KR20110081760A - Composition for inhibiting differentiation and accumulation of fat cells comprising fermented extract of purple sweet potato as an active ingredient - Google Patents
Composition for inhibiting differentiation and accumulation of fat cells comprising fermented extract of purple sweet potato as an active ingredient Download PDFInfo
- Publication number
- KR20110081760A KR20110081760A KR1020100134573A KR20100134573A KR20110081760A KR 20110081760 A KR20110081760 A KR 20110081760A KR 1020100134573 A KR1020100134573 A KR 1020100134573A KR 20100134573 A KR20100134573 A KR 20100134573A KR 20110081760 A KR20110081760 A KR 20110081760A
- Authority
- KR
- South Korea
- Prior art keywords
- sweet potato
- purple sweet
- extract
- composition
- accumulation
- Prior art date
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- A61P3/04—Anorexiants; Antiobesity agents
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P1/00—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes
- C12P1/02—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes by using fungi
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- 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
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/10—Preparation or pretreatment of starting material
- A61K2236/19—Preparation or pretreatment of starting material involving fermentation using yeast, bacteria or both; enzymatic treatment
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/331—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using water, e.g. cold water, infusion, tea, steam distillation, decoction
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
Abstract
Description
본 발명은 자색고구마 추출물 발효액을 이용하는 지방 생성 및 축적 저해용 조성물, 또는 이를 이용한 건강기능식품 및 의약에 관한 것이다.
The present invention relates to a composition for inhibiting fat production and accumulation using a purple sweet potato extract fermentation broth, or a health functional food and medicine using the same.
최근 식생활의 서구화에 따라 영양과다 등의 원인으로 비만이 증가하고 있는데 비만은 동맥경화증의 위험인자의 하나이고, 또한 당뇨병이나 고혈압 등과도 관련이 있어, 심각한 문제가 되고 있다. 비만은 신체에 지방이 과잉되게 축적된 상태이며, 지방이 체내에 축적되는 원인은 당질의 과잉섭취 또는 지방을 과잉섭취 함으로써 음식물 중에 함유되는 당질이 소화되어 단당이 되고 소장을 통해 체내로 흡수되며, 혈당이 상승하여 그 자극으로 분비되는 인슐린이 지방세포에 작용해서 혈액 중의 단당을 지방세포에 받아들이게 해서 지방으로 바꾸는 것이다. 또한 식품성분 중에서 가장 고칼로리인 지방은 췌장 리파아제에 의해 분해되어 소장을 통해 흡수되는 것이며, 섭취 칼로리의 과잉은 저장 칼로리를 증가하도록 작용하여, 저장 칼로리가 증가되는 결과 과잉 지방섭취에 의해 비만에 달하게 된다. Recently, obesity is increasing due to overnourishment due to westernization of diet, obesity is one of the risk factors of atherosclerosis, and also associated with diabetes or hypertension, which is a serious problem. Obesity is a state in which excess fat is accumulated in the body, and the cause of fat accumulation in the body is excessive ingestion of sugar or excess intake of fat, the sugars contained in food are digested into monosaccharides and absorbed into the body through the small intestine, Increasing blood sugar and insulin secreted by the stimulus acts on fat cells, which allows the sugars in the blood to be taken up by the fat cells and converted into fat. In addition, fat, the highest calorie among the food ingredients, is broken down by pancreatic lipase and absorbed through the small intestine, and the excess of calories intake acts to increase stored calories, resulting in increased stored calories, leading to obesity due to excess fat intake. do.
이러한 비만을 억제하기 위해서, 비만에 달하는 경로의 일부분을 저해함으로써, 항비만 작용을 발생시키려는 생각을 바탕으로 현재 각종 항비만제에 관한 연구가 진행되고 있다. 즉, 당질 과잉섭취로 비만에 달하는 경로를 저해하는, 당질분해 소화요소 저해작용, 혈당상승 억제작용, 또는 단당흡수 억제작용에 의해, 혹은 지방 과잉 섭취에 의해 비만에 달하는 경로를 저해하는, 콜산 흡착 배설작용, 콜레스테롤 지하작용, 혈중 트리글리세리드 저하작용, 또는 리파아제 저해작용에 의해 비만을 예방, 개선할 수 있다고 생각되어, 이들 작용을 가지는 의약성분의 연구가 수 없이 행해지고 있다.In order to suppress such obesity, studies on various anti-obesity agents are currently being conducted based on the idea of inhibiting a part of the pathway leading to obesity and causing anti-obesity action. That is, cholic acid adsorption, which inhibits the path leading to obesity due to excessive ingestion of glucose, inhibits the path leading to obesity by inhibiting glycolysis digestion factor, inhibiting blood glucose increase, or inhibiting monosaccharide absorption, or by excessive fat intake. It is thought that obesity can be prevented and improved by excretion, cholesterol underground action, blood triglyceride lowering action, or lipase inhibitory action, and many researches on pharmaceutical ingredients having these actions have been conducted.
포화지방산과 콜레스테롤의 섭취량이 많은 구미제국이면서도, 프랑스 사람은 다른 구미제국의 사람들보다 심장질환 발병률이 낮으며, 이 현상을 프렌치 파라독스로 알려져 있다. 프렌치 파라독스 요인의 하나는 프랑스 사람에게서 볼 수 있는 적와인의 높은 섭취량에 의한다는 것이 제기되어 그 성분의 탐색이 활발하게 행하여졌다. 동맥경화와 밀접하게 관련하는 LDL의 산화가 적와인에 함유된 폴리페놀에 의하여 억제된다는 것이 발견되어 백와인에 비하여 적와인의 높은 폴리페놀 함량이 LDL 의 산화억제와 밀접하게 관련된 것이 발견되었다. 나아가서 폴리페놀로서의 안토시아닌 그중 마루빈, 델피니딘, 사이니딘, 페라루고니딘 등이 LDL 산화를 억제한다는 것이 발견되어 지금까지 거의 상세한 연구가 행하여지지 않은 안토시아닌의 생체 내에서의 생리기능에 많은 관심이 모아지고 있다.
Despite the fact that Western countries have high intakes of saturated fatty acids and cholesterol, French people have a lower incidence of heart disease than other Western countries. This phenomenon is known as French paradox. One of the factors of French paradox was that it was based on the high intake of red wine found in French people, and the exploration of the ingredient was actively conducted. It was found that the oxidation of LDL, which is closely related to atherosclerosis, was inhibited by polyphenols contained in red wine, and the higher polyphenol content of red wine was found to be closely related to the oxidation inhibition of LDL compared to white wine. Furthermore, it has been found that anthocyanins as polyphenols, such as marvin, delphinidin, cinidine, and ferragonidine, inhibit LDL oxidation. Thus, much attention has been paid to the physiological functions of anthocyanins in vivo, which have not been studied so far. ought.
한편, 자색고구마(자미고구마; Purple-Fleshed sweet potato)는 1993년 가공용 다수성 품종 건미를 모본으로하고 자색 색소를 함유한 야마까와무라사끼를 부본으로 인공 교배하여 생산력검정시험과 지역적응시험에서 다수성이 확인되어 품종교배를 통해 지난 1998년 자미, 2000년 보라미, 2001년에 신자미 등 3가지 신품종으로 육성되었다. 이들 신품종은 야생 고구마의 고유 성분은 모두 간직하고 있으면서 크기는 일반 고구마와 비슷한 반면 수확량은 야생보다 2배 이상 많고, 고구마 고유의 당질과 섬유소를 가지고 있다는 장점을 지니고 있다. 자색고구마는 색소추출물을 다량 함유하고 있어 표피층뿐만 아니라 육질 전체가 진한 자색을 띠고 있는데, 이 기능성 색소함량이 크린베리 보다는 10 ~ 60배, 포도보다는 5 ~ 7배 높아 천연 기능성 색소 원뿐만 아니라 기능성 식품소재로서 가치가 우수하다.On the other hand, purple sweet potato (Purple-Fleshed Sweet Potato) was artificially crossed in 1993 using a variety of dried varieties of processed varieties of dry brown rice and purple pigment containing Yamakawamurasaki in the production test and local adaptation test. Multiplicity was confirmed, and breeding was carried out in three new varieties: Jami in 1998, Borami in 2000, and Shinjami in 2001. These new varieties have all the constituents of wild sweet potatoes and are similar in size to regular sweet potatoes, while yielding more than twice the yield of wild sweet potatoes. The purple sweet potato contains a large amount of pigment extract, so that not only the epidermal layer but also the entire flesh has a deep purple color.The functional pigment content is 10 to 60 times higher than a cream berry and 5 to 7 times higher than a grape berry, as well as a functional food source. Excellent value as a material.
이러한 자색고구마의 주요 활성성분은 플라보노이드계의 색소인 안토시아닌으로 항산화작용 (J Agric. Food Chem. 51, pp 628-633, 2003; J Nat. Prod. 62, p 802, 1999; J Med. Food. 4 pp 211-218, 2001), 항 당뇨(J Agric Food Chem. 49, pp 1948-1951, 2001), 항암작용(Cancer Lett. in press pp 1-12, 2005) 효과 등이 있는 것으로 보고되어 있다. 또한 최근 아세트아미노펜과 같은 간 손상을 유발하는 화학물질의 대사에 관여된 간의 약물대사 효소인 시토크롬 P450의 효소들 (P450 1A1, P450 2E1)의 활성을 억제하고, 간손상을 유발시키는 화학물질의 대사체의 포합반응을 통한 체외 방출을 촉진하는 해독효소를 증가시킴으로써, 간질환의 예방 및 치료에 유용하게 사용될 수 있다는 보고도 있다(한국공개특허 제2007-0079637호).
The main active ingredient of these purple sweet potatoes is anthocyanin, a flavonoid pigment (J Agric. Food Chem. 51, pp 628-633, 2003; J Nat. Prod. 62, p 802, 1999; J Med. Food. 4 pp 211-218, 2001), anti-diabetic (J Agric Food Chem. 49, pp 1948-1951, 2001), and anticancer effects (Cancer Lett. In press pp 1-12, 2005). . It also inhibits the activity of cytochrome P450 enzymes (P450 1A1, P450 2E1), a liver metabolic enzyme involved in the metabolism of liver-damaging chemicals such as acetaminophen, and metabolism of chemicals that cause liver damage. There is also a report that by increasing the detoxification enzymes that promote the release of the body through the body reaction reaction can be useful for the prevention and treatment of liver disease (Korean Patent Publication No. 2007-0079637).
본 발명은 자색고구마의 추출물 발효액을 이용한 지방 생성 및 축적 저해용 조성물을 제공하는 것이다.
The present invention provides a composition for inhibiting fat formation and accumulation using the extract fermentation broth of purple sweet potato.
본 발명의 지방 생성 및 축적 저해용 조성물은 자색고구마 추출물을 효모로 발효시켜 얻은 발효액을 유효성분으로 함유하는 것을 특징으로 한다.The composition for inhibiting fat production and accumulation of the present invention is characterized by containing a fermentation broth obtained by fermenting the purple sweet potato extract as yeast as an active ingredient.
본 발명의 지방 생성 및 축적 저해용 조성물에서 상기 자색고구마 추출물은 열수추출물 또는 탄소수 2 ~ 4의 저급 알코올 수용액의 추출물인 것을 특징으로 한다.The purple sweet potato extract in the composition for inhibiting fat production and accumulation of the present invention is characterized in that the hot water extract or an extract of a lower alcohol aqueous solution of 2 to 4 carbon atoms.
본 발명의 지방 생성 및 축적 저해용 조성물에서 상기 열수추출물은 80 ~ 105 ℃에서 0.5 ~ 24 시간 추출한 것을 특징으로 한다.In the composition for inhibiting fat formation and accumulation of the present invention, the hot water extract is characterized in that extracted for 0.5 to 24 hours at 80 ~ 105 ℃.
본 발명의 지방 생성 및 축적 저해용 조성물에서 상기 효모는 이사켄키아 오리엔탈리스(Issatchenkia orientalis)인 것을 특징으로 한다.In the composition for inhibiting fat production and accumulation of the present invention, the yeast is characterized in that Issatchenkia orientalis .
본 발명의 지방 생성 및 축적 저해용 조성물에서 상기 자색고구마 추출물에 효모를 접종한 후 25 ~ 40 ℃에서 18 ~ 78 시간 발효시키는 것을 특징으로 한다.After inoculating yeast in the purple sweet potato extract in the composition for inhibiting fat production and accumulation of the present invention, the fermentation is carried out at 25 to 40 ° C. for 18 to 78 hours.
본 발명의 지방 생성 및 축적 저해용 조성물은 자색고구마를 80 ~ 105 ℃에서 0.5 ~ 24 시간 열수추출하고, 상기 열수추출물에 이사첸키아 오리엔탈리스(Issatchenkia orientalis)를 접종한 후 25 ~ 40 ℃에서 18 ~ 78 시간 발효시킨 발효액을 유효성분으로 포함하는 것을 특징으로 한다.In the composition for inhibiting fat formation and accumulation of the present invention, purple sweet potato is extracted with hot water at 80 to 105 ° C. for 0.5 to 24 hours, and the hot water extract is inoculated with Issatchenkia orientalis at 18 to 25 to 40 ° C. It is characterized in that it comprises a fermentation broth fermented ˜78 hours as an active ingredient.
본 발명의 지방 생성 및 축적 저해용 조성물에서 상기 자색고구마는 야마까와무라사끼, 연자미, 보라미 및 신자미 중에서 선택된 어느 하나 이상의 품종의 고구마인 것을 특징으로 한다.The purple sweet potato in the composition for inhibiting fat production and accumulation of the present invention is characterized in that the sweet potato of any one or more varieties selected from Yamakawamurasaki, Yeonmi, Borami and Shinjami.
본 발명의 지방 생성 및 축적 저해용 조성물에서 상기 지방 생성 및 축적 저해용 조성물은 총 중량을 기준으로 발효액이 0.01 ~ 50 중량%로 함유된 것을 특징으로 한다.In the composition for inhibiting fat production and accumulation of the present invention, the composition for inhibiting fat production and accumulation is characterized in that the fermentation broth is contained in an amount of 0.01 to 50% by weight based on the total weight.
본 발명의 지방 생성 및 축적 저해용 조성물은 의약인 것을 특징으로 한다.The composition for inhibiting fat production and accumulation of the present invention is characterized by being a medicament.
본 발명의 건강기능식품은 자색고구마 추출물을 효모로 발효시켜 얻은 발효액을 유효성분으로 함유하는 것을 특징으로 한다.
The health functional food of the present invention is characterized by containing the fermentation broth obtained by fermenting the purple sweet potato extract with yeast as an active ingredient.
본 발명의 자색고구마 추출물 발효액은 아지방세포를 이용한 항비만 실험에서 지방세포로의 전환과 지방 축적을 강하게 억제하고, 지방 세포 분화에 관련된 PPARγ와 C/EBPα의 발현을 감소시키고, 마우스 동물모델에서 고지방식이로 인한 체중 증가 및 피하지방의 증가를 억제하고, 간조직의 무게, 간조직내 트리글리세라이드 함량을 낮추며, 혈중 글루코오스 농도를 저하시키고, 간조직내 지방합성효소의 활성을 낮추고 AMPK 활성을 증대시킨다.
The purple sweet potato extract fermentation broth of the present invention strongly inhibits the conversion to adipocytes and fat accumulation in anti-obesity experiments using sub-fat cells, reduces the expression of PPARγ and C / EBPa related to adipocyte differentiation, and is known in mouse animal models. It suppresses the weight gain and subcutaneous fat increase caused by the diet, lowers the weight of liver tissue, triglyceride content in liver tissue, lowers blood glucose concentration, lowers the activity of fat synthase in liver tissue and increases AMPK activity. Let's do it.
도 1은 본 발명에서 분리한 YPD Large균의 18s rDNA partial sequencing 계통 분석표이다.
도 2는 본 발명에서 분리한 YPD Small균의 18s rDNA partial sequencing 계통 분석표이다.
도 3은 제조예 2에서 발효시간에 따른 발효액의 pH 변화를 나타낸 그래프이다.
도 4는 제조예 2에서 발효시간에 따른 530 nm에서의 흡광도 변화를 나타낸 그래프이다.
도 5는 본 발명의 실험예 2의 지방축적 억제 효과 확인시험 과정의 개략도이다.
도 6은 본 발명의 실시예 1, 비교예 1 및 대조군을 첨가하여 아지방세포를 지방세포로 분화시킨 후 지방세포를 오일레드-O 염색하고, 탈색시켜 500 nm에서 상대 흡광도를 비교한 그래프이다.
도 7은 본 발명의 실시예 1 및 대조군과 함께 사카로미세스 세레비지에와 다른 이사켄키아 오리엔탈리스(KCTC 17696)으로 실시예1과 동일하게 발효시킨 발효액 분말을 첨가하여 아지방세포를 지방세포로 분화시킨 후 지방세포를 오일레드-O 염색하고, 탈색시켜 500 nm에서 상대 흡광도를 비교한 그래프이다.
도 8은 아지방세포의 지방세포 분화에 관여하는 전사인자의 흐름을 나타낸 개략도이다.
도 9는 본 발명의 실시예 1 및 비교예 1의 C/EBPα의 mRNA 발현량을 비교한 그래프이다.
도 10은 본 발명의 실시예 1 및 비교예 1의 PPARγ의 mRNA 발현량을 비교한 그래프이다.
도 11은 고농도 포도당으로 유도된 인슐린 저항성 세포주의 세포내 지질을 나일레드로 염색한 사진(좌측)과, 대조군과 인슐린 저항성 세포주의 나일레드에 의한 580 nm/ 535 nm의 비를 나타낸 그래프(우측)이다.
도 12는 β-엑틴과 지방합성효소의 발현량을 인슐린 저항성 세포주의 세포분해물의 웨스턴 블랏으로 확인한 사진이다.
도 13은 β-엑틴에 대한 지방합성효소의 상대 발현 정도를 나타낸 그래프(우측)이다.
도 14는 본 발명의 실험예 5의 고지방식이 마우스에서의 항비만 효과를 확인하는 실험 과정의 개략도이다.
도 15는 본 발명의 실험예 5의 각 실험군의 4주 후 마우스의 체중을 나타낸 그래프이다.
도 16은 본 발명의 실험예 5의 각 실험군의 4주 후 마우스의 간조직의 무게를 나타낸 그래프이다.
도 17은 본 발명의 실험예 5의 각 실험군의 4주 후 마우스의 희생전의 사진과 희생 후 적출한 간조직을 보여주는 사진이다.
도 18은 본 발명의 실험예 5의 각 실험군의 4주 후 마우스의 피하지방의 모습을 나타낸 사진이다.
도 19는 본 발명의 실험예 5의 각 실험군의 4주 후 마우스의 간조직내 트리글리세라이 함량을 나타낸 그래프이다.
도 20은 본 발명의 실험예 5의 각 실험군의 4주 후 마우스의 간조직내 총 콜레스테롤 함량을 나타낸 그래프이다.
도 21은 본 발명의 실험예 5의 각 실험군의 4주 후 마우스의 혈중 글루코오스 농도를 나타낸 그래프이다.
도 22는 본 발명의 실험예 5의 각 실험군의 4주 후 마우스의 지방합성효소(Fatty acid synthase; FAS) 및 β-엑틴 발현을 웨스턴 블랏으로 확인한 결과이다.
도 23은 본 발명의 실험예 5의 각 실험군의 4주 후 마우스의 p-AMPK 및 β-엑틴 발현을 웨스턴 블랏으로 확인한 결과이다.1 is a 18s rDNA partial sequencing phylogenetic table of YPD Large bacteria isolated in the present invention.
Figure 2 is a 18s rDNA partial sequencing phylogenetic table of the YPD Small bacteria isolated in the present invention.
Figure 3 is a graph showing the pH change of the fermentation broth with fermentation time in Preparation Example 2.
Figure 4 is a graph showing the change in absorbance at 530 nm with the fermentation time in Preparation Example 2.
Figure 5 is a schematic diagram of the process of confirming the fat accumulation inhibitory effect of Experimental Example 2 of the present invention.
Figure 6 is a graph comparing the relative absorbance at 500 nm after the differentiation of sub-fat cells into adipocytes by the addition of Example 1, Comparative Example 1 and the control group of the adipocytes with oil red-O staining, bleaching .
7 is a fat cell by adding a fermentation broth powder fermented in the same manner as in Example 1 with Saccharomyces cerevisiae and another isacyanchia orientalis (KCTC 17696) together with Example 1 and the control group of the present invention. After differentiation, the adipocytes were stained with oil red-O and decolorized to compare relative absorbance at 500 nm.
8 is a schematic diagram showing the flow of transcription factors involved in adipocyte differentiation of sub-fat cells.
9 is a graph comparing mRNA expression levels of C / EBPa of Example 1 and Comparative Example 1 of the present invention.
10 is a graph comparing mRNA expression levels of PPARγ in Example 1 and Comparative Example 1 of the present invention.
FIG. 11 is a photograph (left) of staining intracellular lipids of a high glucose-induced insulin resistant cell line with Nile red, and a graph showing the ratio of 580 nm / 535 nm by Nile red of the control and insulin resistant cell lines (right). to be.
12 is a photograph confirming the expression level of β-actin and lipase by Western blot of the cell lysate of insulin resistant cell line.
Figure 13 is a graph (right) showing the relative expression level of liposynthase to β-actin.
14 is a schematic diagram of an experimental procedure for confirming the anti-obesity effect in the high fat diet of Experimental Example 5 of the present invention.
15 is a graph showing the body weight of the mouse after 4 weeks of each experimental group of Experimental Example 5 of the present invention.
16 is a graph showing the weight of liver tissue of the mouse after 4 weeks of each experimental group of Experimental Example 5 of the present invention.
FIG. 17 is a photograph showing the pre-sacrifice picture of the mouse and the liver tissue extracted after the sacrifice after 4 weeks of each experimental group of Experimental Example 5 of the present invention.
18 is a photograph showing the subcutaneous fat of the mouse after 4 weeks of each experimental group of Experimental Example 5 of the present invention.
19 is a graph showing triglyceride content in liver tissues of mice after 4 weeks of each experimental group of Experimental Example 5 of the present invention.
20 is a graph showing the total cholesterol content in the liver tissue of mice after 4 weeks of each experimental group of Experimental Example 5 of the present invention.
Figure 21 is a graph showing the blood glucose concentration of the mouse after 4 weeks of each experimental group of Experimental Example 5 of the present invention.
FIG. 22 shows the results of Western blot analysis of the expression of Fatty Acid Synthase (FAS) and β-actin in mice after 4 weeks of each experimental group of Experimental Example 5 of the present invention. FIG.
23 shows the results of Western blot expression of p-AMPK and β-actin of mice after 4 weeks of each experimental group of Experimental Example 5 of the present invention.
본 발명의 지방 생성 및 축적 저해용 조성물은 자색고구마 추출물을 효모로 발효시켜 얻은 발효액을 유효성분으로 함유한다.The composition for inhibiting fat formation and accumulation of the present invention contains a fermentation broth obtained by fermenting the purple sweet potato extract with yeast as an active ingredient.
본 발명에서 자색고구마는 일반 고구마에 비해 안토시아닌 함량이 높은 것으로, 특별히 한정하지는 않으나 야마까와무라사끼, 연자미, 보라미 또는 신자미가 바람직하고, 더욱 바람직하게는 야마까와무라사끼 또는 신자미가 바람직하며, 가장 바람직하게는 신자미이다.In the present invention, purple sweet potato is higher in anthocyanin content than ordinary sweet potato, and is not particularly limited, but is preferably Yamakawamurasaki, lotus, purple or shinjami, and more preferably Yamakawamurasaki or shinjami. And most preferably it is deity.
본 발명의 자색고구마 추출물은 효모에 의해 이용되는 기질로서 당분과 함께 안토시아닌이 함량이 높은 추출물이 바람직하다.The purple sweet potato extract of the present invention is preferably an extract having a high content of anthocyanin together with sugar as a substrate used by the yeast.
본 발명의 자색고구마 추출물은 열수추출물이 바람직하다. 예를 들어, 80 ~ 105 ℃, 바람직하게는 90 ~ 100 ℃의 물에서 0.5 ~ 24 시간, 바람직하게는 1 ~ 6 시간 추출할 수 있다. 상기 추출온도 하한치 미만에서는 안토시아닌의 추출효율이 저하되고 잡균에 대한 살균 효과를 충분히 달성할 수 없고, 이로 인하여 추출시간을 길게하면 오히려 잡균이 증식할 수 있다는 문제가 있고, 상기 추출온도 상한치를 초과하면 잡균의 살균 효율은 증대되지만 안토시아닌이 파괴되어 추출물의 색이 탁해진다. 상기 추출시간의 하한치 미만에서는 충분한 안토시아닌의 추출이 어려워 원재료의 손실과 최종산물의 수율저하라는 문제가 발생하고, 상기 추출시간을 증대하는 경우에는 더 이상 추출효율은 증대되지 않으면서 안토시아닌의 파괴를 가져올 수 있다. 자색고구마의 안토시아닌 추출효율을 증대시키기 위하여 산을 첨가하여 물의 pH를 2 ~ 6, 바람직하게는 pH 3 ~ 5 정도의 산성화시키는 것이 바람직하다. 물의 pH를 낮추기 위하여 특별히 한정하지는 않지만 구연산, 젖산, 호박산, 사과산, 말산, 초산 등의 유기산을 사용할 수 있다.Purple sweet potato extract of the present invention is preferably hot water extract. For example, it can be extracted in water of 80 to 105 ℃, preferably 90 to 100 ℃ 0.5 to 24 hours, preferably 1 to 6 hours. If the extraction temperature is lower than the lower limit, the extraction efficiency of anthocyanin is lowered, and the bactericidal effect on various bacteria cannot be sufficiently achieved. Therefore, if the extraction time is increased, various bacteria can grow. If the extraction temperature is exceeded, The sterilization efficiency of various bacteria is increased, but anthocyanins are destroyed and the color of the extract becomes cloudy. If the anthocyanin is less than the lower limit of the extraction time, it is difficult to extract enough anthocyanin, resulting in a loss of raw materials and a lower yield of the final product, and when the extraction time is increased, the extraction efficiency is not increased any more, but anthocyanin is destroyed. Can be. In order to increase the anthocyanin extraction efficiency of the purple sweet potato, it is preferable to acidify the pH of the water to 2 to 6, preferably
본 발명의 자색고구마 추출물로 탄소수 2 ~ 4의 저급 알코올 수용액에 의한 추출물이 사용될 수 있다. 예를 들어, 에탄올, 메탄올, 이소프로판올 등의 알코올 수용액, 바람직하게는 20 ~ 80 중량%의 알코올 수용액, 더욱 바람직하게는 50 ~ 70 중량%의 알코올 수용액으로 추출한다. 상기 알코올 수용액의 안토시아닌 추출 효율을 증대시키기 위해 열수추출과 마찬가지로 알코올에 산을 첨가할 수 있다. 알코올 추출물을 사용하는 경우 효모를 접종하기 전 먼저 알코올 추출물의 알코올을 기화시켜 알코올 함량을 낮춘 농축액 또는 알코올 추출물을 농축 및 건조시킨 후 물에 용해한 후 사용한다. 또한 바람직하게는 잡균의 번식을 억제하기 위하여 알코올 추출물을 80 ~ 105 ℃, 바람직하게는 90 ~ 100 ℃에서 0.5 ~ 24 시간,바람직하게는 1 ~ 6 시간 살균한 후 사용한다.As a purple sweet potato extract of the present invention, an extract by a lower alcohol aqueous solution having 2 to 4 carbon atoms may be used. For example, it is extracted with an aqueous alcohol solution such as ethanol, methanol, isopropanol, preferably 20 to 80% by weight of an aqueous alcohol solution, more preferably 50 to 70% by weight of an aqueous alcohol solution. In order to increase anthocyanin extraction efficiency of the aqueous alcohol solution, an acid may be added to the alcohol as in hot water extraction. In the case of using an alcohol extract, the alcohol of the alcohol extract is first vaporized before inoculating the yeast, and the concentrated or alcohol extract having a lower alcohol content is concentrated and dried, and then dissolved in water. In addition, in order to suppress the propagation of various bacteria, the alcohol extract is used after sterilization for 0.5 to 24 hours and preferably 1 to 6 hours at 80 to 105 ° C, preferably 90 to 100 ° C.
본 발명의 자색고구마 추출물의 발효를 위해 접종되는 균주는 효모, 바람직하게는 이사켄키아 오리엔탈리스(Issatchenkia orientalis) 효모이다. 효모의 접종량은 1 × 107 cell/ml 효모액을 기준으로 자색고구마 추출액 100 중량부에 대하여 0.001 ~ 10 중량부, 바람직하게는 0.01 ~ 5 중량부, 더욱 바람직하게는 0.1 ~ 3 중량부 사용한다.The strain inoculated for the fermentation of the purple sweet potato extract of the present invention is yeast, preferably Issatchenkia orientalis ) yeast. The amount of inoculation of yeast is 0.001 to 10 parts by weight, preferably 0.01 to 5 parts by weight, and more preferably 0.1 to 3 parts by weight based on 100 parts by weight of the purple sweet potato extract based on 1 × 10 7 cell / ml yeast solution. .
본 발명의 지방 생성 및 축적 저해용 조성물에서 상기 자색고구마 추출물에 효모를 접종한 후 25 ~ 40 ℃, 바람직하게는 30 ~ 35 ℃에서 18 ~ 78 시간, 바람직하게는 48 ~ 72 시간 발효한다.After inoculating yeast with the purple sweet potato extract in the composition for inhibiting fat production and accumulation of the present invention, the fermentation is carried out at 25 to 40 ° C., preferably at 30 to 35 ° C. for 18 to 78 hours, preferably 48 to 72 hours.
본 발명의 지방 생성 및 축적 저해용 조성물에서 상기 자색고구마 추출물 발효액은 총 중량을 기준으로 발효액이 0.01 ~ 50 중량%(고형분 기준)로 함유된 것을 특징으로 한다.In the composition for inhibiting fat formation and accumulation of the present invention, the purple sweet potato extract fermentation broth is characterized in that the fermentation broth is contained in an amount of 0.01 to 50% by weight (based on solids).
본 발명의 조성물은 지방 생성 및 축적 저해용 또는 비만억제용 건강기능식품으로 다양하게 이용될 수 있다. 본 발명의 자색고구마 추출물 발효액을 유효성분으로 포함하는 건강기능식품은 각종 식품류 예를 들어, 음료, 껌, 차, 비타민 복합제, 분말, 과립, 정제, 캡슐 등의 형태로 사용할 수 있다.The composition of the present invention can be used in various ways as a dietary supplement for inhibiting fat production and accumulation or for preventing obesity. Health functional food containing the purple sweet potato extract fermentation broth of the present invention as an active ingredient can be used in the form of various foods, for example, beverages, gum, tea, vitamin complex, powder, granules, tablets, capsules.
본 발명의 자색고구마 추출물 발효액이 건강기능식품에 포함될 때 그 양은 건조물 또는 고형인 경우 전체 식품 중량의 0.01 내지 15 중량%로 가할 수 있으며, 음료인 경우 100 ㎖를 기준으로 0.02 내지 10 g, 바람직하게는 0.3 내지 1 g의 비율로 가할 수 있다. When the purple sweet potato extract fermentation broth of the present invention is included in the health functional food, the amount may be added as 0.01 to 15% by weight of the total food weight when dry or solid, and 0.02 to 10 g based on 100 ml, preferably for beverages. Can be added in a ratio of 0.3 to 1 g.
본 발명의 건강기능식품이 음료 형태로 제조되는 경우 지시된 비율로 필수 성분으로서 상기 자색고구마 추출물 발효액을 함유하는 것 외에 액체성분에는 특별한 제한점은 없으며 통상의 음료와 같이 여러 가지 향미제 또는 천연 탄수화물 등을 추가 성분으로서 함유할 수 있다. 상술한 천연 탄수화물의 예는 모노사카라이드, 예를 들어, 포도당, 과당 등의 디사카라이드, 예를 들어 말토스, 슈크로스 등의 및 폴리사카라이드, 예를 들어 덱스트린, 시클로덱스트린 등과 같은 통상적인 당 및 자일리톨, 소르비톨, 에리트리톨 등의 당알콜이다. 상술한 것 이외의 향미제로서 천연 향미제(타우마틴, 스테비아 추출물(예를 들어 레바우디오시드 A, 글리시르히진등)) 및 합성 향미제(사카린, 아스파르탐 등)를 유리하게 사용할 수 있다. 상기 천연 탄수화물의 비율은 본 발명의 음료 100 ㎖당 일반적으로 약 1 내지 20g, 바람직하게는 약 5 내지 12g이다.When the health functional food of the present invention is prepared in the form of a beverage in addition to containing the purple sweet potato extract fermentation broth as an essential ingredient in the indicated ratio, there are no particular limitations on the liquid components and various flavors or natural carbohydrates, such as ordinary drinks. May be contained as an additional component. Examples of the above-mentioned natural carbohydrates include monosaccharides such as disaccharides such as glucose and fructose such as maltose, sucrose and the like and polysaccharides such as dextrin, cyclodextrin and the like Sugar, and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents other than those mentioned above, natural flavoring agents (tauumatin, stevia extract (e.g., Rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. have. The proportion of said natural carbohydrates is generally about 1 to 20 g, preferably about 5 to 12 g per 100 ml of the beverage of the present invention.
상기 외에 본 발명의 건강기능식품으로 사용되는 지방 생성 및 축적 저해용 또는 비만억제용 조성물은 여러 가지 영양제, 비타민, 광물(전해질), 합성 풍미제 및 천연 풍미제 등의 풍미제, 착색제 및 중진제(치즈, 초콜릿 등), 펙트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알콜, 탄산 음료에 사용되는 탄산화제 등을 함유할 수 있다. 그밖에 본 발명의 건강기능식품으로 사용되는 비만억제용 조성물들은 천연 과일 쥬스 및 과일 쥬스 음료 및 야채 음료의 제조를 위한 과육을 함유할 수 있다. 이러한 성분은 독립적으로 또는 조합하여 사용할 수 있다. 이러한 첨가제의 비율은 그렇게 중요하진 않지만 본 발명의 조성물 100 중량부 당 0 내지 약 20 중량부의 범위에서 선택되는 것이 일반적이다.In addition to the above, the composition for inhibiting fat formation and accumulation or preventing obesity, which is used as a health functional food of the present invention, may include various nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavoring agents such as flavoring agents, coloring agents and neutralizing agents. (Cheese, chocolate, etc.), pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated drinks, and the like. . In addition, the composition for inhibiting obesity used as a health functional food of the present invention may contain a flesh for preparing natural fruit juice and fruit juice beverage and vegetable beverage. These components can be used independently or in combination. The proportion of such additives is not so critical, but is generally selected in the range of 0 to about 20 parts by weight per 100 parts by weight of the composition of the present invention.
본 발명의 조성물은 식용으로 사용하는 자색고구마의 추출물을 발효시켜 제조된 것으로 독성 및 부작용이 거의 없이 예방 또는 치료 목적으로 장기간 복용 시에도 안심하고 사용할 수 있다. The composition of the present invention is prepared by fermenting the extract of purple sweet potato to be used for edible and can be used with confidence even during long-term use for prophylaxis or treatment with little toxicity and side effects.
본 발명은 자색고구마 추출물 발효액을 유효성분으로 포함하는 지방 생성 및 축적 저해용 또는 비만억제용 의약을 제공한다.The present invention provides a medicament for inhibiting fat formation and accumulation or inhibiting obesity, comprising a purple sweet potato extract fermentation broth as an active ingredient.
본 발명의 의약은, 의약 총 중량에 대하여 상기 자색고구마 추출물 발효액을 0.1 내지 50% 중량(고형분 기준)으로 포함한다. 그러나 상기와 같은 조성은 반드시 이에 한정되는 것은 아니고, 환자의 상태 및 질환의 종류 및 진행 정도에 따라 변할 수 있다.The medicine of the present invention comprises the purple sweet potato extract fermentation broth in an amount of 0.1 to 50% by weight, based on the total weight of the medicine. However, the composition is not limited thereto, and may vary depending on the condition of the patient, the type of disease, and the progress of the disease.
본 발명의 의약은 의약품 또는 약학 조성물의 제조에 통상적으로 사용하는 적절한 담체, 부형제 및 희석제를 더 포함할 수 있다.The medicament of the present invention may further comprise suitable carriers, excipients and diluents commonly used in the manufacture of a medicament or pharmaceutical composition.
본 발명에 따른 의약은, 각각 통상의 방법에 따라 산제, 과립제, 정제, 캡슐제, 현탁액, 에멀젼, 시럽, 에어로졸 등의 경구형 제형, 외용제, 좌제 및 멸균 주사용액의 형태로 제형화하여 사용될 수 있으며, 의약에 포함될 수 있는 담체, 부형제 및 희석제로는 락토즈, 덱스트로즈, 수크로스, 솔비톨, 만니톨, 자일리톨, 에리스리톨, 말티톨, 전분, 아카시아 고무, 알지네이트, 젤라틴, 칼슘 포스페이트, 칼슘 실리케이트, 셀룰로즈, 메틸 셀룰로즈, 미정질 셀룰로스, 폴리비닐 피롤리돈, 물, 메틸히드록시벤조에이트, 프로필히드록시벤조에이트, 탈크, 마그네슘 스테아레이트 및 광물유를 들 수 있다. 제제화할 경우에는 보통 사용하는 충진제, 증량제, 결합제, 습윤제, 붕해제, 계면활성제 등의 희석제 또는 부형제를 사용하여 조제된다. 경구투여를 위한 고형제제에는 정제, 환제, 산제, 과립제, 캡슐제 등이 포함되며, 이러한 고형제제는 상기 추출물에 적어도 하나 이상의 부형제 예를 들면, 전분, 칼슘카보네이트(calcium carbonate), 수크로스(sucrose) 또는 락토오스(lactose), 젤라틴 등을 섞어 조제된다. 또한 단순한 부형제 이외에 마그네슘 스티레이트 탈크 같은 윤활제들도 사용된다. 경구를 위한 액상제제로는 현탁제, 내용액제, 유제, 시럽제 등이 해당되는데 흔히 사용되는 단순희석제인 물, 리퀴드 파라핀 이외에 여러 가지 부형제, 예를 들면 습윤제, 감미제, 방향제, 보존제 등이 포함될 수 있다. 비경구 투여를 위한 제제에는 멸균된 수용액, 비수성용제, 현탁제, 유제, 동결건조제제, 좌제가 포함된다. 비수성용제, 현탁제로는 프로필렌글리콜 (propylene glycol), 폴리에틸렌 글리콜, 올리브 오일과 같은 식물성 기름, 에틸올레이트와 같은 주사 가능한 에스테르 등이 사용될 수 있다. 좌제의 기제로는 위텝솔(witepsol), 마크로골, 트윈 (tween) 61, 카카오지, 라우린지, 글리세로제라틴 등이 사용될 수 있다.The medicaments according to the invention can be used in the form of oral formulations, external preparations, suppositories, and sterile injectable solutions, such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, etc., according to conventional methods, respectively. Carriers, excipients and diluents that may be included in the medicament include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose , Methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. When formulated, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, and surfactants are usually used. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and the solid preparations may include at least one excipient such as starch, calcium carbonate, sucrose in the extract. ) Or lactose, gelatin and the like are mixed. In addition to simple excipients, lubricants such as magnesium styrate talc are also used. Examples of the liquid preparation for oral use include suspensions, solutions, emulsions, and syrups. In addition to water and liquid paraffin, simple diluents commonly used, various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included . Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. As the non-aqueous solvent and suspending agent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like can be used. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.
본 발명의 의약의 바람직한 투여량은 환자의 상태 및 체중, 질병의 정도, 약물형태, 투여경로 및 기간에 따라 다르지만, 당업자에 의해 적절하게 선택될 수 있다. 그러나 바람직한 효과를 위해서, 본 발명의 자색고구마 추출물 발효액 고형분을 기준으로 1일 0.01 mg/kg 내지 10 g/kg으로, 바람직하게는 1 mg/kg 내지 1 g/kg으로 투여하는 것이 좋다. 투여는 하루에 한번 투여할 수도 있고, 수회 나누어 투여할 수 있다. 따라서, 상기 투여량은 어떠한 면으로든 본 발명의 범위를 한정하는 것은 아니다.Preferred dosages of the medicaments of the present invention vary depending on the condition and weight of the patient, the extent of the disease, the form of the drug, the route of administration and the duration, and may be appropriately selected by those skilled in the art. However, for the desired effect, it is preferable to administer at 0.01 mg / kg to 10 g / kg, preferably 1 mg / kg to 1 g / kg per day based on the purple sweet potato extract fermentation broth of the present invention. The administration may be carried out once a day or divided into several doses. Thus, the dosage amounts are not intended to limit the scope of the invention in any manner.
본 발명의 의약은 쥐, 생쥐, 가축, 인간 등의 포유동물에 다양한 경로로 투여될 수 있다. 투여의 모든 방식은 예상될 수 있는데, 예를 들면, 경구, 직장 또는 정맥, 근육, 피하, 자궁내 경막 또는 뇌혈관내 (intracerebroventricular) 주사에 의해 투여될 수 있다.
The medicine of the present invention can be administered to mammals such as rats, mice, livestock, humans, and the like by various routes. All modes of administration can be expected, for example by oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dural or intracerebroventricular injection.
이하, 본 발명을 실시예, 비교예, 실험예 및 제제예를 통하여 보다 상세히 설명한다. 하기 실시예는 본 발명을 설명하기 위한 예시적인 것일 뿐 이에 의해 본 발명의 기술적 사상의 범위가 한정되는 것은 아니다.
Hereinafter, the present invention will be described in more detail through Examples, Comparative Examples, Experimental Examples, and Formulation Examples. The following examples are only illustrative for the purpose of illustrating the present invention, and the scope of the technical spirit of the present invention is not limited thereto.
비교예 1: 자색고구마 물추출물의 제조Comparative Example 1: Preparation of Purple Sweet Potato Water Extract
수확 후 1개월 이내의 신자미 품종의 자색고구마를 1 cm 두께로 절단한 후, 자색고구마 중량 3배의 물로 90 ℃, 60분간 추출하고, 추출물을 25 ℃로 냉각시킨 후 여과하여 여액과 잔사를 분리하여 얻어진 추출 여액은 냉각흡입기가 장착된 진공농축기를 사용하여 35℃에서 감압 농축하여 농축물을 얻고, 농축물을 냉동진공건조기를 사용하여 자색고구마 물추출물 분말을 제조하였다.
After cutting the purple sweet potato of Shinjami varieties within 1 month after harvesting to 1 cm thickness, extracted with
제조예 1: 고구마 발효음료로부터 발효균주의 분리Preparation Example 1 Isolation of Fermented Strain from Sweet Potato Fermented Beverage
고구마 발효음료 생산 혼합종균으로부터 발효 주원인균을 분리하였다. YPD agar에서 25℃, 24시간 호기배양 시킨 배지에서 두 종류의 콜로니(YPD Large 및 YPD Small)를 분리하였고, 한국미생물보존센터 (KCCM)에 동정을 의뢰한 결과, 이들 YPD Large균과 Small균은 동일 균주(Issatchenkia orientalis)로 확인되었다(도 1 및 도 2).
The main causative agents of fermentation of sweet potato fermented beverage production were isolated. Two kinds of colonies (YPD Large and YPD Small) were isolated from the media incubated at 25 ° C for 24 hours in YPD agar.They were identified by the Korea Microorganism Conservation Center (KCCM). The same strain ( Issatchenkia orientalis ) was identified (FIGS. 1 and 2).
제조예 2: 자색고구마 물추출물 발효액의 제조Preparation Example 2 Preparation of Purple Sweet Potato Water Extract Fermentation Solution
수확 후 1개월 이내의 신자미 품종의 자색고구마를 1 cm 두께로 절단한 후, 자색고구마 중량 3배의 물로 90 ℃, 60분간 추출하고, 추출물을 25 ℃로 냉각시킨 후 1 × 107 cell/ml 효모(이사켄키아 오리엔탈리스)를 1 중량%로 접종한 후, 30 ℃에서 24시간 내지 96시간 발효시켰다. 발효액은 여과하여 여액과 잔사를 분리하여 얻어진 발효 여액은 냉각흡입기가 장착된 진공농축기를 사용하여 35℃에서 감압 농축하여 농축물을 얻고, 농축물을 냉동진공건조기를 사용하여 자색고구마 물추출물 발효액 분말을 제조하였다.After cutting the purple sweet potato of Shinjami varieties within 1 month after harvesting to 1 cm thickness, extracted with
발효시간에 따른 발효액의 pH 변화를 도 3에 나타내었고, 안소시아닌 색소를 검출하는 530 nm에서의 흡광도 변화를 도 4에 나타내었다.The pH change of the fermentation broth according to the fermentation time is shown in FIG. 3, and the change in absorbance at 530 nm for detecting ansocyanine pigment is shown in FIG. 4.
발효시간 0시간(비교예 1과 동일)에 비해 24시간 이상 발효시켰을 때 pH가 급격히 감소되고 흡광도가 증가됨을 확인할 수 있었고, 24시간 내지 72시간에서는 큰 변화가 없다가, 96시간 발효시 pH가 더욱 낮아지면서 흡광도가 더욱 증가하였다. 예비실험결과 24시간 내지 72시간 발효시킨 발효액의 지방축적 억제효과가 뛰어난 것으로 나타나 이후의 실험은 72시간 발효시킨 것을 실시예 1로 하여 비교예 1과 대비하였다.
When the fermentation time was 0 hours (the same as Comparative Example 1), the pH was rapidly decreased and the absorbance was increased more than 24 hours.Therefore, there was no significant change in the 24-hour to 72-hour fermentation. As it became lower, the absorbance increased further. Preliminary experiments showed that the effect of inhibiting fat accumulation of the fermentation broth fermented from 24 hours to 72 hours was excellent, and the subsequent experiments were compared with Comparative Example 1 with Example 1 being fermented for 72 hours.
실험예 1: 일반성분 분석Experimental Example 1: General Component Analysis
비교예 1 및 실시예 1에서 농축 및 건조 전의 추출 여액과 발효 여액을 시료로 식품공전법에 따라 총질소, 조지방, 수분, 회분 함량을 측정하고 탄수화물 함량을 계산하여 표 1에 나타내었다.In Comparative Example 1 and Example 1, the extraction filtrate and the fermentation filtrate before concentration and drying were measured as a sample according to the Food Code, and the total nitrogen, crude fat, moisture, and ash content were measured and carbohydrate content was calculated and shown in Table 1.
(중량%)Total nitrogen
(weight%)
(중량%)Crude fat
(weight%)
(중량%)moisture
(weight%)
(중량%)Ash
(weight%)
(중량%)carbohydrate
(weight%)
실시예1의 발효 여액은 비교예 1의 추출 여액에 비해 회분, 지방, 그리고 탄수화물의 함량이 감소하는 것을 확인하였는데 이는 발효에 사용된 효모가 자색고구마 물추출물에 함유된 무기질, 지방, 당 등을 영양원으로 사용하여 이들 성분을 분해한 것으로 보인다.
The fermentation filtrate of Example 1 was confirmed to reduce the content of ash, fat, and carbohydrates compared to the extraction filtrate of Comparative Example 1, which means that the yeast used in the fermentation of minerals, fats, sugars, etc. contained in the purple sweet potato water extract It appears to have broken down these components using it as a nutrient source.
실험예 2: 지방축적 억제 효과Experimental Example 2: fat accumulation inhibitory effect
마우스 배아에서 유래된 3T3-L1 아지방 세포는 항비만 효과 확인을 위한 생체 외 실험에서 가장 많이 이용되고 있는 세포로서 4.5g/L, 10% 우태혈청, 페니실린 및 스트렙토마이신(100㎍/㎕㎖ penicillin and 100㎍/㎕㎖ streptomycin in 0.85% saline), 1% 100 μM 소디움 파루베이트를 함유한 DMEM으로 37℃, 5% CO2 배양기에서 배양하였다. 3T3-L1 아지방 세포의 지방세포로의 유도는 100% 합류(confluent)시키고서, 이틀 후 100 μM 3-이소부틸-1-메틸잔신, 250 μM 덱사메타손, 170 μM 인슐린을 함유한 MDI 배지로 유도하였으며, 3일 후에 MDI 배지는 10% 우태혈청과 170 nM 인슐린을 함유한 DMEM배지로 각 실험 날짜까지 이틀에 한번 교환하였다. 이상의 실험 과정의 개략도는 도 5에 나타내었다.3T3-L1 subfat cells derived from mouse embryos are the most frequently used in vitro experiments for anti-obesity effects. 4.5g / L, 10% fetal calf serum, penicillin and streptomycin (100µg / µl penicillin) and 100 μg / μl streptomycin in 0.85% saline) and DMEM containing 1% 100 μM sodium paruvate were incubated in a 37 ° C., 5% CO 2 incubator. Induction of 3T3-L1 subadipocytes into adipocytes was 100% confluent followed by two days later with MDI medium containing 100 μM 3-isobutyl-1-methylzansin, 250 μM dexamethasone, 170 μM insulin. After 3 days, MDI medium was exchanged once every two days by DMEM medium containing 10% fetal calf serum and 170 nM insulin. A schematic diagram of the above experimental procedure is shown in FIG. 5.
비교예 1와 실시예 1의 시료 및 음성대조군으로 증류수를 각각 40 ㎍/ml 씩 MDI 배지에 첨가하여(0일째), 3T3-L1 아지방 세포의 지방 세포로의 분화를 유도하였고 7일 째 되는 날 오일레드-O 용액으로 세포의 지방을 염색시키고 100% 이소프로판올로 탈색시켜 500 nm에서 흡광도를 측정한 결과를 음성대조군에 대한 상대흡광도로 도 6에 나타내었다.Distilled water was added to the MDI medium by 40 μg / ml of the sample and negative control of Comparative Example 1 and Example 1 (day 0) to induce differentiation of 3T3-L1 subfat cells into adipocytes. The fats of cells were stained with raw oil red-O solution and destained with 100% isopropanol to measure absorbance at 500 nm. The results are shown in FIG. 6 as the relative absorbance of the negative control group.
본원발명의 실시예 1은 음성대조군이나 비교예 1에 비하여 20% 정도의 지방축적을 저해하였다.Example 1 of the present invention inhibited the fat accumulation of about 20% compared to the negative control group or Comparative Example 1.
한편, 본 발명에서 분리한 균주의 효과를 다른 효모와 비교하기 위하여 상업용 사카로미세스 세레비지에와 이사켄키아 오리엔탈리스(KCTC 17696)를 이용하여 제조예 1과 동일한 방식으로 자색고구마 물추출물 발효액 분말을 제조하여 지방축적 억제 효과를 비교하여 도 7에 나타내었다. 이사켄키아 오리엔탈리스를 사용하여 발효시킨 발효추출물의 지방축적 억제 효능이 가장 좋았으며, 같은 효모이지만 사카로마이세스 세레비지에를 사용하여 발효시킨 추출물은 지방축적 억제효과가 크지 않음을 확인하였다.
On the other hand, in order to compare the effect of the strain isolated in the present invention with other yeast purple sweet potato water extract fermentation powder in the same manner as in Preparation Example 1 using commercial Saccharomyces cerevisiae and Isakenchia orientalis (KCTC 17696) To prepare and compare the effect of inhibiting fat accumulation is shown in Figure 7. The effect of fermentation extract fermented with Isakenkia orientalis was the best inhibiting effect, and the same yeast, but the extract fermented with Saccharomyces cerevisiae was confirmed that the effect of inhibiting fat accumulation.
실험예 3: C/EBPα 및 PPARγ 발현 억제 효과Experimental Example 3: Inhibitory Effect of C / EBPa and PPARγ Expression
3T3-L1 아지방세포의 지방세포로의 분화에 있어서 많은 전사인자들이 필요하며 분화됨에 따라 지방세포로부터 여러 지방세포 특이 유전자들이 발현된다(도 8). 시료의 농도에 따른 C/EBPα의 mRNA 발현량을 리얼타임 RT-PCR을 통해 분석하여 그 결과를 표 2 및 도 9에 나타내었다.Many transcription factors are required for the differentiation of 3T3-L1 subadipocytes into adipocytes and as they differentiate, several adipocyte specific genes are expressed from adipocytes (Fig. 8). The mRNA expression level of C / EBPa according to the concentration of the sample was analyzed by real-time RT-PCR, and the results are shown in Table 2 and FIG. 9.
C/EBPα는 PPARγ와 결합하여 adipsin, aP2, SCD-1, GLUT4, PEPCK, Leptin, CD36, insulin recepter 등과 같은 지방세포 특이 유전자들을 활성화시키는 중요한 전사인자로서, 비교예 1의 물추출물은 C/EBPα의 mRNA 발현량을 오히려 증가시키고 특히 20 ㎍/ml에서는 4배이상 증가됨을 알 수 있었다. 그러나 본 발명의 발효액은 농도와 관계없이 모두 C/EBPα의 mRNA 발현량을 크게 감소시킴을 확인하였다.C / EBPa is an important transcription factor that binds to PPARγ to activate adipocyte-specific genes such as adipsin, aP2, SCD-1, GLUT4, PEPCK, Leptin, CD36, insulin recepter, etc. The water extract of Comparative Example 1 is C / EBPa Rather, the mRNA expression level of the rather increased, especially at 20 ㎍ / ml was found to increase more than four times. However, it was confirmed that the fermentation broth of the present invention greatly reduced the amount of mRNA expression of C / EBPa regardless of the concentration.
또한, 시료의 농도에 따른 PPARγ의 mRNA 발현량을 리얼타임 RT-PCR을 통해 분석하여 그 결과를 표 3 및 도 10에 나타내었다.In addition, the mRNA expression amount of PPARγ according to the concentration of the sample was analyzed by real-time RT-PCR and the results are shown in Table 3 and FIG. 10.
sample
기존의 보고에 의하면 3T3-L1이 지방세포로 분화가 유도되면 PPARγ는 분화되기 전보다 mRNA 발현량이 10배 이상 증가한다고 한다. 도 10에서 보여주는 바와 같이 비교예 1과 실시예 1의 시료는 모두 농도 의존적으로 PPARγ의 mRNA 발현량을 감소시켰고, 그 발현양은 실시예 1에서 현저히 억제됨을 확인할 수 있었고, 특히 낮은 농도에서 그 차이가 더욱 크게 나타났다.
According to the existing report, when 3T3-L1 is induced to differentiate into adipocytes, PPARγ is expressed by 10 times or more than before the differentiation. As shown in FIG. 10, the samples of Comparative Example 1 and Example 1 reduced the mRNA expression level of PPARγ in a concentration-dependent manner, and the amount of expression was remarkably suppressed in Example 1, especially at low concentrations. Even greater.
실험예 4: 지방합성효소 발현 억제 효과Experimental Example 4: Effect of inhibiting the expression of fat synthase
인체 간세포인 HepG2 세포에 고농도의 포도당 30 mM을 24시간 이상 처리하여 인슐린 저항성 세포주를 만들어 사용하였으며, 세포주의 확립은 세포주에서 지질의 생성량을 나일레드 분석법을 이용하여 확인하였다. 도 11에서 세포내 지질은 나일레드로 염색되어 나타내어지고, 나일레드에 의한 580 nm/ 535 nm의 비는 지방의 축적을 나타낸 것으로 인슐린 저항성 세포주의 지방축적을 확인할 수 있었다.HepG2 cells, which are human hepatocytes, were treated with high concentrations of 30 mM glucose for more than 24 hours to make insulin-resistant cell lines. The establishment of cell lines was confirmed by the Nilered assay for the production of lipids in cell lines. In FIG. 11, the intracellular lipids were stained with Nile red, and the ratio of 580 nm / 535 nm by Nile red indicates the accumulation of fat, indicating the fat accumulation of the insulin resistant cell line.
인슐린 저항성 세포주에서 지방합성효소(Fatty acid synthase; FAS) 발현에 대한 영향을 측정하기 위해서 세포 분해물(cell lysate) 내의 지방합성효소와 β-엑틴 발현량을 웨스턴 블랏으로 확인하고(도 12), β-엑틴에 대한 지방합성효소의 상대 발현정도를 표 4 및 도 13에 나타내었다.In order to determine the effect on the expression of Fatty acid synthase (FAS) in insulin resistant cell lines, the amount of lipase and β-actin expression in the cell lysate was confirmed by Western blot (Fig. 12). Relative expression of fat synthase relative to actin is shown in Table 4 and FIG. 13.
division
비교예1에 비해 실시예1에서 지방합성효소의 상대 발현정도가 감소함을 확인하였다.
Compared with Comparative Example 1, it was confirmed in Example 1 that the relative expression level of fat synthase decreased.
실험예 5: 고지방식이 마우스에서의 항비만 효과Experimental Example 5: Anti-obesity effect in high fat diet mice
4 주령의 수컷 C57BL/6J mice에 고지방식이를 4 주간 투여한 후, 몸무게를 측정하여 몸무게가 증가한 마우스를 선별하여, 선별된 마우스를 4 그룹, 즉 음성대조군: 일반식이(NA 또는 control), 양성 대조군: 고지방식이(HFD), 고지방식이에 실시예 1의 자색고구마 물추출물 발효액 50 mg/kg 첨가한 실험군(HFD+FPSP 50), 고지방식이에 실시예 1의 자색고구마 물추출물 발효액 200 mg/kg 첨가한 실험군(HFD+FPSP 200)으로 나누어 4주 동안 구강 투여하고, 각 실험군 당 5마리를 사용하였다. 도 14에 실험 과정의 개략도를 나타내었다. Four-week-old male C57BL / 6J mice were given high-fat diet for 4 weeks, and then weighed to select mice with increased weight, and the selected mice were divided into four groups, namely, negative control group (NA or control), Positive control group: high-fat diet (HFD), the experimental group (HFD + FPSP 50) added the purple sweet potato water extract fermentation solution of Example 1 to a high-fat diet (Example 1) purple sweet potato water extract fermentation solution of Example 1 Oral administration for 4 weeks was divided into the experimental group (HFD + FPSP 200) added 200 mg / kg, 5 animals were used for each experimental group. 14 shows a schematic diagram of the experimental procedure.
마우스의 체중의 변화를 살펴본 결과, 고지방식이에 의해 체중이 현저히 증가되었으나, 자색고구마 물추출물 발효액을 50 mg/kg 첨가한 경우 체중이 감소하고, 200 mg/kg 첨가한 경우 더욱 현저히 감소함을 확인할 수 있었다(도 15).As a result of examining the change of body weight of the mouse, the weight gain was significantly increased by the high fat diet, but when the 50 mg / kg of purple sweet potato water extract fermentation broth was added, the weight was decreased, and when the 200 mg / kg was added, the weight was significantly decreased. It could be confirmed (Fig. 15).
마우스의 간조직의 무게의 변화 여부를 살펴본 결과, 자색고구마 물추출물 발효액을 50 mg/kg 첨가한 경우 간조직의 무가가 감소하고, 200 mg/kg 첨가한 경우 더욱 현저히 감소함을 확인할 수 있었다(도 16). 마우스의 희생전의 사진과 희생 후 적출한 간조직의 사진을 도 17에 나타내었다.As a result of examining the change in the weight of the liver tissue of the mouse, it was confirmed that the addition of 50 mg / kg of purple sweet potato water extract fermentation decreased the no-cost of liver tissue, and further significantly decreased the addition of 200 mg / kg ( 16). The photograph of the mouse before sacrifice and the liver tissue extracted after sacrifice are shown in FIG.
마우스의 피하지방의 축적 정도를 살펴본 결과, 고지방식이에 의해 피하지방이 현저히 증가하였음을 확인할 수 있고, 자색고구마 물추출물 발효액을 200 mg/kg 첨가한 경우 피하지방의 양이 현저히 감소했음을 확인할 수 있었다(도 18).As a result of examining the accumulation of subcutaneous fat in mice, it was confirmed that the subcutaneous fat increased significantly by the high fat diet, and that the amount of subcutaneous fat decreased significantly when 200 mg / kg of purple sweet potato water extract fermentation broth was added. (FIG. 18).
마우스의 간조직내 트리글리세라이드 함량을 살펴본 결과, 고지방식이에 의해 간조직내 트리글리세라이드 함량이 현저히 증가하였다가, 자색고구마 물추출물 발효액을 200 mg/kg 첨가한 경우 일반식이와 같은 수준으로 현저히 감소했음을 확인할 수 있었다(도 19).The triglyceride content in the liver of the mouse showed that the triglyceride content of the liver was significantly increased by the high fat diet. However, when 200 mg / kg of the purple sweet potato water extract fermentation broth was added, it significantly decreased to the same level as the general diet. It could be confirmed that (Fig. 19).
마우스의 간조직내 총 콜레스테롤 함량을 살펴본 결과, 고지방식이에 의해 간조직내 총 콜레스테롤 함량이 현저히 증가하였다가, 자색고구마 물추출물 발효액을 200 mg/kg 첨가한 경우 현저히 감소했음을 확인할 수 있었다(도 20).As a result of examining the total cholesterol content in the liver of the mouse, it was confirmed that the total cholesterol content in the liver tissue was significantly increased by the high fat diet, but significantly decreased when 200 mg / kg of the purple sweet potato water extract fermentation broth was added (Fig. 20).
마우스의 혈중 글루코오스 농도를 살펴본 결과, 고지방식이에 의해 혈중 글루코오스 농도가 현저히 증가하였다가, 자색고구마 물추출물 발효액을 50 mg/kg 및 200 mg/kg첨가한 경우 혈중 글루코오스 농도가 현저히 감소했음을 확인할 수 있었다(도 21).As a result of examining the blood glucose concentration in the mouse, it was confirmed that the blood glucose concentration was significantly increased by the high fat diet, but the blood glucose concentration was significantly decreased when 50 mg / kg and 200 mg / kg of purple sweet potato water extract were added. (FIG. 21).
고지방식이 마우스 동물모델에서 간조직의 지방합성효소(Fatty acid synthase; FAS) 활성 억제 여부 및 AMPK 단백질 발현을 웨스턴 블랏으로 확인한 결과, 자색고구마 물추출물 발효액을 200 mg/kg 첨가한 경우 β-엑틴 발현량에 대한 FAS의 상대 발현량이 현저히 감소했음을 확인할 수 있었고(도 22), β-엑틴 발현량에 대한 p-AMPK의 상대 발현량이 현저히 증가했음을 확인할 수 있었다(도 23).
Western blot confirmed the inhibition of Fatty acid Synthase (FAS) activity and AMPK protein expression in liver tissues of high-fat mouse animal model. When 200 mg / kg of purple sweet potato water extract was added, β-actin It was confirmed that the relative expression level of FAS relative to the expression amount was significantly reduced (FIG. 22), and the relative expression level of p-AMPK was significantly increased (β 23).
하기에 본 발명의 자색고구마 추출물 발효액을 함유하는 제제예를 설명하나, 본 발명은 이를 한정하고자 함이 아닌 단지 구체적으로 설명하고자 함이다.Hereinafter, an example of the preparation containing the purple sweet potato extract fermentation broth of the present invention will be described, but the present invention is not intended to be limited thereto, but is intended to be described in detail.
제제예 1. 산제의 제조(단위: 중량%) Formulation Example 1 Preparation of Powder (Unit: Weight%)
실시예 1의 자색고구마 추출물 발효액 66.0Purple Sweet Potato Extract Fermentation Solution of Example 1 66.0
유당 34.0Lactose 34.0
상기의 성분을 혼합하고 기밀포에 충진하여 산제를 제조한다.
The above ingredients are mixed and filled in an airtight cloth to prepare a powder.
제제예 2. 정제의 제조(단위: 중량%) Formulation Example 2 Preparation of Tablet (Unit: wt%)
실시예 1의 자색고구마 추출물 발효액 33.0Purple Sweet Potato Extract Fermentation Solution of Example 1 33.0
유당 33.0Lactose 33.0
옥수수전분 33.3Corn Starch 33.3
스테아린산 마그네슘 0.7Magnesium Stearate 0.7
상기의 성분을 혼합한 후 통상의 정제 제조방법에 따라서 타정하여 정제를 제조한다.
After mixing the above components and tableting according to a conventional tablet production method to produce a tablet.
제제예 3. 선식의 제조(단위: 중량%) Formulation Example 3 Preparation of Wire (Unit: Weight%)
현미 40%, 보리 30%, 율무 20%, 알파미분 10%를 배전한 후 분쇄기로 입도 110메쉬의 곡물류 분말을 만든다. 검정콩 40%, 검정깨 30%, 들깨 30%를 배전한 후 분쇄기로 입도 110메쉬의 종실류 분말을 만든다. 상기 곡물류 75%, 종실류 20% 및 실시예 1의 자색고구마 추출물 발효액 분말 5%를 배합후 과립화하여 선식을 제조한다.
After roasting 40% brown rice, 30% barley, 20% barley, and 10% alpha fine powder, grains of grain size of 110 mesh are made with a grinder. After roasting 40% black beans, 30% black sesame seeds, and 30% perilla sesame seeds, a seed grain powder having a particle size of 110 mesh is prepared. 75% of the grains, 20% of the seeds, and 5% of the purple sweet potato extract fermentation broth powder of Example 1 were granulated and then granulated.
제제예 4. 츄잉검의 제조(단위: 중량%) Formulation Example 4 Preparation of Chewing Gum (Unit: wt%)
껌 베이스 20%, 설탕 76%, 향료 1.5% 및 물 2%와 실시예 1의 자색고구마 추출물 발효액 분말 0.5%를 배합하여 통상의 방법으로 츄잉껌을 제조한다.
Chewing gum is prepared by a conventional method by combining
제제예 5. 음료의 제조(단위: 중량%) Formulation Example 5 Preparation of Beverage (Unit: Weight%)
꿀 5중량%, 과당 3%, 염산리보플라빈나트륨 0.0001%, 염산피리독신 0.0001%, 및 물 91.9998%와 실시예 1의 자색고구마 추출물 발효액 1%를 배합하여 통상의 방법으로 건강 음료를 제조한다.
5% by weight of honey, 3% fructose, 0.0001% of sodium riboflavin hydrochloride, 0.0001% of pyridoxine hydrochloride, and 91.9998% of water and 1% of the purple sweet potato extract fermentation broth of Example 1 were combined to prepare a healthy beverage.
본 발명의 자색고구마 추출물 발효액은 지방세포의 형성 및 지방 축적을 억제할 수 있는 활성성분을 포함하고 있으므로 향후 체중감소, 비만방지, 체지방감소, 혈중 지방감소 등에 효능이 있는 건강기능식품 또는 의약으로 사용이 가능할 것이다.Since the purple sweet potato extract fermentation broth of the present invention contains an active ingredient that can inhibit the formation of fat cells and fat accumulation, it is used as a health functional food or a medicine that is effective in weight loss, obesity prevention, body fat reduction, blood fat reduction, etc. This will be possible.
Claims (9)
A composition for inhibiting fat formation and accumulation containing a fermentation broth obtained by fermenting a purple sweet potato extract as an active ingredient.
The composition of claim 1, wherein the purple sweet potato extract is a hot water extract or an extract of a lower alcohol aqueous solution having 2 to 4 carbon atoms.
The composition for inhibiting fat production and accumulation according to claim 2, wherein the hot water extract is extracted at 80 to 105 ° C for 0.5 to 24 hours.
The composition for inhibiting fat production and accumulation according to claim 1, wherein the fermentation broth is a fermentation broth obtained by inoculating purple sweet potato extract with Issatchenkia orientalis yeast.
The composition for inhibiting fat production and accumulation according to claim 1, wherein the purple sweet potato extract is inoculated with yeast and fermented at 25 to 40 ° C. for 18 to 78 hours.
The method of claim 1, wherein the purple sweet potato is extracted with hot water at 80 to 105 ℃ for 0.5 to 24 hours, and the hot water extract is inoculated with Issatchenkia orientalis and then fermented at 25 to 40 ℃ for 18 to 78 hours. Fat production and accumulation inhibition composition comprising a fermentation broth as an active ingredient.
The composition for inhibiting fat production and accumulation according to claim 1, wherein the purple sweet potato is sweet potato of any one or more varieties selected from Yamakawamurasaki, Yeonmi, Borami, and Shinjami.
The composition for inhibiting fat production and accumulation according to claim 1, which is a pharmaceutical composition.
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CN103190613A (en) * | 2013-03-18 | 2013-07-10 | 石发军 | Preparation method of purple sweet potato health product composition |
KR101386727B1 (en) * | 2012-02-06 | 2014-04-24 | 재단법인 전남생물산업진흥원 | A Phamaceutical Composition Comprising a Fermented Purple Sweet Potato for Treating or Preventing Liver Disease |
KR101422248B1 (en) * | 2012-09-12 | 2014-07-24 | 안동대학교 산학협력단 | Issatchenkia orientalis ms-1 and its use |
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KR101422066B1 (en) * | 2013-12-30 | 2014-07-24 | 한국인삼농업법인(주) | Making method of blueberry beverage |
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US20060234957A1 (en) * | 2003-03-03 | 2006-10-19 | Takanori Tsuda | Adiponectin expression promoter |
MX2010008879A (en) * | 2008-02-14 | 2010-10-20 | Barley & Oats Co Ltd | Method for producing fermented product using natural material, and food or medicine containing fermented product made from same. |
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KR101386727B1 (en) * | 2012-02-06 | 2014-04-24 | 재단법인 전남생물산업진흥원 | A Phamaceutical Composition Comprising a Fermented Purple Sweet Potato for Treating or Preventing Liver Disease |
KR101422248B1 (en) * | 2012-09-12 | 2014-07-24 | 안동대학교 산학협력단 | Issatchenkia orientalis ms-1 and its use |
CN103190613A (en) * | 2013-03-18 | 2013-07-10 | 石发军 | Preparation method of purple sweet potato health product composition |
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