KR102217834B1 - Composition for Preventing or Treating Obesity Comprising Heat-killed Lactic Acid Bacteria from Kefir and Grape Seed Flour - Google Patents
Composition for Preventing or Treating Obesity Comprising Heat-killed Lactic Acid Bacteria from Kefir and Grape Seed Flour Download PDFInfo
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- KR102217834B1 KR102217834B1 KR1020180139442A KR20180139442A KR102217834B1 KR 102217834 B1 KR102217834 B1 KR 102217834B1 KR 1020180139442 A KR1020180139442 A KR 1020180139442A KR 20180139442 A KR20180139442 A KR 20180139442A KR 102217834 B1 KR102217834 B1 KR 102217834B1
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- South Korea
- Prior art keywords
- lactic acid
- kefir
- acid bacteria
- grape seed
- dead cells
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Abstract
본 발명은 포도씨 분말 및 케피어 유래 유산균을 포함하는 비만의 예방 또는 치료용 조성물에 관한 것이다. 본 발명에 따른 케피어 유래 유산균 사균체 및 포도씨 분말을 포함하는 조성물은 포도씨 분말이 프리바이오틱스로 작용하고 케피어에서 분리한 유산균 사균체의 복합 균체가 프로바이오틱스로 작용하여 비만을 예방 또는 치료할 수 있다. 본 발명의 조성물은 기능성 식품 또는 약학 조성물의 형태로 섭취되어 체중 감소, 지방 세포 무게 및 크기 감소, 콜레스테롤 수치 감소, 지방 형성 유전자 발현 감소 감소 등의 효과를 발휘할 수 있다.The present invention relates to a composition for preventing or treating obesity comprising grape seed powder and kefir-derived lactic acid bacteria. In the composition comprising the kefir-derived lactic acid bacteria dead cells and grape seed powder according to the present invention, the grape seed powder acts as a prebiotic, and the complex cells of the lactic acid bacteria dead cells isolated from the kefir act as probiotics, thereby preventing or treating obesity. . The composition of the present invention may be ingested in the form of a functional food or pharmaceutical composition to exert effects such as weight loss, fat cell weight and size reduction, cholesterol level reduction, and reduction in fat-forming gene expression.
Description
본 발명은 케피어 유래 유산균의 사균체 및 포도씨 분말을 포함하는 비만의 개선, 예방 또는 치료용 기능성 식품 및 약학 조성물에 관한 것이다. The present invention relates to functional foods and pharmaceutical compositions for improving, preventing or treating obesity, including dead cells of kefir-derived lactic acid bacteria and grape seed powder.
음식 섭취와 에너지 소비의 불균형에 의한 대사 장애인 비만(obesity)은 가장 일반적이고 심각한 전세계적인 건강 문제 중 하나이다. 비만은 선진국과 개발도상국 모두에게 막대한 의료비용을 발생시킬 수 있는 고혈압, 심혈관계 질환, 제2형 당뇨병, 호흡기 질환, 암 등을 포함하는 다양한 만성 질병의 중요한 위험 요소로 여겨지고 있다. Obesity, a metabolic disorder caused by an imbalance between food intake and energy expenditure, is one of the most common and serious global health problems. Obesity is considered an important risk factor for a variety of chronic diseases, including high blood pressure, cardiovascular disease,
비만을 치료하기 위한 다양한 접근법이 개발되고 있지만, 이러한 치료법들은 혈압을 증가시키거나, 간부전, 췌장염, 두통, 구강건조증, 월경과다증 등의 부작용들을 갖는 것으로 보고되고 있다. 이러한 이유로 비만을 치료할 수 있는 천연 제품들이 더 안전하고 더 효과적인 항-비만 치료제로서 기대되고 있다. Various approaches have been developed to treat obesity, but these treatments have been reported to have side effects such as increased blood pressure, liver failure, pancreatitis, headache, dry mouth, and hypermenorrhea. For this reason, natural products that can treat obesity are expected as safer and more effective anti-obesity treatments.
최근, 많은 연구들이 프로바이오틱스(probiotics)의 천연 항-비만 치료제로서의 잠재력을 강조하고 있다. 프로바이오틱스(probiotics)는 건강에 도움이 되는 장내 세균총 즉, 숙주의 건강에 이로움을 제공하는 살아있는 미생물 즉 생균류를 의미한다. 일반적으로 프로바이오틱스는 요거트 등의 발효식품의 일부 또는 식이보충제로서 소비된다. 프로바이오틱스로 알려진 미생물에는 유산균(lactic acid bacteria; LAB), 비피도박테리아(bifidobacteria), 바실러스 등이 있다. 상기 프로바이오틱스 중 일부는 비만, 인슐린 저항, 비알코올성 지방간 등에 치료 효과가 있는 것으로 알려져 있다.Recently, many studies have highlighted the potential of probiotics as natural anti-obesity treatments. Probiotics refer to the intestinal flora that is beneficial to health, that is, living microorganisms, or live fungi, that provide benefits to the health of the host. In general, probiotics are consumed as part of fermented foods such as yogurt or as dietary supplements. Microorganisms known as probiotics include lactic acid bacteria (LAB), bifidobacteria, and Bacillus. Some of the probiotics are known to have therapeutic effects on obesity, insulin resistance, and non-alcoholic fatty liver.
케피어(Kefir)는 코카서스 산간 지방에서 유래한 발효유의 일종으로, 티벳지방 승려들이 건강을 위해 음용하던 것이 대중화된 음료이다. 케피어의 구성영양소로는 단백질 및 다당체가 포함되어 있고, 비타민 A, 비타민 B1, 비타민B2, 비타민B6, 비타민D, 비타민K2, 엽산, 니코틴산 및 칼슘, 철분, 요오드 등을 고루 포함하고 있다. 이러한 케피어는 유산균과 효모 복합체인 케피어 그레인으로 불리우는 균종(케피어 과립균이라고도 한다)을 개시제로 하여 우유를 발효시켜 얻어진 요구르트상의 음료이다. Kefir is a kind of fermented milk derived from the mountainous regions of the Caucasus, and it is a popular drink that Tibetan monks drink for health. Constituent nutrients of kefir include proteins and polysaccharides, and they evenly contain vitamin A, vitamin B1, vitamin B2, vitamin B6, vitamin D, vitamin K2, folic acid, nicotinic acid and calcium, iron, and iodine. Such kefir is a yogurt-like beverage obtained by fermenting milk using a fungus called kefir grain (also referred to as kefir granule bacteria), which is a complex of lactic acid bacteria and yeast, as an initiator.
케피어는 유산균(Lactobacillus, Lactococcus, Leuconostoc, Acetobacter, 및 Streptococcus 종), 초산균 및 효모(Saccharomyces, Kluyveromyces, Torula 및 Candida 종)를 포함하는 다양한 미생물을 함유하고 있다. 케피어는 항돌연변이성, 콜레스테롤 저하, 항균성, 면역 조절 및 항염증 효과를 갖는 것이 증명되었지만, 아직까지 열처리에 의해 불활성화 된 케피어 및 불활성화된 사균체 유산균 및 효모의 항비만에 대해서는 구체적으로 알려진 바가 없다. Kefir contains various microorganisms including lactic acid bacteria ( Lactobacillus, Lactococcus, Leuconostoc, Acetobacter , and Streptococcus species), acetic acid bacteria and yeast ( Saccharomyces, Kluyveromyces, Torula and Candida species). Kefir has been proven to have anti-mutagenic, cholesterol-lowering, antimicrobial, immune-modulating and anti-inflammatory effects, but it has been specifically for the anti-obesity of kefir and inactivated dead cell lactic acid bacteria and yeasts inactivated by heat treatment. Nothing is known.
일반적으로, 프로바이오틱스 함유 기능성 식품의 경우, 생균 또는 사균의 제품군으로 나눌 수 있는데, 생균으로 섭취할 경우 보통 생균을 냉동 건조시킨 가루 형태나 생균을 함유한 발효유 형태 등으로 시중에 판매되고 있다. 사균의 경우 생균에 비해 생리활성이 밝혀진 것이 거의 없고 장내 유산균의 먹이가 되어 생장을 촉진시켜주는 프리바이오틱스(prebiotics)의 개념으로 섭취되고 있다. In general, functional foods containing probiotics can be divided into live bacteria or dead bacteria. When consumed as live bacteria, they are usually sold in the form of freeze-dried powder or fermented milk containing live bacteria. In the case of dead bacteria, physiological activity is hardly revealed compared to live bacteria, and it is consumed as a concept of prebiotics that promote growth as food for intestinal lactic acid bacteria.
이러한 종래의 프로바이오틱스 소재의 경우, 생리활성을 유지하기 위해 생균 상태로 가공식품에 함유시키려면 살균 등의 가공 과정에서 사멸할 수 있어 가루나 발효유 외의 다양한 가공 형태로 응용하기에 어려움이 있고, 한편으로 사균으로 함유시키기에는 사균의 생리활성이 밝혀진 바가 없고 생균에 비해 활성이 떨어지는 것으로 알려져 아직 식품 또는 의약 분야에 적용되기 어려움이 있다. In the case of such a conventional probiotic material, it is difficult to apply it in various processing forms other than powder or fermented milk, as it can be killed in processing such as sterilization in order to contain it in processed foods in a state of living bacteria to maintain physiological activity. It is known that the physiological activity of the dead cells has not been found to be contained as dead cells, and is known to be less active than the live cells, and it is still difficult to be applied to the field of food or medicine.
사균체를 활용한 식품 또는 의약 분야 특허로는, 대한민국 등록특허공보 제10-1472190호에서 식물성 유산균 락토바실러스 플란타룸 피엠오08 사균체를 유효성분으로 포함하는 변비 예방, 치료 또는 개선용 조성물을 개시하고 있고, 대한민국 등록특허공보 제10-1369219호에서는 항균용 유산균 사균체 및 이의 제조방법에 대해서 개시한 바 있다.As a patent in the field of food or medicine using dead cells, a composition for preventing, treating or improving constipation containing the plant lactic acid bacteria Lactobacillus plantarum PMO08 dead cells as an active ingredient in Korean Patent Publication No. 10-1472190 It has been disclosed, and Korean Patent Publication No. 10-1369219 discloses an antibacterial lactic acid bacteria dead cell and a method for producing the same.
프로바이오틱스와 유사한 개념으로, 프리바이오틱스(prebiotics)는 장내 유익한 균의 생장을 돕는 난소화성 성분으로서 프로바이오틱스(probiotics)의 영양원이 되어 장내 환경을 개선하는 데 도움을 주는 물질을 말한다. 어떠한 식품 성분이, 대장 내 유용 미생물에 의해 이용되어 미생물의 생육이나 활성을 촉진함으로써 숙주 건강에 좋은 효과를 나타내게 하는 프리바이오틱스로 활용되기 위해서는, 위장관의 상부에서 소화 또는 흡수되지 않아야 하고 대장 내 미생물 중 유용 세균을 선택적으로 활성화시키고 병원균 등의 유해균은 억제할 수 있어야 한다.In a similar concept to probiotics, prebiotics are indigestible ingredients that help the growth of beneficial bacteria in the intestine, and they refer to substances that help improve the intestinal environment by becoming a nutrient source of probiotics. In order for any food ingredient to be used as a prebiotic that is used by useful microorganisms in the large intestine to promote the growth or activity of microorganisms, thereby exhibiting a good effect on host health, it must not be digested or absorbed in the upper part of the gastrointestinal tract It should be able to selectively activate useful bacteria and suppress harmful bacteria such as pathogens.
한편, 포도에는 식물에서 발견되는 항산화물질인 폴리페놀(polyphenol) 계열의 레스베라트롤(resveratrol)이 많이 함유되어 있으며, 레스베라트롤은 항균, 항암, 항바이러스, 항알레르기, 및 항염증 감소효과를 지니며 강력한 항산화 활성의 기능이 있는 물질로 포도부산물인 과피, 씨, 송이 가지에 다량 함유되어 있으므로 포도부산물을 그대로 폐기할 경우 농산물에 함유된 유용한 성분을 손실하게 되는 것은 물론이고, 환경오염의 문제가 되므로 포도부산물의 효용가치를 높이는 연구가 활발하게 진행되고 있다. On the other hand, grapes contain a lot of resveratrol, a polyphenol-based antioxidant found in plants, and resveratrol has antibacterial, anticancer, antiviral, antiallergic, and anti-inflammatory effects, and is a powerful antioxidant. As a substance with an active function, it is contained in a large amount in grape by-products such as peels, seeds, and pine branches, so if the grape by-products are discarded as they are, the useful ingredients contained in the agricultural products will be lost as well as environmental pollution. Research to increase the utility value of the company is actively being conducted.
본 발명자들은 케피어 유래 유산균의 사균체 및 포도씨 분말이 각각 프로바이오틱스 및 프리바이오틱스로 작용하여 비만의 예방 또는 치료에 효과적이라는 것을 확인함으로써, 본 발명을 완성하였다.The present inventors have completed the present invention by confirming that the dead cells of kefir-derived lactic acid bacteria and grape seed powder are effective in preventing or treating obesity by acting as probiotics and prebiotics, respectively.
이와 같은 과제를 해결하기 위한 본 발명의 목적은 케피어 유래 유산균의 사균체 및 포도씨 분말을 포함하는 비만 개선용 식품을 제공하는 것이다. An object of the present invention for solving such a problem is to provide a food for improving obesity, including dead cells of kefir-derived lactic acid bacteria and grape seed powder.
본 발명의 다른 목적은 케피어 유래 유산균의 사균체 및 포도씨 분말을 포함하는 비만의 예방 또는 치료용 약학 조성물을 제공하는 것이다.Another object of the present invention is to provide a pharmaceutical composition for preventing or treating obesity, including dead cells of kefir-derived lactic acid bacteria and grape seed powder.
상기 목적을 달성하기 위해, 본 발명은 케피어(Kefir) 유래 유산균 및 포도씨 분말을 유효성분으로 포함하는 비만 개선용 기능성 식품을 제공한다. In order to achieve the above object, the present invention provides a functional food for improving obesity, including kefir-derived lactic acid bacteria and grape seed powder as an active ingredient.
본 발명에 있어서, 상기 케피어 유래 유산균은 류코노스톡 메센테로이데스(Leuconostoc mesenteroides), 락토바실러스 케피리(Lactobacillus kefiri) 또는 이들의 복합 균체일 수 있으며, 바람직하게는 류 류코노스톡 메센테로이데스 LCM4(leuconostoc mesenteroides LCM4, 기탁번호: KCCM12117P) 및 락토바실러스 케피리 DH5(Lactobacillus kefiri DH5, 기탁번호: KCCM11837P)의 복합 균체일 수 있다. In the present invention, the kefir-derived lactic acid bacteria may be Leuconostoc mesenteroides , Lactobacillus kefiri or a complex cell thereof, preferably Leuconostoc mesenteroides LCM4 ( leuconostoc mesenteroides LCM4, accession number: KCCM12117P) and Lactobacillus kefiri DH5 ( Lactobacillus kefiri DH5, accession number: KCCM11837P) may be a complex cell.
본 발명에서, 상기 사균체는 케피어 유래 유산균을 80℃ 이상에서 열처리 사멸시킨 것일 수 있다. In the present invention, the dead cells may be killed by heat treatment at 80 ℃ or higher derived from kefir lactic acid bacteria.
본 발명에서, 상기 사균체는 케피어 유래 유산균을 열처리 사멸 후 파쇄한 현탁액의 형태일 수 있다. In the present invention, the dead cells may be in the form of a suspension obtained by crushing the kefir-derived lactic acid bacteria after heat treatment and death.
본 발명에서, 상기 사균체는 케피어 유래 유산균을 열처리 사멸 후 파쇄한 현탁액의 수용성 분획일 수 있다.In the present invention, the dead cells may be an aqueous fraction of a suspension obtained by crushing kefir-derived lactic acid bacteria after heat treatment and death.
본 발명에 있어서, 상기 복합 균체는 류코노스톡 메센테로이데스 LCM4 및 락토바실러스 케피리 DH5를 5:1 내지 1:5의 농도비로 포함할 수 있다.In the present invention, the complex cells may contain Leukonostock mecenteroides LCM4 and Lactobacillus kepiri DH5 in a concentration ratio of 5:1 to 1:5.
본 발명에 있어서, 상기 케피어 유래 유산균 사균체는 0.1x108 내지 20x108 유산균/gram 포도씨분말로 포함될 수 있다. In the present invention, the kefir-derived lactic acid bacteria dead cells may be included as 0.1x10 8 to 20x10 8 lactic acid bacteria / gram grape seed powder.
본 발명에 있어서, 상기 포도씨 분말은 와인 제조 부산물일 수 있으며, 샤르도네(Chardonnay) 품종의 포도로 제조된 것일 수 있다. In the present invention, the grape seed powder may be a wine production by-product, and may be prepared from grapes of Chardonnay (Chardonnay) varieties.
본 발명에서, 상기 기능성 식품은 지방 세포 형성 유전자의 발현을 억제시키는 것을 특징으로 할 수 있다. In the present invention, the functional food may be characterized in that it suppresses the expression of adipocyte-forming genes.
본 발명에서, 상기 기능성 식품은 발효식품일 수 있다. In the present invention, the functional food may be a fermented food.
본 발명은 또한, 케피어 유래 유산균 및 포도씨 분말을 유효성분으로 포함하는 비만의 예방 또는 치료용 약학 조성물을 제공한다.The present invention also provides a pharmaceutical composition for preventing or treating obesity, comprising kefir-derived lactic acid bacteria and grape seed powder as active ingredients.
본 발명에 따른 케피어 유래 유산균 사균체 및 포도씨 분말을 포함하는 조성물은 포도씨 분말이 프리바이오틱스로 작용하고 케피어에서 분리한 유산균 사균체의 복합 균체가 프로바이오틱스로 작용하여 비만을 예방 또는 치료할 수 있다. 본 발명의 조성물은 기능성 식품 또는 약학 조성물의 형태로 섭취되어 체중 감소, 지방 세포 무게 및 크기 감소, 콜레스테롤 수치 감소, 지방 형성 유전자 발현 감소 감소 등의 효과를 발휘할 수 있다. In the composition comprising the kefir-derived lactic acid bacteria dead cells and grape seed powder according to the present invention, the grape seed powder acts as a prebiotic, and the complex cells of the lactic acid bacteria dead cells isolated from the kefir act as probiotics, thereby preventing or treating obesity. . The composition of the present invention can be ingested in the form of a functional food or pharmaceutical composition to exert effects such as weight loss, fat cell weight and size reduction, cholesterol level reduction, and reduction in fat-forming gene expression.
도 1(a)는 식이 종류에 따른 실험쥐의 총 에너지 섭취량을 나타낸 그래프이다.
도 1(b)는 식이 종류에 따른 실험쥐의 체중 증가량을 나타낸 그래프이다.
도 2는 식이 종류에 따른 실험쥐의 지방 조직 무게를 나타낸 그래프이다.
도 3은 식이 종류에 따른 실험쥐의 지방 세포 이미지를 나타낸다.
도 4는 식이 종류에 따른 실험쥐의 TG 및 TC 수치를 나타낸 그래프이다.
도 5는 식이 종류에 따른 실험쥐의 FASN 유전자 발현량을 나타낸 그래프이다.Figure 1 (a) is a graph showing the total energy intake of mice according to the type of diet.
1(b) is a graph showing the amount of weight gain in mice according to diet type.
2 is a graph showing the weight of adipose tissues of experimental mice according to the type of diet.
3 shows images of adipocytes of experimental mice according to diet types.
4 is a graph showing the TG and TC values of the experimental mice according to the type of diet.
5 is a graph showing the expression level of FASN gene in experimental mice according to diet type.
다른 식으로 정의되지 않는 한, 본 명세서에서 사용된 모든 기술적 및 과학적 용어들은 본 발명이 속하는 기술 분야에서 숙련된 전문가에 의해서 통상적으로 이해되는 것과 동일한 의미를 갖는다. 일반적으로, 본 명세서에서 사용된 명명법은 본 기술 분야에서 잘 알려져 있고 통상적으로 사용되는 것이다.Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by an expert skilled in the art to which the present invention belongs. In general, the nomenclature used in this specification is well known and commonly used in the art.
본 발명은 케피어 유래 유산균의 사균체 및 포도씨 분말을 유효성분으로 함유하는 비만의 개선용 조성물에 관한 것이다.The present invention relates to a composition for improving obesity containing dead cells of kefir-derived lactic acid bacteria and grape seed powder as an active ingredient.
본 발명에서 “프로바이오틱스(probiotics)”란, 섭취되어 장에 도달하였을 때에 장내 환경에 유익한 작용을 하는 균주를 의미한다. In the present invention, "probiotics" refers to strains that have a beneficial effect on the intestinal environment when ingested and reach the intestine.
본 발명에서 "프리바이오틱스(Prebiotics)"란, 장내 유익한 박테리아의 생장을 돕는 난소화성 성분으로서 프로바이오틱스(probiotics)의 영양원이 되어 장내 환경을 개선하는 데 도움을 주는 물질을 말한다.In the present invention, "Prebiotics" refers to substances that help improve the intestinal environment by serving as a nutrient source for probiotics as an indigestible component that helps the growth of beneficial bacteria in the intestine.
본 발명에서, 케피어(kefir)는 코카서스 산간 지방에서 유래한 발효유의 일종으로, 케피어는 유산균과 효모 복합체인 케피어 그레인으로 불리우는 균종을 개시제로 하여 우유를 발효시켜 얻어진 요구르트상의 음료를 의미한다.In the present invention, kefir is a kind of fermented milk derived from mountainous Caucasus, and kefir refers to a yogurt-like beverage obtained by fermenting milk using a species called kefir grain, which is a complex of lactic acid bacteria and yeast, as an initiator. .
본 발명에 있어서, “사균체”란 생균의 반대되는 개념으로서 발효를 통해 얻어진 생균과 대사산물들을 열처리 등에 의해 균의 성장이 일어나지 못하도록 한 형태를 의미한다. 사균체는 세포질(cytoplasm), 세포벽(cell wall), 박테리오신(bacteriocin) 등의 항균활성 물질, 다당류(polysaccharide), 유기산 등을 포함할 수 있다. In the present invention, the term "dead cells" is the opposite concept of live cells and refers to a form in which the growth of the bacteria is prevented by heat treatment or the like obtained through fermentation. Dead cells may include cytoplasm, cell wall, antibacterial active substances such as bacteriocin, polysaccharides, organic acids, and the like.
사균체 제품은 생균 제품과 비교하여 높은 안정성을 가지고 있다. 특히 내열성이 우수하며, 외부 환경에 대한 안정성이 높아 기존 생균 제품보다 보관이 용이하고 유통기간을 늘일 수 있다는 장점을 가지고 있다. 또한, 항생제 사용에 대한 규제가 강화되고 있기 때문에 대체제로서의 활용성과 아직 사균체 제품 생산에 본격적으로 뛰어든 업체가 손에 꼽을 정도이기 때문에 시장성과 성장가능성이 매우 크다.Dead cell products have high stability compared to live cells. In particular, it has the advantage that it has excellent heat resistance and high stability against the external environment, so it is easier to store and extend the shelf life than existing live bacteria products. In addition, as the regulations on the use of antibiotics are strengthening, the marketability and growth potential are very high, as there are many companies that have been able to use them as substitutes and are still in full swing in the production of dead cells.
본 발명에서는, 케피어로부터 분리된 유산균이 열처리(heat-killed)에 의해 불활성화된 사균체의 형태로 섭취되는 경우에도 항-비만 활성을 발휘한다는 것을 발견하였다. 따라서, 가공 식품의 살균 가공 과정에서도 항-비만 활성을 잃지 않고 다양한 가공 형태로 응용될 수 있는 가능성이 있다.In the present invention, it was found that the lactic acid bacteria isolated from kefir exhibit anti-obesity activity even when ingested in the form of dead cells inactivated by heat-killed. Therefore, there is a possibility that it can be applied in various processing forms without losing anti-obesity activity even in the sterilization process of processed foods.
본 발명에서, 항-비만 활성을 갖는 사균체로서 적용될 수 있는 케피어 유래 유산균은 락토바실러스 케피리(Lactobacillus kefiri), 류코노스톡 메센테로이데스(leuconostoc mesenteroides), 또는 이들의 복합 균체일 수 있다.In the present invention, kefir-derived lactic acid bacteria that can be applied as dead cells having anti-obesity activity may be Lactobacillus kefiri , leuconostoc mesenteroides , or a complex cell thereof.
바람직하게는, 본 발명에 사용가능한 케피어 유래 유산균은 락토바실러스 케피리 DH5(Lactobacillus kefiri DH5, 기탁기관: 한국미생물보존센터, 수탁번호: KCCM11837P, 수탁일자: 2016년 4월 29일), 류코노스톡 메센테로이데스 LCM4(leuconostoc mesenteroides LCM4, 기탁기관: 한국미생물보존센터, 수탁번호: KCCM12117P, 수탁일자: 2017년 9월 21일), 또는 이들의 복합 균체일 수 있다. 가장 바람직하게는, 락토바실러스 케피리 DH5 및 류코노스톡 메센테로이데스 LCM4의 복합 균체를 사용하는 것이 비만의 예방 또는 치료에 적합하다. Preferably, the kefir-derived lactic acid bacteria usable in the present invention is Lactobacillus kefiri DH5 ( Lactobacillus kefiri DH5, deposit institution: Korea Microorganism Conservation Center, accession number: KCCM11837P, accession date: April 29, 2016), Leukono It may be a stock mesenteroides LCM4 ( leuconostoc mesenteroides LCM4, deposit institution: Korea Microorganism Conservation Center, accession number: KCCM12117P, accession date: September 21, 2017), or a complex cell thereof. Most preferably, it is suitable for the prevention or treatment of obesity to use complex cells of Lactobacillus kepiri DH5 and Leukonostock mecenteroides LCM4.
상기 복합 균체는 락토바실러스 케피리 DH5 및 류코노스톡 메센테로이데스 LCM4를 10:1 내지 1:10의 농도로 혼합하여 사용할 수 있으며, 바람직하게는 5:1 내지 1:5의 농도로 혼합하여 사용할 수 있다.The complex cells may be used by mixing Lactobacillus kepiri DH5 and Leukonostock mecenteroides LCM4 at a concentration of 10:1 to 1:10, preferably mixed at a concentration of 5:1 to 1:5. I can.
본 발명에 따른 케피어 유래 유산균의 사균체는 케피어 유래 유산균을 80℃ 이상에서 열처리 사멸시킨 것일 수 있다. 또는, 상기 사균체는 케피어 유래 유산균을 열처리 사멸시킨 후 파쇄한 현탁액의 형태일 수 있다. 또는, 상기 사균체는 케피어 유래 유산균을 열처리 사멸 후 파쇄한 현탁액의 수용성 분획의 형태일 수 있다.The dead cells of kefir-derived lactic acid bacteria according to the present invention may be killed by heat treatment at 80° C. or higher of kefir-derived lactic acid bacteria. Alternatively, the dead cells may be in the form of a suspension obtained by crushing the kefir-derived lactic acid bacteria by heat treatment. Alternatively, the dead cells may be in the form of an aqueous fraction of a suspension obtained by crushing kefir-derived lactic acid bacteria after heat treatment and killing.
본 발명자들은 또한, 포도씨 분말이 케피어 유래 유산균 사균체의 프로바이오틱스 활성에 유익한 영향을 미치는 프리바이오틱스(prebiotics)로서 작용한다는 것을 확인하였다.The present inventors also confirmed that the grape seed powder acts as prebiotics having a beneficial effect on the probiotic activity of dead cells of kefir-derived lactic acid bacteria.
본 발명에 이용되는 포도의 품종, 산지 또는 수확시기 등에는 특별한 제한은 없으나, 샤르도네(Chardonnay) 포도를 이용하는 것이 가장 바람직하다.There is no particular limitation on the variety, production area, or harvest time of grapes used in the present invention, but it is most preferable to use Chardonnay grapes.
본 발명의 포도씨 분말은 포도를 재료로 하는 음료나 주류의 제조과정에서 발생되는 부산물로부터 생성될 수 있다. 바람직하게는, 상기 포도씨 분말은 포도주 또는 와인(wine)의 제조과정에서 발생되는 부산물로부터 생성될 수 있다.The grape seed powder of the present invention may be produced from by-products generated during the manufacturing process of beverages or alcoholic beverages using grapes. Preferably, the grape seed powder may be produced from by-products generated during the manufacturing process of wine or wine.
여기서, "부산물"은 포도를 원료로 하여 파쇄 및 발효 공정을 거친 후 증류 공정에서 생성되는 와인 또는 포도주 증류액을 제외한 나머지 생성물을 총칭한다.Here, "by-product" refers to the remaining products excluding wine or wine distillate produced in a distillation process after crushing and fermentation using grapes as a raw material.
통상적으로, 와인 또는 포도주 제조시 발생되는 부산물들은 폐기되고 있다. 그러나 이러한 폐기물은 원료 포도의 40% 정도에 이르러 경제적으로 큰 손실이 아닐 수 없다. 본 발명에 따르면, 와인 제조시 발생되는 부산물을 프리바이오틱스로 활용할 수 있기 때문에, 환경적, 경제적으로도 큰 이점이 있다.Typically, wine or by-products generated during wine production are being discarded. However, such waste amounts to about 40% of the raw grapes, which is a huge economic loss. According to the present invention, since by-products generated during wine production can be utilized as prebiotics, there is a great advantage in environmental and economic terms.
본 발명의 일 실시예에 따른 "포도씨 분말"(Grape seed flour, GSF)은 와인을 생산하고 남은 부산물로부터 포도씨를 분리, 건조한 뒤, "냉각프레스", "열프레스" 기법 등을 이용하여 포도씨로부터 오일을 추출한 다음, 가루로 분쇄함으로써 생성될 수 있다. 포도씨 분말의 분리, 건조, 압축, 분쇄 방법은 종래 알려진 기술들에 의해 다양한 형태로 수행될 수 있다."Grape seed flour" (GSF) according to an embodiment of the present invention separates grape seeds from by-products remaining after producing wine, dried, and then from grape seeds using a "cooling press", "heat press" technique, etc. It can be produced by extracting the oil and then grinding it into a powder. Separation, drying, compression, and pulverization of the grape seed powder may be performed in various forms by conventionally known techniques.
포도씨 분말의 입자 크기는 분쇄 환경에 따라 다양하게 조절할 수 있으며, 더 작은 입자를 가진 분말을 생성하기 위해 체로 거르는 작업을 추가할 수 있다. 포도씨 분말의 입자 크기는 식품, 의약품 등 활용 용도와 목적에 맞게 조절 가능하다.The particle size of the grape seed powder can be variously adjusted according to the grinding environment, and sieving can be added to produce a powder with smaller particles. The particle size of grape seed powder can be adjusted according to the purpose and use of food and medicine.
본 발명에서는 포도씨 분말 및 케피어 유래 유산균의 사균체를 섭취하는 경우 고지방식이로 인한 비만 개선에 우수한 효과를 나타낸다는 것을 발견하였다. 특히, 포도씨 분말 및 케피어 유래 유산균 사균체는 함께 섭취되었을 때 시너지 효과를 발휘하여 이들을 각각 섭취하였을 경우로부터 예측하기 어려운 현저한 효과를 나타낸다는 것을 확인하였다.In the present invention, it was found that the ingestion of dead cells of grape seed powder and kefir-derived lactic acid bacteria exhibits excellent effects in improving obesity due to a high fat diet. In particular, it was confirmed that grape seed powder and kefir-derived lactic acid bacteria dead cells exert a synergistic effect when ingested together, and show a remarkable effect that is difficult to predict from the case of ingesting them respectively.
구체적으로, 포도씨 분말 및 케피어 유래 유산균 사균체를 함께 섭취하는 경우 고지방식이로 인한 체중 증가와 지방 세포 무게의 증가를 억제할 수 있다. 본 발명의 일 실시예에서는 고지방식이로 유도된 비만 실험쥐에게 포도씨 분말 및 케피어 유래 유산균 사균체를 고지방식이와 함께 섭취시킨 경우 체중 증가와 지방 세포 무게의 증가가 크게 감소할 수 있다는 것을 확인하였다. Specifically, when ingesting grape seed powder and kefir-derived lactic acid bacteria dead cells together, weight gain and fat cell weight increase due to a high fat diet can be suppressed. In one embodiment of the present invention, when the obese mice induced by a high fat diet are ingested with grape seed powder and kefir-derived lactic acid bacteria dead cells together with a high fat diet, weight gain and an increase in fat cell weight can be significantly reduced. Confirmed.
또한, 본 발명의 포도씨 분말 및 케피어 유래 유산균 사균체를 섭취한 실험쥐는 이들을 각각 섭취한 실험쥐에 비하여 총콜레스테롤 수치가 더욱 감소되었으며, 지방을 생성하는데 관여하는 유전제 발현도 감소시킬 수 있다는 것을 확인하였다. In addition, the mice that ingested the grape seed powder and kefir-derived lactic acid bacteria dead cells of the present invention further reduced the total cholesterol level compared to the mice that ingested them respectively, and that the expression of genetic agents involved in fat production can also be reduced. Confirmed.
상기 케피어 유래 유산균 사균체는 포도씨분말의 중량에 대하여, 0.1x108 내지 20x108 유산균/포도씨분말 gram으로 혼합되는 것이 바람직하며, 1.0x108 내지 10x108 유산균/포도씨분말 gram이 더욱 바람직하고, 약 8x108 유산균/포도씨분말 gram으로 혼합되는 것이 가장 바람직하다. The kefir four-derived lactic acid bacterium cells is based on the weight of the grape seed powder, 0.1x10 8 to 20x10 8 preferably mixed with lactic acid / grape seed powder, and gram, 1.0x10 8 to 10x10 8 lactic acid bacteria and / grape seed powder gram is more preferred, from about It is most preferable to mix 8x10 8 lactic acid bacteria/grams of grape seed powder.
본 발명의 포도씨 분말 및 케피어 유래 유산균 사균체를 함유하는 조성물은 비만 개선용 기능성 식품으로서 활용될 수 있다.The composition containing the grape seed powder and kefir-derived lactic acid bacteria dead cells of the present invention can be utilized as a functional food for improving obesity.
상기 기능성 식품은 발효식품인 것이 바람직하지만, 이에 제한되지는 않는다. 예를 들어서, 기능성 식품은 음료, 껌, 차, 비타민 복합제, 건강보조 식품류일 수 있으며, 더욱 자세히는, 유제품, 제과물, 조미료, 음료 및 드링크제, 스낵, 캔디류, 젤리류, 아이스크림 및 냉동용 디저트, 아침 곡물류, 영양바, 스낵 바 초콜렛 제품, 가공 식품, 곡물 제품 및 파스타, 스프, 소스 및 드레싱, 과자 제품, 오일 및 지방 제품, 유제품 음료 (dairy drink) 및 우유 음료, 차, 두유 및 콩 유제품 (soy dairy-like product), 냉동식품, 조리 음식 및 대체 음식, 육류 제품, 치즈, 요구르트, 빵 및 롤빵, 케이크, 쿠키 및 크래커로 이루어진 군에서 선택된 어느 하나일 수 있다. The functional food is preferably a fermented food, but is not limited thereto. For example, functional foods may be beverages, gums, teas, vitamin complexes, dietary supplements, and more specifically, dairy products, confectionery, seasonings, beverages and drinks, snacks, candies, jellies, ice cream and frozen desserts. , Breakfast cereals, nutrition bars, snack bars chocolate products, processed foods, grain products and pasta, soups, sauces and dressings, confectionery products, oil and fat products, dairy drinks and milk beverages, tea, soy milk and soy milk products (soy dairy-like product), frozen foods, cooked foods and alternative foods, meat products, cheese, yogurt, bread and rolls, cakes, cookies, and may be any one selected from the group consisting of crackers.
또한, 상기 기능성 식품은 케피어 유래 유산균 사균체 및 포도씨 분말을 포함하는 캡슐, 정제, 분말, 액상 현탁액, 환제 및 과립제 등의 제형으로 제조될 수 있다.In addition, the functional food may be prepared in a dosage form such as capsules, tablets, powders, liquid suspensions, pills and granules including kefir-derived lactic acid bacteria dead cells and grape seed powder.
또한, 상기 기능성 식품은 유효성분으로서 케피어 유래 유산균 사균체 및 포도씨 분말 뿐만 아니라, 식품 제조 시에 통상적으로 첨가되는 성분을 추가로 포함할 수 있다.In addition, the functional food may further include, as an active ingredient, not only kefir-derived lactic acid bacteria dead cells and grape seed powder, but also ingredients commonly added during food production.
본 발명의 기능성 식품은 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, 알코올, 탄산 음료에 사용되는 탄산화제 등을 함유할 수 있으며, 과일주스나 야채 음료 제조를 위한 과육을 함유할 수 있다.Functional foods of the present invention include various nutrients, vitamins, minerals (electrolytes), flavoring agents such as synthetic flavoring agents and natural flavoring agents, coloring agents and thickeners (cheese, chocolate, etc.), pectic acid and salts thereof, alginic acid and salts thereof. , Organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonates used in carbonated beverages, and the like, and may contain pulp for the production of fruit juice or vegetable beverages.
본 발명의 포도씨 분말 및 케피어 유래 유산균 사균체를 함유하는 조성물은 또한, 비만의 예방 또는 치료용 약학 조성물로서 활용될 수 있다.The composition containing the grape seed powder and kefir-derived lactic acid bacteria dead cells of the present invention can also be utilized as a pharmaceutical composition for preventing or treating obesity.
본 발명에 따른 약학 조성물은 케피어 유래 유산균 사균체 및 포도씨 분말을 유효성분으로서 함유하며, 통상의 방법에 따른 적절한 담체, 부형제 또는 희석제를 추가로 포함할 수 있다. 상기 약제학적으로 허용되는 담체는 제제시에 통상적으로 이용되는 것으로서, 락토스, 덱스트로스, 수크로스, 솔비톨, 만니톨, 전분, 아카시아 고무, 인산 칼슘, 알기네이트, 젤라틴, 규산 칼슘, 미세결정성 셀룰로스, 폴리비닐피롤리돈, 셀룰로스, 물, 시럽, 메틸 셀룰로스, 메틸히드록시벤조에이트, 프로필히드록시벤조에이트, 활석, 스테아르산 마그네슘 및 미네랄 오일 등을 포함하나, 이에 한정되는 것은 아니다. The pharmaceutical composition according to the present invention contains kefir-derived lactic acid bacteria dead cells and grape seed powder as active ingredients, and may further include an appropriate carrier, excipient, or diluent according to a conventional method. The pharmaceutically acceptable carrier is commonly used in the formulation, lactose, dextrose, sucrose, sorbitol, mannitol, starch, gum acacia, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, Polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, mineral oil, and the like, but are not limited thereto.
본 발명의 약학 조성물은 상기 성분들 이외에 윤활제, 습윤제, 감미제, 향미제, 유화제, 현탁제, 보존제 등을 추가로 포함할 수 있다. 적합한 약제학적으로 허용되는 담체 및 제제화에 관해서는 레밍턴의 문헌[Remington's Pharmaceutical Sciences (19th ed., 1995)]에 개시되어 있는 방법을 이용하여 각 성분에 따라 바람직하게 제제화할 수 있다.The pharmaceutical composition of the present invention may further include a lubricant, a wetting agent, a sweetening agent, a flavoring agent, an emulsifying agent, a suspending agent, a preservative, and the like in addition to the above components. Regarding suitable pharmaceutically acceptable carriers and formulations, it can be preferably formulated according to each component using the method disclosed in Remington's Pharmaceutical Sciences (19th ed., 1995).
본 발명의 약학 조성물은 경구 또는 비경구 투여 모두 가능하며, 비경구 투여는 정맥내 주입, 피하 주입, 근육 주입, 복강 주입, 경피 투여 등을 포함한다. The pharmaceutical composition of the present invention may be administered orally or parenterally, and parenteral administration includes intravenous injection, subcutaneous injection, intramuscular injection, intraperitoneal injection, transdermal administration, and the like.
경구 투여용 제형으로는 예를 들면 정제, 환제, 경질, 연질 캅셀제, 액제, 현탁제, 유화제, 시럽제, 과립제 등이 있는데, 이들 제형은 유효성분 이외에 희석제(예: 락토즈, 덱스트로즈, 수크로즈, 만니톨, 솔비톨, 셀룰로즈 및/또는 글리신), 활택제(예: 실리카, 탈크, 스테아르산 및 그의 마그네슘 또는 칼슘염 및/ 또는 폴리에틸렌 글리콜)를 추가로 포함할 수 있다. 또한, 상기 정제는 마그네슘 알루미늄 실리케이트, 전분 페이스트, 젤라틴, 트라가칸스, 메틸셀룰로즈, 나트륨 카복시메틸셀룰로즈 및/또는 폴리비닐피롤리딘과 같은 결합제를 함유할 수 있으며, 경우에 따라 전분, 한천, 알긴산 또는 그의 나트륨 염과 같은 붕해제 또는 비등 혼합물 및/또는 흡수제, 착색제, 향미제 및 감미제를 함유할 수 있다. 상기 제형은 통상적인 혼합, 과립화 또는 코팅 방법에 의해 제조될 수 있다.Formulations for oral administration include, for example, tablets, pills, hard, soft capsules, solutions, suspensions, emulsifiers, syrups, and granules. These formulations include diluents (e.g., lactose, dextrose, water) in addition to the active ingredients. Krose, mannitol, sorbitol, cellulose and/or glycine), lubricants (eg, silica, talc, stearic acid and magnesium or calcium salts thereof and/or polyethylene glycol). In addition, the tablet may contain a binder such as magnesium aluminum silicate, starch paste, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose and/or polyvinylpyrrolidine, in some cases starch, agar, alginic acid Or disintegrants or boiling mixtures and/or absorbents, colorants, flavors and sweeteners such as sodium salts thereof. The formulation can be prepared by conventional mixing, granulating or coating methods.
또한, 비경구 투여용 제형의 대표적인 것은 주사용 제제이며, 주사용 제제의 용매로서 물, 링거액, 등장성 생리 식염수 또는 현탁액을 들 수 있다. 상기 주사용 제제의 멸균 고정 오일은 용매 또는 현탁 매질로서 사용할 수 있으며 모노-, 디-글리세라이드를 포함하여 어떠한 무자극성 고정오일도 이러한 목적으로 사용될 수 있다. 또한, 상기 주사용 제제는 올레산과 같은 지방산을 사용할 수 있다.In addition, a typical formulation for parenteral administration is a formulation for injection, and as a solvent for the formulation for injection, water, Ringer's solution, isotonic physiological saline or suspension may be mentioned. The sterile fixed oil of the injectable preparation can be used as a solvent or suspension medium, and any non-irritating fixed oil including mono- and di-glycerides can be used for this purpose. In addition, the injectable formulation may use a fatty acid such as oleic acid.
본 발명에 따른 조성물은 약학적으로 유효한 양으로 투여한다. 본 발명에 있어서, “약학적으로 유효한 양”은 의학적 치료에 적용 가능한 합리적인 수혜/위험 비율로 질환을 치료하기에 충분한 양을 의미하며, 유효 용량 수준은 환자의 질환의 종류, 중증도, 약물의 활성, 약물에 대한 민감도, 투여 시간, 투여 경로 및 배출 비율, 치료기간, 동시 사용되는 약물을 포함한 요소 및 기타 의학 분야에 잘 알려진 요소에 따라 결정될 수 있다. 본 발명에 따른 조성물은 개별 치료제로 투여하거나 다른 치료제와 병용하여 투여될 수 있고 종래의 치료제와는 순차적 또는 동시에 투여될 수 있으며, 단일 또는 다중 투여될 수 있다. 상기한 요소들을 모두 고려하여 부작용 없이 최소한의 양으로 최대 효과를 얻을 수 있는 양을 투여하는 것이 중요하며, 이는 통상의 지식을 가진 기술자에 의해 용이하게 결정될 수 있다.The composition according to the present invention is administered in a pharmaceutically effective amount. In the present invention, “pharmaceutically effective amount” means an amount sufficient to treat a disease at a reasonable benefit/risk ratio applicable to medical treatment, and the effective dose level is the type of disease, severity, and drug activity of the patient. , Sensitivity to drugs, time of administration, route of administration and rate of excretion, duration of treatment, factors including drugs used concurrently, and other factors well known in the medical field. The composition according to the present invention may be administered as an individual therapeutic agent or administered in combination with other therapeutic agents, may be administered sequentially or simultaneously with a conventional therapeutic agent, and may be administered single or multiple. It is important to administer an amount capable of obtaining the maximum effect in a minimum amount without side effects in consideration of all the above factors, and this can be easily determined by a person skilled in the art.
구체적으로, 본 발명에 따른 조성물의 유효량은 환자의 나이, 성별, 체중에 따라 달라질 수 있으며, 일반적으로는 체중 1kg 당 0.001 내지 150mg, 바람직하게는 0.01 내지 100mg을 매일 또는 격일 투여하거나 1일 1 내지 3회로 나누어 투여할 수 있다. 그러나 투여 경로, 성별, 체중, 연령 등에 따라서 증감될 수 있으므로 상기 투여량이 어떠한 방법으로도 본 발명의 범위를 한정하는 것은 아니다.Specifically, the effective amount of the composition according to the present invention may vary depending on the age, sex, and weight of the patient, and in general, 0.001 to 150 mg, preferably 0.01 to 100 mg per 1 kg of body weight, daily or every other day, or 1 to 1 day It can be administered in 3 divided doses. However, since it may increase or decrease according to the route of administration, sex, weight, age, etc., the dosage amount does not limit the scope of the present invention in any way.
실시예Example
이하, 실시예를 통하여 본 발명을 더욱 상세히 설명하고자 한다. 이들 실시예는 오로지 본 발명을 예시하기 위한 것으로, 본 발명의 범위가 이들 실시예에 의해 제한되는 것으로 해석되지는 않는다는 것은 당업계에서 통상의 지식을 가진 자에게 있어서 자명할 것이다.Hereinafter, the present invention will be described in more detail through examples. These examples are for illustrative purposes only, and it will be apparent to those of ordinary skill in the art that the scope of the present invention is not to be construed as being limited by these examples.
모든 실시예의 실험 결과는 평균(mean)±표준오차(SEM)으로 표시하였고, 각 균주간의 통계적 유의성 검증은 Staticstical Package for Social Sience(SPSS) version 21.0을 이용하여 p<0.05의 수준에서 one-way analysis of variance(ANOVA) 및 Duncan's multiple range test를 통해 검증하였다.Experimental results of all examples were expressed as mean ± standard error (SEM), and statistical significance between each strain was verified using Staticstical Package for Social Sience (SPSS) version 21.0, and one-way analysis at a level of p<0.05. of variance (ANOVA) and Duncan's multiple range test.
제조예 1: 열처리 사균체의 제조Preparation Example 1: Preparation of heat-treated dead cells
락토바실러스 케피리 DH5 및 류코노스톡 메센테로이데스 LCM4를 MRS 액체 배지에서 30℃로 24시간 동안 배양하였다.Lactobacillus kepiri DH5 and Leukonostock mecenteroides LCM4 were incubated in MRS liquid medium at 30° C. for 24 hours.
배양된 각 균을 9,190×g로 5분간 4℃에서 원심분리한 후 멸균생리식염수에 2회, 멸균증류수에 1회 세척하고, 95℃에서 5분간 열처리 사멸(heat-killed)시켰다. Each cultured bacteria was centrifuged at 4°C for 5 minutes at 9,190×g, washed twice in sterile physiological saline and once in sterile distilled water, and heat-killed for 5 minutes at 95°C.
사균 처리된 두 균주의 샘플을 1:1의 비율로 잘 섞어 튜브식(gavage) 투여를 통해 최종적으로 실험쥐에게 투여하였다.Samples of the two strains treated with dead bacteria were mixed well in a ratio of 1:1 and finally administered to the experimental mice through gavage administration.
제조예 2: 포도씨 분말 준비Preparation Example 2: Grape seed powder preparation
샤르도네(Chardonnay) 포도씨 가루를 미국 ApresVin Enterprises, Inc.에서 구매하여 사용하였다. Chardonnay grape seed powder was purchased and used from ApresVin Enterprises, Inc., USA.
상기 포도씨 가루는 샤르도네(Chardonnay) 포도를 이용하여 와인을 만들고 남은 부산물을 냉압착(cold-pressed)하여, 포도씨에서 기름을 제거하고, 남은 생성물을 열풍 건조하고, air blow를 이용하여 껍질 부분을 제거한 다음, 남은 생성물을 100메쉬(150마이크론) 사이즈를 가진 체를 이용하여 거른 고운 입자의 포도씨 분말(Grape seed flour, GSF)의 형태이다.The grape seed powder is made of wine using Chardonnay grapes, and the remaining by-products are cold-pressed to remove oil from the grape seeds, and the remaining products are hot air dried, and the skin part is removed using an air blow. Next, the remaining product is in the form of fine grained grape seed flour (GSF), which is filtered using a sieve having a size of 100 mesh (150 microns).
실험예 1: 식이에 따른 실험쥐의 체중 변화 분석Experimental Example 1: Analysis of body weight change in mice according to diet
C57BL/6J 실험쥐 110마리에게 아래의 표 1에 나타낸 바와 같은 4가지 종류의 식이를 9주 동안 섭취하도록 하여 비만 질환을 유도하였다.Obesity disease was induced by allowing 110 C57BL/6J mice to consume four types of diets as shown in Table 1 below for 9 weeks.
(HFrD)(HFrD)
2.5% GSF2.5% GSF
2.5% GSF +2.5% GSF +
HLABHLAB
유산균 사균체(HLAB)Lactobacillus dead cells (HLAB)
도 1(a)는 식이 종류에 따른 실험쥐의 총 에너지 섭취량을 나타내며, 도 1(b)는 식이 종류에 따른 실험쥐의 체중 증가량을 나타낸다.Figure 1 (a) shows the total energy intake of the mice according to the type of diet, Figure 1 (b) shows the weight gain of the mice according to the type of diet.
도 1(a) 및 (b)에서, 총 에너지 섭취량이 실험군 사이에 유의적인 차이가 없었음에도 불구하고, 포도씨 가루 및/또는 유산균 사균체를 섭취한 군에서 유의적으로 낮은 체중 증가가 발생한 것을 확인할 수 있다. In Figures 1 (a) and (b), although there was no significant difference in total energy intake between the experimental groups, it was confirmed that a significantly lower weight gain occurred in the group ingesting grape seed powder and/or lactic acid bacteria dead cells. I can.
특히, 포도씨 가루와 케피어 유산균 사균체를 함께 섭취한 실험군은 이들을 각각 섭취한 실험군에 비하여 현저히 낮은 체중 증가를 나타내어, 포도씨 가루와 케피어 유산균 사균체가 시너지 효과를 발휘한다는 것을 알 수 있다.In particular, the experimental group ingesting grape seed powder and kefir lactic acid bacteria together showed significantly lower weight gain compared to the experimental group ingesting them, indicating that grape seed powder and kefir lactic acid bacteria dead cells exert a synergistic effect.
실험예 2: 식이에 따른 지방 조직 무게 변화Experimental Example 2: Changes in fat tissue weight according to diet
도 2는 식이 종류에 따른 지방 조직 무게의 변화를 나타낸다.2 shows the change in the weight of adipose tissue according to the type of diet.
각 실험군에 따른 총 에너지 섭취량이 유의적인 차이가 없었음에도 불구하고, 포도씨 가루 및/또는 유산균 사균체를 섭취한 군에서 유의적으로 지방 조직 무게 감소가 발생하였으며, 포도씨 가루와 케피어 유산균 사균체를 함께 섭취한 실험군은 이들을 각각 섭취한 실험군에 비하여 현저히 낮은 지방 조직 무게 감소를 나타내었다. Although there was no significant difference in total energy intake according to each experimental group, a significant decrease in the weight of adipose tissue occurred in the group that ingested grape seed powder and/or lactic acid bacteria dead cells. The experimental group ingested together showed significantly lower adipose tissue weight reduction compared to the experimental group ingesting them.
실험예 3: 지방 조직 크기 분석Experimental Example 3: Adipose tissue size analysis
상기 실험예 1에서 9주간 식이를 섭취한 실험쥐에 대하여, 지방 조직을 적출하고 10% 포르말린에 고정시킨 후 헤마톡실린-에오신 2중 염색(H&E stain)을 수행하였다. 그 결과를 도 3에 나타내었다. In Experimental Example 1, the adipose tissue was excised and fixed in 10% formalin, and then hematoxylin-eosin double staining (H&E stain) was performed for the mice that had been fed for 9 weeks. The results are shown in FIG. 3.
도 3에서, 포도씨 분말과 케피어 유래 유산균 사균체를 함께 섭취한 실험군의 지방 세포 크기가 이들을 각각 섭취한 실험군의 지방 세포 크기보다 훨씬 작은 것을 확인할 수 있다. In Figure 3, it can be seen that the fat cell size of the experimental group in which the grape seed powder and the kefir-derived lactic acid bacteria dead cells were taken together was much smaller than the fat cell size of the experimental group that ingested them.
실험예 4: 콜레스테롤 분석Experimental Example 4: Cholesterol Analysis
상기 실험예 1의 실험쥐의 간 조직으로부터 생화학자동분석기(7020, HITACHI)를 이용하여, 트리글리세라이드(triglyceride: TG, mg/dL) 및 총콜레스테롤(total cholesterol: T.C, mg/dL)을 측정한 결과를 도 4에 나타내었다. Triglyceride (TG, mg/dL) and total cholesterol (total cholesterol: TC, mg/dL) were measured using an automated biochemical analyzer (7020, HITACHI) from the liver tissue of the experimental mouse of Experimental Example 1 The results are shown in FIG. 4.
도 4에서, TG와 TC 모두 포도씨 분말 및 케피어 유래 유산균 사균체를 함께 섭취한 실험군의 실험쥐가 이들을 각각 섭취한 실험군의 평균값보다 현저히 낮은 값을 나타낸다는 것을 확인할 수 있다.In FIG. 4, it can be seen that both TG and TC showed a significantly lower value than the average value of the experimental group in which the experimental mice ingested grape seed powder and kefir-derived lactic acid bacteria dead cells together.
실험예 5: 유전자 발현 분석Experimental Example 5: Analysis of gene expression
상기 실험예 1의 실험쥐의 지방 조직(epididymal adipose tissue)에서 RNA를 분리한 후 RT-PCR을 통하여 FASN(fatty acid synthase) 유전자 발현량을 측정하였다. 측정한 결과를 도 5에 나타내었다. After RNA was isolated from the epididymal adipose tissue of the experimental mouse of Experimental Example 1, the expression level of the FASN (fatty acid synthase) gene was measured through RT-PCR. The measurement results are shown in FIG. 5.
실험 결과, 포도씨 가루와 케피어 유래 유산균 사균체를 섭취한 실험군의 FASN 유전자의 발현량이 유의적으로 감소하였으며, 포도씨 가루와 케피어 유래 유산균 사균체를 함께 섭취한 실험군의 FASN 유전자 발현량이 가장 많이 감소한 것을 확인할 수 있다. As a result of the experiment, the expression level of FASN gene in the experimental group that ingested grape seed powder and kefir-derived lactic acid bacteria dead cells significantly decreased, and the FASN gene expression level in the experimental group that ingested grape seed powder and kefir-derived lactic acid bacteria dead cells was the most decreased Can be confirmed.
이상으로 본 발명의 내용의 특정부분을 상세히 기술하였는 바, 당업계의 통상의 지식을 가진 자에게 있어서, 이러한 구체적 기술은 단지 바람직한 실시양태일 뿐이며, 이에 의해 본 발명의 범위가 제한되는 것이 아닌 점은 명백할 것이다. 따라서, 본 발명의 실질적인 범위는 첨부된 청구항들과 그것들의 등가물에 의하여 정의된다고 할 것이다.As described above, a specific part of the content of the present invention has been described in detail, and for those of ordinary skill in the art, this specific technology is only a preferred embodiment, and the scope of the present invention is not limited thereby It will be obvious. Accordingly, it will be said that the substantial scope of the present invention is defined by the appended claims and their equivalents.
Claims (15)
상기 케피어 유래 유산균의 사균체가 0.1x108 내지 20x108 유산균/포도씨분말 gram으로 포함되고,
상기 사균체가 케피어 유래 유산균을 열처리 사멸 후 파쇄한 현탁액의 수용성 분획의 형태인, 비만 개선용 기능성 식품.
Kefir (Kefir) containing dead cells of lactic acid bacteria and grape seed powder as an active ingredient,
The dead cells of the kefir-derived lactic acid bacteria are contained in 0.1x10 8 to 20x10 8 lactic acid bacteria/grams of grape seed powder,
Functional food for improving obesity, wherein the dead cells are in the form of a water-soluble fraction of a suspension obtained by crushing kefir-derived lactic acid bacteria after heat treatment and killing.
상기 케피어 유래 유산균이 락토바실러스 케피리(Lactobacillus kefiri), 류코노스톡 메센테로이데스(Leuconostoc mesenteroides), 또는 이들의 복합 균체인 것을 특징으로 하는, 비만 개선용 기능성 식품.
The method of claim 1,
The kefir-derived lactic acid bacteria Lactobacillus kefiri ( Lactobacillus kefiri ), Leuconostoc mesenteroides ( Leuconostoc mesenteroides ), or a complex microorganism thereof, characterized in that, functional food for improving obesity.
상기 복합 균체가 락토바실러스 케피리 및 류코노스톡 메센테로이데스를 5:1 내지 1:5의 농도비로 포함하는 것을 특징으로 하는, 비만 개선용 기능성 식품.
The method of claim 2,
Functional food for improving obesity, characterized in that the complex cells contain Lactobacillus kepiri and Leukonostock mecenteroides in a concentration ratio of 5:1 to 1:5.
상기 케피어 유래 유산균이 락토바실러스 케피리 DH5(Lactobacillus kefiri DH5, 기탁번호: KCCM11837P) 및 류코노스톡 메센테로이데스 LCM4(Leuconostoc mesenteroides LCM4, 기탁번호: KCCM12117P)의 복합 균체인 것을 특징으로 하는, 비만 개선용 기능성 식품.
The method of claim 1,
Improvement of obesity, characterized in that the kefir-derived lactic acid bacteria are complex bacteria of Lactobacillus kefiri DH5 ( Lactobacillus kefiri DH5, accession number: KCCM11837P) and Leuconostoc mesenteroides LCM4 ( Leuconostoc mesenteroides LCM4, accession number: KCCM12117P) For functional food.
상기 사균체는 케피어 유래 유산균을 80℃ 이상에서 열처리 사멸시킨 것임을 특징으로 하는, 비만 개선용 기능성 식품.
The method of claim 1,
The dead cells are functional foods for improving obesity, characterized in that the kefir-derived lactic acid bacteria are killed by heat treatment at 80°C or higher.
지방 세포 형성 유전자의 발현을 억제시키는 것을 특징으로 하는, 비만 개선용 기능성 식품.
The method of claim 1,
Functional food for improving obesity, characterized in that it suppresses the expression of adipocyte-forming genes.
상기 포도씨 분말이 와인 제조 부산물인 것을 특징으로 하는, 비만 개선용 기능성 식품.
The method of claim 1,
Functional food for improving obesity, characterized in that the grape seed powder is a wine production by-product.
상기 포도씨 분말이 샤르도네(Chardonnay) 품종의 포도로 제조된 것을 특징으로 하는, 비만 개선용 기능성 식품.
The method of claim 1,
Functional food for improving obesity, characterized in that the grape seed powder is made of grapes of Chardonnay (Chardonnay) varieties.
상기 케피어 유래 유산균의 사균체가 0.1x108 내지 20x108 유산균/포도씨분말 gram으로 포함되고,
상기 사균체가 케피어 유래 유산균을 열처리 사멸 후 파쇄한 현탁액의 수용성 분획의 형태인, 비만의 예방 또는 치료용 약학 조성물.
Including the dead cells of kefir-derived lactic acid bacteria and grape seed powder as active ingredients,
The dead cells of the kefir-derived lactic acid bacteria are contained in 0.1x10 8 to 20x10 8 lactic acid bacteria/grams of grape seed powder,
A pharmaceutical composition for preventing or treating obesity, wherein the dead cells are in the form of a water-soluble fraction of a suspension obtained by crushing kefir-derived lactic acid bacteria after heat treatment and killing.
상기 케피어 유래 유산균이 락토바실러스 케피리(Lactobacillus kefiri), 류코노스톡 메센테로이데스(Leuconostoc mesenteroides), 또는 이들의 복합 균체인 것을 특징으로 하는, 비만의 예방 또는 치료용 약학 조성물.
The method of claim 12,
The kefir-derived lactic acid bacteria Lactobacillus kefiri ( Lactobacillus kefiri ), Leuconostoc mesenteroides ( Leuconostoc mesenteroides ), or a pharmaceutical composition for the prevention or treatment of obesity, characterized in that the compound thereof.
상기 케피어 유래 유산균이 락토바실러스 케피리 DH5(Lactobacillus kefiri DH5, 기탁번호: KCCM11837P) 및 류코노스톡 메센테로이데스 LCM4(Leuconostoc mesenteroides LCM4, 기탁번호: KCCM12117P)의 복합 균체인 것을 특징으로 하는, 비만의 예방 또는 치료용 약학 조성물.
The method of claim 12,
The kefir-derived lactic acid bacteria are Lactobacillus kefiri DH5 ( Lactobacillus kefiri DH5, accession number: KCCM11837P) and Leuconostoc mesenteroides LCM4 ( Leuconostoc mesenteroides LCM4, accession number: KCCM12117P). Pharmaceutical composition for prophylaxis or treatment.
상기 사균체는 케피어 유래 유산균을 80℃ 이상에서 열처리 사멸시킨 것임을 특징으로 하는, 비만의 예방 또는 치료용 약학 조성물.
The method of claim 12,
The dead cells are characterized in that the killing of kefir-derived lactic acid bacteria by heat treatment at 80° C. or higher, a pharmaceutical composition for preventing or treating obesity.
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