA-N 0.000 description 2
- 235000020673 eicosapentaenoic acid Nutrition 0.000 description 2
- 229960005135 eicosapentaenoic acid Drugs 0.000 description 2
- 235000010350 erythorbic acid Nutrition 0.000 description 2
- 239000000796 flavoring agent Substances 0.000 description 2
- 235000019634 flavors Nutrition 0.000 description 2
- 229930182470 glycoside Natural products 0.000 description 2
- 150000002338 glycosides Chemical class 0.000 description 2
- 229940026239 isoascorbic acid Drugs 0.000 description 2
- 235000014655 lactic acid Nutrition 0.000 description 2
- 239000004310 lactic acid Substances 0.000 description 2
- 201000001441 melanoma Diseases 0.000 description 2
- 235000010384 tocopherol Nutrition 0.000 description 2
- 229930003799 tocopherol Natural products 0.000 description 2
- 229960001295 tocopherol Drugs 0.000 description 2
- 239000011732 tocopherol Substances 0.000 description 2
- 231100000419 toxicity Toxicity 0.000 description 2
- 230000001988 toxicity Effects 0.000 description 2
- GVJHHUAWPYXKBD-IEOSBIPESA-N α-tocopherol Chemical compound OC1=C(C)C(C)=C2O[C@@](CCC[C@H](C)CCC[C@H](C)CCCC(C)C)(C)CCC2=C1C GVJHHUAWPYXKBD-IEOSBIPESA-N 0.000 description 2
- 241000776564 Acetobacter cerevisiae Species 0.000 description 1
- 206010007269 Carcinogenicity Diseases 0.000 description 1
- 208000002177 Cataract Diseases 0.000 description 1
- 206010008805 Chromosomal abnormalities Diseases 0.000 description 1
- 208000031404 Chromosome Aberrations Diseases 0.000 description 1
- ZZZCUOFIHGPKAK-UHFFFAOYSA-N D-erythro-ascorbic acid Natural products OCC1OC(=O)C(O)=C1O ZZZCUOFIHGPKAK-UHFFFAOYSA-N 0.000 description 1
- MYMOFIZGZYHOMD-UHFFFAOYSA-N Dioxygen Chemical compound O=O MYMOFIZGZYHOMD-UHFFFAOYSA-N 0.000 description 1
- 229930091371 Fructose Natural products 0.000 description 1
- 239000005715 Fructose Substances 0.000 description 1
- RFSUNEUAIZKAJO-ARQDHWQXSA-N Fructose Chemical compound OC[C@H]1O[C@](O)(CO)[C@@H](O)[C@@H]1O RFSUNEUAIZKAJO-ARQDHWQXSA-N 0.000 description 1
- 241000589232 Gluconobacter oxydans Species 0.000 description 1
- 244000286779 Hansenula anomala Species 0.000 description 1
- 235000014683 Hansenula anomala Nutrition 0.000 description 1
- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-N 0.000 description 1
- UPYKUZBSLRQECL-UKMVMLAPSA-N Lycopene Natural products CC(=C/C=C/C=C(C)/C=C/C=C(C)/C=C/C1C(=C)CCCC1(C)C)C=CC=C(/C)C=CC2C(=C)CCCC2(C)C UPYKUZBSLRQECL-UKMVMLAPSA-N 0.000 description 1
- JEVVKJMRZMXFBT-XWDZUXABSA-N Lycophyll Natural products OC/C(=C/CC/C(=C\C=C\C(=C/C=C/C(=C\C=C\C=C(/C=C/C=C(\C=C\C=C(/CC/C=C(/CO)\C)\C)/C)\C)/C)\C)/C)/C JEVVKJMRZMXFBT-XWDZUXABSA-N 0.000 description 1
- 229940124200 Melanin inhibitor Drugs 0.000 description 1
- 241000235062 Pichia membranifaciens Species 0.000 description 1
- 208000012641 Pigmentation disease Diseases 0.000 description 1
- 239000012980 RPMI-1640 medium Substances 0.000 description 1
- OUUQCZGPVNCOIJ-UHFFFAOYSA-M Superoxide Chemical compound [O-][O] OUUQCZGPVNCOIJ-UHFFFAOYSA-M 0.000 description 1
- 102000003425 Tyrosinase Human genes 0.000 description 1
- 108060008724 Tyrosinase Proteins 0.000 description 1
- 229930003268 Vitamin C Natural products 0.000 description 1
- 229930003427 Vitamin E Natural products 0.000 description 1
- 244000195452 Wasabia japonica Species 0.000 description 1
- 235000000760 Wasabia japonica Nutrition 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- OENHQHLEOONYIE-UKMVMLAPSA-N all-trans beta-carotene Natural products CC=1CCCC(C)(C)C=1/C=C/C(/C)=C/C=C/C(/C)=C/C=C/C=C(C)C=CC=C(C)C=CC1=C(C)CCCC1(C)C OENHQHLEOONYIE-UKMVMLAPSA-N 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 235000010208 anthocyanin Nutrition 0.000 description 1
- 239000004410 anthocyanin Substances 0.000 description 1
- 229930002877 anthocyanin Natural products 0.000 description 1
- 150000004636 anthocyanins Chemical class 0.000 description 1
- 230000000845 anti-microbial effect Effects 0.000 description 1
- 235000013734 beta-carotene Nutrition 0.000 description 1
- 239000011648 beta-carotene Substances 0.000 description 1
- TUPZEYHYWIEDIH-WAIFQNFQSA-N beta-carotene Natural products CC(=C/C=C/C=C(C)/C=C/C=C(C)/C=C/C1=C(C)CCCC1(C)C)C=CC=C(/C)C=CC2=CCCCC2(C)C TUPZEYHYWIEDIH-WAIFQNFQSA-N 0.000 description 1
- 229960002747 betacarotene Drugs 0.000 description 1
- 210000004204 blood vessel Anatomy 0.000 description 1
- 230000007670 carcinogenicity Effects 0.000 description 1
- 231100000260 carcinogenicity Toxicity 0.000 description 1
- 235000021466 carotenoid Nutrition 0.000 description 1
- 150000001747 carotenoids Chemical class 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 208000026106 cerebrovascular disease Diseases 0.000 description 1
- 235000015140 cultured milk Nutrition 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 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 1
- 230000002500 effect on skin Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 210000002615 epidermis Anatomy 0.000 description 1
- 239000004318 erythorbic acid Substances 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 238000005562 fading Methods 0.000 description 1
- 229930003935 flavonoid Natural products 0.000 description 1
- 235000017173 flavonoids Nutrition 0.000 description 1
- 150000002215 flavonoids Chemical class 0.000 description 1
- 235000013373 food additive Nutrition 0.000 description 1
- 239000002778 food additive Substances 0.000 description 1
- 235000013569 fruit product Nutrition 0.000 description 1
- SJWWTRQNNRNTPU-ABBNZJFMSA-N fucoxanthin Chemical compound C[C@@]1(O)C[C@@H](OC(=O)C)CC(C)(C)C1=C=C\C(C)=C\C=C\C(\C)=C\C=C\C=C(/C)\C=C\C=C(/C)C(=O)C[C@]1(C(C[C@H](O)C2)(C)C)[C@]2(C)O1 SJWWTRQNNRNTPU-ABBNZJFMSA-N 0.000 description 1
- AQLRNQCFQNNMJA-UHFFFAOYSA-N fucoxanthin Natural products CC(=O)OC1CC(C)(C)C(=C=CC(=CC=CC(=CC=CC=C(/C)C=CC=C(/C)C(=O)CC23OC2(C)CC(O)CC3(C)C)C)CO)C(C)(O)C1 AQLRNQCFQNNMJA-UHFFFAOYSA-N 0.000 description 1
- 235000013376 functional food Nutrition 0.000 description 1
- WIGCFUFOHFEKBI-UHFFFAOYSA-N gamma-tocopherol Natural products CC(C)CCCC(C)CCCC(C)CCCC1CCC2C(C)C(O)C(C)C(C)C2O1 WIGCFUFOHFEKBI-UHFFFAOYSA-N 0.000 description 1
- 231100000722 genetic damage Toxicity 0.000 description 1
- 229940094952 green tea extract Drugs 0.000 description 1
- 235000020688 green tea extract Nutrition 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- OUUQCZGPVNCOIJ-UHFFFAOYSA-N hydroperoxyl Chemical compound O[O] OUUQCZGPVNCOIJ-UHFFFAOYSA-N 0.000 description 1
- TUJKJAMUKRIRHC-UHFFFAOYSA-N hydroxyl Chemical compound [OH] TUJKJAMUKRIRHC-UHFFFAOYSA-N 0.000 description 1
- 208000019423 liver disease Diseases 0.000 description 1
- 230000005976 liver dysfunction Effects 0.000 description 1
- 239000006210 lotion Substances 0.000 description 1
- 235000012661 lycopene Nutrition 0.000 description 1
- 239000001751 lycopene Substances 0.000 description 1
- OAIJSZIZWZSQBC-GYZMGTAESA-N lycopene Chemical compound CC(C)=CCC\C(C)=C\C=C\C(\C)=C\C=C\C(\C)=C\C=C\C=C(/C)\C=C\C=C(/C)\C=C\C=C(/C)CCC=C(C)C OAIJSZIZWZSQBC-GYZMGTAESA-N 0.000 description 1
- 229960004999 lycopene Drugs 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 210000002752 melanocyte Anatomy 0.000 description 1
- 210000002780 melanosome Anatomy 0.000 description 1
- 206010027175 memory impairment Diseases 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- 230000000813 microbial effect Effects 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
- 230000007659 motor function Effects 0.000 description 1
- 230000007886 mutagenicity Effects 0.000 description 1
- 231100000299 mutagenicity Toxicity 0.000 description 1
- 230000035772 mutation Effects 0.000 description 1
- 239000013642 negative control Substances 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 235000014593 oils and fats Nutrition 0.000 description 1
- 230000001590 oxidative effect Effects 0.000 description 1
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N phenol group Chemical group C1(=CC=CC=C1)O ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 1
- 150000002989 phenols Chemical class 0.000 description 1
- 235000021110 pickles Nutrition 0.000 description 1
- 210000004694 pigment cell Anatomy 0.000 description 1
- 230000019612 pigmentation Effects 0.000 description 1
- 238000006116 polymerization reaction Methods 0.000 description 1
- 235000013824 polyphenols Nutrition 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- 230000001603 reducing effect Effects 0.000 description 1
- 206010039073 rheumatoid arthritis Diseases 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
- 150000005846 sugar alcohols Chemical class 0.000 description 1
- 239000013589 supplement Substances 0.000 description 1
- 235000013616 tea Nutrition 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 150000003568 thioethers Chemical class 0.000 description 1
- ZCIHMQAPACOQHT-ZGMPDRQDSA-N trans-isorenieratene Natural products CC(=C/C=C/C=C(C)/C=C/C=C(C)/C=C/c1c(C)ccc(C)c1C)C=CC=C(/C)C=Cc2c(C)ccc(C)c2C ZCIHMQAPACOQHT-ZGMPDRQDSA-N 0.000 description 1
- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 0.000 description 1
- 230000007371 visceral function Effects 0.000 description 1
- 235000019154 vitamin C Nutrition 0.000 description 1
- 239000011718 vitamin C Substances 0.000 description 1
- 235000019165 vitamin E Nutrition 0.000 description 1
- 239000011709 vitamin E Substances 0.000 description 1
- 229940046009 vitamin E Drugs 0.000 description 1
- 230000037303 wrinkles Effects 0.000 description 1
- OENHQHLEOONYIE-JLTXGRSLSA-N β-Carotene Chemical compound CC=1CCCC(C)(C)C=1\C=C\C(\C)=C\C=C\C(\C)=C\C=C\C=C(/C)\C=C\C=C(/C)\C=C\C1=C(C)CCCC1(C)C OENHQHLEOONYIE-JLTXGRSLSA-N 0.000 description 1
Landscapes
- Food Preservation Except Freezing, Refrigeration, And Drying (AREA)
- Non-Alcoholic Beverages (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Medicines Containing Plant Substances (AREA)
- Anti-Oxidant Or Stabilizer Compositions (AREA)
- Fodder In General (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Cosmetics (AREA)
Abstract
Description
化粧料、飲料、食品又は飼料等の原料として、あるいは、これら製品又は原料に添加することにより、メラニン生成抑制及びそれらの酸化による品質低下防止や、それらを利用する人又は動物の生体内に発生する活性酸素やフリーラジカルの発生を消去もしくは抑制する天然物由来の抗酸化能を有する微生物培養物であって、緑茶、黒茶及びヨモギ抽出液を主成分とする培地で培養することにより得られる微生物培養物に関する。さらに、本発明はそれらを用いた化粧料、飲料、食品又は飼料に関する。 As a raw material for cosmetics, beverages, foods, feeds, etc., or by adding to these products or raw materials, it suppresses melanin production and prevents quality deterioration due to their oxidation, and occurs in the living body of humans or animals using them. Microbial culture having antioxidative ability derived from natural products that eliminates or suppresses generation of active oxygen and free radicals, and is obtained by culturing in a medium mainly composed of green tea, black tea and mugwort extract Relates to microbial cultures. Furthermore, this invention relates to the cosmetics, drink, foodstuff, or feed using them.
日焼け等による肌の色黒化、シミ、ソバカス等は、黒褐色無定形の色素であるメラニンの生成によるものと考えられている。そこで、化粧品には、メラニン生成抑制物質が添加されていることが多い。 Skin darkening due to sunburn or the like, spots, freckles, etc. are thought to be due to the production of melanin, an amorphous pigment of blackish brown color. Therefore, melanin production-suppressing substances are often added to cosmetics.
このような目的のために、コウジ酸及びその誘導体、アスコルビン酸及びその誘導体、ハイドロキノン及びその配糖体等、動植物からの抽出物を成分とするメラニン抑制物質が知られている。 For such purposes, melanin-inhibiting substances containing extracts from animals and plants such as kojic acid and its derivatives, ascorbic acid and its derivatives, hydroquinone and its glycosides are known.
また、昨今、悪性新生物（がん等）、脳血管疾患や糖尿病などの生活習慣病が死亡原因の上位を占めている。その生活習慣病を引き起こす大きな原因として、活性酸素やフリーラジカルがある。抗酸化物質には、化粧料、飲料、食品又は飼料などに添加し、品質低下の防止及び品質保持の目的で使用されるものと、生体内に発生する活性酸素やフリーラジカルの発生を消去もしくは抑制する目的で使用されるものとがある。 In addition, malignant neoplasms (cancer, etc.), lifestyle-related diseases such as cerebrovascular diseases and diabetes are the top causes of death. There are active oxygen and free radicals as major causes of lifestyle-related diseases. Antioxidants can be added to cosmetics, beverages, foods, feeds, etc. and used for the purpose of preventing quality deterioration and maintaining quality, and eliminate the generation of active oxygen and free radicals generated in the body. Some are used for control purposes.
化粧料、飲料、食品又は飼料などに含まれている油脂類が酸化されると、製品の色や風味が悪くなるばかりでなく、褐変や退色、栄養価の低下の原因になる。そこで、化粧料、飲料、食品又は飼料には、品質保持のために抗酸化物質が添加されていることが多い。 Oxidation of fats and oils contained in cosmetics, beverages, foods, feeds, etc. not only deteriorates the color and flavor of the product, but also causes browning, fading and a decrease in nutritional value. Therefore, antioxidants are often added to cosmetics, beverages, foods or feeds to maintain quality.
このような目的のために、飲料、食品又は飼料などの分野で使用されている抗酸化物質としては、例えば、Ｌ−アスコルビン酸（ビタミンＣ、Ｖ．Ｃ）、エリソルビン酸（イソアスコルビン酸）、トコフェロール（ビタミンＥ、Ｖ．Ｅ）などの天然抗酸化物質やジブチルヒドロキシトルエン（ＢＨＴ）、ブチルヒドロキシアニソール（ＢＨＡ）などのフェノール系合成抗酸化物質が知られている。 For such purposes, as antioxidants used in the field of beverages, foods or feeds, for example, L-ascorbic acid (vitamin C, VC), erythorbic acid (isoascorbic acid), Natural antioxidants such as tocopherol (vitamin E, VE) and phenolic synthetic antioxidants such as dibutylhydroxytoluene (BHT) and butylhydroxyanisole (BHA) are known.
従来、皮膚美白効果を有する化粧料（化粧水）としては、コウジ酸及びその誘導体、アスコルビン酸及びその誘導体、ハイドロキノン及びその配糖体等、動植物からの抽出物を成分とするものであったが、人体への悪影響や肌荒れ等の心配があった。また、充分に美白効果が得られるものではなかった。 Conventionally, cosmetics (skin lotions) having a skin-whitening effect are composed of extracts from animals and plants such as kojic acid and derivatives thereof, ascorbic acid and derivatives thereof, hydroquinone and glycosides thereof, and the like. There were concerns about adverse effects on the human body and rough skin. Further, a sufficient whitening effect was not obtained.
一方、日焼け等による肌の色黒化、シミ、ソバカス等は、黒褐色無定形の色素であるメラニンの生成によるものと考えられている。
このメラニンは表皮基底層に存在するメラノサイトと呼ばれる色素細胞内のメラノソームにおいて、チロシナーゼによってチロシンから各種メラニン中間代謝産物を経て、酸化重合して生成されると考えられている。
そして、紫外線等により生成された活性酸素がこの反応系を活性化させるためにメラニンが過剰に生成され、皮膚への色素沈着を促進していると考えられている。
On the other hand, skin darkening due to sunburn, stains, freckles, etc. are thought to be due to the production of melanin, a black-brown amorphous pigment.
This melanin is considered to be produced by oxidative polymerization in melanosomes in pigment cells called melanocytes existing in the basal layer of the epidermis by various melanin intermediate metabolites from tyrosine by tyrosinase.
And it is thought that the active oxygen produced | generated by the ultraviolet-ray etc. activates this reaction system, melanin is produced | generated excessively, and the pigmentation to the skin is accelerated | stimulated.
抗酸化物質のＬ−アスコルビン酸は、水に溶けやすく酸性で強い還元作用があるので良く利用されているが、油脂へは使用しにくく酸性（酸味）が食品の味をかえてしまうことから、果実加工品、漬物などに限られる。また、トコフェロールは油脂成分の不必要な酸化を防ぐのでこれも良く用いられているが、抗酸化能があまり高くないため他の抗酸化物質と併用する必要がある。 The antioxidant substance L-ascorbic acid is often used because it is easily soluble in water and has an acidic and strong reducing action, but it is difficult to use for oils and fats, so acidity changes the taste of food. Limited to processed fruit products and pickles. In addition, tocopherol is also often used because it prevents unnecessary oxidation of fat and oil components, but it has to be used in combination with other antioxidants because its antioxidant ability is not so high.
一方、天然抗酸化物質は合成抗酸化物質と比較してその抗酸化能が劣るという欠点がある。しかし、合成抗酸化物質のジブチルヒドロキシトルエン（ＢＨＴ）やブチルヒドロキシアニソールは、抗酸化能が強いが遺伝子損傷、変異原性、染色体異常などによる発がん性を誘発する疑わしい食品添加剤とされ、安全性が疑問視されるようになってきた。 On the other hand, natural antioxidants have the disadvantage that their antioxidant ability is inferior to synthetic antioxidants. However, synthetic antioxidants such as dibutylhydroxytoluene (BHT) and butylhydroxyanisole, which have strong antioxidant ability, are regarded as suspicious food additives that induce carcinogenicity due to genetic damage, mutagenicity, chromosomal abnormalities, etc. Has come to be questioned.
化粧品としても、油脂類の酸化防止の為に天然抗酸化物質や合成抗酸化物質が用いられているが、皮膚への影響や肌荒れなどの原因とされ、安全性が疑問視されるようになってきた。
このようなことから、抗酸化能が強く、しかも、汎用性が高く、さらに、天然物由来の安全性の高い抗酸化物質の開発が望まれていた。
As cosmetics, natural antioxidants and synthetic antioxidants are used to prevent the oxidation of fats and oils, but it is considered to be a cause of skin effects and rough skin, and safety is questioned. I came.
For these reasons, it has been desired to develop an antioxidant having strong antioxidant ability, high versatility, and high safety derived from natural products.
天然の抗酸化物質としては、褐変物質、アミノ酸、糖アルコール、果糖、果皮、わさびやカテキンなどが報告されている。また、微生物培養物の抗酸化物質としては、乳酸菌の抽出物に抗酸化効果があることが示されてきている。 As natural antioxidants, browning substances, amino acids, sugar alcohols, fructose, pericarp, wasabi and catechins have been reported. Moreover, it has been shown that the extract of lactic acid bacteria has an antioxidant effect as an antioxidant substance of a microorganism culture.
これらの抗酸化物質は、従来、化粧料、飲料、食品又は飼料などに添加し、品質低下の防止及び品質保持の目的で使用されてきたが、生体内に発生する活性酸素やフリーラジカルを消去もしくは抑制することが明らかになり、生体内抗酸化能が強い天然抽出物の探索が行われるようになってきた。 These antioxidants have been added to cosmetics, beverages, foods, feeds, etc., and have been used for the purpose of preventing quality degradation and maintaining quality, but they eliminate active oxygen and free radicals generated in the living body. Or it became clear that it suppresses, and the search of the natural extract with strong antioxidative ability in a living body has come to be performed.
生体内酸化により発生する活性酸素には、一重項酸素、スーパーオキシド、ヒドロキシルラジカル、過酸化水素の４種、また、フリーラジカルには、アルキルヒドロペルオキシド、ヒドロペルオキシルラジカルなど１０種以上が知られている。抗酸化能は、一般的に１,１−ジフェニル−２−ピクリルヒドラジル（ＤＰＰＨ）を用いたラジカル消去活性を測定することで判定できる。 There are four types of active oxygen generated by in-vivo oxidation: singlet oxygen, superoxide, hydroxyl radical, and hydrogen peroxide, and more than 10 types of free radicals such as alkyl hydroperoxide and hydroperoxyl radical. ing. Antioxidant ability can generally be determined by measuring radical scavenging activity using 1,1-diphenyl-2-picrylhydrazyl (DPPH).
生体内酸化により生体への影響として、老化、生活習慣病及びがんが引き起こされると考えられている。老化は、運動機能や内臓の機能の低下、物忘れ、眼の白内障、皮膚のしわなど、病気とまではいかない衰えのほか、血管が衰えて動脈硬化などにかかりやすくなる。生活習慣病としては、特に関係が深いと考えられているのが動脈硬化と糖尿病で、他にも肝機能障害、リウマチ性関節炎など挙げられる。 It is thought that aging, lifestyle-related diseases and cancer are caused by in vivo oxidation as an effect on the living body. Aging is not limited to illness such as decreased motor function and visceral function, forgetfulness, eye cataracts, skin wrinkles, etc., and blood vessels tend to suffer from arteriosclerosis. As lifestyle-related diseases, arteriosclerosis and diabetes are considered to be particularly closely related, and other examples include liver dysfunction and rheumatoid arthritis.
また、がんについては、酸化によってＤＮＡを構成する物質を変質させて傷をつけ、別の細胞が再生される際に突然変異を起こす原因の一つとされている。このような疾患を防止する上で、生体内酸化を防止し、あるいは、抑制する抗酸化物質が注目され、機能性食品、特定保健用食品やサプリメントして研究開発されている。 In addition, cancer is considered to be one of the causes of mutation when a substance constituting DNA is altered and damaged by oxidation and another cell is regenerated. In order to prevent such diseases, antioxidant substances that prevent or suppress in-vivo oxidation have attracted attention and have been researched and developed as functional foods, foods for specified health use and supplements.
このような用途の抗酸化物質として、カロチノイド類（ベータカロチン、リコペン、フコキサンチン）、フェノール化合物類（フラボノイド、アントシアニン、カテキン）、含硫化物やベータ・ジケトン類などの植物由来、微生物培養物ではＤＨＡ（ドコサヘキサエン酸）、ＥＰＡ（エイコサペンタエン酸）などがある。 Antioxidants for such applications include carotenoids (beta-carotene, lycopene, fucoxanthin), phenolic compounds (flavonoids, anthocyanins, catechins), plant-derived sulfides and beta-diketones, and microbial cultures. Examples include DHA (docosahexaenoic acid) and EPA (eicosapentaenoic acid).
本発明の第一の目的は、このような天然抽出物であって安全性が高く、抗酸化能が強いメラニン生成抑制効果に優れ、皮膚美白効果が高い化粧料の提供及び様々な飲料、食品又は飼料など、汎用性が高く、製品本来の味や風味が維持でき、また、生体内抗酸化能にも効果のある緑茶、黒茶及びヨモギ抽出液を主成分とする培地で培養することにより得られる微生物培養物を提供することにある。また、本発明の第二の目的は、それらを用いた様々な化粧料、飲料、食品又は飼料を合わせて提供することにある。 The first object of the present invention is such a natural extract that is highly safe, has a strong antioxidant ability, is excellent in melanin production inhibitory effect, has a high skin whitening effect, and provides various cosmetics and various beverages and foods. Or by culturing in a medium mainly composed of green tea, black tea and mugwort extract, which is highly versatile, can maintain the original taste and flavor of the product, and is also effective for in vivo antioxidant capacity The object is to provide the resulting microbial culture. The second object of the present invention is to provide various cosmetics, beverages, foods or feeds using them.
コーカサス地方が発祥の地と考えられている伝統的な乳酸菌と酵母の複合醗酵乳ケフィア（Ｋｅｆｉｒ）は、そのスターターであるケフィア粒を牛乳にて培養することにより製造される。
本発明者は、天然物であるこのケフィア粒を化粧料、飲料、食品又は飼料などに利用した場合、人、動物にとって安全であることを見出し、さらに、様々な利用方法について鋭意研究を重ねてきた。そして、ケフィア粒を緑茶のエキスにて培養し、その微生物培養物から菌体を除去した培養液が優れた皮膚美白効果を有することを見出し、また、ケフィア粒に抗酸化能を有することを見出し、その培養液を化粧料として用いることにした（特許文献１参照）。
The present inventor has found that when this kefir grain, which is a natural product, is used in cosmetics, beverages, foods, feeds, etc., it has been found that it is safe for humans and animals. It was. Then, the kefir grains are cultured with green tea extract, and the culture solution obtained by removing the cells from the microorganism culture is found to have an excellent skin whitening effect, and the kefir grains are found to have an antioxidant ability. The culture solution was used as a cosmetic (see Patent Document 1).
また、ケフィア粒から分離された微生物とこの微生物あるいはそれを含む微生物群を培養することにより得られる微生物培養物ならびにそれらを用いた製品に関しても、すでに出願を済ませている。（特願２００４−１７５９６９、出願日：平成１６年６月１４日）。
その他、ケフィア粒から分離された微生物とこの微生物あるいはそれを含む微生物群を、ヨモギ抽出液を主成分とする培地で培養することにより得られる微生物培養物ならびにそれらを用いた製品に関しても、すでに出願を済ませている。（特願２００４−３１１９８３、出願日：平成１６年１０月２７日）。
An application has already been filed for a microorganism isolated from kefir grains, a microorganism culture obtained by culturing this microorganism or a group of microorganisms containing the microorganism, and a product using them. (Japanese Patent Application No. 2004-175969, filing date: June 14, 2004).
In addition, we have already filed applications for microorganism cultures obtained by culturing microorganisms isolated from kefir grains and microorganisms containing these microorganisms or microorganisms containing these microorganisms in a medium mainly composed of mugwort extract and products using them. Have finished. (Japanese Patent Application No. 2004-311983, filing date: October 27, 2004).
そして、さらに研究を重ねた結果、ケフィア粒から分離された微生物とこの微生物あるいはそれを含む微生物群を、緑茶、黒茶及びヨモギ抽出液を主成分とする培地で培養することにより得られる微生物培養物を、飲料、食品、化粧品又は飼料等の原料として、あるいは、これら製品又は原料に添加することにより、メラニン生成抑制効果及びそれらの酸化による品質低下防止や、それらを利用する人又は動物の生体内に発生する活性酸素やフリーラジカルの発生を消去もしくは抑制することを見出した。そして、この天然物由来の抗酸化能を有する微生物培養物であって、緑茶、黒茶及びヨモギ抽出液を主成分とする培地で培養することにより得られる微生物培養物を用いることにより、上記の目的を達成し得ることを見出した。さらに、本発明者はそれらを化粧料、飲料、食品又は飼料に添加することによって、メラニン生成抑制効果、抗酸化能を有する製品を開発するに至ったものである。 As a result of further research, the microorganism culture obtained by culturing the microorganism isolated from kefir grains and the microorganism or a group of microorganisms containing the microorganism in a medium mainly composed of green tea, black tea and mugwort extract. By adding the product as a raw material for beverages, foods, cosmetics, feeds, etc., or by adding to these products or raw materials, the melanin production inhibitory effect and the quality degradation due to their oxidation can be prevented. It has been found that the generation of active oxygen and free radicals generated in the body is eliminated or suppressed. And, by using a microorganism culture having an antioxidant ability derived from this natural product, the microorganism culture obtained by culturing in a medium mainly composed of green tea, black tea and mugwort extract, We have found that the objective can be achieved. Furthermore, this inventor came to develop the product which has a melanin production inhibitory effect and antioxidant ability by adding them to cosmetics, a drink, food, or feed.
本発明は、以下の構成からなる発明である。すなわち、
(1) ケフィア（Ｋｅｆｉｒ）粒から分離された抗酸化能を有するラクトバシラシー（Lactobacillaceae）に属する新種の微生物とこの微生物あるいはそれを含む微生物群を、緑茶、黒茶及びヨモギ抽出液を主成分とする培地で培養することにより得られる微生物培養物であって、メラニン生成抑制効果を有する微生物培養物に関するものである。
(2) ケフィア（Ｋｅｆｉｒ）粒から分離されたラクトバシラシー（Lactobacillaceae）に属する新種の微生物とこの微生物あるいはそれを含む微生物群を、緑茶、黒茶及びヨモギ抽出液を主成分とする培地で培養することにより得られる微生物培養物であって、抗酸化能を有する微生物培養物に関するものである。
(3) ケフィア（Ｋｅｆｉｒ）粒から分離されたラクトバシラシー（Lactobacillaceae）に属する新種の微生物とこの微生物あるいはそれを含む微生物群を、緑茶、黒茶及びヨモギ抽出液を主成分とする培地で培養することにより得られる微生物培養物を添加した、メラニン生成抑制効果及び抗酸化作用を有する化粧料に関するものである。
(4) ケフィア（Ｋｅｆｉｒ）粒から分離されたラクトバシラシー（Lactobacillaceae）に属する新種の微生物とこの微生物あるいはそれを含む微生物群を、緑茶、黒茶及びヨモギ抽出液を主成分とする培地で培養することにより得られる微生物培養物を添加した、抗酸化作用を有する飲料に関するものである。
(5) ケフィア（Ｋｅｆｉｒ）粒から分離されたラクトバシラシー（Lactobacillaceae）に属する新種の微生物とこの微生物あるいはそれを含む微生物群を、緑茶、黒茶及びヨモギ抽出液を主成分とする培地で培養することにより得られる微生物培養物を添加した、抗酸化作用を有する食品に関するものである。
(6) ケフィア（Ｋｅｆｉｒ）粒から分離されたラクトバシラシー（Lactobacillaceae）に属する新種の微生物とこの微生物あるいはそれを含む微生物群を、緑茶、黒茶及びヨモギ抽出液を主成分とする培地で培養することにより得られる微生物培養物を添加した、抗酸化作用を有する飼料に関するものである。
The present invention is an invention having the following configuration. That is,
(1) A new kind of microorganism belonging to Lactobacillaceae with anti-oxidant ability isolated from kefir grains, and this microorganism or a group of microorganisms containing it, mainly consisting of green tea, black tea and mugwort extract The present invention relates to a microbial culture obtained by culturing in a culture medium having a melanin production inhibitory effect.
(2) Culture of a new microorganism belonging to Lactobacillaceae isolated from kefir grains and this microorganism or a group of microorganisms containing this microorganism in a medium mainly composed of green tea, black tea and mugwort extract The present invention relates to a microbial culture obtained by carrying out antimicrobial activity.
(3) A new type of microorganism belonging to Lactobacillaceae isolated from kefir grains and this microorganism or a group of microorganisms containing it are cultured in a medium mainly composed of green tea, black tea and mugwort extract. It is related with the cosmetics which have the melanin production inhibitory effect and antioxidant activity which added the microbial culture obtained by doing.
(4) Culture of a new species of microorganism belonging to Lactobacillaceae isolated from kefir grains and this microorganism or a group of microorganisms containing the same in a medium mainly composed of green tea, black tea and mugwort extract The present invention relates to a beverage having an antioxidative effect, to which a microbial culture obtained by doing so is added.
(5) A new type of microorganism belonging to Lactobacillaceae isolated from kefir (Kefir) grains and this microorganism or a group of microorganisms containing it are cultured in a medium mainly composed of green tea, black tea and mugwort extract It is related with the foodstuff which has the anti-oxidation effect which added the microorganism culture obtained by doing.
(6) Culture of a new microorganism belonging to Lactobacillaceae isolated from kefir grains and this microorganism or a group of microorganisms containing this microorganism in a medium mainly composed of green tea, black tea and mugwort extract It is related with the feed which has the antioxidant effect | action which added the microorganism culture obtained by doing.
ここに、「メラニン生成抑制効果」とは、紫外線や老化により皮膚に生じるシミ・ソバカスの原因物質であるメラニン色素の生成を抑制する効果をいう。 Here, the “melanin production inhibitory effect” refers to an effect of inhibiting the production of melanin pigment, which is a causative substance of spots and freckles generated in the skin by ultraviolet rays and aging.
ここに、「抗酸化能」とは、製品の品質を保持する抗酸化能力と、飲料、食品又は飼料を人、動物に投与した際に、生体内に発生する活性酸素やフリーラジカルを消去もしくは抑制する生体内酸化防止能力をいう。また、「微生物培養物」とは、ケフィア粒から分離された微生物群又は分離された微生物単独で、緑茶、黒茶及びヨモギ抽出液を主成分とする培地で培養することにより得られる培養物をいう。 Here, the “antioxidant ability” means an antioxidant ability that maintains the quality of the product, and eliminates active oxygen and free radicals generated in the living body when a beverage, food or feed is administered to humans or animals. It refers to the ability to inhibit in vivo oxidation. The “microorganism culture” refers to a culture obtained by culturing a group of microorganisms isolated from kefir grains or isolated microorganisms alone in a medium mainly composed of green tea, black tea and mugwort extract. Say.
請求項１記載の発明によれば、ケフィア（Ｋｅｆｉｒ）粒から分離されたラクトバシラシー（Lactobacillaceae）に属する新種の微生物とこの微生物あるいはそれを含む微生物群を、緑茶、黒茶及びヨモギ抽出液を主成分とする培地で培養することによって得られる微生物培養物であって、メラニン生成抑制効果を有する微生物培養物を提供できる効果がある。そして、その培養物は毒性や副作用がなく安全であり、化粧料として利用できる。 According to the first aspect of the present invention, a new type of microorganism belonging to Lactobacillaceae isolated from kefir grains and this microorganism or a group of microorganisms containing the microorganism are mixed with green tea, black tea and mugwort extract. It is a microorganism culture obtained by culturing in a medium containing the main component, and has an effect of providing a microorganism culture having a melanin production inhibitory effect. The culture is safe without toxicity and side effects, and can be used as a cosmetic.
請求項２記載の発明によれば、ケフィア（Ｋｅｆｉｒ）粒から分離されたラクトバシラシー（Lactobacillaceae）に属する新種の微生物とこの微生物あるいはそれを含む微生物群を、緑茶、黒茶及びヨモギ抽出液を主成分とする培地で培養することにより得られる微生物培養物であって、抗酸化能を有する微生物培養物を提供できる効果がある。そして、その培養物は毒性や副作用がなく安全であり、化粧料、飲料、食品又は飼料等の原料、添加物として様々な分野で利用できる。 According to the invention described in claim 2, a new type of microorganism belonging to Lactobacillaceae isolated from kefir grains and the microorganism or a group of microorganisms containing the microorganism, green tea, black tea and mugwort extract are added. It is a microorganism culture obtained by culturing in a medium having a main component, and has an effect of providing a microorganism culture having antioxidant ability. The culture is safe without toxicity and side effects, and can be used in various fields as a raw material or additive for cosmetics, beverages, foods or feeds.
請求項３記載の発明によれば、ケフィア（Ｋｅｆｉｒ）粒から分離されたラクトバシラシー（Lactobacillaceae）に属する新種の微生物とこの微生物あるいはそれを含む微生物群を、緑茶、黒茶及びヨモギ抽出液を主成分とする培地で培養することにより得られる微生物培養物を添加した、メラニン生成抑制効果及び抗酸化作用を有する化粧料を提供できる効果がある。 According to the invention described in claim 3, a new kind of microorganism belonging to Lactobacillaceae isolated from kefir grains and the microorganism or a group of microorganisms containing the microorganism are mixed with green tea, black tea and mugwort extract. There is an effect capable of providing a cosmetic material having an inhibitory effect on melanin production and an antioxidative effect, to which a microbial culture obtained by culturing in a medium containing the main component is added.
請求項４〜６のいずれかに記載の発明によれば、ケフィア（Ｋｅｆｉｒ）粒から分離されたラクトバシラシー（Lactobacillaceae）に属する新種の微生物とこの微生物あるいはそれを含む微生物群を、緑茶、黒茶及びヨモギ抽出液を主成分とする培地で培養することにより得られる微生物培養物を添加した、抗酸化作用を有する飲料、食品又は飼料を提供できる効果がある。 According to the invention described in any one of claims 4 to 6, a new kind of microorganism belonging to Lactobacillaceae isolated from kefir grains and the microorganism or a group of microorganisms containing the microorganism are mixed with green tea, black There is an effect of providing a beverage, food or feed having an antioxidant action, to which a microbial culture obtained by culturing in a medium mainly composed of tea and mugwort extract is added.
本発明の好ましい実施の形態について、詳細に説明する。
ケフィア粒から分離された微生物群には、菌株アセトバクター セレヴィシアエ（Acetobacter cerevisiae）ＳＩＩＤ１７１９−２ｂ、菌株グルコノバクター オキシダンス（Gluconobacter oxydans）ＳＩＩＤ１７１９−３ｂ、菌株ラクトバシラシー（Lactobacillaceae）科の新種であるラクトバシラス属（Lactobaccilus sp.）ＳＩＩＤ１７１９−６ｂ、菌株ピヒアメンブラニ ファシエンス（Pichiamembrani faciens）ＳＩＩＤ１７１９−１ｙ、菌株サッカロマイセス セレヴィシアエ（Saccharomyces cerevisiae）ＳＩＩＤ１７１９−４ｙ及び菌株ピヒアアノマラ（Pichia anomala）ＳＩＩＤ１７１９−５ｙが存在している。
A preferred embodiment of the present invention will be described in detail.
Microorganisms isolated from kefir grains include strain Acetobacter cerevisiae SIID1719-2b, strain Gluconobacter oxydans SIID1719-3b, and a new species of the family Lactobacillaceae. The genus (Lactobaccilus sp.) SIID1719-6b, the strain Pichiamembrani faciens SIID1719-1y, the strain Saccharomyces cerevisiae SIID1719-4y, and the strain Pichia anomala SID1719-1 are present.
ケフィア粒から分離された微生物群及び菌株ラクトバシラシー（Lactobacillaceae）科の新種であるラクトバシラス属（Lactobacillus sp.）ＳＩＩＤ１７１９−６ｂを、緑茶、黒茶及びヨモギ抽出液を主成分とする培地で単独培養することにより得られた微生物培養物が、メラニン生成抑制効果及び抗酸化能を有するか、以下菌株の評価試験を行った。その結果、微生物群及びラクトバシラス属（Lactobacillus sp.）ＳＩＩＤ１７１９−６ｂ単独でメラニン生成抑制効果及び抗酸化能を有していた。さらに、本発明では、ラクトバシラス属（Lactobacillus sp.）ＳＩＩＤ１７１９−６ｂと安全性の確認できている微生物を共生し、同様の作用を示すものであれば、いずれも使用できる。
なお、ラクトバシラシー（Lactobacillaceae）科の新種であるラクトバシラス属（Lactobacillus sp.）ＳＩＩＤ１７１９−６ｂ（ＦＥＲＭ ＡＰ−２００６２）は、すでに独立行政法人産業技術総合研究所特許生物寄託センターに寄託済みである。
Lactobacillus sp. SIID1719-6b, a new species of the microbial community isolated from kefir grains and the strain Lactobacillaceae, is cultured alone in a medium mainly composed of green tea, black tea and mugwort extract Whether the microbial culture obtained by doing so has a melanin production inhibitory effect and an antioxidant ability was tested for the following strains. As a result, the microorganism group and Lactobacillus sp. SIID1719-6b alone had a melanin production inhibitory effect and an antioxidant ability. Furthermore, in the present invention, any microorganism may be used as long as it coexists with Lactobacillus sp. SIID1719-6b and a microorganism whose safety has been confirmed and exhibits a similar action.
In addition, Lactobacillus sp. SIID1719-6b (FERM AP-20062), a new species of the Lactobacillaceae family, has already been deposited at the National Institute of Advanced Industrial Science and Technology Patent Organism Depositary.
代表例として、菌株ラクトバシラシー（Lactobacillaceae）科の新種であるラクトバシラス属（Lactobacillus sp.）ＳＩＩＤ１７１９−６ｂの緑茶、黒茶及びヨモギ抽出液を主成分とする培地での培養及び、メラニン生成抑制効果及び抗酸化評価試験方法を次のように実施した。 As a representative example, cultivation of Lactobacillus sp. SIID1719-6b, a new species of the strain Lactobacillaceae, in a medium mainly composed of green tea, black tea, and mugwort extract, and melanin production inhibitory effect And the antioxidant evaluation test method was implemented as follows.
（メラニン生成抑制効果試験）
先ず、Ｂ１６ｍｏｕｓｅ ｍｅｌａｎｏｍａ ｃｅｌｌを２．０×１０5ｃｅｌｌ／ｗｅｌｌとなるように細胞培養用ｗｅｌｌに添加し、ＲＰＭＩ１６４０培地で培養した。２４時間後にＳＩＩＤ１７１９−６ｂを緑茶、黒茶、ヨモギ抽出液が６．６７：１．６６５：１．６６５の割合で調製した培地で４８時間培養した培養物（Ｂ３溶液）を、Ｂ１６ｍｏｕｓｅ ｍｅｌａｎｏｍａ ｃｅｌｌが良好な生育の確認された終濃度を全培養液の１／５０となるように添加した。また、ポジティブコントロールとして、代表的なメラニン抑制物質であるコウジ酸を終濃度１００μｇ／ｍｌとなるように、ネガティブコントロールとして、蒸留水を１０倍希釈となるように添加し、３日間培養した。次に、１０％ＤＭＳＯ−１ＮＮａＯＨを用いて細胞からメラニンを溶かし出し、４７０ｎｍの吸光度を測定することでメラニンを定量した。各サンプルのメラニン生成抑制効果は、１／５０Ｂ３溶液、コウジ酸で培養した細胞のメラニン量（Ｓ）と、蒸留水（Ｃ）で得られた細胞のメラニン量を比較することで求めた（次式(1)参照）。その結果を、図１のグラフにメラニン生成抑制効果として示す。
メラニン生成抑制効果で知られているコウジ酸よりも、Ｂ３溶液の全培養液の終濃度を１／５０にした１／５０Ｂ３溶液培地の方が、メラニン生成抑制効果が高まることが確認された。
(Melanin production inhibitory effect test)
First, B16mouse melanoma cell was added to a cell culture well so as to be 2.0 × 10 5 cells / well and cultured in RPMI1640 medium. After 24 hours, a culture (B3 solution) obtained by cultivating SIID1719-6b with green tea, black tea, mugwort extract at a ratio of 6.67: 1.665: 1.665 for 48 hours was used as B16mouse melanoma cell. The final concentration confirmed to have good growth was added to 1/50 of the total culture solution. As a positive control, kojic acid, which is a typical melanin inhibitor, was added to a final concentration of 100 μg / ml, and distilled water was added to a 10-fold dilution as a negative control, and cultured for 3 days. Next, melanin was quantified by dissolving melanin from the cells using 10% DMSO-1N NaOH and measuring the absorbance at 470 nm. The melanin production inhibitory effect of each sample was determined by comparing the amount of melanin (S) of cells cultured with 1 / 50B3 solution and kojic acid and the amount of melanin of cells obtained with distilled water (C) (next) (See Equation (1)). The result is shown as a melanin production inhibitory effect in the graph of FIG.
It was confirmed that the 1/50 B3 solution medium in which the final concentration of the whole culture solution of the B3 solution was 1/50 was more effective than the kojic acid known for the melanin production suppressing effect.
（抗酸化能試験）
抗酸化能試験は次のように実施した。
菌株ラクトバシラシー（Lactobacillaceae）科の新種であるラクトバシラス属（Lactobacillus sp.）ＳＩＩＤ１７１９−６ｂを緑茶、黒茶、ヨモギ抽出液が６．６７：１．６６５：１．６６５の割合で調製した培地で４８時間培養し、この培養物（Ｂ３溶液）とコントロールとしてＳＩＩＤ１７１９−６ｂを添加していない培地のみ（Ｂ３培地）を終濃度０．２％となるように添加し、ＤＰＰＨラジカル消去率を確認した。Ｂ３溶液の抗酸化能を、ブランクとして蒸留水を用いて比較した。
(Antioxidant ability test)
Antioxidant ability test was carried out as follows.
Lactobacillus sp. SIID1719-6b, a new species of the family Lactobacillaceae, is prepared with a green tea, black tea, mugwort extract at a ratio of 6.67: 1.665: 1.665. After culturing for 48 hours, this culture (B3 solution) and only a medium without addition of SIID1719-6b (B3 medium) as a control were added to a final concentration of 0.2%, and the DPPH radical scavenging rate was confirmed. . The antioxidant capacity of the B3 solution was compared using distilled water as a blank.
先ず、９６ｗｅｌｌマイクロプレートの各ｗｅｌｌに、１９９．６μｌの０．１ｍＭ ＤＰＰＨ（５０％エタノールに溶解）を添加する。次に、Ｂ３溶液、Ｂ３培地、ブランクの各０．４μｌをそれぞれｗｅｌｌに添加（終濃度０．２％）して、ラジカル消去反応を開始させた。添加直後（０分）及び３０分後のλ＝５１７ｎｍの吸光度をマイクロプレートリーダーで測定し、Ｂ３溶液、Ｂ３培地とブランクの吸光度より、ラジカル消去率を求めた。その結果を、図２のグラフに抗酸化能として示す。
Ｂ３培地と比較してＢ３溶液の抗酸化能がおおよそ３．２倍高かったことから、菌株を添加することにより高い抗酸化能が得られることが確認できた。
First, 199.6 μl of 0.1 mM DPPH (dissolved in 50% ethanol) is added to each well of a 96-well microplate. Next, 0.4 μl of each of B3 solution, B3 medium, and blank was added to each well (final concentration 0.2%) to initiate radical scavenging reaction. The absorbance at λ = 517 nm immediately after addition (0 minutes) and after 30 minutes was measured with a microplate reader, and the radical scavenging rate was determined from the absorbance of the B3 solution, B3 medium and blank. The results are shown as antioxidant ability in the graph of FIG.
Since the antioxidant capacity of the B3 solution was approximately 3.2 times higher than that of the B3 medium, it was confirmed that a high antioxidant capacity was obtained by adding the strain.
Claims (6)
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2005091624A JP4085141B2 (en) | 2005-03-28 | 2005-03-28 | Microbial culture obtained on medium containing green tea, black tea and mugwort extract |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2005091624A JP4085141B2 (en) | 2005-03-28 | 2005-03-28 | Microbial culture obtained on medium containing green tea, black tea and mugwort extract |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JP2006271220A true JP2006271220A (en) | 2006-10-12 |
| JP4085141B2 JP4085141B2 (en) | 2008-05-14 |
Family
ID=37206566
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2005091624A Active JP4085141B2 (en) | 2005-03-28 | 2005-03-28 | Microbial culture obtained on medium containing green tea, black tea and mugwort extract |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JP4085141B2 (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1787651A1 (en) * | 2005-11-22 | 2007-05-23 | K.K. Yonezawa Biru Shisutemu Sabisu | Bacterial culture having effects of suppressing dermatitis development and accelarating skin wound healing, and product using the same |
| WO2010023716A1 (en) * | 2008-09-01 | 2010-03-04 | 株式会社 カロッツェリアジャパン | Microbial culture having anticancer effect and product using the microbial culture |
| CN102120957A (en) * | 2010-12-07 | 2011-07-13 | 张健 | Preparation device, application and preparation method of complex agent of lactic acid bacteria |
| CN112940529A (en) * | 2021-03-20 | 2021-06-11 | 山西医科大学 | Method for extracting melanin from melanoma cells |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2002330725A (en) * | 2001-03-07 | 2002-11-19 | Ryukyu Bio Resource Kaihatsu:Kk | Fermented food material, method for producing the same, drink or food, and extract |
| JP2003210133A (en) * | 2002-01-18 | 2003-07-29 | Oubiken:Kk | Production and use of fermentation product |
| WO2003072119A1 (en) * | 2002-02-28 | 2003-09-04 | Nihon Kefir Co., Ltd. | External compositions for the skin |
| JP2004149442A (en) * | 2002-10-30 | 2004-05-27 | Yonezawa Biru System Service:Kk | Cosmetic |
-
2005
- 2005-03-28 JP JP2005091624A patent/JP4085141B2/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2002330725A (en) * | 2001-03-07 | 2002-11-19 | Ryukyu Bio Resource Kaihatsu:Kk | Fermented food material, method for producing the same, drink or food, and extract |
| JP2003210133A (en) * | 2002-01-18 | 2003-07-29 | Oubiken:Kk | Production and use of fermentation product |
| WO2003072119A1 (en) * | 2002-02-28 | 2003-09-04 | Nihon Kefir Co., Ltd. | External compositions for the skin |
| JP2004149442A (en) * | 2002-10-30 | 2004-05-27 | Yonezawa Biru System Service:Kk | Cosmetic |
Non-Patent Citations (3)
| Title |
|---|
| 月刊フードケミカル, vol. 6, no. 8, JPN4007009545, 1990, pages 106 - 112, ISSN: 0000857167 * |
| 月刊フードケミカル, vol. 6, no. 8, JPN6007011883, 1990, pages 106 - 112, ISSN: 0000940846 * |
| 月刊フードケミカル, vol. 6, no. 8, JPNX007052343, 1990, pages 106 - 112, ISSN: 0000898805 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1787651A1 (en) * | 2005-11-22 | 2007-05-23 | K.K. Yonezawa Biru Shisutemu Sabisu | Bacterial culture having effects of suppressing dermatitis development and accelarating skin wound healing, and product using the same |
| WO2010023716A1 (en) * | 2008-09-01 | 2010-03-04 | 株式会社 カロッツェリアジャパン | Microbial culture having anticancer effect and product using the microbial culture |
| JP5325223B2 (en) * | 2008-09-01 | 2013-10-23 | 株式会社カロッツェリアジャパン | Microbial culture having anticancer activity and product using the microorganism culture |
| CN102120957A (en) * | 2010-12-07 | 2011-07-13 | 张健 | Preparation device, application and preparation method of complex agent of lactic acid bacteria |
| CN112940529A (en) * | 2021-03-20 | 2021-06-11 | 山西医科大学 | Method for extracting melanin from melanoma cells |
| CN112940529B (en) * | 2021-03-20 | 2022-06-07 | 山西医科大学 | Method for extracting melanin from melanoma cells |
Also Published As
| Publication number | Publication date |
|---|---|
| JP4085141B2 (en) | 2008-05-14 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Lee et al. | Antioxidant properties of three extracts from Pleurotus citrinopileatus | |
| Takyar et al. | Evaluation of antioxidant properties of Chlorella vulgaris and Spirulina platensis and their application in order to extend the shelf life of rainbow trout (Oncorhynchus mykiss) fillets during refrigerated storage | |
| JP4085140B2 (en) | Microbial culture obtained in medium containing green tea and black tea | |
| Wu et al. | Antioxidant properties of Lactobacillus-fermented and non-fermented Graptopetalum paraguayense E. Walther at different stages of maturity | |
| EP1607476B1 (en) | A microorganism separated from Kefir grains, a microorganism culture obtained by culturing said microorganism or microorganisms including it, and a product using such microorganisms or microorganism cultures | |
| JP4085141B2 (en) | Microbial culture obtained on medium containing green tea, black tea and mugwort extract | |
| JP7458660B2 (en) | Profilaggrin mRNA expression promoter, serine palmitoyltransferase mRNA expression promoter, and hyaluronic acid synthase 3 mRNA expression promoter | |
| JP4798973B2 (en) | Antibacterial composition | |
| Manzoor et al. | Banana Peel: Characteristics and consideration of its extract for use in meat products preservation: A review | |
| JP5226297B2 (en) | Method for producing at least one of 8-hydroxyliquiritigenin and 3'-hydroxyisoquiritigenin | |
| JP7227647B2 (en) | Anti-obesity agents, cosmetics, and food and drink | |
| JP2006124290A (en) | Microorganism cultured material obtained by culture medium containing mugwort extract as main ingredient and product using the same | |
| RU2475052C2 (en) | Application of at least one derivative of cholest-4-en-3-one oxime as antioxidant | |
| Chedea et al. | Antibacterial action of an aqueous grape seed polyphenolic extract | |
| Manzoor et al. | Recent progress in natural seaweed pigments: Green extraction, health-promoting activities, techno-functional properties and role in intelligent food packaging | |
| JP5055540B2 (en) | Tyrosinase inhibitor | |
| KR102298782B1 (en) | Polygonum multiflorum Thunberg fermented Extract composition and manufacturing method thereof | |
| Baek et al. | Antioxidant activity of the halophyte Ligustrum japonicum | |
| KR102330009B1 (en) | Lactobacillus paracasei JSKIU 12-9 (KCTC18805P) from fermented local food in Jeju and composition with the function of anti-aging, anti-cancer and anti-dental caries thereof | |
| JP5832714B2 (en) | Tyrosinase activity inhibitor and melanin production inhibitor | |
| KR20200070503A (en) | Polygonum multiflorum Thunberg fermented Extract composition and manufacturing method thereof | |
| JP5422137B2 (en) | Whitening agent, tyrosinase activity inhibitor, melanin production inhibitor, and skin external preparation for whitening | |
| KR101693782B1 (en) | Antibacterial and antioxidant composition comprising sea-urchins roe extract | |
| KR101761607B1 (en) | Antioxidant Composition and Antibacterial Composition Using an Extract of Zonaria diesingiana | |
| JP5499352B2 (en) | Antioxidants, food and drink, cosmetics, quasi drugs, and whitening agents |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20070528 |
|
| A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20070710 |
|
| A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A821 Effective date: 20070710 |
|
| A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20070925 |
|
| A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20071106 |
|
| TRDD | Decision of grant or rejection written | ||
| A01 | Written decision to grant a patent or to grant a registration (utility model) |
Free format text: JAPANESE INTERMEDIATE CODE: A01 Effective date: 20071210 |
|
| A711 | Notification of change in applicant |
Free format text: JAPANESE INTERMEDIATE CODE: A711 Effective date: 20071225 |
|
| A61 | First payment of annual fees (during grant procedure) |
Free format text: JAPANESE INTERMEDIATE CODE: A61 Effective date: 20080108 |
|
| A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A821 Effective date: 20071225 |
|
| FPAY | Renewal fee payment (event date is renewal date of database) |
Free format text: PAYMENT UNTIL: 20110228 Year of fee payment: 3 |
|
| R150 | Certificate of patent or registration of utility model |
Ref document number: 4085141 Country of ref document: JP Free format text: JAPANESE INTERMEDIATE CODE: R150 Free format text: JAPANESE INTERMEDIATE CODE: R150 |
|
| FPAY | Renewal fee payment (event date is renewal date of database) |
Free format text: PAYMENT UNTIL: 20110228 Year of fee payment: 3 |
|
| S111 | Request for change of ownership or part of ownership |
Free format text: JAPANESE INTERMEDIATE CODE: R313117 |
|
| FPAY | Renewal fee payment (event date is renewal date of database) |
Free format text: PAYMENT UNTIL: 20110228 Year of fee payment: 3 |
|
| R360 | Written notification for declining of transfer of rights |
Free format text: JAPANESE INTERMEDIATE CODE: R360 |
|
| FPAY | Renewal fee payment (event date is renewal date of database) |
Free format text: PAYMENT UNTIL: 20110228 Year of fee payment: 3 |
|
| R360 | Written notification for declining of transfer of rights |
Free format text: JAPANESE INTERMEDIATE CODE: R360 |
|
| R371 | Transfer withdrawn |
Free format text: JAPANESE INTERMEDIATE CODE: R371 |
|
| FPAY | Renewal fee payment (event date is renewal date of database) |
Free format text: PAYMENT UNTIL: 20110228 Year of fee payment: 3 |
|
| S111 | Request for change of ownership or part of ownership |
Free format text: JAPANESE INTERMEDIATE CODE: R313113 |
|
| FPAY | Renewal fee payment (event date is renewal date of database) |
Free format text: PAYMENT UNTIL: 20110228 Year of fee payment: 3 |
|
| R350 | Written notification of registration of transfer |
Free format text: JAPANESE INTERMEDIATE CODE: R350 |
|
| FPAY | Renewal fee payment (event date is renewal date of database) |
Free format text: PAYMENT UNTIL: 20120229 Year of fee payment: 4 |
|
| R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
| FPAY | Renewal fee payment (event date is renewal date of database) |
Free format text: PAYMENT UNTIL: 20120229 Year of fee payment: 4 |
|
| FPAY | Renewal fee payment (event date is renewal date of database) |
Free format text: PAYMENT UNTIL: 20130228 Year of fee payment: 5 |
|
| R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
| FPAY | Renewal fee payment (event date is renewal date of database) |
Free format text: PAYMENT UNTIL: 20140228 Year of fee payment: 6 |
|
| R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
| R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
| R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
| R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
| R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
| R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
| R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
| R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
| R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
| R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
| R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
| R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |