KR20030097264A - Fermented soymilk and method for preparation of same - Google Patents
Fermented soymilk and method for preparation of same Download PDFInfo
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- KR20030097264A KR20030097264A KR1020020034557A KR20020034557A KR20030097264A KR 20030097264 A KR20030097264 A KR 20030097264A KR 1020020034557 A KR1020020034557 A KR 1020020034557A KR 20020034557 A KR20020034557 A KR 20020034557A KR 20030097264 A KR20030097264 A KR 20030097264A
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- South Korea
- Prior art keywords
- bifidobacterium
- soymilk
- int
- soybean
- milk
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- 235000013322 soy milk Nutrition 0.000 title claims description 71
- 238000000034 method Methods 0.000 title claims description 6
- 238000002360 preparation method Methods 0.000 title description 2
- 235000010469 Glycine max Nutrition 0.000 claims abstract description 39
- 244000068988 Glycine max Species 0.000 claims abstract description 39
- 241000131482 Bifidobacterium sp. Species 0.000 claims abstract description 32
- 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 claims abstract description 23
- 235000008696 isoflavones Nutrition 0.000 claims abstract description 23
- 229930182470 glycoside Natural products 0.000 claims abstract description 11
- 238000004519 manufacturing process Methods 0.000 claims abstract description 11
- 241000894006 Bacteria Species 0.000 claims abstract description 10
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 claims abstract description 6
- 239000007858 starting material Substances 0.000 claims abstract description 6
- XUJNEKJLAYXESH-UHFFFAOYSA-N cysteine Natural products SCC(N)C(O)=O XUJNEKJLAYXESH-UHFFFAOYSA-N 0.000 claims abstract description 4
- 239000004310 lactic acid Substances 0.000 claims abstract description 3
- 235000014655 lactic acid Nutrition 0.000 claims abstract description 3
- 235000015140 cultured milk Nutrition 0.000 claims description 28
- 238000000855 fermentation Methods 0.000 claims description 24
- 230000004151 fermentation Effects 0.000 claims description 24
- 150000002515 isoflavone derivatives Chemical class 0.000 claims description 7
- 150000002338 glycosides Chemical class 0.000 claims description 5
- 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 abstract description 10
- -1 isoflavone glycosides Chemical class 0.000 abstract description 10
- 235000013336 milk Nutrition 0.000 abstract description 9
- 239000008267 milk Substances 0.000 abstract description 9
- 210000004080 milk Anatomy 0.000 abstract description 9
- GOMNOOKGLZYEJT-UHFFFAOYSA-N isoflavone Chemical compound C=1OC2=CC=CC=C2C(=O)C=1C1=CC=CC=C1 GOMNOOKGLZYEJT-UHFFFAOYSA-N 0.000 abstract description 7
- TZBJGXHYKVUXJN-UHFFFAOYSA-N genistein Natural products C1=CC(O)=CC=C1C1=COC2=CC(O)=CC(O)=C2C1=O TZBJGXHYKVUXJN-UHFFFAOYSA-N 0.000 abstract description 6
- 235000006539 genistein Nutrition 0.000 abstract description 6
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- 230000001766 physiological effect Effects 0.000 abstract description 5
- ZQSIJRDFPHDXIC-UHFFFAOYSA-N daidzein Chemical compound C1=CC(O)=CC=C1C1=COC2=CC(O)=CC=C2C1=O ZQSIJRDFPHDXIC-UHFFFAOYSA-N 0.000 abstract description 4
- TWCMVXMQHSVIOJ-UHFFFAOYSA-N Aglycone of yadanzioside D Natural products COC(=O)C12OCC34C(CC5C(=CC(O)C(O)C5(C)C3C(O)C1O)C)OC(=O)C(OC(=O)C)C24 TWCMVXMQHSVIOJ-UHFFFAOYSA-N 0.000 abstract description 2
- PLMKQQMDOMTZGG-UHFFFAOYSA-N Astrantiagenin E-methylester Natural products CC12CCC(O)C(C)(CO)C1CCC1(C)C2CC=C2C3CC(C)(C)CCC3(C(=O)OC)CCC21C PLMKQQMDOMTZGG-UHFFFAOYSA-N 0.000 abstract description 2
- 235000018417 cysteine Nutrition 0.000 abstract description 2
- 235000007240 daidzein Nutrition 0.000 abstract description 2
- 239000000796 flavoring agent Substances 0.000 abstract description 2
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- 239000001963 growth medium Substances 0.000 abstract description 2
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- 208000019790 abdominal distention Diseases 0.000 abstract 1
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- 230000000694 effects Effects 0.000 description 24
- 102000006995 beta-Glucosidase Human genes 0.000 description 19
- 108010047754 beta-Glucosidase Proteins 0.000 description 19
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- CJPNHKPXZYYCME-UHFFFAOYSA-N Genistin Natural products OCC1OC(Oc2ccc(O)c3OC(=CC(=O)c23)c4ccc(O)cc4)C(O)C(O)C1O CJPNHKPXZYYCME-UHFFFAOYSA-N 0.000 description 4
- YCUNGEJJOMKCGZ-UHFFFAOYSA-N Pallidiflorin Natural products C1=CC(OC)=CC=C1C1=COC2=CC=CC(O)=C2C1=O YCUNGEJJOMKCGZ-UHFFFAOYSA-N 0.000 description 4
- 238000010521 absorption reaction Methods 0.000 description 4
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- 102000004169 proteins and genes Human genes 0.000 description 4
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- 239000000243 solution Substances 0.000 description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 4
- BTJIUGUIPKRLHP-UHFFFAOYSA-N 4-nitrophenol Chemical compound OC1=CC=C([N+]([O-])=O)C=C1 BTJIUGUIPKRLHP-UHFFFAOYSA-N 0.000 description 3
- 241000186000 Bifidobacterium Species 0.000 description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 3
- 108010044467 Isoenzymes Proteins 0.000 description 3
- 235000010627 Phaseolus vulgaris Nutrition 0.000 description 3
- 244000046052 Phaseolus vulgaris Species 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 238000004458 analytical method Methods 0.000 description 3
- 230000001580 bacterial effect Effects 0.000 description 3
- 230000009286 beneficial effect Effects 0.000 description 3
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 description 3
- 230000002950 deficient Effects 0.000 description 3
- 230000012010 growth Effects 0.000 description 3
- 238000006460 hydrolysis reaction Methods 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 235000016709 nutrition Nutrition 0.000 description 3
- 241001608472 Bifidobacterium longum Species 0.000 description 2
- 208000024172 Cardiovascular disease Diseases 0.000 description 2
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 2
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 description 2
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 description 2
- 239000007621 bhi medium Substances 0.000 description 2
- 210000001072 colon Anatomy 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 230000036541 health Effects 0.000 description 2
- 238000004128 high performance liquid chromatography Methods 0.000 description 2
- 230000007062 hydrolysis Effects 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- 230000000968 intestinal effect Effects 0.000 description 2
- 239000008101 lactose Substances 0.000 description 2
- 230000035764 nutrition Effects 0.000 description 2
- 239000008213 purified water Substances 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 208000024891 symptom Diseases 0.000 description 2
- 235000013618 yogurt Nutrition 0.000 description 2
- JLIDVCMBCGBIEY-UHFFFAOYSA-N 1-penten-3-one Chemical class CCC(=O)C=C JLIDVCMBCGBIEY-UHFFFAOYSA-N 0.000 description 1
- GUBGYTABKSRVRQ-XLOQQCSPSA-N Alpha-Lactose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@H](O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-XLOQQCSPSA-N 0.000 description 1
- 102100026189 Beta-galactosidase Human genes 0.000 description 1
- 241001486432 Bifidobacterium adolescentis ATCC 15703 Species 0.000 description 1
- 241000186016 Bifidobacterium bifidum Species 0.000 description 1
- 241000286268 Bifidobacterium bifidum ATCC 29521 = JCM 1255 = DSM 20456 Species 0.000 description 1
- 241000226932 Bifidobacterium bifidum BGN4 Species 0.000 description 1
- 206010006187 Breast cancer Diseases 0.000 description 1
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- 241001107116 Castanospermum australe Species 0.000 description 1
- 229920000742 Cotton Polymers 0.000 description 1
- KYQZWONCHDNPDP-UHFFFAOYSA-N Daidzoside Natural products OC1C(O)C(O)C(CO)OC1OC1=CC=C2C(=O)C(C=3C=CC(O)=CC=3)=COC2=C1 KYQZWONCHDNPDP-UHFFFAOYSA-N 0.000 description 1
- 101100379080 Emericella variicolor andB gene Proteins 0.000 description 1
- 101710154606 Hemagglutinin Proteins 0.000 description 1
- 102000004157 Hydrolases Human genes 0.000 description 1
- 108090000604 Hydrolases Proteins 0.000 description 1
- IFQSXNOEEPCSLW-DKWTVANSSA-N L-cysteine hydrochloride Chemical compound Cl.SC[C@H](N)C(O)=O IFQSXNOEEPCSLW-DKWTVANSSA-N 0.000 description 1
- 108010059881 Lactase Proteins 0.000 description 1
- 102000004856 Lectins Human genes 0.000 description 1
- 108090001090 Lectins Proteins 0.000 description 1
- 102000003820 Lipoxygenases Human genes 0.000 description 1
- 108090000128 Lipoxygenases Proteins 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 102000014171 Milk Proteins Human genes 0.000 description 1
- 108010011756 Milk Proteins Proteins 0.000 description 1
- 208000001132 Osteoporosis Diseases 0.000 description 1
- 101710093908 Outer capsid protein VP4 Proteins 0.000 description 1
- 101710135467 Outer capsid protein sigma-1 Proteins 0.000 description 1
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- 101710176177 Protein A56 Proteins 0.000 description 1
- 102000004142 Trypsin Human genes 0.000 description 1
- 108090000631 Trypsin Proteins 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 239000003513 alkali Substances 0.000 description 1
- 230000003698 anagen phase Effects 0.000 description 1
- 230000001093 anti-cancer Effects 0.000 description 1
- 235000004458 antinutrient Nutrition 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 108010005774 beta-Galactosidase Proteins 0.000 description 1
- 235000021279 black bean Nutrition 0.000 description 1
- 208000024330 bloating Diseases 0.000 description 1
- 210000004556 brain Anatomy 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 230000010261 cell growth Effects 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 235000012000 cholesterol Nutrition 0.000 description 1
- 230000015271 coagulation Effects 0.000 description 1
- 238000005345 coagulation Methods 0.000 description 1
- 208000029742 colonic neoplasm Diseases 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
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- 238000011161 development Methods 0.000 description 1
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- 230000029087 digestion Effects 0.000 description 1
- 230000001079 digestive effect Effects 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 239000003797 essential amino acid Substances 0.000 description 1
- 235000020776 essential amino acid Nutrition 0.000 description 1
- 235000020774 essential nutrients Nutrition 0.000 description 1
- 229940011871 estrogen Drugs 0.000 description 1
- 239000000262 estrogen Substances 0.000 description 1
- 239000004744 fabric Substances 0.000 description 1
- 210000003608 fece Anatomy 0.000 description 1
- 239000000706 filtrate Substances 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 210000001035 gastrointestinal tract Anatomy 0.000 description 1
- 230000007407 health benefit Effects 0.000 description 1
- 239000000185 hemagglutinin Substances 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 230000028993 immune response Effects 0.000 description 1
- ADFCQWZHKCXPAJ-UHFFFAOYSA-N indofine Natural products C1=CC(O)=CC=C1C1CC2=CC=C(O)C=C2OC1 ADFCQWZHKCXPAJ-UHFFFAOYSA-N 0.000 description 1
- 238000001802 infusion Methods 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 229910052742 iron Inorganic materials 0.000 description 1
- 229940116108 lactase Drugs 0.000 description 1
- 239000002523 lectin Substances 0.000 description 1
- 208000029565 malignant colon neoplasm Diseases 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 235000013384 milk substitute Nutrition 0.000 description 1
- 238000006386 neutralization reaction Methods 0.000 description 1
- 150000002482 oligosaccharides Chemical class 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 239000003075 phytoestrogen Substances 0.000 description 1
- XYKIUTSFQGXHOW-UHFFFAOYSA-N propan-2-one;toluene Chemical compound CC(C)=O.CC1=CC=CC=C1 XYKIUTSFQGXHOW-UHFFFAOYSA-N 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 229910000029 sodium carbonate Inorganic materials 0.000 description 1
- 239000012064 sodium phosphate buffer Substances 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 235000012424 soybean oil Nutrition 0.000 description 1
- 239000003549 soybean oil Substances 0.000 description 1
- 230000004936 stimulating effect Effects 0.000 description 1
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- 239000000758 substrate Substances 0.000 description 1
- 239000013076 target substance Substances 0.000 description 1
- 238000004448 titration Methods 0.000 description 1
- 239000012588 trypsin Substances 0.000 description 1
- 235000021122 unsaturated fatty acids Nutrition 0.000 description 1
- 150000004670 unsaturated fatty acids Chemical class 0.000 description 1
- 235000019156 vitamin B Nutrition 0.000 description 1
- 239000011720 vitamin B Substances 0.000 description 1
- 238000005406 washing Methods 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
- A23L11/00—Pulses, i.e. fruits of leguminous plants, for production of food; Products from legumes; Preparation or treatment thereof
- A23L11/60—Drinks from legumes, e.g. lupine drinks
- A23L11/65—Soy drinks
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
- A23C11/00—Milk substitutes, e.g. coffee whitener compositions
- A23C11/02—Milk substitutes, e.g. coffee whitener compositions containing at least one non-milk component as source of fats or proteins
- A23C11/10—Milk substitutes, e.g. coffee whitener compositions containing at least one non-milk component as source of fats or proteins containing or not lactose but no other milk components as source of fats, carbohydrates or proteins
- A23C11/103—Milk substitutes, e.g. coffee whitener compositions containing at least one non-milk component as source of fats or proteins containing or not lactose but no other milk components as source of fats, carbohydrates or proteins containing only proteins from pulses, oilseeds or nuts, e.g. nut milk
- A23C11/106—Addition of, or treatment with, microorganisms
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L11/00—Pulses, i.e. fruits of leguminous plants, for production of food; Products from legumes; Preparation or treatment thereof
- A23L11/50—Fermented pulses or legumes; Fermentation of pulses or legumes based on the addition of microorganisms
-
- 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
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/51—Bifidobacterium
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Botany (AREA)
- Agronomy & Crop Science (AREA)
- Microbiology (AREA)
- Health & Medical Sciences (AREA)
- Nutrition Science (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Biotechnology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Dairy Products (AREA)
- Beans For Foods Or Fodder (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
Abstract
Description
본 발명은 두유 발효유의 제조 방법 및 제조된 두유 발효유에 관한 것으로, 더욱 상세하게는 비피도박테리움 아이엔티-57 (Bifidobacteriumsp. Int-57)을 이용하여, 생리 활성이 높은 이소플라본 비배당체 (isoflavone aglycone)를 함유한 두유 발효유의 제조 방법 및 제조된 대두 발효유에 관한 것이다.The present invention relates to a method for producing soy milk fermented milk and to a manufactured soy milk fermented milk, and more particularly, using a Bifidobacterium sp. It relates to a method for producing soymilk fermented milk containing isoflavone aglycone) and to a soybean fermented milk produced.
대두는 필수 영양소를 골고루 함유하고 있는 고단백질 식품으로, 필수 아미노산의 조성이 우수하며, 지방의 대부분이 불포화 지방산으로 구성되어 있어 심혈관계 질환을 포함한 각종 질환의 예방 및 치료 효과가 있어 현대에 들어와서 그 이용성이 증대되고 있다.Soybean is a high protein food that contains essential nutrients. Its composition of essential amino acids is excellent, and most of the fat is composed of unsaturated fatty acids, so it is effective in preventing and treating various diseases including cardiovascular diseases. Its usability is increasing.
대두에는 단백질 가수분해효소인 트립신의 저해제와 적혈구 응집소로 렉틴이라고 불리는 단백질이 함유되어 있기는 하나, 이러한 항영양성 물질은 열에 의해서 대부분 파괴되기 때문에 식용에는 별문제가 없다. 대두에는 단백질, 비타민 B군, 철분 이외에 이소플라본 (isoflavone)이라는 식물성 호르몬 (phytoestrogen)이 함유되어 있는데, 이는 여성호르몬인 에스트로겐과 유사한 역할을 하는 대체물질로서, 유방암, 대장암, 전립선암 등의 암질환, 심혈관질환 및 골다공증 등에 유익한 물질이다.Soybean contains a protein hydrolase, an inhibitor of trypsin and a protein called lectin as a hemagglutinin, but since these anti-nutrients are mostly destroyed by heat, there is no problem with food. In addition to proteins, B vitamins, and iron, soybeans contain a phytoestrogen called isoflavone, which is a substitute for estrogen, a female hormone, and is a cancer of breast, colon and prostate cancers. It is a beneficial substance for diseases, cardiovascular diseases and osteoporosis.
이러한 이소플라본은 대두 중에 주로 배당체 형태인 다이드진 (daidzin) 및 제니스틴 (genistin)으로 존재하고 있으며, 이는 대장내의 박테리아가 생성하는 베타-글루코시다제 (β-glucosidase)에 의하여 가수분해되어 이소플라본 비배당체 형태인 다이드제인 (daidzein) 및 제니스테인 (genistein)전환된 후 흡수된다.These isoflavones exist mainly in the glycoside form of diadzin and genistin, which are hydrolyzed by beta-glucosidase produced by bacteria in the colon and isoflavones. The nonglycoside forms of daidzein and genistein are converted and then absorbed.
이소플라본 배당체는 그들의 비배당체 형태에 비하여 그 생리 활성이 떨어지고 흡수율 또한 매우 낮다.Isoflavone glycosides are less physiologically active and have a very low absorption rate compared to their nonglycoside forms.
이러한 대두를 이용한 식품 중 가장 손쉽게 섭취할 수 있는 식품으로는 두유를 들 수 있는데, 두유는 대두의 물 추출물로서 고단백질 식품이나 콜레스테롤 또는 유당은 함유하지 않으므로, 락타제 (lactase)가 결핍되어 우유 중의 유당을 소화시키지 못하는 사람들에게 우유 대체식품으로 이용되어 왔다.Soy milk is one of the most easily consumed foods using soybeans. Soymilk is a water extract of soybean and does not contain high protein foods or cholesterol or lactose, so it is deficient in lactase. It has been used as a milk substitute for people who cannot digest lactose.
그러나, 두유는 특유의 콩비린내 (beany flavor)와 더불어, 인체 내에서 소화되지 않는 알파-디-갈락토실 올리고사카라이드 (α-D-galactosyl oligosaccharides)를 함유하고 있어 복부팽만감을 유발할 수 있어 생으로 대두를 섭취하는 것은 쉽지 않았으며, 이러한 문제점을 극복하고 영양가를 높이기 위하여 발효의 방법이 도입되기도 하였으나, 이를 완전히 극복하지는 못하였다.However, soy milk contains a unique beany flavor and contains indigestible alpha-di-galactosyl oligosaccharides, which can lead to bloating. Ingesting soybeans was not easy, and fermentation methods were introduced to overcome these problems and increase nutritional value, but did not completely overcome them.
또한, 두유에 함유되어 있는 이소플라본 배당체인 다이드진(daidzin) 및 제니스틴 (genistin)을 생리 활성이 높고 소화 흡수가 잘되는 그들의 비배당체 형태인 다이드제인 (daidzein) 및 제니스테인 (genistein)으로 변환하기 위한 시도로서 종래에는 산과 알칼리 가수분해 방법이나 정제된 효소를 이용하였으나, 목적 물질의 파괴 및 고비용 등의 단점을 지니고 있었고, 장내 상주 박테리아를 이용한 생전환 (bioconversion)의 개념을 이용한 방법은 시도되지 않았다.In addition, converting the isoflavone glycosides of diaidzin and genistin in soymilk into their non-glycoside forms, diaidzein and genistein, which are highly physiologically active and well digested and absorbed. In the past, acid and alkali hydrolysis methods or purified enzymes were used, but they had disadvantages such as destruction of target substance and high cost, and no method using bioconversion using intestinal resident bacteria was attempted. .
따라서, 본 발명은 이러한 종래 기술의 문제점을 개선하기 위하여 발명된 것으로서, 두유에 함유되어 있는 이소플라본 배당체가 생리 활성이 높은 비배당체 형태로 100% 전환된 기능성 두유 발효유 제조방법 및 제조된 두유발효유를 제공하고자 한다.Therefore, the present invention has been invented to improve the problems of the prior art, a method for producing a functional soymilk fermented milk 100% isoflavone glycosides contained in soy milk is converted to a non-glycoside form of high physiological activity and the soy milk fermented milk prepared To provide.
본 발명의 또 다른 목적은 두유 특유의 콩비린내와 복부 팽만감 증상이 제거된 두유 발효유의 제조방법 및 제조된 발효유를 제공하고자 한다.Still another object of the present invention is to provide a method for producing soybean milk fermented milk and soybean milk fermented milk which has been eliminated soybean milk specific symptoms and abdominal bloating symptoms.
도 1 은 37℃에서 비피도박테리움 아이엔티-57 (Bifidobacteriumsp. Int-57) 배양시 배양시간에 따른 베타-글루코시다제 (β-glucosidase) 활성 및 세포성장을 도시한 그래프이다.1 is a graph showing beta-glucosidase activity and cell growth according to incubation time when Bifidobacterium sp. Int-57 was incubated at 37 ° C.
도 2 는 37℃에서 두유를 비피도박테리움 아이엔티-57 (Bifidobacteriumsp. Int-57)로 접종시 발효시간에 따른 생균수를 도시한 그래프이다.Figure 2 is a graph showing the viable cell number according to the fermentation time when inoculated with soy milk at 37 ℃ Bifidobacterium sp. Int-57 ( Bifidobacterium sp. Int-57).
도 3 은 37℃에서 두유를 비피도박테리움 아이엔티-57 (Bifidobacteriumsp. Int-57)로 접종시 발효시간에 따른 pH 변화를 도시한 그래프이다.Figure 3 is a graph showing the pH change with the fermentation time when inoculated with soy milk at 37 ℃ Bifidobacterium sp. Int-57.
도 4 는 37℃에서 두유를 비피도박테리움 아이엔티-57 (Bifidobacteriumsp. Int-57)로 접종시 발효시간에 따른 베타-글루코시다제 (β-glucosidase) 활성을 도시한 그래프이다.Figure 4 is a graph showing the beta-glucosidase (β-glucosidase) activity according to the fermentation time when inoculated with soy milk at 37 ℃ Bifidobacterium sp. Int-57.
본 발명은 이러한 목적을 달성하기 위하여, 베타-글루코시다제 (β-glucosidase) 활성 및 알파-갈락토시다제 (α-galactosidase) 활성을 갖는 비피도박테리움 아이엔티-57 (Bifidobacteriumsp. Int-57)을 이용함으로써 두유 내의 이소플라본이 모두 비배당체 형태로 존재하는 생리 활성이 우수하고 복부 팽만감 증상이 해소된 발효된 두유를 제공한다.In order to achieve this object, the present invention provides a Bifidobacterium sp. Int- with beta-glucosidase activity and alpha-galactosidase activity. 57) provides fermented soymilk with excellent physiological activity in which all of the isoflavones in the soymilk are present in nonglycoside form and relieving abdominal bloating.
비피도박테리움 아이엔티-57 (Bifidobacteriumsp. Int-57)은 편성 혐기성균으로 항온 동물의 장내에 서식하면서, 장내 균총을 개선하며, 혈청 콜레스테롤 감소, 잠재적인 병원균의 성장 억제, 면역반응 및 항암 활성을 자극함으로써 건강을 증진시키는 유익한 균이다. Bifidobacterium sp. Int-57 is a organized anaerobic bacterium that lives in the intestinal tract of incubated animals, improves the intestinal flora, reduces serum cholesterol, inhibits the growth of potential pathogens, immune responses and anticancer It is a beneficial bacterium that promotes health by stimulating activity.
본 발명에 따르면, 두유의 제조는 기존의 방법을 일부 변형하여 대두를 정수에 수침시킨 후 정수와 혼합하여 마쇄한 후 비지를 여과하여 제거하고 살균하는 기존의 일반적인 발효조건으로 행할 수 있다.According to the present invention, the production of soy milk can be carried out under the conventional general fermentation conditions in which some of the conventional methods are soaked in soybeans and purified, mixed with purified water, and ground.
발효의 방법은 통상의 발효 조건으로 행할 수 있고, 두유를 발효시키기 위하여 스타터로 비피도박테리움 아이엔티-57 (Bifidobacteriumsp. Int-57)을 접종한 후 pH 5-7, 30-40℃, 바람직하게는 37℃에서 배양시킴으로써 발효된 두유를 얻는다. 이때, 균주가 편성 혐기성임을 감안하여 혐기적인 조건에서 배양하여야 한다.The fermentation method can be carried out under normal fermentation conditions, and after inoculating Bifidobacterium sp. Int-57 with starter to ferment soy milk, pH 5-7, 30-40 ° C., Preferably fermented soymilk is obtained by incubating at 37 ° C. At this time, the strain should be cultured under anaerobic conditions in consideration of being anaerobic.
비피도박테리움 아이엔티-57 (Bifidobacteriumsp. Int-57)은 BHI(brain heart infusion) 배지에서 배양시키나, 편성 혐기성이므로, 시스틴 (cystein)이 첨가된 변형된 BHI 배지에서 배양하는 것이 바람직하다. Bifidobacterium sp. Int-57 is incubated in BHI (brain heart infusion) medium, but is preferably anaerobic, and therefore cultured in modified BHI medium with cysteine added.
두유에 접종된 비피도박테리움 아이엔티-57 (Bifidobacteriumsp. Int-57)은 베타-글루코시다제 (β-glucosidase) 활성을 가지므로, 대두 발효유 내의 이소플라본 배당체를 그들의 비배당체 형태로 100%의 전환율로 전환시킬 뿐만 아니라, 알파-갈락토시다제(α-galactosidase)활성을 가지므로, 대두에 함유된 복부 팽만감의 원인이 되었던 비소화성당인 알파-디 가락토실 올리고사카라이드(α-D-galactosyl oligosacharides)를 분해하여, 결국, 생리 활성이 우수하며, 영양가 풍부하고, 소화 흡수가 용이한 고기능성 대두 발효유를 공급할 수 있다. Bifidobacterium sp. Int-57 in soymilk has beta-glucosidase activity, so 100% of isoflavone glycosides in soybean fermented milk in their nonglycoside form In addition to being converted to a conversion rate of, it has alpha-galactosidase activity, and thus alpha-digaractyl oligosaccharide (α-D), a non-digestible sugar that has caused abdominal bloating in soybeans. -Galactosyl oligosacharides) can be decomposed to provide high functional soybean fermented milk with good physiological activity, nutrition, and easy digestion and absorption.
또한, 대두 중 진품콩 1호는 대두 특유의 콩비린내를 유발하는 에틸비닐케톤류를 생성하는 세 종류의 리폭시게나제 이소자임 (lipoxigenase isozyme)중에서 2 종류(L-2 및 L-3)가 결여되어 있고, 진품콩 2호는 모든 리폭시게나제 이소자임(lipoxigenase isozyme)(L-1, L-2 및 L-3)이 결핍되어 있으므로, 이들을 사용시 대두 제품의 콩비린내를 제거할 수 있으므로, 더욱 바람직하다.In addition, genuine soybean No. 1 in soybeans lacks two kinds (L-2 and L-3) among three kinds of lipoxigenase isozymes that produce ethyl vinyl ketones that cause soybean-specific soybean odor. Genuine soybean 2 is deficient in all lipoxygenase isozymes (L-1, L-2 and L-3). Do.
하기의 실시예에서 본 발명을 더욱 상세하게 설명하고자 한다.In the following Examples will be described the present invention in more detail.
[실시예 1]Example 1
미생물 및 배지조건Microorganisms and Medium Conditions
다섯 종류의 상업적인 비피더스균 (Bifidobacterium animalisATCC 25527,B. adolescentisATCC 15703,B.longum ATCC15707,B. bifidumATCC 29521 및B. infantisATCC 15697, American type Culture Collection에서 구입) 및 건강한 한국인의 분변에서 분리한 10 종류의 비피더스균 (Bifidobacterium sp.Int-57,B.bifidumBGN 2,B. bifidumBGN 3,B. bifidumBGN 4,Bifidobacterium sp.RD 13,Bifidobacteriumsp. SJ 32,Bifidobacteriumsp. RD 54,Bifidobacteriumsp. RD 3,Bifidobacteriumsp. RD 65 및Bifidobacteriumsp. SH5, 서울대학교 식품 영양학과 지근억 박사님으로부터 분양)을 준비하였다.Five types of commercial bifidus (Bifidobacterium animalisATCC 25527,B. adolescentisATCC 15703,B.longum ATCC15707,B. bifidumATCC 29521 andB. infantisATCC 15697, purchased from the American type Culture Collection) and 10 types of bifidus bacteria isolated from the feces of healthy Koreans (Bifidobacterium sp.Int-57,B.bifidumBGN 2,B. bifidumBGN 3,B. bifidumBGN 4,Bifidobacterium sp.RD 13,Bifidobacteriumsp. SJ 32,Bifidobacteriumsp. RD 54,Bifidobacteriumsp. RD 3,Bifidobacteriumsp. RD 65 andBifidobacteriumsp. SH5, Department of Food and Nutrition, Seoul National University).
비피더스균의 배양은 BHI 액체배지(미국 Difco사에서 구입)에 5% L-cystein-HCl(w/w)(미국 Sigma Chemical Co.에서 구입)을 첨가한 변형된 BHI 액체배지를 이용하여 37℃에서 24시간 동안 혐기적 조건 하에서 2회 계대 배양 후 사용하였다. 상기 15종류의 비피더스균을 BHI 액체배지에 1% 되도록 접종한 후 6시간 간격으로 균주의 성장과 β-글루코시다제 활성을 측정하였다.Cultivation of bifidus bacteria was performed at 37 ° C. using a modified BHI liquid medium in which 5% L-cystein-HCl (w / w) (obtained from Sigma Chemical Co., USA) was added to BHI liquid medium (purchased from Difco, USA). It was used after two passages under anaerobic conditions for 24 hours at. After inoculating 15 kinds of Bifidobacteria in 1% BHI liquid medium, the growth of the strain and β-glucosidase activity were measured at 6 hour intervals.
[실시예 2]Example 2
β-글루코시다제 활성의 분석Assay of β-glucosidase Activity
이소플라본 배당체의 대사에 관여하는 β-글루코시다제 활성이 가장 높은 비피더스균을 선별하기 위하여, 이 효소의 활성을 분석하였다.In order to select bifidus bacteria having the highest β-glucosidase activity involved in metabolism of isoflavone glycosides, the activity of this enzyme was analyzed.
분석 방법은 파라-니트로 페닐-베타-글루코피라노사이드 (p-nitrophenyl-β-glucopyranoside)의 가수분해 정도 즉, 생성된 파라-니트로페놀 (p-nitrophenol)을 측정함으로서 분석되었다.The analytical method was analyzed by measuring the degree of hydrolysis of p-nitrophenyl-β-glucopyranoside, that is, the resulting para-nitrophenol.
비피더스균 배양액을 4℃에서 15분 동안 11,000rpm의 속도로 원심분리하고, 0.2M 인산염나트륨 완충액(pH 6.0)으로 두 번 세척하고 동일한 완충액에 재현탁시킴으로써 균체를 회수하였다. 베타-글루코시다제 (β-glucosidase) 활성은 아세톤-톨루엔 용액(부피 비로 9:1) 및 1mM의 파라-니트로페닐-베타-글루코피라노사이드 (p-nitrophenyl-β-glucopyranoside)를 첨가하여 그 혼합물을 45℃에서 반응시킨 후 0.5 M 탄산염나트륨 용액을 첨가하여 반응을 종결시켰다.Cells were recovered by centrifugation of the bifidus culture medium at 11,000 rpm for 15 minutes at 4 ° C, washed twice with 0.2 M sodium phosphate buffer (pH 6.0) and resuspended in the same buffer. Beta-glucosidase activity was determined by adding acetone-toluene solution (volume ratio 9: 1) and 1 mM para-nitrophenyl-beta-glucopyranoside (p-nitrophenyl-β-glucopyranoside). The mixture was reacted at 45 ° C. and then 0.5 M sodium carbonate solution was added to terminate the reaction.
반응액의 상층액에 유리된 파라-니트로페놀의 함량은 모델 DU 530 스펙트로미터 (미국 Beckman에서 구입)을 사용하여 400nm에서 측정되었다. 효소의 1 단위는 1분당 기질로부터 1μmol의 파라-니트로페놀을 유리시킨 효소의 양으로 정의하였다. 각 실험은 3반복 실시하였으며, 그 값은 이들 결과의 평균값으로 나타내었다.The content of free para-nitrophenol in the supernatant of the reaction solution was measured at 400 nm using a model DU 530 spectrometer (purchased from Beckman, USA). One unit of enzyme was defined as the amount of enzyme that liberated 1 μmol of para-nitrophenol from the substrate per minute. Each experiment was repeated three times, and the value is shown as the average value of these results.
실험 결과, 비피도박테리움 아이엔티-57 (Bifidobacteriumsp. Int-57)에서 효소활성이 가장 높게 나타났으므로, 이 균주를 본 발명의 두유 발효시 이용하였다.As a result of the experiment, Bifidobacterium sp. Int-57 showed the highest enzymatic activity, so the strain was used for fermenting soy milk of the present invention.
변형된 BHI 배지에서 베타-글루코시다제 (β-glucosidase) 활성은 도 1에 도시된 바와 같이, 균주의 성장 정지기에 해당하는 18-36 시간 배양 후 최고치에 도달하였으며, 그 후 사멸기 동안은 감소하였다.Beta-glucosidase activity in modified BHI medium reached its highest after 18-36 hours of incubation, which corresponds to the growth stop phase of the strain, as shown in FIG. It was.
또한, 도 1에 도시된 바와 같이, 비피도박테리움 아이엔티-57 (Bifidobacteriumsp. Int-57)의 베타-글루코시다제 (β-glucosidase) 활성은 배양 6시간 후 24.40 mU/ml 이었고, 대수증식기에 갑자기 증가한 후 최고치인 45.70mU/ml에 도달한 후 빠르게 감소하였다. 그러나, 비피도박테리움 아이엔티-57 (Bifidobacteriumsp. Int-57)은 배양기의 마지막에도 높은 베타-글루코시다제 (β-glucosidase) 활성인 15.17 mU/ml를 나타냄을 알 수 있다.In addition, as shown in FIG. 1, beta-glucosidase activity of Bifidobacterium sp. Int-57 was 24.40 mU / ml after 6 hours of culture, It suddenly increased during the growth phase and then rapidly declined after reaching its peak of 45.70 mU / ml. However, it can be seen that Bifidobacterium sp. Int-57 shows 15.17 mU / ml of high beta-glucosidase activity even at the end of the incubator.
[실시예3]Example 3
두유의 제조 및 발효Preparation and Fermentation of Soy Milk
두 종류의 황색 대두, 진품콩 1호 (이하, JP#1라 칭함)와 진품콩 2호 (이하, JP#2라 칭함) 및 두 종류의 검은콩, 검정콩 1호 (이하, BL#1라 칭함)와 검정콩 2호 (이하, BL#2라 칭함)는 한국농촌 진흥청 작물시험장에서 분양 받았다.Two types of yellow soybeans, Genuine Bean No. 1 (hereinafter referred to as JP # 1) and Genuine Bean No.2 (hereinafter referred to as JP # 2) and two types of black and black bean No.1 (hereinafter referred to as BL # 1) ) And Black Bean No. 2 (hereinafter referred to as BL # 2) were distributed by the Korea Rural Development Administration.
상기한 대두를 수세한 후 20℃에서 8시간동안 정수에 수침시킨 후 물은 따라 버리고 수침 시킨 대두를 원료 무게의 10배에 해당하는 정수를 이용하여 분쇄하였다. 면포를 이용하여 여과시킨 후 121℃에서 15분간 가압 멸균하여 두유를 준비하였다.After washing the soybeans, the soaked soybeans were immersed in purified water at 20 ° C. for 8 hours, followed by crushing the soybeans. After filtering using a cotton cloth, soymilk was prepared by autoclaving at 121 ° C. for 15 minutes.
그 후 두유에 비피도박테리움 아이엔티-57 (Bifidobacteriumsp. Int-57)을 106cfu/ml 농도로 접종하여 37℃에서 발효시켰다.Thereafter, Bifidobacterium sp. Int-57 was inoculated in soymilk at a concentration of 10 6 cfu / ml and fermented at 37 ° C.
시료를 6시간 간격으로 24시간 동안 취하여 비피도박테리움 아이엔티-57 (Bifidobacteriumsp. Int-57)의 생균수를 BL 고체배지 (미국 Difco사에서 구입)를 이용하여 측정하였다. 혐기적 배양을 위하여 혐기성 용기 (Anaerobic jar) (미국 Difco사에서 구입)와 가스팩 시스템 (영국 OXOID Ltd에서 구입한 AnaeroGenTMCompact)을 이용하여 37℃에서 2-3일간 배양하였다.Samples were taken at 6 hour intervals for 24 hours and the viable bacterial count of Bifidobacterium sp. Int-57 was measured using a BL solid medium (purchased from Difco, USA). For anaerobic cultivation, an anaerobic jar (purchased from Difco, USA) and a gas pack system (AnaeroGen ™ Compact, purchased from OXOID Ltd, UK) were incubated at 37 ° C. for 2-3 days.
도 2는 각각의 두유 (BL#1 두유, BL#2 두유, JP#1 두유, JP#2 두유)에서 균주의 생균수를 나타낸다. 상기한 도면을 검토 시 12시간의 발효 후 모든 두유에서 비피도박테리움 아이엔티-57 (Bifidobacteriumsp. Int-57)의 생균수는 108cfu/ml 이상이었으며, 이 후 발효 24시간 후에도 뚜렷한 변화를 보이지 않았다.Figure 2 shows the viable bacterial count of each soymilk (BL # 1 soymilk, BL # 2 soymilk, JP # 1 soymilk, JP # 2 soymilk). After 12 hours of fermentation, the viable bacterial count of Bifidobacterium sp. Int-57 was more than 10 8 cfu / ml in all soymilk after 12 hours of fermentation. Did not look.
Shulter-Malyoth 등에 의하면 발효유의 섭취를 통한 건강상의 유익한 효과를 얻기 위해서는 발효유 섭취시 1 ml 당 106-108cfu의 비피더스균을 함유하고 있어야 한다고 지적하고 있고, 비피도박테리움 아이엔티-57 (Bifidobacteriumsp. Int-57)을 이용하여 제조한 본 발명의 두유 발효유 섭취시 이러한 효과를 볼 수 있으므로, 본 균주는 건강상의 유익한 효과를 제공할 수 있는 대두 발효유 스타터임을 알 수 있다.Shulter-Malyoth et al. Pointed out that in order to obtain a health benefit from the consumption of fermented milk, it should contain 10 6 -10 8 cfu of bifidus bacteria per 1 ml of fermented milk, and Bifidobacterium ENT-57 ( Bifidobacterium sp. Int-57) can see this effect when ingesting the soy milk fermented milk of the present invention prepared using the present invention, it can be seen that this strain is a soybean fermented milk starter that can provide a beneficial effect on health.
[실시예 4]Example 4
두유의 pH 및 적정산도PH and titratable acidity of soymilk
각 시료의 pH는 pH 미터로 측정되었다. 적정산도는 0.1N 수산화나트륨 용액을 이용한 중화적정법을 이용하여 측정하였으며, 젖산(lactic acid) %로 나타내었다. 각 실험은 3반복 실시하였으며 결과는 도 3과 같다.The pH of each sample was measured with a pH meter. Titratable acidity was measured by neutralization titration with 0.1 N sodium hydroxide solution and expressed as% lactic acid. Each experiment was repeated three times, and the results are shown in FIG. 3.
두유의 pH는 발효 6시간 후부터 급속히 감소하기 시작하여 18시간 후에는 유의적인 변화를 보이지 않았다. 두유 발효 중의 pH 감소는 JP#1 두유가 2.45 pH 단위 감소로 가장 크게 감소하였고, BL#2 두유, JP#2 두유 및 BL#1 두유에서 각각 2.38, 2.32 및 2.29 pH 단위 감소하였다. Kamaly는B. longum및B. bifidum에 의한 pH 변화가 각각 1.01 및 1.30 pH 단위 감소함을 보고한 바 있는데, 비피도박테리움 아이엔티-57 (Bifidobacteriumsp. Int-57)을 이용한 발효의 경우 pH 감소가 크게 나타남을 알 수 있다.The pH of soymilk began to decrease rapidly after 6 hours of fermentation and did not change significantly after 18 hours. The decrease in pH during soymilk fermentation was the greatest decrease in JP # 1 soymilk with a 2.45 pH unit reduction and 2.38, 2.32 and 2.29 pH units decrease in BL # 2 soymilk, JP # 2 soymilk and BL # 1 soymilk, respectively. Kamaly reported that the pH change by B. longum and B. bifidum decreased by 1.01 and 1.30 pH units, respectively.For fermentation with Bifidobacterium sp. Int-57, It can be seen that the decrease is large.
또한, Angeles 및 Marth는 약 pH 5.7에서 두유가 커드(curd)를 형성하며,Park 및 Lee는 상업적인 요구르트의 pH는 4.2 내지 4.4의 범위인 것을 보고한 바 있는 데, 본 발명의 경우 두유 발효시 pH는 18시간의 발효 후 두유 단백질의 응고로 커드를 형성하는 pH의 범위에 도달할 뿐 만 아니라, 상업적인 요구르트의 pH에 도달하므로, 비피도박테리움 아이엔티-57 (Bifidobacteriumsp. Int-57)를 이용한 대두 발효유는 발효유로서 적합한 pH를 갖는다는 것을 알 수 있다.In addition, Angeles and Marth, soy milk forms a curd (curd) at about pH 5.7, Park and Lee reported that the pH of commercial yogurt ranges from 4.2 to 4.4, in the case of the present invention, soybean milk pH Bifidobacterium sp. Int-57, which not only reaches the pH range that forms curds after coagulation of soy milk protein after 18 hours of fermentation, but also reaches the pH of commercial yogurt. It can be seen that the soybean fermented milk used has a pH suitable as fermented milk.
적정산도의 경우, BL#1 두유, BL#2 두유 및 JP#1 두유의 경우 초기에는 0.18%에서 발효의 종반에는 각각 1.21, 1.15, 및 1.08%로 증가하였다. 또한 JP#2 두유의 적정산도는 24시간 발효 후 비교적 낮은 0.9%로 증가하였다.In the case of the titratable acidity, BL # 1 soymilk, BL # 2 soymilk, and JP # 1 soymilk increased from 0.18% initially to 1.21, 1.15, and 1.08% at the end of fermentation, respectively. Also, the titratable acidity of JP # 2 soymilk increased to 0.9% after 24 hours fermentation.
[실시예 5]Example 5
두유 발효중의 이소플라본의 분석 및 베타-글루코시다제 활성Analysis of Isoflavones and Beta-glucosidase Activity in Soymilk Fermentation
이소플라본의 분석은 HPLC를 이용하여 실시하였으며, 제니스틴, 다이드제인 및 제니스테인의 표준품은 미국 Sigma Chemical Co.에서 구입하였으며, 다이드진의 표준품은 미국의 Indofine 으로부터 구입하였다.Analysis of isoflavones was performed using HPLC, and standard of Genistin, Dyzedein and Genistein were purchased from Sigma Chemical Co., USA, and Standard of Dydzin was purchased from Indofine, USA.
상기한 각 두유 발효유는 80% 에탄올을 이용하여 50℃에서 24시간동안 추출한 후 상층액을 필터 시스템으로 여과한 후 그 여과액을 HPLC로 분석하였다.Each of the soy milk fermented milk was extracted with 80% ethanol at 50 ° C. for 24 hours, and then the supernatant was filtered through a filter system, and the filtrate was analyzed by HPLC.
이동상은 0.1%의 아세트산을 함유한 아세토니트릴 및 0.1%의 아세트산을 함유한 3차 증류수를 이용하였다. 시료 20㎕를 주입 후 아세토니트릴을 15%에서 35%로 50분에 걸쳐 증가시키고 10분 동안 35%에서 유지시켰다. 이동상의 유속은 1㎖/분으로 하였으며, 이소플라본은 254nm에서 측정되었다. 두유 발효중의 이소플라본 함량 및 조성 변화는 하기 표 1과 같다.The mobile phase used acetonitrile containing 0.1% acetic acid and tertiary distilled water containing 0.1% acetic acid. After injection of 20 μl acetonitrile was increased from 15% to 35% over 50 minutes and held at 35% for 10 minutes. The flow rate of the mobile phase was 1 ml / min, and isoflavones were measured at 254 nm. The isoflavone content and composition change during soymilk fermentation are shown in Table 1 below.
* n.d.: 발견되지 않음* n.d .: not found
상기한 표에서 알 수 있는 바와 같이, 두유에 함유되어 있는 이소플라본 배당체인 다이드진 및 제니스틴은 비교적 짧은 18시간의 발효 후 비피도박테리움 아이엔티-57 (Bifidobacteriumsp. Int-57)에 의하여 생성된 베타-글루코시다제 (β-glucosidase)에 의한 가수분해 반응에 의하여, 100% 완전히 가수분해되어 다이드제인 및 제니스테인으로 전환됨을 알 수 있다.As can be seen from the above table, the isoflavone glycosides Dydzin and Genistin contained in soy milk were treated by Bifidobacterium sp. Int-57 after a relatively short 18-hour fermentation. By hydrolysis reaction with the resulting beta-glucosidase, it can be seen that 100% is completely hydrolyzed to be converted to dydzein and genistein.
또한, 도 4에 도시된 바와 같이, 두유 발효 중의 베타-글루코시다제 (β-glucosidase) 활성은 12시간 발효 후 급격히 증가되었고 24시간 발효 후 BL#1 두유, BL#2 두유, JP#2 두유에서 베타-글루코시다제 (β-glucosidase) 활성은 각각 59.528, 40.643, 70.844 및 56.962mU/ml로 증가되었다.In addition, as shown in Figure 4, beta-glucosidase activity in soymilk fermentation was rapidly increased after 12 hours fermentation and BL # 1 soy milk, BL # 2 soy milk, JP # 2 soy milk after 24 hours fermentation Β-glucosidase activity was increased to 59.528, 40.643, 70.844 and 56.962mU / ml, respectively.
이상에서 살펴본 바와 같이, 비피도박테리움 아이엔티-57 (Bifidobacteriumsp. Int-57)은 두유 발효를 위한 스타터 균주로 적합하고, 이를 이용한 두유 발효유는 하기와 같은 효과를 갖는다.As described above, Bifidobacterium sp. Int-57 is suitable as a starter strain for fermenting soy milk, and fermented soy milk using the same has the following effects.
첫째, 비피도박테리움 아이엔티-57 (Bifidobacteriumsp. Int-57)은 높은 베타-글루코시다제 (β-glucosidase) 활성을 가지므로, 대두 내의 이소플라본 배당체를 생리 활성이 우수하고, 체내 흡수율이 높은 이소플라본 비배당체 형태로 100% 전환시켜, 기능성 두유 발효유를 공급하게 된다.First, Bifidobacterium sp. Int-57 has high beta-glucosidase activity, so isoflavone glycosides in soybean have excellent physiological activity and absorption rate in the body. 100% conversion to the high isoflavone nonglycoside form provides a functional soymilk fermented milk.
둘째, 비피도박테리움 아이엔티-57 (Bifidobacteriumsp. Int-57)은 알파-갈락토시다제 (α-galactosidase) 활성을 가지므로, 두유 섭취시 문제가 될 수 있는 난소화성당인 알파-디-갈락토실 올리고사카라이드 (α-D-galactosyl-oligosaccharides)를 분해 시켜 소화 흡수가 용이한 두유 발효유를 제공하게 된다.Second, Bifidobacterium sp. Int-57 has alpha-galactosidase activity, so alpha-di, an indigestible sugar that can be a problem when ingesting soy milk. Decomposes galactosyl oligosaccharides (α-D-galactosyl-oligosaccharides) to provide soymilk fermented milk that is easy to digest and absorb.
셋째, 진품콩 1호 및 진품콩 2호는 두유 특유의 콩비린내를 유발하는 리폭시게나제 이소자임(lipoxigenase isozyme)이 결핍되어 있으므로, 이를 사용하여 제조한 두유 발효유는 대두 특유의 비릿한 맛 또한 없앨 수 있는 효과를 가진다.Third, since Genuine Beans 1 and 2 are deficient in lipoxigenase isozyme, which causes soybean oil-specific soybean odor, soy milk fermented milk prepared using it can also eliminate soybean's peculiar taste. Has the effect.
따라서, 본 발명은 실험을 통하여, 콩비린내를 유발하는 효소가 결핍된 진품콩으로 두유를 제조하고, 비피도박테리움 아이엔티-57 (Bifidobacteriumsp. Int-57)을 접종하여 이 균주가 갖는 높은 베타-글루코시다제 (β-glucosidase) 활성, 알파-갈락토시다제 (α-galactosidase) 활성 및 발효유 제조시 스타터로서의 적합성을 이용하여 두유 발효유를 제조함으로서, 두유 중의 이소플라본 배당체를 생리활성이 높은 이소플라본 비배당체로 완전 전환시키고 소화흡수력이 뛰어난 고기능성 두유 발효유를 공급하는 효과를 갖는다.Therefore, the present invention through the experiment, soybean milk prepared with genuine soybeans lacking the enzyme causing soybeany, and inoculated with Bifidobacterium sp. Int-57 ( Bifidobacterium sp. Int-57) high Soymilk fermented milk is prepared by using beta-glucosidase activity, alpha-galactosidase activity, and suitability as a starter in fermented milk production. It is completely converted into isoflavone non-glycoside and has a high functional soymilk fermented milk with excellent digestive absorption ability.
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CN103229838A (en) * | 2013-04-01 | 2013-08-07 | 邵爱珍 | Hawthorn kernel papaya fermented soybean milk and preparation method thereof |
CN103229841A (en) * | 2013-04-01 | 2013-08-07 | 邵爱珍 | Hawthorn kernel red date fermented soybean milk and preparation method thereof |
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KR101146102B1 (en) * | 2009-09-10 | 2012-05-16 | 재단법인 강릉과학산업진흥원 | method for manufacturing whole soybean milk containing high content of active isoflavon |
CN103229838A (en) * | 2013-04-01 | 2013-08-07 | 邵爱珍 | Hawthorn kernel papaya fermented soybean milk and preparation method thereof |
CN103229841A (en) * | 2013-04-01 | 2013-08-07 | 邵爱珍 | Hawthorn kernel red date fermented soybean milk and preparation method thereof |
CN103229837A (en) * | 2013-04-01 | 2013-08-07 | 邵爱珍 | Hawthorn kernel mulberry fermented soybean milk and preparation method thereof |
CN113558193A (en) * | 2021-06-24 | 2021-10-29 | 苏州大学 | Preparation method of fermented beverage rich in soybean isoflavone aglycone |
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