JP6785324B2 - Bacillus licheniformis NY1505 strain that mass-produces α-glucosidase inhibitors - Google Patents
Bacillus licheniformis NY1505 strain that mass-produces α-glucosidase inhibitors Download PDFInfo
- Publication number
- JP6785324B2 JP6785324B2 JP2018565720A JP2018565720A JP6785324B2 JP 6785324 B2 JP6785324 B2 JP 6785324B2 JP 2018565720 A JP2018565720 A JP 2018565720A JP 2018565720 A JP2018565720 A JP 2018565720A JP 6785324 B2 JP6785324 B2 JP 6785324B2
- Authority
- JP
- Japan
- Prior art keywords
- strain
- bacillus licheniformis
- spores
- glucosidase
- glucosidase inhibitor
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 229940077274 Alpha glucosidase inhibitor Drugs 0.000 title claims description 48
- 239000003888 alpha glucosidase inhibitor Substances 0.000 title claims description 48
- 241000194108 Bacillus licheniformis Species 0.000 title claims description 39
- 210000000936 intestine Anatomy 0.000 claims description 29
- 235000013305 food Nutrition 0.000 claims description 22
- 239000000203 mixture Substances 0.000 claims description 16
- 244000068988 Glycine max Species 0.000 claims description 8
- 235000010469 Glycine max Nutrition 0.000 claims description 8
- 108020004465 16S ribosomal RNA Proteins 0.000 claims description 4
- 210000004215 spore Anatomy 0.000 description 44
- 108010028144 alpha-Glucosidases Proteins 0.000 description 16
- 102100024295 Maltase-glucoamylase Human genes 0.000 description 15
- 230000000694 effects Effects 0.000 description 15
- 230000002401 inhibitory effect Effects 0.000 description 13
- 235000005911 diet Nutrition 0.000 description 11
- 241000894006 Bacteria Species 0.000 description 10
- 230000037213 diet Effects 0.000 description 10
- 235000009200 high fat diet Nutrition 0.000 description 10
- 238000004519 manufacturing process Methods 0.000 description 10
- 241000699670 Mus sp. Species 0.000 description 9
- 230000037396 body weight Effects 0.000 description 9
- 244000005700 microbiome Species 0.000 description 9
- 239000000047 product Substances 0.000 description 9
- 235000013557 nattō Nutrition 0.000 description 7
- 241000193830 Bacillus <bacterium> Species 0.000 description 6
- 241001465754 Metazoa Species 0.000 description 6
- LXBIFEVIBLOUGU-JGWLITMVSA-N duvoglustat Chemical compound OC[C@H]1NC[C@H](O)[C@@H](O)[C@@H]1O LXBIFEVIBLOUGU-JGWLITMVSA-N 0.000 description 6
- 241000699666 Mus <mouse, genus> Species 0.000 description 5
- 210000003608 fece Anatomy 0.000 description 5
- 239000003112 inhibitor Substances 0.000 description 5
- 230000000968 intestinal effect Effects 0.000 description 5
- 244000063299 Bacillus subtilis Species 0.000 description 4
- 235000014469 Bacillus subtilis Nutrition 0.000 description 4
- 241000196324 Embryophyta Species 0.000 description 4
- 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 4
- 238000004458 analytical method Methods 0.000 description 4
- -1 aza sugars Chemical class 0.000 description 4
- 239000008280 blood Substances 0.000 description 4
- 210000004369 blood Anatomy 0.000 description 4
- 210000004027 cell Anatomy 0.000 description 4
- 239000008103 glucose Substances 0.000 description 4
- 239000002609 medium Substances 0.000 description 4
- 239000000243 solution Substances 0.000 description 4
- 235000019786 weight gain Nutrition 0.000 description 4
- 230000004580 weight loss Effects 0.000 description 4
- LXBIFEVIBLOUGU-UHFFFAOYSA-N Deoxymannojirimycin Natural products OCC1NCC(O)C(O)C1O LXBIFEVIBLOUGU-UHFFFAOYSA-N 0.000 description 3
- 102000004366 Glucosidases Human genes 0.000 description 3
- 108010056771 Glucosidases Proteins 0.000 description 3
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 3
- 230000008901 benefit Effects 0.000 description 3
- 150000001720 carbohydrates Chemical class 0.000 description 3
- 235000014633 carbohydrates Nutrition 0.000 description 3
- 238000000855 fermentation Methods 0.000 description 3
- 230000004151 fermentation Effects 0.000 description 3
- 210000001035 gastrointestinal tract Anatomy 0.000 description 3
- 230000012010 growth Effects 0.000 description 3
- 238000005259 measurement Methods 0.000 description 3
- 235000019645 odor Nutrition 0.000 description 3
- 238000012545 processing Methods 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 235000000346 sugar Nutrition 0.000 description 3
- 230000004584 weight gain Effects 0.000 description 3
- 102000004190 Enzymes Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- 208000013016 Hypoglycemia Diseases 0.000 description 2
- SIKJAQJRHWYJAI-UHFFFAOYSA-N Indole Chemical compound C1=CC=C2NC=CC2=C1 SIKJAQJRHWYJAI-UHFFFAOYSA-N 0.000 description 2
- 240000000249 Morus alba Species 0.000 description 2
- 235000008708 Morus alba Nutrition 0.000 description 2
- 208000008589 Obesity Diseases 0.000 description 2
- 240000007594 Oryza sativa Species 0.000 description 2
- 235000007164 Oryza sativa Nutrition 0.000 description 2
- 239000013543 active substance Substances 0.000 description 2
- 238000009395 breeding Methods 0.000 description 2
- 230000001488 breeding effect Effects 0.000 description 2
- 230000010261 cell growth Effects 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- ZQSIJRDFPHDXIC-UHFFFAOYSA-N daidzein Chemical compound C1=CC(O)=CC=C1C1=COC2=CC(O)=CC=C2C1=O ZQSIJRDFPHDXIC-UHFFFAOYSA-N 0.000 description 2
- XBDQKXXYIPTUBI-UHFFFAOYSA-N dimethylselenoniopropionate Natural products CCC(O)=O XBDQKXXYIPTUBI-UHFFFAOYSA-N 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 235000011949 flavones Nutrition 0.000 description 2
- 229930182470 glycoside Natural products 0.000 description 2
- 230000002218 hypoglycaemic effect Effects 0.000 description 2
- 230000001939 inductive effect Effects 0.000 description 2
- 230000005764 inhibitory process Effects 0.000 description 2
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 2
- 238000000034 method Methods 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 230000000877 morphologic effect Effects 0.000 description 2
- 229930014626 natural product Natural products 0.000 description 2
- 235000020824 obesity Nutrition 0.000 description 2
- 229920001542 oligosaccharide Polymers 0.000 description 2
- 150000002482 oligosaccharides Chemical class 0.000 description 2
- 125000000636 p-nitrophenyl group Chemical group [H]C1=C([H])C(=C([H])C([H])=C1*)[N+]([O-])=O 0.000 description 2
- 235000009566 rice Nutrition 0.000 description 2
- 229930000044 secondary metabolite Natural products 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- 239000010902 straw Substances 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- XUFXOAAUWZOOIT-SXARVLRPSA-N (2R,3R,4R,5S,6R)-5-[[(2R,3R,4R,5S,6R)-5-[[(2R,3R,4S,5S,6R)-3,4-dihydroxy-6-methyl-5-[[(1S,4R,5S,6S)-4,5,6-trihydroxy-3-(hydroxymethyl)-1-cyclohex-2-enyl]amino]-2-oxanyl]oxy]-3,4-dihydroxy-6-(hydroxymethyl)-2-oxanyl]oxy]-6-(hydroxymethyl)oxane-2,3,4-triol Chemical compound O([C@H]1O[C@H](CO)[C@H]([C@@H]([C@H]1O)O)O[C@H]1O[C@@H]([C@H]([C@H](O)[C@H]1O)N[C@@H]1[C@@H]([C@@H](O)[C@H](O)C(CO)=C1)O)C)[C@@H]1[C@@H](CO)O[C@@H](O)[C@H](O)[C@H]1O XUFXOAAUWZOOIT-SXARVLRPSA-N 0.000 description 1
- DYARIVMCYYQNNQ-UHFFFAOYSA-N 7-hydroxy-2-(3-hydroxyphenyl)chromen-4-one Chemical compound OC1=CC=CC(C=2OC3=CC(O)=CC=C3C(=O)C=2)=C1 DYARIVMCYYQNNQ-UHFFFAOYSA-N 0.000 description 1
- 241000187844 Actinoplanes Species 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- 241000723920 Bacillus subtilis B2 Species 0.000 description 1
- 102000016938 Catalase Human genes 0.000 description 1
- 108010053835 Catalase Proteins 0.000 description 1
- 206010021033 Hypomenorrhoea Diseases 0.000 description 1
- 239000006142 Luria-Bertani Agar Substances 0.000 description 1
- 229910002651 NO3 Inorganic materials 0.000 description 1
- NHNBFGGVMKEFGY-UHFFFAOYSA-N Nitrate Chemical compound [O-][N+]([O-])=O NHNBFGGVMKEFGY-UHFFFAOYSA-N 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- 241000187747 Streptomyces Species 0.000 description 1
- 229920006328 Styrofoam Polymers 0.000 description 1
- 241000282887 Suidae Species 0.000 description 1
- GAMYVSCDDLXAQW-AOIWZFSPSA-N Thermopsosid Natural products O(C)c1c(O)ccc(C=2Oc3c(c(O)cc(O[C@H]4[C@H](O)[C@@H](O)[C@H](O)[C@H](CO)O4)c3)C(=O)C=2)c1 GAMYVSCDDLXAQW-AOIWZFSPSA-N 0.000 description 1
- 239000007983 Tris buffer Substances 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 229960002632 acarbose Drugs 0.000 description 1
- XUFXOAAUWZOOIT-UHFFFAOYSA-N acarviostatin I01 Natural products OC1C(O)C(NC2C(C(O)C(O)C(CO)=C2)O)C(C)OC1OC(C(C1O)O)C(CO)OC1OC1C(CO)OC(O)C(O)C1O XUFXOAAUWZOOIT-UHFFFAOYSA-N 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 239000008186 active pharmaceutical agent Substances 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 230000000996 additive effect Effects 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 230000009604 anaerobic growth Effects 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 210000004666 bacterial spore Anatomy 0.000 description 1
- 230000000975 bioactive effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 229940041514 candida albicans extract Drugs 0.000 description 1
- 239000005018 casein Substances 0.000 description 1
- BECPQYXYKAMYBN-UHFFFAOYSA-N casein, tech. Chemical compound NCCCCC(C(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(CC(C)C)N=C(O)C(CCC(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(C(C)O)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(COP(O)(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(N)CC1=CC=CC=C1 BECPQYXYKAMYBN-UHFFFAOYSA-N 0.000 description 1
- 235000021240 caseins Nutrition 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 230000000052 comparative effect Effects 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 238000012136 culture method Methods 0.000 description 1
- 239000012228 culture supernatant Substances 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 235000007240 daidzein Nutrition 0.000 description 1
- 238000000354 decomposition reaction Methods 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 206010012601 diabetes mellitus Diseases 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 230000000378 dietary effect Effects 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 238000000635 electron micrograph Methods 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 230000001747 exhibiting effect Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 229930003944 flavone Natural products 0.000 description 1
- 235000013312 flour Nutrition 0.000 description 1
- 230000037406 food intake Effects 0.000 description 1
- 235000021474 generally recognized As safe (food) Nutrition 0.000 description 1
- 235000021473 generally recognized as safe (food ingredients) Nutrition 0.000 description 1
- ZCOLJUOHXJRHDI-CMWLGVBASA-N genistein 7-O-beta-D-glucoside Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1=CC(O)=C2C(=O)C(C=3C=CC(O)=CC=3)=COC2=C1 ZCOLJUOHXJRHDI-CMWLGVBASA-N 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- PZOUSPYUWWUPPK-UHFFFAOYSA-N indole Natural products CC1=CC=CC2=C1C=CN2 PZOUSPYUWWUPPK-UHFFFAOYSA-N 0.000 description 1
- RKJUIXBNRJVNHR-UHFFFAOYSA-N indolenine Natural products C1=CC=C2CC=NC2=C1 RKJUIXBNRJVNHR-UHFFFAOYSA-N 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- JEIPFZHSYJVQDO-UHFFFAOYSA-N iron(III) oxide Inorganic materials O=[Fe]O[Fe]=O JEIPFZHSYJVQDO-UHFFFAOYSA-N 0.000 description 1
- 230000001788 irregular Effects 0.000 description 1
- GOMNOOKGLZYEJT-UHFFFAOYSA-N isoflavone Chemical compound C=1OC2=CC=CC=C2C(=O)C=1C1=CC=CC=C1 GOMNOOKGLZYEJT-UHFFFAOYSA-N 0.000 description 1
- CJWQYWQDLBZGPD-UHFFFAOYSA-N isoflavone Natural products C1=C(OC)C(OC)=CC(OC)=C1C1=COC2=C(C=CC(C)(C)O3)C3=C(OC)C=C2C1=O CJWQYWQDLBZGPD-UHFFFAOYSA-N 0.000 description 1
- 235000008696 isoflavones Nutrition 0.000 description 1
- 239000004310 lactic acid Substances 0.000 description 1
- 235000014655 lactic acid Nutrition 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 150000002772 monosaccharides Chemical class 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 239000008363 phosphate buffer Substances 0.000 description 1
- 230000000291 postprandial effect Effects 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 235000019260 propionic acid Nutrition 0.000 description 1
- IUVKMZGDUIUOCP-BTNSXGMBSA-N quinbolone Chemical compound O([C@H]1CC[C@H]2[C@H]3[C@@H]([C@]4(C=CC(=O)C=C4CC3)C)CC[C@@]21C)C1=CCCC1 IUVKMZGDUIUOCP-BTNSXGMBSA-N 0.000 description 1
- 235000021067 refined food Nutrition 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- 238000010993 response surface methodology Methods 0.000 description 1
- 230000000717 retained effect Effects 0.000 description 1
- 230000001953 sensory effect Effects 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 210000000813 small intestine Anatomy 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 239000008261 styrofoam Substances 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 150000008163 sugars Chemical class 0.000 description 1
- 230000001629 suppression Effects 0.000 description 1
- 230000009897 systematic effect Effects 0.000 description 1
- 229940124597 therapeutic agent Drugs 0.000 description 1
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 1
- 239000012137 tryptone Substances 0.000 description 1
- 241001148471 unidentified anaerobic bacterium Species 0.000 description 1
- 125000000391 vinyl group Chemical group [H]C([*])=C([H])[H] 0.000 description 1
- 229920002554 vinyl polymer Polymers 0.000 description 1
- VHBFFQKBGNRLFZ-UHFFFAOYSA-N vitamin p Natural products O1C2=CC=CC=C2C(=O)C=C1C1=CC=CC=C1 VHBFFQKBGNRLFZ-UHFFFAOYSA-N 0.000 description 1
- 239000012138 yeast extract Substances 0.000 description 1
Classifications
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2200/00—Function of food ingredients
- A23V2200/30—Foods, ingredients or supplements having a functional effect on health
- A23V2200/328—Foods, ingredients or supplements having a functional effect on health having effect on glycaemic control and diabetes
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2200/00—Function of food ingredients
- A23V2200/30—Foods, ingredients or supplements having a functional effect on health
- A23V2200/332—Promoters of weight control and weight loss
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/07—Bacillus
- C12R2001/10—Bacillus licheniformis
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Biotechnology (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Wood Science & Technology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Medicinal Chemistry (AREA)
- Biomedical Technology (AREA)
- Virology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Nutrition Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
Description
本発明は、α−グルコシダーゼ阻害剤を多量生産する新菌株であるバチルス・リケニフォルミスＮＹ１５０５菌株（Ｂａｃｉｌｌｕｓ ｌｉｃｈｅｎｉｆｏｒｍｉｓ ＮＹ１５０５）に関する。 The present invention relates to the Bacillus licheniformis NY1505 strain, which is a new strain that mass-produces α-glucosidase inhibitors.
α−グルコシダーゼ阻害剤（α−ｇｌｕｃｏｓｉｄａｓｅ ｉｎｈｉｂｉｔｏｒｓ）は、一部の植物や微生物に分布し、オリゴ糖の単糖類への分解を抑制する作用を行う物質である。 α-Glucosidase inhibitors (α-glucosidase inhibitors) are substances that are distributed in some plants and microorganisms and have an action of suppressing the decomposition of oligosaccharides into monosaccharides.
α−グルコシダーゼ活性の阻害は、腸内で糖類の円滑な吸収を阻害することにより、食後血糖上昇抑制はもとより、肥満治療のような付随的な効果が期待されるので、α−グルコシダーゼ阻害剤を経済的に量産するための必要性は次第に高まりつつある。 Inhibition of α-glucosidase activity is expected to have ancillary effects such as treatment of obesity as well as suppression of postprandial blood glucose elevation by inhibiting smooth absorption of sugars in the intestine. The need for economical mass production is increasing.
植物由来のα−グルコシダーゼ阻害剤としては、桑の葉に含有された１−デオキシノジリマイシン（１−ｄｅｏｘｙｎｏｊｉｒｉｍｙｃｉｎ）のようなアザ糖（ａｚａ ｓｕｇａｒ）類、大豆に含有されたゲニステイン（ｇｅｎｉｓｔｅｉｎ）、ダイゼイン（ｄａｉｄｚｅｉｎ）のようなイソフラボン（ｉｓｏｆｌａｖｏｎｅ）、一部のフラボン配糖体（ｆｌａｖｏｎｅ ｇｌｙｃｏｓｉｄｅ）などが知られており、微生物由来のα−グルコシダーゼ阻害剤としては、アクチノプラネス属（Ａｃｔｉｎｏｐｌａｎｅｓ）菌株由来の類似オリゴ糖（ｐｓｅｕｄｏｏｌｉｇｏｓａｃｃｈａｒｉｄｅ）、ストレプトミセス属（Ｓｔｒｅｐｔｏｍｙｃｅｓ）またはバチルス属（Ｂａｃｉｌｌｕｓ）などの微生物が生産する１−デオキシノジリマイシン、トリス塩（ｔｒｉｓ ｂａｓｅ）などが知られており、医薬品としては、アカルボース（ａｃａｒｂｏｓｅ）、ボグリボース（ｖｏｇｌｉｂｏｓｅ）のようなα−グルコシダーゼ阻害剤が既に糖尿病治療剤として販売されている。 Plant-derived α-glucosidase inhibitors include aza sugars such as 1-deoxynojirimycin contained in mulberry leaves, genistine contained in soybeans, and daizein. Isoflavone such as (daidzein), some flavon glycosides (flavone glycoside), etc. are known, and as an α-glucosidase inhibitor derived from a microorganism, a similarity derived from a strain of Actinoplanes is known. 1-Deoxynojirimycin, tris base, etc. produced by microorganisms such as oligosaccharide (pseudoglucosidase), Streptomyces or Bacillus are known, and acarbose (acrabose) is known as a pharmaceutical. ), Α-Glucosidase inhibitors such as boglibose are already on the market as therapeutic agents for diabetes.
最近、血糖降下に効果があるα−グルコシダーゼ阻害剤を食品として摂取するために、食用可能な天産物についての研究が活発に進められている。食品として使われる天産物由来のα−グルコシダーゼ阻害剤は、植物の場合、桑の葉、微生物の場合、一部の納豆を代表的な例として挙げられる。これらの１次加工品または抽出物は、量産されているが、共通してにおい、色沢のような嗜好性を落とす因子を有しているので、製品の多様化に制限がある。また、これらは、植物または微生物が生産した二次代謝産物を利用する場合なので、一日に数回一定量を服用して初めて、所期の目的を果たすことができるという短所がある。 Recently, research on edible natural products has been actively promoted in order to ingest an α-glucosidase inhibitor that is effective in lowering blood glucose as a food. Typical examples of α-glucosidase inhibitors derived from natural products used as foods are mulberry leaves in the case of plants and some natto in the case of microorganisms. Although these primary processed products or extracts are mass-produced, there is a limit to the diversification of products because they have a common odor and a factor that reduces the taste such as color. In addition, since these are cases where secondary metabolites produced by plants or microorganisms are used, there is a disadvantage that the intended purpose can be achieved only after taking a certain amount several times a day.
一方、乳酸菌のような腸内細菌は、腸内で生息しながら増殖する特性を有している。したがって、α−グルコシダーゼ阻害剤のような二次代謝産物を生産する腸内細菌は、腸内で増殖しながら切れ目なく腸内環境にα−グルコシダーゼ阻害剤を生産して供給すると予想される。しかし、従来に知られたバチルス・サブチリス（Ｂａｃｉｌｌｕｓ ｓｕｂｔｉｌｉｓ）のような一部の納豆菌は、α−グルコシダーゼ阻害剤を生産すると知られているが、バチルス・サブチリスは、好気性菌で腸内で生息しにくいために、バチルス・サブチリスの増殖でα−グルコシダーゼ阻害剤を腸内で供給されることは難しく、納豆の摂取を通じてα−グルコシダーゼ阻害剤を供給されるためには、多量摂取しなければならないという問題点がある。 On the other hand, intestinal bacteria such as lactic acid bacteria have the property of growing while inhabiting the intestines. Therefore, gut flora that produce secondary metabolites such as α-glucosidase inhibitors are expected to produce and supply α-glucosidase inhibitors seamlessly into the intestinal environment while growing in the intestine. However, while some Bacillus subtilis, previously known, such as Bacillus subtilis, are known to produce α-glucosidase inhibitors, Bacillus subtilis is an aerobic bacterium in the intestine. It is difficult to supply α-glucosidase inhibitor in the intestine by the growth of Bacillus subtilis because it is difficult to inhabit, and in order to supply α-glucosidase inhibitor through ingestion of natto, it is necessary to take a large amount. There is a problem that it does not become.
したがって、腸内で生息が可能であり、α−グルコシダーゼ阻害剤を多量生成することができて、体重減量と血糖降下などのための目的として効果的に活用可能な新たな菌株についての研究及び開発が必要である。 Therefore, research and development of a new strain that can live in the intestine, can produce a large amount of α-glucosidase inhibitor, and can be effectively used for purposes such as weight loss and hypoglycemia. is required.
本発明者らは、α−グルコシダーゼ阻害活性が強い腸内微生物を選抜するために鋭意研究した結果、α−グルコシダーゼ阻害活性を示す微生物のうちから通性嫌気性であり、胞子を形成し、腸内で増殖が可能なＧＲＡＳ菌としてα−グルコシダーゼ阻害剤を多量で生産する新規なバチルス・リケニフォルミスＮＹ１５０５菌株を分離・同定し、前記微生物の発酵物または胞子を含んだ栄養細胞は、体重減量と血糖降下とを目的として食品、医薬品、飼料などに有用に使われうるということを確認した。 As a result of diligent research to select intestinal microorganisms having strong α-glucosidase inhibitory activity, the present inventors are facultative anaerobic, form spores, and intestines from among microorganisms exhibiting α-glucosidase inhibitory activity. A novel Bacillus likeniformis NY1505 strain that produces a large amount of α-glucosidase inhibitor as a GRAS bacterium capable of growing within the strain was isolated and identified, and the vegetative cells containing the fermented product or spores of the microorganism were used for weight loss and blood glucose. It was confirmed that it can be usefully used in foods, medicines, feeds, etc. for the purpose of descent.
これにより、本発明では、α−グルコシダーゼ阻害剤を多量生産する新菌株であるバチルス・リケニフォルミスＮＹ１５０５菌株に関する技術内容を提供することをその目的とする。 Accordingly, it is an object of the present invention to provide a technical content concerning a Bacillus licheniformis NY1505 strain, which is a new strain that mass-produces an α-glucosidase inhibitor.
前記のような技術的課題を果たすために、本発明は、α−グルコシダーゼ阻害剤を高生産するバチルス・リケニフォルミスＮＹ１５０５菌株（受託番号ＫＣＴＣ１３０２１ＢＰ）を提供する。 In order to fulfill the above technical problems, the present invention provides a Bacillus licheniformis NY1505 strain (accession number KCTC13021BP) that highly produces an α-glucosidase inhibitor.
また、前記菌株は、配列番号１の１６ｓ ＲＮＡを含むことを特徴とする。 In addition, the strain is characterized by containing 16s RNA of SEQ ID NO: 1.
また、前記菌株は、嫌気性であるか、または、好気性であり得る。 Also, the strain can be anaerobic or aerobic.
また、前記菌株は、胞子を形成し、前記菌株が投与された後、生物体から２週以内に排泄されうる。 In addition, the strain forms spores and can be excreted from the organism within 2 weeks after the strain is administered.
また、前記菌株は、動物の腸内でα−グルコシダーゼ阻害剤を生産し、腸内で生産されたα−グルコシダーゼ阻害剤は、前記菌株が投与された後、生物体から２週以内に排泄されうる。 In addition, the strain produces an α-glucosidase inhibitor in the intestine of an animal, and the α-glucosidase inhibitor produced in the intestine is excreted from the organism within 2 weeks after the administration of the strain. sell.
また、本発明は、前記菌株を含む食品組成物を提供する。 The present invention also provides a food composition containing the strain.
また、前記食品組成物には、前記菌株が胞子の形態で含まれ、１ｋｇ当たり１０６〜１０１０個の胞子で含まれうる。 Moreover, the food composition, the strain is included in the form of spores can be included in 10 6 to 10 10 spores per 1 kg.
また、前記食品組成物は、前記菌株の発酵物をさらに含み、前記発酵物は、大豆発酵物であり得る。 In addition, the food composition further contains a fermented product of the strain, and the fermented product may be a fermented soybean product.
本発明によるバチルス・リケニフォルミスＮＹ１５０５菌株は、嫌気性条件でも生息が可能な新菌株であって、腸内でα−グルコシダーゼ阻害剤を生産して供給するので、有効濃度を一定に保持して、優れた体重減量及び血糖降下の効果を示し、腸内に固着せず、一時的に生息する特性を示して、必要に応じて投与された菌株または菌株が生産したα−グルコシダーゼ阻害剤は、排出が可能であって、安全に使用することができる。 The Bacillus licheniformis NY1505 strain according to the present invention is a new strain that can live even under anaerobic conditions, and since it produces and supplies an α-glucosidase inhibitor in the intestine, it is excellent in keeping the effective concentration constant. The strains administered as needed or the α-glucosidase inhibitors produced by the strains, which show the effects of weight loss and hypoglycemia, do not stick to the intestine, and show the property of temporarily inhabiting, are excreted. It is possible and can be used safely.
また、α−グルコシダーゼ阻害剤の生成のための別途の発酵コスト及び加工費が不要であって、生産コストを節減し、発酵臭などの問題が発生せず、多様な形態の食品に適用が可能である。 In addition, there is no need for separate fermentation costs and processing costs for the production of α-glucosidase inhibitors, which reduces production costs, does not cause problems such as fermented odors, and can be applied to various forms of food. Is.
以下、本発明を詳しく説明する。 Hereinafter, the present invention will be described in detail.
本発明は、α−グルコシダーゼ阻害剤を高生産するバチルス・リケニフォルミスＮＹ１５０５菌株（受託番号ＫＣＴＣ１３０２１ＢＰ）を提供する。 The present invention provides a Bacillus licheniformis NY1505 strain (accession number KCTC13021BP) that produces a high α-glucosidase inhibitor.
前記菌株は、稲わら及び乾草を試料として耐熱性胞子を形成する菌を分離し、生体内で生育が可能であり、α−グルコシダーゼ阻害剤を高生産する菌株を選別することで得られる。 The strain can be obtained by separating strains that form heat-resistant spores using rice straw and hay as samples, and selecting a strain that can grow in vivo and produces a high α-glucosidase inhibitor.
前記菌株は、下記の配列番号１の１６ｓ ＲＮＡを含むことを特徴とする。 The strain is characterized by containing the 16s RNA of SEQ ID NO: 1 below.
配列番号１：
ＡＧＧＧＣＴＴＣＣＣＣＴＴＣＧＧＧＧＧＣＡＧＡＧＴＧＡＣＡＧＧＴＧＧＴＧＣＡＴＧＧＴＴＧＴＣＧＴＣＡＧＣＴＣＧＴＧＴＣＧＴＧＡＧＡＴＧＴＴＧＧＧＴＴＡＡＧＴＣＣＣＧＣＡＡＣＧＡＧＣＧＣＡＡＣＣＣＴＴＧＡＴＣＴＴＡＧＴＴＧＣＣＡＧＣＡＴＴＣＡＧＴＴＧＧＧＣＡＣＴＣＴＡＡＧＧＴＧＡＣＴＧＣＣＧＧＴＧＡＣＡＡＡＣＣＧＧＡＧＧＡＡＧＧＴＧＧＧＧＡＴＧＡＣＧＴＣＡＡＡＴＣＡＴＣＡＴＧＣＣＣＣＴＴＡＴＧＡＣＣＴＧＧＧＣＴＡＣＡＣＡＣＧＴＧＣＴＡＣＡＡＴＧＧＧＣＡＧＡＡＣＡＡＡＧＧＧＣＡＧＣＧＡＡＧＣＣＧＣＧＡＧＧＣＴＡＡＧＣＣＡＡＴＣＣＣＡＣＡＡＡＴＣＴＧＴＴＣＴＣＡＧＴＴＣＧＧＡＴＣＧＣＡＧＴＣＴＧＣＡＡＣＴＣＧＡＣＴＧＣＧＴＧＡＡＧＣＴＧＧＡＡＴＣＧＣＴＡＧＴＡＡＴＣＧＣＧＧＡＴＣＡＧＣＡＴＧＣＣＧＣＧＧＴＧＡＡＴＡＣＧＴＴＣＣＣＧＧＧＣＣＴＴＧＴＡＣＡＣＡＣＣＧＣＣＣＧＴＣＡＣＡＣＣＡＣＧＡＧＡＧＴＴＴＧＴＡＡＣＡＣＣＣＧＡＡＧＴＣＧＧＴＧＡＧＧＴＡＡＣＣＴＴＴＴＧＧＡＧＣＣＡＧＣＣＧＣＣＧＡＡＧＧＴＧＧＧＡＣＡＧＡＴＧＡＴＴＧＧＧＧＴＧＡＡＧＴＣＧＴＡＣＡＧＧＧＡＡＡＡ
SEQ ID NO: 1:
AGGGCTTCCCCTTCGGGGGCAGAGTGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTGATCTTAGTTGCCAGCATTCAGTTGGGCACTCTAAGGTGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAATCATCATGCCCCTTATGACCTGGGCTACACACGTGCTACAATGGGCAGAACAAAGGGCAGCGAAGCCGCGAGGCTAAGCCAATCCCACAAATCTGTTCTCAGTTCGGATCGCAGTCTGCAACTCGACTGCGTGAAGCTGGAATCGCTAGTAATCGCGGATCAGCATGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCACGAGAGTTTGTAACACCCGAAGTCGGTGAGGTAACCTTTTGGAGCCAGCCGCCGAAGGTGGGACAGATGATTGGGGTGAAGTCGTACAGGGAAAA
前記菌株は、α−グルコシダーゼ阻害剤を腸内で高収率で生産可能な嫌気性特性と生体外でもα−グルコシダーゼ阻害剤を生産可能な好気的特性とを同時に有しうる。 The strain may simultaneously have an anaerobic property capable of producing an α-glucosidase inhibitor in high yield in the intestine and an aerobic property capable of producing an α-glucosidase inhibitor in vitro.
前記菌株は、胞子を形成し、腸内に固着せず、一時的に生息する特性を示して、必要に応じて投与された菌株の除去が可能であって、安全に使用することができる。望ましくは、前記菌株は、生物体に投与された後、２週経過する時点に生物体外に排泄され、より望ましくは、１週経過する時点に排泄されうる。 The strain forms spores, does not adhere to the intestine, exhibits the property of temporarily inhabiting, and can remove the administered strain as needed, and can be used safely. Desirably, the strain can be excreted from the organism two weeks after being administered to the organism, and more preferably, it can be excreted one week later.
また、前記菌株は、腸内で前記α−グルコシダーゼ阻害剤を生産することができて、α−グルコシダーゼ阻害剤の有効濃度を一定に保持し、腸内で生産した前記α−グルコシダーゼ阻害剤は、排泄が可能であって、安全に使用することができる。望ましくは、前記α−グルコシダーゼ阻害剤は、菌株が生物体に投与された後、２週経過する時点に生物体外に排泄されうる。 In addition, the strain can produce the α-glucosidase inhibitor in the intestine, keep the effective concentration of the α-glucosidase inhibitor constant, and the α-glucosidase inhibitor produced in the intestine can be used. It can be excreted and can be used safely. Desirably, the α-glucosidase inhibitor can be excreted from the organism two weeks after the strain is administered to the organism.
これにより、前記菌株を含む食品を摂取する方法によって投与して、腸内でα−グルコシダーゼ阻害剤を高収率で生産することができて、生物体外に排出が可能であって、体重減量と血糖降下との目的として効果的に使用することができる。 As a result, the α-glucosidase inhibitor can be produced in a high yield in the intestine by ingesting a food containing the strain, and can be excreted from the organism, resulting in weight loss. It can be effectively used for the purpose of lowering blood glucose.
本発明は、前記菌株を含む食品組成物を提供する。 The present invention provides a food composition containing the strain.
前記食品組成物は、通用されるバチルス・リケニフォルミスの培養方法によって培養された菌株、菌株の胞子または菌株の発酵物を含みうる。 The food composition may include a strain, spores of the strain or a fermented product of the strain cultivated by a commonly used Bacillus licheniformis culture method.
一例として、前記食品組成物は、前記菌株の胞子を含み、前記食品組成物は、１ｋｇ当たり１０６〜１０１０個の胞子であって、前記菌株を含んで前記菌株が、腸内でα−グルコシダーゼ阻害剤を多量生産することができる。 As an example, the food composition comprises spores of the strain, the food composition is a 10 6 to 10 10 spores per 1 kg, the strain comprising said strain, in the intestines α- A large amount of glucosidase inhibitor can be produced.
さらに他の例として、前記食品組成物は、前記菌株の発酵物をさらに含み、前記発酵物は、前記菌株を大豆に接種して発酵した大豆発酵物であり得る。 As yet another example, the food composition may further comprise a fermented product of the strain, which may be a fermented soybean product fermented by inoculating soybean with the strain.
以下、本発明を実施例を挙げてより詳しく説明する。 Hereinafter, the present invention will be described in more detail with reference to examples.
提示された実施例は、本発明の具体的な例示であり、本発明の範囲を制限するためのものではない。 The examples presented are specific examples of the present invention and are not intended to limit the scope of the present invention.
＜実施例＞
１．菌株の探索
韓国、中国、日本、米国など世界各地から収集した約５００余点の稲わら及び乾草を試料として微生物を分離した。各試料を滅菌食塩水に少量加えて懸濁させた後、８０℃恒温水槽で２０分間熱処理して得た胞子液を２％寒天を含有するＬＢ平板培地（１％トリプトン、０．２％砂糖、０．５％酵母抽出物及び０．５％ ＮａＣｌ、ｐＨ７．０）に塗抹し、５５℃培養器で２日間嫌気的に培養し、培養後、菌集落を形成する菌体を分離した。α−グルコシダーゼ阻害剤の生産量を調査するために、分離された菌株を５ｍｌの５％大豆粉を懸濁した培地に接種し、３７℃で２４時間培養した。培養液を遠心分離して、上澄み液からα−グルコシダーゼ阻害剤活性を測定した（表１の好気的培養）。前記の過程で約６０種の通性嫌気性細菌をα−グルコシダーゼ阻害剤生産菌として１次選抜した後、５０℃で嫌気的に生育が可能であり、プロピオン酸（ｐｒｏｐｉｏｎｉｃ ａｃｉｄ）を磁化する菌株を選抜して、α−グルコシダーゼ阻害活性が高い３種を２次選抜し、培養液を遠心分離して、上澄み液からα−グルコシダーゼ阻害剤活性を測定した（表１の嫌気的培養）。前記３種の菌株は、バージェイズマニュアル（Ｂｅｒｇｅｙ’ｓ Ｍａｎｕａｌ ｏｆ Ｓｙｓｔｅｍａｔｉｃ Ｂａｃｔｅｒｉｏｌｏｇｙ、２００２）による分類学的特性の分析方法によって分類し、そのうち、１種が食用可能な菌であるバチルス・リケニフォルミス（Ｂａｃｉｌｌｕｓ ｌｉｃｈｅｎｉｆｏｒｍｉｓ）に同定されてバチルス・リケニフォルミスＮＹ１５０５と名付け、前記菌株が、下記の配列番号１の１６Ｓ ＲＮＡ配列を有するということを確認し、前記配列を韓国生命工学研究院生物資源センター（ＫＣＴＣ）に２０１６年５月３日付で寄託した（寄託番号：ＫＣＴＣ１３０２１ＢＰ）。
<Example>
1. 1. Search for strains Microorganisms were isolated from about 500 rice straws and hay collected from all over the world such as Korea, China, Japan, and the United States. After adding a small amount of each sample to sterile saline and suspending it, the spore solution obtained by heat-treating in a constant temperature water bath at 80 ° C. for 20 minutes is LB plate medium (1% tryptone, 0.2% sugar) containing 2% agar. , 0.5% yeast extract and 0.5% NaCl, pH 7.0), and anaerobically cultured in a 55 ° C. incubator for 2 days. After culturing, the cells forming the bacterial colony were separated. In order to investigate the production of α-glucosidase inhibitor, the isolated strain was inoculated into a medium in which 5 ml of 5% soybean flour was suspended and cultured at 37 ° C. for 24 hours. The culture broth was centrifuged and the α-glucosidase inhibitor activity was measured from the supernatant (aerobic culture in Table 1). After primary selection of about 60 facultative anaerobic bacteria as α-glucosidase inhibitor-producing bacteria in the above process, a strain capable of anaerobic growth at 50 ° C. and magnetizing propionic acid. , Three species having high α-glucosidase inhibitory activity were secondarily selected, the culture solution was centrifuged, and the α-glucosidase inhibitor activity was measured from the supernatant (anaerobic culture in Table 1). The three strains are classified by the method of analyzing taxonomic characteristics according to the Bergey's Manual of Systematic Bacteriology (2002), and one of them is an edible bacterium, Bacillus licheniformis. ), Named Bacillus licheniformis NY1505, confirmed that the strain has the 16S RNA sequence of SEQ ID NO: 1 below, and transferred the sequence to the Biological Resources Center (KCTC) of the Korea Institute of Biotechnology in 2016. Deposited on May 3 (deposit number: KCTC13021BP).
配列番号１：
ＡＧＧＧＣＴＴＣＣＣＣＴＴＣＧＧＧＧＧＣＡＧＡＧＴＧＡＣＡＧＧＴＧＧＴＧＣＡＴＧＧＴＴＧＴＣＧＴＣＡＧＣＴＣＧＴＧＴＣＧＴＧＡＧＡＴＧＴＴＧＧＧＴＴＡＡＧＴＣＣＣＧＣＡＡＣＧＡＧＣＧＣＡＡＣＣＣＴＴＧＡＴＣＴＴＡＧＴＴＧＣＣＡＧＣＡＴＴＣＡＧＴＴＧＧＧＣＡＣＴＣＴＡＡＧＧＴＧＡＣＴＧＣＣＧＧＴＧＡＣＡＡＡＣＣＧＧＡＧＧＡＡＧＧＴＧＧＧＧＡＴＧＡＣＧＴＣＡＡＡＴＣＡＴＣＡＴＧＣＣＣＣＴＴＡＴＧＡＣＣＴＧＧＧＣＴＡＣＡＣＡＣＧＴＧＣＴＡＣＡＡＴＧＧＧＣＡＧＡＡＣＡＡＡＧＧＧＣＡＧＣＧＡＡＧＣＣＧＣＧＡＧＧＣＴＡＡＧＣＣＡＡＴＣＣＣＡＣＡＡＡＴＣＴＧＴＴＣＴＣＡＧＴＴＣＧＧＡＴＣＧＣＡＧＴＣＴＧＣＡＡＣＴＣＧＡＣＴＧＣＧＴＧＡＡＧＣＴＧＧＡＡＴＣＧＣＴＡＧＴＡＡＴＣＧＣＧＧＡＴＣＡＧＣＡＴＧＣＣＧＣＧＧＴＧＡＡＴＡＣＧＴＴＣＣＣＧＧＧＣＣＴＴＧＴＡＣＡＣＡＣＣＧＣＣＣＧＴＣＡＣＡＣＣＡＣＧＡＧＡＧＴＴＴＧＴＡＡＣＡＣＣＣＧＡＡＧＴＣＧＧＴＧＡＧＧＴＡＡＣＣＴＴＴＴＧＧＡＧＣＣＡＧＣＣＧＣＣＧＡＡＧＧＴＧＧＧＡＣＡＧＡＴＧＡＴＴＧＧＧＧＴＧＡＡＧＴＣＧＴＡＣＡＧＧＧＡＡＡＡ
SEQ ID NO: 1:
AGGGCTTCCCCTTCGGGGGCAGAGTGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTGATCTTAGTTGCCAGCATTCAGTTGGGCACTCTAAGGTGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAATCATCATGCCCCTTATGACCTGGGCTACACACGTGCTACAATGGGCAGAACAAAGGGCAGCGAAGCCGCGAGGCTAAGCCAATCCCACAAATCTGTTCTCAGTTCGGATCGCAGTCTGCAACTCGACTGCGTGAAGCTGGAATCGCTAGTAATCGCGGATCAGCATGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCACGAGAGTTTGTAACACCCGAAGTCGGTGAGGTAACCTTTTGGAGCCAGCCGCCGAAGGTGGGACAGATGATTGGGGTGAAGTCGTACAGGGAAAA
２．納豆の製造
大豆を常温で１２時間ふやかした後、１２１℃で４０分間煮た。煮た大豆を冷却する前、ｇ当たり１０３匹の胞子を接種した後、５０ｇずつ納豆製造用発泡スチレン容器に入れ、ビニールカバーを被せた後、３７℃で２０時間培養した。該培養された納豆を１２時間冷蔵保管した後、α−グルコシダーゼ阻害活性を測定及び官能評価を行った。（表１の納豆）。
2. Production of natto Soybeans were soaked at room temperature for 12 hours and then boiled at 121 ° C. for 40 minutes. Before cooling the boiled soybean, following inoculation with g per 10 Three spores, 50g each placed in natto manufacturing styrofoam container, after covered with vinyl cover, and 20 hours at 37 ° C.. After refrigerating the cultured natto for 12 hours, the α-glucosidase inhibitory activity was measured and sensory evaluation was performed. (Natto in Table 1).
３．α−グルコシダーゼ阻害活性の測定
α−グルコシダーゼは、豚の小腸から抽出して部分精製したものを酵素源として使用した。α−グルコシダーゼ酵素液は、２４ｕｎｉｔ／ｍｌになるように０．５Ｍリン酸塩緩衝溶液（ｐｏｔａｓｓｉｕｍ ｐｈｏｓｐｈａｔｅ ｂｕｆｆｅｒ、ｐＨ７．０）に適当に希釈して使用し、基質は、ｐ−ニトロフェニルα−Ｄ−グルコピラノシド（ｐ−ｎｉｔｒｏｐｈｅｎｙｌ α−Ｄ−ｇｌｕｃｏｐｙｒａｎｏｓｉｄｅ、ｐＮＰＧ）を利用した。試験管に培養上澄み液３０μｌと酵素５０μｌとを混ぜて３７℃で１０分間予備保温を行った後、３ｍＭ ｐＮＰＧ ５０μｌを添加し、３７℃で２０分間反応させた。反応液に０．１Ｍ Ｎａ２ＣＯ３を加えて反応を停止し、４０５ｎｍで吸光度を測定して、下記数式１で阻害活性を測定し、α−グルコシダーゼ阻害活性の測定結果を下記の表１に示した。この際、α−グルコシダーゼ阻害単位（α−ｇｌｕｃｏｓｉｄａｓｅ ｉｎｈｉｂｉｔｉｏｎ ｕｎｉｔ、ＡＧＩ ｕｎｉｔ）は、下記数式１で活性測定に使われたα−グルコシダーゼ（α−ｇｌｕｃｏｓｉｄａｓｅ）を１００％阻害する時の阻害剤の量と定義した。
3. 3. Measurement of α-glucosidase inhibitory activity α-glucosidase was extracted from the small intestine of pigs and partially purified, and used as an enzyme source. The α-glucosidase enzyme solution is appropriately diluted with 0.5 M phosphate buffer (pH 7.0) so as to be 24 units / ml, and the substrate is p-nitrophenyl α-D. -Glucopyranoside (p-nitrophenyl α-D-glucosidase, pNPG) was used. 30 μl of the culture supernatant and 50 μl of the enzyme were mixed in a test tube and preheated at 37 ° C. for 10 minutes, then 50 μl of 3 mM pNPG was added, and the mixture was reacted at 37 ° C. for 20 minutes. 0.1 M Na 2 CO 3 was added to the reaction solution to stop the reaction, the absorbance was measured at 405 nm, the inhibitory activity was measured by the following formula 1, and the measurement results of the α-glucosidase inhibitory activity are shown in Table 1 below. Indicated. At this time, the α-glucosidase inhibitory unit (α-glucosidase inhibition unit, AGI unit) is the amount of the inhibitor when 100% of the α-glucosidase (α-glucosidase) used for the activity measurement in the following formula 1 is inhibited. Defined.
表１に示されたように、酸素の供給有無に関係なく、均一なＡＧＩ阻害活性を示して、バチルス・リケニフォルミスが通性嫌気性細菌なので、嫌気的な条件でも生育が可能であるという事実を確認することができた。したがって、動物の腸管内でバチルス・リケニフォルミスは、増殖を行い、有用な生理活性物質を生産することができると確認された。 As shown in Table 1, the fact that Bacillus licheniformis is a facultative anaerobic bacterium, showing uniform AGI inhibitory activity with or without oxygen supply, can grow even under anaerobic conditions. I was able to confirm. Therefore, it was confirmed that Bacillus licheniformis can proliferate and produce useful bioactive substances in the intestinal tract of animals.
４．バチルス・リケニフォルミスＮＹ１５０５菌株の食餌実験
バチルス・リケニフォルミスＮＹ１５０５菌株が、動物の腸管内で有用物質を生産するか否かを確認するために、マウスの飼料にバチルス・リケニフォルミスＮＹ１５０５胞子を混合して投与することにより、バチルス・リケニフォルミスＮＹ１５０５菌株の腸内増殖有無、腸内でのα−グルコシダーゼ阻害剤の生成有無などを検討した。
4. Dietary experiment of Bacillus licheniformis NY1505 strain In order to confirm whether the Bacillus licheniformis NY1505 strain produces useful substances in the intestinal tract of animals, the Bacillus licheniformis NY1505 spores are mixed and administered to the mouse feed. The presence or absence of intestinal growth of the Bacillus licheniformis NY1505 strain and the presence or absence of α-glucosidase inhibitor production in the intestine were examined.
まず、α−グルコシダーゼ阻害剤を生産するバチルス・リケニフォルミスＮＹ１５０５菌株の胞子を混合した飼料（１０８胞子／ｋｇ飼料）をマウスに給与し、体重の変化を観察した。このために、販売される標準飼料と水とを自在に給与し、雄マウス（００週齢）を１週間予備飼育した後に使用した。明暗サイクルは、午前０８：００時から午後２０：００を明期、午後２０：００から午前０８：００を暗期にし、２２．５±０．５℃の温度及び５０〜６０％の湿度が保持される環境下で飼育した。肥満誘導式で下記表２及び表３の高炭水化物食餌と高脂肪食餌とにし、バチルス・リケニフォルミスＮＹ１５０５菌株の胞子が含有された飼料は、それぞれの肥満誘導式１ｋｇ当たり１０８胞子を混合して製造した。マウスは、５個の群（各８匹）に分けて、対照群、高炭水化物食餌群、高炭水化物食餌＋胞子群、高脂肪食餌群、高脂肪食餌＋胞子群に分けて、それぞれ７週間飼育した。 First, alpha-feed by mixing spores of Bacillus licheniformis NY1505 producing strain glucosidase inhibitor (10 8 spores / kg diet) and salary mice were observed changes in body weight. For this purpose, the standard feed and water to be sold were freely fed, and male mice (00 weeks old) were used after pre-breeding for 1 week. The light-dark cycle is from 08:00 to 20:00 in the light period and from 20:00 to 08:00 in the dark period, with a temperature of 22.5 ± 0.5 ° C and a humidity of 50 to 60%. Breeding in a retained environment. High carbohydrate diet and high-fat diet and west in the following Table 2 and Table 3 in obese inductive, feed spores of Bacillus licheniformis NY1505 strain is contained, produced by mixing the respective obesity inductive 1kg per 10 8 spores did. Mice were divided into 5 groups (8 animals each) and divided into a control group, a high-carbohydrate diet group, a high-carbohydrate diet + spore group, a high-fat diet group, and a high-fat diet + spore group, and bred for 7 weeks each. did.
（１）高炭水化物食餌の給与時に、バチルス・リケニフォルミスＮＹ１５０５菌胞子の投与がマウスの体重に及ぼす影響分析
高炭水化物食餌給与されたマウスの体重（ｂｏｄｙ ｗｅｉｇｈｔ、単位ｇｒａｍ）にバチルス・リケニフォルミスＮＹ１５０５菌胞子の投与が及ぼす影響を分析した結果、図１に示されたように、高炭水化物飼料群は、対照群に比べて、明らかな体重増加を示すが、バチルス・リケニフォルミスＮＹ１５０５菌株の胞子が混合された高炭水化物飼料群では、対照群と同様の体重増加の態様を示すことを確認することができた。
(1) Analysis of the effect of administration of Bacillus licheniformis NY1505 spores on the body weight of mice when fed a high-carbohydrate diet. Analysis of the body weight (body weight, unit gram) of Bacillus likeniformis NY1505 spores As a result of analyzing the effect of administration, as shown in FIG. 1, the high-carbohydrate feed group showed a clear weight gain as compared with the control group, but the spores of the Bacillus likeniformis NY1505 strain were mixed and high. It was confirmed that the carbohydrate feed group showed the same aspect of weight gain as the control group.
（２）高脂肪食餌の給与時に、バチルス・リケニフォルミスＮＹ１５０５菌胞子の投与がマウスの体重に及ぼす影響
高脂肪食餌給与されたマウスの体重にバチルス・リケニフォルミスＮＹ１５０５菌胞子の投与がマウスの体重に及ぼす影響を分析した結果、図２に示されたように、高脂肪飼料群は、対照群に比べて、明らかな体重増加を示すが、バチルス・リケニフォルミスＮＹ１５０５菌株の胞子が混合された高脂肪飼料群では、対照群と同様の体重増加の態様を示すことを確認することができた。
(2) Effect of administration of Bacillus licheniformis NY1505 spores on body weight of mice fed with high-fat diet Effect of administration of Bacillus licheniformis NY1505 spores on body weight of mice fed with high-fat diet As a result of the analysis, as shown in FIG. 2, the high-fat feed group showed a clear weight gain as compared with the control group, but in the high-fat feed group mixed with spores of the Bacillus licheniformis NY1505 strain. , It was confirmed that the body weight gain mode was similar to that of the control group.
（３）糞便で排泄されるバチルス・リケニフォルミスＮＹ１５０５菌数の変化
バチルス・リケニフォルミスＮＹ１５０５菌胞子が投与されたマウスの便に含まれた胞子数を測定して、便で排泄される菌株の変化を分析した結果、図３に示されたように、飼料１ｇ当たり１０５匹の胞子が混合されており、一匹当たり一日に約５ｇの飼料を摂取するので、一日に約５ｘ１０５匹の胞子を摂取する。胞子投与３週後には、糞便１ｇ当たり約１０７匹のバチルス・リケニフォルミスＮＹ１５０５が検出されるので、腸内で本菌株が旺盛に増殖して排出されることが分かった。胞子を７週間投与した場合、胞子を投与していない１週間である８週次には、菌の数字が格段に減少するので、腸内に固着せず、一時的に生息すると判断することができた。したがって、本菌株の投与を中断すれば、腸内菌叢の変化が予想されるので、必要に応じて腸内で本菌株の除去も可能な長所があるということを確認することができた。
(3) Changes in the number of Bacillus licheniformis NY1505 excreted in feces The number of spores contained in the spores of mice to which Bacillus licheniformis NY1505 spores were administered was measured to analyze changes in the strains excreted in the stool. as a result, as shown in FIG. 3, 10 five spores per feed 1g are mixed, since intake of feed of approximately 5g per day per animal, about 5x10 five spores per day Ingest. After spore administered three weeks, since about 107 animals of Bacillus licheniformis NY1505 per feces 1g is detected, it was found that this strain in the intestine is discharged by vigorous proliferation. When spores are administered for 7 weeks, the number of bacteria decreases dramatically after 8 weeks, which is one week when spores are not administered, so it can be judged that they do not stick to the intestines and inhabit temporarily. did it. Therefore, if the administration of this strain is discontinued, changes in the intestinal flora are expected, and it was confirmed that there is an advantage that the strain can be removed in the intestine as needed.
（４）便で排泄されるα−グルコシダーゼ阻害活性の変化
バチルス・リケニフォルミスＮＹ１５０５菌胞子が投与されたマウスの便に含まれたα−グルコシダーゼの阻害活性を分析した結果、図４に示されたように、バチルス・リケニフォルミスＮＹ１５０５菌株は、便で排出される程度に腸内で旺盛に増殖し、α−グルコシダーゼ阻害剤を生産することを確認することができた。胞子が投与されたマウスの２週経過した時点の便で腸内で増殖したバチルス・リケニフォルミスＮＹ１５０５菌株が便で排出されて活性が検出された。また、４週次からは、７週までは排出されるバチルス・リケニフォルミスＮＹ１５０５菌株の量が安定化される時期に排出されるα−グルコシダーゼ阻害剤の量も一定していることを確認することができた。すなわち、α−グルコシダーゼ阻害剤は、常に腸内で生産されるので、腸内での濃度が一定量保持され、効率が高く作用する長所を有し、生産された阻害剤の一定量が排出されることが分かる。糞便で排出されるα−グルコシダーゼ阻害剤の活性を見れば、バチルス・リケニフォルミスＮＹ１５０５菌株で製造した納豆の活性（表１）と比べると、相当量のα−グルコシダーゼ阻害剤が腸内で生産され、その一部が排出されるという事実を確認することができた。
(4) Changes in α-glucosidase inhibitory activity excreted in stool As a result of analyzing the inhibitory activity of α-glucosidase contained in the stool of mice to which Bacillus licheniformis NY1505 bacterial spores were administered, as shown in FIG. In addition, it was confirmed that the Bacillus licheniformis NY1505 strain proliferates vigorously in the intestine to the extent that it is excreted by stool and produces an α-glucosidase inhibitor. The Bacillus licheniformis NY1505 strain grown in the intestine was excreted in the stool after 2 weeks from the mice to which the spores were administered, and the activity was detected. From the 4th week onward, it can be confirmed that the amount of α-glucosidase inhibitor excreted during the period when the amount of Bacillus licheniformis NY1505 strain excreted until the 7th week is stabilized is also constant. did it. That is, since the α-glucosidase inhibitor is always produced in the intestine, it has the advantage that the concentration in the intestine is maintained in a constant amount and acts with high efficiency, and a certain amount of the produced inhibitor is discharged. You can see that. Looking at the activity of the α-glucosidase inhibitor excreted in feces, a considerable amount of the α-glucosidase inhibitor was produced in the intestine as compared with the activity of natto produced by the Bacillus licheniformis NY1505 strain (Table 1). We were able to confirm the fact that some of it was discharged.
前記のような結果を通じて、本発明によるバチルス・リケニフォルミスＮＹ１５０５菌株が発酵によって生成された生理活性物質を使用することではなく、体内で菌株が生理活性物質を連続して作って供給するので、α−グルコシダーゼ阻害剤の生成のための別途の発酵コスト及び加工費が不要であって、生産コストを節減し、体内で連続して生産することにより、有効な濃度を一定に保持できるという長所があるということを確認することができ、有用菌の胞子を利用することにより、発酵臭などの問題が発生せず、多様な加工食品に添加物として使用することができるので、多様な形態の食品に適用することができるという事実を確認することができた。 Through the above results, the Bacillus licheniformis NY1505 strain according to the present invention does not use the physiologically active substance produced by fermentation, but the strain continuously produces and supplies the physiologically active substance in the body. There is no need for additional fermentation costs and processing costs for the production of glucosidase inhibitors, and there is the advantage that the effective concentration can be kept constant by reducing the production cost and continuously producing in the body. By using the spores of useful bacteria, it is possible to use it as an additive in various processed foods without causing problems such as fermented odor, so it can be applied to various forms of food. I was able to confirm the fact that it can be done.
５．分離された菌株の形態学的及び生化学的特性の究明
（１）分離された菌株の形態学的特性
前記のように分離されたバチルス・リケニフォルミスＮＹ１５０５菌株は、ＬＢ寒天平板培地でコロニーの中央部分は薄いさび色を示し、縁部は不規則な外形を示した（図５）。菌の生育が可能な温度範囲は、２５〜５０℃の範囲で成長が良好であり、６０℃以上では、菌体成長が確認されていない。ＬＢ液体培地を利用した菌体成長の対数期で顕微鏡観察した結果、グラム陽性の短い棒状を示せれば、比較菌株であるバチルス・リケニフォルミスと類似した。また、分離した菌株は、下記表４に示されたように、グラム陽性であり、細胞のサイズは、ほぼ０．５〜０．７×１．５〜１．８μｍであることを確認することができ、前記結果を総合して、バチルス・リケニフォルミスＮＹ１５０５菌株の分類学上の位置を図６に示した。
5. Investigation of morphological and biochemical characteristics of the isolated strain (1) Morphological characteristics of the isolated strain The Bacillus licheniformis NY1505 strain isolated as described above is the central portion of the colony on the LB agar plate medium. Showed a light rust color, and the edges showed an irregular outline (Fig. 5). The temperature range in which the bacteria can grow is in the range of 25 to 50 ° C., and the growth is good, and above 60 ° C., cell growth has not been confirmed. As a result of microscopic observation in the logarithmic phase of cell growth using LB liquid medium, if a short gram-positive rod was shown, it was similar to the comparative strain Bacillus licheniformis. In addition, it should be confirmed that the isolated strains are Gram-positive and the cell size is approximately 0.5 to 0.7 × 1.5 to 1.8 μm, as shown in Table 4 below. The above results were combined, and the taxonomic position of the Bacillus licheniformis NY1505 strain is shown in FIG.
（２）分離された菌株の生化学的特性
バージェイズマニュアルによるバチルス属菌株の分類学的特性の分析方法とＡＰＩ５０ＣＨＢキット（ＢｉｏＭｅｒｉｅｕｘ社、Ｆｒａｎｃｅ）とを使用して、分離された菌株の生化学的特性などを調査した結果、分離した菌株は、グラム陽性の円筒状であって、胞子は細胞中央に形成され、硝酸塩の還元力は陽性であり、インドール形成は陰性であった。また、カゼインと澱粉とを分解し、カタラーゼ陽性であり、好気的条件及び嫌気的条件で成長すると確認され、下記表５に示すような生化学的特性（ＡＰＩ ｔｅｓｔ結果）を有することを確認することができた。
(2) Biochemical characteristics of the isolated strain The biochemical of the isolated strain using the analysis method of the taxonomic characteristics of the Bacillus strain according to the Verjay's Manual and the API50CHB kit (BioMeriaux, France). As a result of investigating the characteristics and the like, the isolated strain had a Gram-positive cylindrical shape, spores were formed in the center of the cell, the reducing power of nitrate was positive, and indole formation was negative. In addition, it was confirmed that casein and starch were decomposed, catalase-positive, and grown under aerobic and anaerobic conditions, and that it had biochemical properties (API test results) as shown in Table 5 below. We were able to.
本発明によるバチルス・リケニフォルミスＮＹ１５０５菌株は、腸内でα−グルコシダーゼ阻害剤を生産して供給するので、食品組成物に使われる。
The Bacillus licheniformis NY1505 strain according to the present invention is used in food compositions because it produces and supplies an α-glucosidase inhibitor in the intestine.
寄託機関名：韓国生命工学研究院韓国生物資源センター（ＫＣＴＣ）
受託番号：ＫＣＴＣ１３０２１ＢＰ
受託日：２０１６０５０３
Depositary name: Korea Research Institute of Bioscience and Biotechnology Korea Bioresource Center (KCTC)
Access number: KCTC13021BP
Contract date: 20160503
（受託証）
(Certificate of acceptance)
Claims (12)
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| KR1020160076424A KR101854705B1 (en) | 2016-06-20 | 2016-06-20 | Novel strains of Bacillus licheniformis NY1505 producing high amount of α-glucosidase inhibitors |
| KR10-2016-0076424 | 2016-06-20 | ||
| PCT/KR2017/001754 WO2017222142A1 (en) | 2016-06-20 | 2017-02-17 | Bacillus licheniformis strain ny1505 for mass producing α-glucosidase inhibitor |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JP2019517810A JP2019517810A (en) | 2019-06-27 |
| JP6785324B2 true JP6785324B2 (en) | 2020-11-18 |
Family
ID=60784253
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2018565720A Active JP6785324B2 (en) | 2016-06-20 | 2017-02-17 | Bacillus licheniformis NY1505 strain that mass-produces α-glucosidase inhibitors |
Country Status (3)
| Country | Link |
|---|---|
| JP (1) | JP6785324B2 (en) |
| KR (1) | KR101854705B1 (en) |
| WO (1) | WO2017222142A1 (en) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR102238733B1 (en) * | 2019-07-09 | 2021-04-09 | 서울여자대학교 산학협력단 | Novel Paenisporosarcina quisquiliarum 17mud 1-1541 strain and its use |
| KR102387028B1 (en) | 2020-10-06 | 2022-04-14 | 이해익 | A novel Bacillus coagulans CC strain having high productivity for alpha glucosidase inhibitor |
| CN114752518B (en) * | 2022-01-21 | 2024-04-16 | 武汉中博绿亚生物科技有限公司 | Bacillus licheniformis XCL-09 and application thereof |
Family Cites Families (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5589381A (en) | 1994-06-30 | 1996-12-31 | Rutgers, The State University Of New Jersey | Bacillus licheniformis producing antifungal agents and uses thereof for control of phytopathogenic fungi |
| KR100708286B1 (en) * | 2005-09-08 | 2007-04-16 | 강원대학교산학협력단 | A strain having high productivity for alpha glucosidase inhibitor and a method for preparing the strain |
| KR101302652B1 (en) * | 2011-03-21 | 2013-09-03 | (주)바이오토피아 | PHARMACEUTICAL COMPOSITION FOR PREVENTING OR TREATING DIABETES MELLITUS COMPRISING α-GLUCOSIDASE INHIBITOR AND BREWER'S DRIED YEAST |
| KR101666268B1 (en) * | 2014-04-07 | 2016-10-13 | 강원대학교산학협력단 | Culturing Methods for the Production of Alpha-Glucosidase Inhibitor with Reduced Fermentation Odor |
-
2016
- 2016-06-20 KR KR1020160076424A patent/KR101854705B1/en active IP Right Grant
-
2017
- 2017-02-17 JP JP2018565720A patent/JP6785324B2/en active Active
- 2017-02-17 WO PCT/KR2017/001754 patent/WO2017222142A1/en active Application Filing
Also Published As
| Publication number | Publication date |
|---|---|
| WO2017222142A1 (en) | 2017-12-28 |
| KR101854705B1 (en) | 2018-05-04 |
| KR20170142635A (en) | 2017-12-28 |
| JP2019517810A (en) | 2019-06-27 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105132322B (en) | One lactobacillus plantarum and application thereof | |
| JP5872104B2 (en) | New Bacillus subtilis {NOVELBACILLUSSUBTILIS} | |
| CN112011481B (en) | Lactobacillus reuteri for preventing and treating bacterial diarrhea of livestock and poultry and application thereof | |
| CN109749957B (en) | Preparation and application of lactobacillus gasseri preparation with aquatic pathogenic bacteria antagonistic property | |
| KR101845709B1 (en) | Lactobacillus plantarum KCC-26 and composition comprising the same | |
| CN105524855B (en) | A kind of bacillus coagulans and its application with aquatic pathogenic bacterium antagonistic properties | |
| CN115094012B (en) | Preparation method and application of bacillus coagulans BC-HYC strain microbial inoculum | |
| CN114134083B (en) | Bacillus bailii and application thereof | |
| JP6785324B2 (en) | Bacillus licheniformis NY1505 strain that mass-produces α-glucosidase inhibitors | |
| JP2023547056A (en) | Novel Bacillus coagulans CC strain producing α-glucosidase inhibitor | |
| CN108522884B (en) | Application of composite microecological preparation in preventing fat deposition of broiler chickens | |
| CN115637240A (en) | Bacillus belgii and application thereof | |
| KR100264361B1 (en) | Lactobacillus plantarum PMO08 (KFCC-11028) with cholesterol lowering ability | |
| KR20190047756A (en) | A composition comprising lactobacillus probiotics and a functional feed additive comprising the same | |
| CN112226389A (en) | Planting culture method of intestinal probiotic groups of Sanhuang young chickens and application of intestinal probiotic groups | |
| CN116286514A (en) | Multifunctional microbial inoculum, lactobacillus reuteri contained therein and application thereof | |
| CN116083272A (en) | Clostridium butyricum and application of biological feed additive thereof | |
| KR20180134128A (en) | Sparassis crispa fermentation product having antioxidative and immunological activity, and preparing method thereof | |
| KR100621657B1 (en) | New Bacillus subtilis UBT-M02 strain which has an acid and bile acid resistances, feed additive composition using it, and feed of animal having thereof | |
| CN114456969A (en) | Probiotic preparation for losing weight and reducing blood sugar as well as preparation method and application thereof | |
| KR102260320B1 (en) | Synbiotic composition comprising lactulose and probiotic | |
| KR101357118B1 (en) | Probiotics with improved stability using jujube extracts | |
| CN111454867B (en) | Phoxinus lagowskii waxy bacillus strain and application thereof | |
| Raju et al. | Isolation, Characterization and Sequencing of Lactobacillus from the Oral and Fecal Samples of Healthy Dogs | |
| CN117016673B (en) | Application of bacillus subtilis in improving intestinal flora or intestinal health or nutrient absorption of laying hens |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| A621 | Written request for application examination |
Free format text: JAPANESE INTERMEDIATE CODE: A621 Effective date: 20181214 |
|
| A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20191126 |
|
| A601 | Written request for extension of time |
Free format text: JAPANESE INTERMEDIATE CODE: A601 Effective date: 20200221 |
|
| A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20200526 |
|
| 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: 20201020 |
|
| A61 | First payment of annual fees (during grant procedure) |
Free format text: JAPANESE INTERMEDIATE CODE: A61 Effective date: 20201026 |
|
| R150 | Certificate of patent or registration of utility model |
Ref document number: 6785324 Country of ref document: JP Free format text: JAPANESE INTERMEDIATE CODE: R150 |
|
| R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |