KR20180040899A - Novel Lactobacillus plantarum Ln4 strain and compositions for the prevention and treatment of obesity containing the same - Google Patents
Novel Lactobacillus plantarum Ln4 strain and compositions for the prevention and treatment of obesity containing the same Download PDFInfo
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- KR20180040899A KR20180040899A KR1020160132778A KR20160132778A KR20180040899A KR 20180040899 A KR20180040899 A KR 20180040899A KR 1020160132778 A KR1020160132778 A KR 1020160132778A KR 20160132778 A KR20160132778 A KR 20160132778A KR 20180040899 A KR20180040899 A KR 20180040899A
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Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
- A61K35/741—Probiotics
- A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
- A61K35/747—Lactobacilli, e.g. L. acidophilus or L. brevis
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
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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
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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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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Abstract
Description
본 발명은 전통발효식품으로부터 분리된 프로바이오틱스인 락토바실러스 플란타룸 Ln4(Lactobacillus plantarum Ln4) 균주 및 이의 용도를 제공하는 것이며, 보다 상세하게는 상기 균주의 항비만 용도를 제공한다.The present invention provides strains of Lactobacillus plantarum Ln4 ( Lactobacillus plantarum Ln4), which is a probiotic isolated from traditional fermented foods, and its use, and more specifically, provides anti-obesity uses of said strains.
비만은 주요 성인병들의 위험요소(risk factor)로서, 고혈압, 당뇨병, 동맥경화증, 뇌졸증, 심장마비 및 각종 종양 등과 같은 성인병의 발생에 관여하고(Wilson et al., 2005, Circulation 112: 3066-3072), 병의 진행을 촉진시키기도 하는데, 비만으로 인해 성인병에 걸릴 위험은 정상인보다 3∼6배 높다고 알려져 있다. 따라서, 비만은 단순히 외모상의 문제만이 아니라, 건강에 직결되는 중대한 문제가 아닐 수 없다. 이러한 비만의 예방은 현 상태의 식사량은 유지하면서 운동량을 늘리는 것이 중요하며, 비만이 되었을 경우에는 원인을 파악하고 거기에 맞는 적절한 식이요법을 통하여 적정 체중을 유지하면서, 신체가 정상적인 신진대사를 할 수 있도록 하는 것이 바람직하다.Obesity is a risk factor for major adult diseases and is involved in the development of adult diseases such as hypertension, diabetes, arteriosclerosis, stroke, heart attack and various tumors (Wilson et al., 2005, Circulation 112: 3066-3072) , And promote the progression of disease, obesity, the risk of adult disease is three to six times higher than normal people are known. Therefore, obesity is not only a matter of appearance but also a serious problem directly linked to health. It is important to prevent this obesity by increasing the amount of exercise while maintaining the amount of food in the current state. When obesity becomes a cause, it is necessary to identify the cause and maintain proper body weight through proper dieting .
한편, 김치, 된장, 청국장 등의 다양한 한국 전통발효식품들에는 프로바이오틱스로 가능성이 있는 유산균들이 많이 존재하고 있으며, 전통발효식품에서 분리한 유산균을 이용하는 연구도 많이 진행되고 있다. 또한 프로바이오틱스를 이용한 제품들이 다양하게 출시되고 있다. 프로바이오틱스(probiotics)란 살아있는 상태로 체내에 들어가서 면역 및 다양한 기능성을 주어 건강에 도움을 주는 유익한 미생물을 말한다. 현재까지 알려진 대부분의 프로바이오틱스는 유산균들이지만 바실러스 속(Bacillus spp.) 또는 효모 등도 포함하고 있다. 프로바이오틱스 균주로서 인정을 받기 위해서는 위산 및 담즙산에서 생존하여 소장까지 도달하여야 한다. 또한 장에서 증식하고 정착하여야 하며 장관 내에서 면역 및 다양한 기능성을 나타내야 하며 안전성이 입증되어야 한다.On the other hand, various traditional Korean fermented foods such as kimchi, soybean paste, and chunggukjang have a lot of lactic acid bacteria that are probiotic and there are many studies using lactic acid bacteria isolated from traditional fermented foods. There are also a variety of products using probiotics. Probiotics (probiotics) is a living microorganism that enters the body and gives immunity and various functions to help health. Most probiotics known to date are lactic acid bacteria but also include Bacillus spp. Or yeast. To be recognized as a probiotic strain, it must survive in gastric and bile acids and reach the small intestine. They should also proliferate and settle in the intestines, demonstrate immunity and diverse functionality within the intestinal tract, and demonstrate safety.
현재까지 연구된 바에 따르면, 장류 또는 김치와 같은 발효식품으로부터 분리된 유산균을 프로바이오틱스로 사용할 수 있으며, 이들의 다양한 생리적 기능을 통해 약학적 치료 용도로서 적용할 수 있음이 공지된 바 있다. 예를 들어, 대한민국 등록특허 제 10-161653호에서는 가자미식혜로부터 분리된 신규한 락토코쿠스 락티스(Lactococcus lactis) KR-W.W-2 균주를 포함하는 프로바이오틱스 조성물을 양식 어류의 병원균에 의해 생기는 질병의 치료에 사용할 수 있음을 개시하고 있고, 대한민국 공개특허 10-2015-0106093호에서는 장류로부터 분리된 바실러스 메틸로트로피쿠스(Bacillus methylotrophicus) C14 균주를 프로바이오틱스 제제로서 사용하여 항균용 조성물로서 사용할 수 있음을 개시하고 있다. 이 외에도 다양한 유산균을 유용 용도로서 사용하기 위한 연구가 계속되고 있다.It has been known to date that lactic acid bacteria isolated from fermented foods such as soybean paste or kimchi can be used as probiotics and can be applied as pharmaceutical therapeutic applications through their various physiological functions. For example, Korean Patent No. 10-161653 discloses a probiotic composition comprising a novel Lactococcus lactis KR-WW-2 strain isolated from flounder sikhye, Korean Patent Laid-Open No. 10-2015-0106093 discloses that Bacillus methylotrophicus C14 strain isolated from soybean can be used as a antibacterial composition by using as a probiotic preparation. . In addition, studies have been continuing to use various lactic acid bacteria as useful applications.
이에, 본 발명자들은 전통발효식품으로부터 프로바이오틱스로서 사용할 수 있는 유산균을 분리하고 이의 용도를 찾기 위해 노력한 결과, 된장, 고추장 및 간장과 같은 전통발효식품으로부터 분리된 락토바실러스 플란타룸 Ln4(Lactobacillus plantarum Ln4) 균주를 생균제로서 사용할 수 있으며, 상기 균주는 지방세포에서 지방의 축적을 억제할 수 있고, 고지방식이로 유도된 비만 마우스 모델에 투여하였을 때 유의적인 체지방 감소 효과를 나타낼 수 있음을 확인함으로써 본 발명을 완성하였다.As a result of efforts to isolate lactic acid bacteria which can be used as probiotics from traditional fermented foods and to find their use, Lactobacillus plantarum Ln4 ( Lactobacillus plantarum Ln4) isolated from traditional fermented foods such as soybean paste, The strain can be used as a probiotic agent and the strain can inhibit accumulation of fat in adipocytes and exhibit a significant effect of reducing body fat when administered to an obesity mouse model induced by high fat diet, .
이에, 본 발명자들은 전통발효식품으로부터 분리된 락토바실러스 플란타룸 Ln4(Lactobacillus plantarum Ln4) 균주를 생균제로서 사용할 수 있으며, 이를 고지방식이로 유도된 비만 마우스 모델에 투여하였을 때 유의적인 항비만 효과를 나타낼 수 있음을 확인함으로써 본 발명을 완성하였다.Accordingly, the present inventors have found that a lactobacillus plantarum Ln4 strain isolated from a conventional fermented food can be used as a probiotic agent and has a significant anti-obesity effect when administered to a high fat diet-induced obesity mouse model The present invention has been completed.
따라서, 본 발명의 목적은 수탁번호 KCCM 11897P로 기탁된 신규한 락토바실러스 플란타룸 Ln4(Lactobacillus plantarum Ln4) 균주를 제공하는 것이다.Accordingly, an object of the present invention is to provide a novel Lactobacillus plantarum Ln4 strain deposited with Accession No. KCCM 11897P.
또한, 본 발명의 또다른 목적은 상기 균주를 포함하는 프로바이오틱스 제제를 제공하는 것이다.Still another object of the present invention is to provide a probiotic preparation containing the strain.
아울러, 본 발명의 또다른 목적은 상기 균주의 약학적 용도를 제공하는 것이다.In addition, another object of the present invention is to provide a pharmaceutical use of the strain.
상기 목적을 달성하기 위하여, 본 발명은 수탁번호 KCCM 11897P로 기탁된 락토바실러스 플란타룸 Ln4(Lactobacillus plantarum Ln4) 균주를 제공한다.In order to achieve the above object, the present invention provides a Lactobacillus plantarum Ln4 strain deposited with Accession No. KCCM 11897P.
또한, 본 발명은 수탁번호 KCCM 11897P로 기탁된 락토바실러스 플란타룸 Ln4 균주, 이의 배양액 또는 이들의 혼합물을 유효성분으로 함유하는 프로바이오틱스 제제를 제공한다.The present invention also provides a probiotic preparation containing as an active ingredient a Lactobacillus plantarum Ln4 strain deposited with Accession No. KCCM 11897P, a culture thereof or a mixture thereof.
또한, 본 발명은 수탁번호 KCCM 11897P로 기탁된 락토바실러스 플란타룸 Ln4 균주, 이의 배양액 또는 이들의 혼합물을 유효성분으로 함유하는 비만의 예방 및 치료용 약학적 조성물을 제공한다.In addition, the present invention provides a pharmaceutical composition for preventing and treating obesity, which comprises Lactobacillus plantarum Ln4 strain deposited with Accession No. KCCM 11897P, a culture thereof or a mixture thereof as an active ingredient.
아울러, 본 발명은 수탁번호 KCCM 11897P로 기탁된 락토바실러스 플란타룸 Ln4 균주, 이의 배양액 또는 이들의 혼합물을 유효성분으로 함유하는 비만의 예방 및 개선용 건강기능식품을 제공한다.In addition, the present invention provides a health functional food for prevention and improvement of obesity containing Lactobacillus plantarum Ln4 strain deposited with Accession No. KCCM 11897P, a culture thereof or a mixture thereof as an active ingredient.
본 발명의 바람직한 일실시예에서, 상기 균주는 서열번호 1로 기재되는 16S rRNA 염기서열을 가지는 것일 수 있다.In one preferred embodiment of the present invention, the strain may have the 16S rRNA base sequence set forth in SEQ ID NO: 1.
본 발명의 또다른 바람직한 일실시예에서, 상기 균주는 내산성, 내담즙성, 장부착능 및 항생제 내성을 가지는 것을 특징한다.In another preferred embodiment of the present invention, the strain is characterized by having acid resistance, bile resistance, intestinal adherence, and antibiotic resistance.
본 발명의 또다른 바람직한 일실시예에서, 상기 락토바실러스 플란타룸 Ln4 균주는 지방축적 억제 및 체지방 감소 효과를 나타내는 것일 수 있다.In another preferred embodiment of the present invention, the Lactobacillus plantarum Ln4 strain may exhibit fat accumulation inhibition and body fat reducing effect.
된장, 고추장 및 간장과 같은 전통발효식품으로부터 분리된 락토바실러스 플란타룸 Ln4(Lactobacillus plantarum Ln4) 균주를 생균제로서 사용할 수 있으며, 이는 지방세포에서 지방의 축적을 억제할 수 있고, 고지방식이로 유도된 비만 마우스 모델에 투여하였을 때 유의적인 체지방 감소 효과를 나타낼 수 있으므로, 본 발명의 락토바실러스 플란타룸 Ln4 균주를 비만의 예방, 치료 및 개선을 위한 조성물의 유효성분으로서 사용할 수 있다. Lactobacillus plantarum Ln4 strain isolated from traditional fermented foods such as soybean paste, doenjang, kochujang and soy sauce can be used as a probiotic, which can inhibit the accumulation of fat in adipocytes, , The Lactobacillus plantarum Ln4 of the present invention can be used as an active ingredient of a composition for prevention, treatment and improvement of obesity.
도 1은 락토바실러스 플란타룸 Ln4(Lactobacillus plantarum Ln4) 균주의 계통도이다.
도 2는 지방세포 내에서 락토바실러스 플란타룸 Ln4 처리에 따른 지방축적 억제 효과를 나타낸다.
도 3은 고지방식이에 의한 비만 및 대사성질환 유도 동물 모델에서 락토바실러스 플란타룸 Ln4 투여에 따른 체중 조절 효과를 나타낸다.
도 4는 고지방식이에 의한 비만 및 대사성질환 유도 동물 모델에서 락토바실러스 플란타룸 Ln4 투여에 따른 체지방 감소 효능을 나타낸다.BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a flow diagram of a strain of Lactobacillus plantarum Ln4.
Fig. 2 shows the effect of inhibiting fat accumulation following treatment with Lactobacillus plantarum Ln4 in adipocytes.
FIG. 3 shows the weight control effect of Lactobacillus plantarum Ln4 administration in an animal model for induction of obesity and metabolic disease by high fat diet.
FIG. 4 shows the effect of reducing body fat according to administration of Lactobacillus plantarum Ln4 in an animal model of induction of obesity and metabolic disease by high fat diet.
이하, 본 발명을 상세하게 설명한다.Hereinafter, the present invention will be described in detail.
본 발명은 수탁번호 KCCM 11897P로 기탁된 락토바실러스 플란타룸 Ln4(Lactobacillus plantarum Ln4) 균주를 제공한다.The present invention provides a Lactobacillus plantarum Ln4 strain deposited with Accession No. KCCM 11897P.
또한, 본 발명은 상기 락토바실러스 플란타룸 Ln4 균주, 이의 배양액 또는 이들의 혼합물을 유효성분으로 함유하는 프로바이오틱스 제제를 제공한다.The present invention also provides a probiotic preparation containing the Lactobacillus plantarum Ln4 strain, a culture thereof or a mixture thereof as an active ingredient.
본 발명의 “락토바실러스 플란타룸 Ln4 균주”는 서열번호 1로 기재되는 16S rRNA 염기서열을 가지는 것이 바람직하다.The "Lactobacillus plantarum Ln4 strain" of the present invention preferably has the 16S rRNA nucleotide sequence shown in SEQ ID NO: 1.
본 발명의 “락토바실러스 플란타룸 Ln4 균주”는 내산성, 내담즙성, 장부착능 및 항생제 내성을 가지는 것이 바람직하나, 이에 한정되지 않는다. The " Lactobacillus plantarum Ln4 strain " of the present invention is preferably, but not limited to, those having acid resistance, biliary properties, adherence and antibiotic resistance.
구체적으로, 상기 내산성은 pH 2 내지 3의 배지에서도 3 내지 5 시간 동안 99% 이상의 생존률을 나타낼 수 있음을 나타내며, 상기 내담즙성은 인간의 담즙과 유사한 환경에서 24 시간 동안 80% 이상의 생존률을 나타낼 수 있음이 바람직하다. 또한, 상기 장부착능을 대장 세포와 유의적인 수준으로 결합할 수 있는 능력을 나타낸다. 본 발명의 락토바실러스 플란타룸 Ln4 균주를 프로바이오틱스 제제로 사용하기 위해, 체내 경구투여시에도 균주의 생존능이 유지되는 것을 통해 효과가 저하 또는 증진될 수 있으므로, 소화기관에서의 환경에서 생존할 수 있도록 내산성, 내담즙성 및 장부착능을 가지는 것에서 유의적인 효과를 나타낼 수 있다. 또한, 상기 항생제 내성은 제타마이신, 시프로플록사신, 가나마이신 및 스트렙토마이신에 대해서 내성을 가지는 것이 바람직하나, 이에 한정되지 않는다.Specifically, the acid resistance indicates a survival rate of 99% or more for 3 to 5 hours even in a medium having a pH of 2 to 3, and the biliary bile acid has a survival rate of 80% or more for 24 hours in an environment similar to human bile . It also exhibits the ability to bind the intestinal adherence to the colon cells at a significant level. In order to use the Lactobacillus plantarum Ln4 strain of the present invention as a probiotic agent, the effect can be lowered or promoted by maintaining the viability of the strain even when administered orally in the body, so that it can survive in the environment in the digestive organs. Acidic, brittle and intestine-adhering ability. In addition, the antibiotic resistance is preferably resistant to zetamycin, ciprofloxacin, kanamycin and streptomycin, but is not limited thereto.
본 발명의 구체적인 실시예에 있어서, 본 발명자들은 전통발효식품으로부터 프로바이오틱스로 사용할 수 있는 유산균을 분리하여, 서열번호 1로 기재되는 염기서열로 구성된 16S rRNA를 포함하는 신규 균주를 분리하였으며, 이의 상동성 분석을 통해 락토바실러스 플란타룸 Ln4의 신규한 균주임을 확인하였고(도 1), 이를 2016년 09월 22일 한국미생물보존센터(KCCM)에 기탁하였다(기탁번호 KCCM 11897P).In a specific embodiment of the present invention, the present inventors isolated a lactic acid bacterium which can be used as a probiotic from a fermented food product of the present invention to isolate a novel strain containing 16S rRNA consisting of the nucleotide sequence shown in SEQ ID NO: 1, Analysis confirmed Lactobacillus plantarum Ln4 as a novel strain (Fig. 1), which was deposited on September 22, 2016 with the Korean Society for Microbiological Protection (KCCM) (Deposit No. KCCM 11897P).
또한, 락토바실러스 플란타룸 Ln4의 특징을 확인한 결과, 락토바실러스 플란타룸 Ln4은 내산성, 내담즙성, 장 부착능 및 일부 항생제에 대한 내성을 나타내는 것을 확인하였고, 이에 비해 발암의 원인이 되는 효소를 생산하지 않고 정상 세포에 대하여 독성 역시 나타내지 않음을 확인하여(표 1 내지 표 5), 이러한 특징을 통해 본 발명의 락토바실러스 플란타룸 Ln4 균주를 생균체 또는 사멸체의 형태로 프로바이오틱스 제제에 사용할 수 있을 것으로 확인하였다.Lactobacillus plantarum Ln4 was found to exhibit resistance to acid resistance, bile resistance, intestinal adherence, and resistance to some antibiotics. In contrast, Lactobacillus plantarum Ln4, an enzyme responsible for carcinogenesis (Table 1 to Table 5). Thus, the Lactobacillus plantarum Ln4 strain of the present invention can be used as a bacterial cell or an apoptotic cell in a probiotic preparation Respectively.
따라서, 본 발명의 락토바실러스 플란타룸 Ln4 균주는 신규한 균주로서 정상 세포에 대하여 안전하고 내산성, 내담즙성 및 장 부착능의 특징을 가지고 있으므로, 상기 균주, 이의 배양액 또는 이들의 혼합물을 프로바이오틱스 제제의 유효성분으로 유용하게 사용할 수 있다.Therefore, the Lactobacillus plantarum Ln4 strain of the present invention is a novel strain and is safe against normal cells and has characteristics of acid resistance, biliary and intestinal adherence. Therefore, the strain, its culture or a mixture thereof is used as a probiotic preparation Can be usefully used as an effective ingredient of the present invention.
또한, 본 발명은 수탁번호 KCCM 11897P로 기탁된 락토바실러스 플란타룸 Ln4 균주, 이의 배양액 또는 이들의 혼합물을 유효성분으로 함유하는 비만의 예방 및 치료용 약학적 조성물을 제공한다.In addition, the present invention provides a pharmaceutical composition for preventing and treating obesity, which comprises Lactobacillus plantarum Ln4 strain deposited with Accession No. KCCM 11897P, a culture thereof or a mixture thereof as an active ingredient.
본 발명의 “락토바실러스 플란타룸 Ln4 균주”는 서열번호 1로 기재되는 16S rRNA 염기서열을 가지는 것이 바람직하다.The "Lactobacillus plantarum Ln4 strain" of the present invention preferably has the 16S rRNA nucleotide sequence shown in SEQ ID NO: 1.
본 발명의 “락토바실러스 플란타룸 Ln4 균주”는 내산성, 내담즙성, 장부착능 및 항생제 내성을 가지는 것이 바람직하나, 이에 한정되지 않는다. The " Lactobacillus plantarum Ln4 strain " of the present invention is preferably, but not limited to, those having acid resistance, biliary properties, adherence and antibiotic resistance.
본 발명의 “락토바실러스 플란타룸 Ln4 균주”는 지방축적억제 및 체지방 감소 효과를 나타내는 것을 통해 비만에 대하여 치료 효과를 나타낼 수 있다.The " Lactobacillus plantarum Ln4 strain " of the present invention can exhibit a therapeutic effect on obesity through the inhibition of fat accumulation and the effect of reducing body fat.
본 발명의 또다른 구체적인 실시예에서, 본 발명자들은 락토바실러스 플란타룸 Ln4 균주의 항비만 효과를 확인하기 위해, 고지방식이를 통해 대사성증후군이 유발된 마우스 모델을 제작하였다. 상기 마우스 모델에 락토바실러스 플란타룸 Ln4 균주를 투여하였을 때 지방세포 내에서 지방의 축적이 억제되었으며, 고지방식이로 유도된 비만 마우스 모델에 투여하였을 때 유의적인 체지방 감소 효과를 나타낼 수 있음을 확인하였다(도 2 내지 도 4).In another specific embodiment of the present invention, the present inventors produced a mouse model in which a metabolic syndrome was induced by a high-fat diet method in order to confirm the anti-obesity effect of Lactobacillus plantarum Ln4 strain. It was confirmed that the accumulation of fat in the adipocyte was inhibited when the Lactobacillus plantarum Ln4 strain was administered to the above mouse model and that a significant fat reduction effect could be obtained when administered to a high fat diet induced mouse model of obesity (Figs. 2 to 4).
따라서, 본 발명의 락토바실러스 플란타룸 Ln4 균주를 생균체로서 투여하였을 때 지방 세포에서 지방의 축적을 억제할 수 있고, 체지방 감소 효과를 나타내어 유의적인 항비만 효과를 나타낼 수 있으므로, 본 발명의 락토바실러스 플란타룸 Ln4 균주를 비만의 예방 및 치료용 약학적 조성물의 유효성분으로 사용할 수 있다.Therefore, when the Lactobacillus plantarum Ln4 strain of the present invention is administered as a live cell, it is possible to inhibit accumulation of fat in adipocytes, exhibit a body fat decreasing effect and exhibit a significant anti-obesity effect, The Bacillus plantarum Ln4 strain can be used as an active ingredient of a pharmaceutical composition for the prevention and treatment of obesity.
본 발명의 미생물 제제는 통상적인 생균제 조성물 제조방법에 따라 제조될 수 있으며, 일반적으로, 배양현탁액이나 건조분말 형태일 수 있다. 또한, 주성분인 상기 락토바실러스 플란타룸 Ln4 균주, 또는 이의 배양액의 유효량에 1종 또는 2종 이상의 약제학적으로 허용 가능한 통상적인 담체 또는 1종 또는 2종 이상의 첨가제를 선택하여 통상적인 제형의 조성물로 제조할 수 있다.The microorganism preparation of the present invention can be produced according to a conventional method for producing a probiotic composition, and can be generally in the form of a culture suspension or a dry powder. In addition, it is also possible to use one or more pharmaceutically acceptable conventional carriers or one or more additives to the effective amount of the Lactobacillus plantarum Ln4 strain or a culture solution thereof, which is the main component, Can be manufactured.
담체는 희석제, 활택제, 결합제, 붕해제, 감미제, 안정제, 방부제 중에서 1종 또는 2종 이상을 선택하여 사용할 수 있으며, 첨가제로는 향료, 비타민류, 항산화제 중에서 1종 또는 2종 이상을 선택하여 사용할 수 있다.The carrier may be selected from one or more selected from among diluents, lubricants, binders, disintegrants, sweeteners, stabilizers and preservatives. Examples of the additives include one or more of flavorings, vitamins and antioxidants Can be used.
본 발명에 있어서, 담체 및 첨가제는 약제학적으로 허용 가능한 것은 모두 사용이 가능하며, 구체적으로는 희석제로는 유당(lactose monohydrate), 트레할로스(Trehalose), 옥수수 전분(cornstarch), 콩기름(soybean oil), 미결정 셀룰로오스(microcrystalline cellulose) 또는 만니톨(D-mannitorl)이 좋고, 활택제로는 스테아린산 마그네슘(magnesiumstearate) 또는 탈크(talc)가 바람직하며, 결합제로는 폴리비닐피롤리돈(PVP: polyvinyipyrolidone) 또는 하이드록시프로필셀룰로오스(HPC: hydroxypropylcellulose) 중에서 선택함이 바람직하다. 또한, 붕해제로는 카르복시메칠셀룰로오스칼슘(Ca-CMC: carboxymethylcellulose calcium), 전분글리콜산나트륨(sodium starchglycolate), 폴라크릴린칼륨(polacrylin potassium) 또는 크로스포비돈(cross-linked polyvinylpyrrolidone)중에서 선택함이 바람직하고, 감미제로는 백당, 과당, 소르비톨(sorbitol) 또는 아스파탐(aspartame) 중에서 선택되고, 안정제로는 카르복시메칠셀룰로오스나트륨(Na-CMC: carboxymethylcellulose sodium), 베타-싸이크로덱스트린(β-cyclodextrin), 백납(white bee's wax) 또는 잔탄검(xanthan gum) 중에서 선택되며, 방부제로는 파라옥시안식향산메칠(methyl p-hydroxy benzoate, methlparaben), 파라옥시안식향산프로필(propyl p-hydroxybenzoate, propylparaben), 또는 소르빈산칼륨(potassium sorbate) 중에서 선택하는 것이 바람직하다.In the present invention, the carrier and additives may be any pharmaceutically acceptable ones. Specific examples of the diluent include lactose monohydrate, trehalose, cornstarch, soybean oil, Microcrystalline cellulose or mannitol may be preferably used and the lubricant may be magnesium stearate or talc. The binder may be polyvinylpyrrolidone (PVP) or hydroxypropyl It is preferable to select from hydroxypropylcellulose (HPC). The disintegrant is preferably selected from carboxymethylcellulose calcium (Ca-CMC), sodium starch glycollate, polacrylin potassium or cross-linked polyvinylpyrrolidone And the sweetening agent is selected from white sugar, fructose, sorbitol or aspartame. Examples of the stabilizer include sodium carboxymethylcellulose sodium (Na-CMC), beta-cyclodextrin (beta -cyclodextrin) white bee's wax or xanthan gum and the preservative is selected from the group consisting of methyl p-hydroxy benzoate, methlparaben, propyl p-hydroxybenzoate, propylparaben, or potassium sorbate potassium sorbate).
아울러, 본 발명은 수탁번호 KCCM 11897P로 기탁된 락토바실러스 플란타룸 Ln4 균주, 이의 배양액 또는 이들의 혼합물을 유효성분으로 함유하는 비만의 예방 및 개선용 건강기능식품을 제공한다.In addition, the present invention provides a health functional food for prevention and improvement of obesity containing Lactobacillus plantarum Ln4 strain deposited with Accession No. KCCM 11897P, a culture thereof or a mixture thereof as an active ingredient.
본 발명의 “락토바실러스 플란타룸 Ln4 균주”는 서열번호 1로 기재되는 16S rRNA 염기서열을 가지는 것이 바람직하다.The "Lactobacillus plantarum Ln4 strain" of the present invention preferably has the 16S rRNA nucleotide sequence shown in SEQ ID NO: 1.
본 발명의 “락토바실러스 플란타룸 Ln4 균주”는 내산성, 내담즙성, 장부착능 및 항생제 내성을 가지는 것이 바람직하나, 이에 한정되지 않는다. The " Lactobacillus plantarum Ln4 strain " of the present invention is preferably, but not limited to, those having acid resistance, biliary properties, adherence and antibiotic resistance.
본 발명의 “락토바실러스 플란타룸 Ln4 균주”는 지방축적억제 및 체지방 감소 효과를 나타내는 것을 통해 비만에 대하여 치료 효과를 나타낼 수 있다.The " Lactobacillus plantarum Ln4 strain " of the present invention can exhibit a therapeutic effect on obesity through the inhibition of fat accumulation and the effect of reducing body fat.
본 발명의 락토바실러스 플란타룸 Ln4 균주를 생균체로서 투여하였을 때 지방세포에서 지방의 축적을 억제할 수 있고, 고지방식이로 유도된 비만 마우스 모델에 투여하였을 때 유의적인 체지방 감소 효과를 나타낼 수 있으므로, 본 발명의 락토바실러스 플란타룸 Ln4 균주를 비만의 예방 및 개선용 건강기능식품의 유효성분으로 사용할 수 있다.When the Lactobacillus plantarum Ln4 strain of the present invention was administered as a live cell, fat accumulation in adipocytes could be inhibited, and when administered to a high fat diet induced mouse model of obesity, Therefore, the Lactobacillus plantarum Ln4 strain of the present invention can be used as an active ingredient of a health functional food for prevention and improvement of obesity.
본 발명에 따른 식품 조성물은 당업계에 공지된 통상적인 방법에 따라 다양한 형태로 제조할 수 있다. 일반 식품으로는 이에 한정되지 않지만 음료(알콜성 음료 포함), 과실 및 그의 가공식품(예: 과일통조림, 병조림, 잼, 마아말레이드 등), 어류, 육류 및 그 가공식품(예: 햄, 소시지 콘비이프 등), 빵류 및 면류(예: 우동, 메밀국수, 라면, 스파게티, 마카로니 등), 과즙, 각종 드링크, 쿠키, 엿, 유제품(예: 버터, 치이즈 등), 식용식물 유지, 마아가린, 식물성 단백질, 레토르트 식품, 냉동식품, 각종 조미료(예: 된장, 간장, 소스 등) 등에 본 발명의 락토바실러스 플란타룸 Ln4 균주을 첨가하여 제조할 수 있다. 또한, 영양보조제로는 이에 한정되지 않지만 캡슐, 타블렛, 환 등에 본 발명의 락토바실러스 플란타룸 Ln4 균주을 첨가하여 제조할 수 있다. 또한, 건강기능식품으로는 이에 한정되지 않지만 예를 들면, 본 발명의 락토바실러스 플란타룸 Ln4 균주 자체를 차, 쥬스 및 드링크의 형태로 제조하여 음용(건강음료)할 수 있도록 액상화, 과립화, 캡슐화 및 분말화하여 섭취할 수 있다. 또한, 본 발명의 락토바실러스 플란타룸 Ln4 균주을 식품 첨가제의 형태로 사용하기 위해서는 분말 또는 농축액 형태로 제조하여 사용할 수 있다. 또한, 본 발명의 락토바실러스 플란타룸 Ln4 균주와 비만의 예방 및 개선 효과가 있다고 알려진 공지의 활성 성분과 함께 혼합하여 조성물의 형태로 제조할 수 있다.The food composition according to the present invention can be prepared in various forms according to a conventional method known in the art. Common foods include but are not limited to beverages (including alcoholic beverages), fruits and processed foods (eg, canned fruits, jam, maamalade, etc.), fish, meat and processed foods (Eg butter, chewing), edible plant oil, margarine, soybean oil, etc., etc.), breads and noodles (eg udon, buckwheat noodles, ramen noodles, spaghetti, macaroni, etc.), juice, various drinks, cookies, Vegetable protein, retort food, frozen food, various seasonings (for example, soybean paste, soy sauce, sauce, etc.) by adding the Lactobacillus plantarum Ln4 strain of the present invention. The nutritional supplement may be prepared by adding the Lactobacillus plantarum Ln4 strain of the present invention to capsules, tablets, rings and the like. For example, the health functional foods include, but are not limited to, lactobacillus plantarum Ln4 strain itself of the present invention in the form of tea, juice, and drink to be liquefied, granulated, Encapsulated and powdered. In order to use the Lactobacillus plantarum Ln4 strain of the present invention in the form of a food additive, it may be used in the form of a powder or a concentrated liquid. The Lactobacillus plantarum Ln4 of the present invention may be mixed with a known active ingredient known to have an effect of preventing and improving obesity, and may be prepared in the form of a composition.
본 발명의 락토바실러스 플란타룸 Ln4 균주을 건강음료로 이용하는 경우, 상기 건강음료 조성물은 통상의 음료와 같이 여러 가지 향미제 또는 천연 탄수화물 등을 추가 성분으로 함유할 수 있다. 상술한 천연 탄수화물은 포도당, 과당과 같은 모노사카라이드; 말토스, 슈크로스와 같은 디사카라이드; 덱스트린, 사이클로덱스트린과 같은 폴리사카라이드; 자일리톨, 소르비톨, 에리트리톨 등의 당알콜일 수 있다. 감미제는 타우마틴, 스테비아 추출물과 같은 천연 감미제; 사카린, 아스파르탐과 같은 합성 감미제 등을 사용할 수 있다. 상기 천연 탄수화물의 비율은 본 발명의 조성물 100 mL 당 일반적으로 약 0.01 ～ 0.04 g, 바람직하게는 약 0.02 ～ 0.03 g 이다.When the Lactobacillus plantarum Ln4 strain of the present invention is used as a health drink, the health beverage composition may contain various flavors or natural carbohydrates as an additional ingredient such as ordinary beverages. The above-mentioned natural carbohydrates include monosaccharides such as glucose and fructose; Disaccharides such as maltose, sucrose; Polysaccharides such as dextrin, cyclodextrin; Xylitol, sorbitol, erythritol, and the like. Sweeteners include natural sweeteners such as tau Martin and stevia extract; Synthetic sweetening agents such as saccharin and aspartame, and the like can be used. The ratio of the natural carbohydrate is generally about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g per 100 mL of the composition of the present invention.
또한, 본 발명의 락토바실러스 플란타룸 Ln4 균주은 비만의 예방 및 개선용 식품 조성물의 유효성분으로 함유될 수 있는데, 그 양은 비만의 예방 및 개선의 작용을 달성하기에 유효한 양으로 특별히 한정되는 것은 아니나, 전체 조성물 총 중량에 대하여 0.01 내지 100 중량%인 것이 바람직하다. 본 발명의 식품 조성물은 락토바실러스 플란타룸 Ln4 균주과 함께 비만의 예방 및 개선용 조성물에 효과가 있는 것으로 알려진 다른 활성 성분과 함께 혼합하여 제조될 수 있다.The Lactobacillus plantarum Ln4 strain of the present invention may be contained as an effective ingredient of a food composition for preventing and improving obesity. The amount of the Lactobacillus strain Ln4 is not particularly limited as long as it is effective for preventing and improving obesity , Preferably 0.01 to 100% by weight based on the total weight of the composition. The food composition of the present invention may be prepared by mixing together with Lactobacillus plantarum Ln4 strain together with other active ingredients known to be effective in compositions for the prevention and improvement of obesity.
상기 외에 본 발명의 건강식품은 여러 가지 영양제, 비타민, 전해질, 풍미제, 착색제, 펙트산, 펙트산의 염, 알긴산, 알긴산의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알코올 또는 탄산화제 등을 함유할 수 있다. 그 밖에 본 발명의 건강식품은 천연 과일주스, 과일주스 음료, 또는 야채 음료의 제조를 위한 과육을 함유할 수 있다. 이러한 성분은 독립적으로 또는 혼합하여 사용할 수 있다. 이러한 첨가제의 비율은 크게 중요하진 않지만 본 발명의 조성물 100 중량부당 0.01 ~ 0.1 중량부의 범위에서 선택되는 것이 일반적이다.In addition to the above, the health food of the present invention may contain various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid, salts of pectic acid, alginic acid, salts of alginic acid, organic acid, protective colloid thickener, pH adjusting agent, Glycerin, an alcohol or a carbonating agent, and the like. In addition, the health food of the present invention may contain natural fruit juice, fruit juice drink, or pulp for the production of vegetable drinks. These components may be used independently or in combination. The proportion of such additives is not critical, but is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.
이하, 실시예를 통하여 본 발명을 더욱 상세히 설명하고자 한다. 이들 실시예는 오로지 본 발명을 예시하기 위한 것으로서, 본 발명의 범위가 이들 실시예에 의해 제한되는 것으로 해석되지 않는 것은 당업계에서 통상의 지식을 가진 자에 있어서 자명할 것이다.Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these examples are for illustrative purposes only and that the scope of the present invention is not construed as being limited by these examples.
전통발효식품으로부터 유용 Useful from traditional fermented foods 프로바이오틱스Probiotics 균주의 분리 및 확보 Isolation and securing of strain
<1-1> <1-1> 전통발효식품 유래Traditional fermented food origin 미생물의 순수 분리 Pure Isolation of Microorganisms
본 발명의 실시예를 수행하기 위해서, 전통발효식품으로부터 미생물을 순수 분리하였다.In order to carry out the embodiment of the present invention, the microorganisms were separated from the conventional fermented food.
구체적으로, 재래시장에서 구입한 된장, 간장 및 고추장의 장류를 구입하여, 이를 증류수에 10-1 내지 10-7의 비율로 단계 희석하여 발효식품 시료를 준비하였다. 그런 다음, 상기 발효식품 시료를 유산균 선택 배지인 락토바실러스 MRS 배지(lactobacilli MRS), 페닐에틸 알콜 배지(phenylethyl alcohol) 및 락토바실러스 선택 한천배지(lactobacillus selective agar)에 도말하였다. 도말한 배지는 37℃에서 정치 배양하여, 생성된 콜로니를 수득하고 형태를 관찰하여, 새로운 배지에 도말하고 미생물을 순수 분리하였다.Specifically, fermented food samples were prepared by purchasing soy sauce, soy sauce, and kochujang purchased from traditional markets and diluting them in distilled water at a ratio of 10 -1 to 10 -7 . The fermented food samples were then plated on lactobacilli MRS medium, lactobacilli MRS medium, phenylethyl alcohol medium and lactobacillus selective agar medium. The stained medium was subjected to stationary culture at 37 DEG C, and the resulting colonies were obtained. The morphology was observed, and the microorganisms were purified by sprinkling on a fresh medium.
<1-2> 분리된 유산균의 동정<1-2> Identification of isolated lactic acid bacteria
유산균 선택 배지를 통해서 순수 분리한 미생물이 어떠한 균주인지 확인하기 위해서, 분리한 미생물을 동정하였다.In order to identify the microorganisms which were purely isolated from the lactic acid bacteria selection medium, the isolated microorganisms were identified.
구체적으로, 상기 실시예 <1-1>에서 분리한 균주에서 유전체 DNA를 추출하고, 염기서열을 분석하여(Bionics 사, 한국에 의뢰) 16S rRNA의 유전자 염기서열을 확보하였다. 상기 16S rRNA의 염기서열은 BLAST 프로그램을 이용하여 데이터베이스에 등록된 다른 표준 균주와 상동성을 분석하고, 98% 이상의 상동성을 나타내는 균주로 동정하였다. 또한 클러스탈 X, 메가 7 프로그램(Clustal X, Mega 7 program)을 사용하여 상동성을 분석하고 계통도를 구축하였다.Specifically, the genomic sequence of 16S rRNA was obtained by extracting the genomic DNA from the strain isolated in Example <1-1> and analyzing the base sequence (Bionics Co., Ltd., Korea). The nucleotide sequence of the 16S rRNA was analyzed by using the BLAST program and the homology with other standard strains registered in the database, and the strain was identified as having a homology of 98% or more. In addition, clustal X, Mega 7 program (Clustal X, Mega 7 program) was used to analyze homology and construct a genealogy diagram.
그 결과, 도 1에서 나타난 바와 같이 전통발효식품으로부터 분리된 유산균은 락토바실러스 플란타룸(Lactobacillus plantarum)으로 보고된 균주들과 상동성을 나타내는 것으로 락토바실러스 플란타룸 Ln4(Lactobacillus plantarum Ln4)로 명명하여, 2016년 09월 22일 한국미생물보존센터(KCCM)에 기탁하였다(기탁번호 KCCM 11897P).As a result, it called the lactic acid bacteria is Lactobacillus Planta room (Lactobacillus plantarum) with Lactobacillus Planta room Ln4 (Lactobacillus plantarum Ln4) showed a homology with the reported strain isolated from a traditional fermented foods, as also shown in the first (KCCM 11897P) deposited on September 22, 2016 with the Korean Center for Microorganism Conservation (KCCM).
락토바실러스Lactobacillus 플란타룸Flora Room Ln4Ln4 균주의 특징 확인 Identification of the characteristics of the strain
<2-1> <2-1> 락토바실러스Lactobacillus 플라타룸Platamum Ln4의Ln4 내산성Acid resistance 및 And 내담즙성My bile 확인 Confirm
전통발효식품으로부터 분리된 락토바실러스 플란타룸 Ln4의 특징을 확인하기 위하여, 내산성 및 내담즙성을 확인하였다.In order to confirm the characteristics of Lactobacillus plantarum Ln4 isolated from traditional fermented foods, acid resistance and biliary properties were confirmed.
구체적으로, 먼저 내산성을 확인하기 위해서 락토바실러스 플란타룸 Ln4 균주를 0.3% 펩신(pepsin) 수용액의 인공 위액 배지(pH 2.5)에 접종하여 3 시간 동안 정치배양하였다. 배양 후, 균을 수득하여 평판배지에 도말하고 37℃에서 48 시간동안 배양한 다음, 생성된 콜로니 수를 계수하여 생존 균수를 측정하여 용매 대조군의 생존 균수에 대한 상대적인 생존 균수를 확인하였다.Specifically, in order to confirm the acid resistance, Lactobacillus plantarum Ln4 strain was inoculated into an artificial gastric juice medium (pH 2.5) containing 0.3% pepsin aqueous solution and cultured for 3 hours. After culturing, the bacteria were obtained, plated on a plate medium, cultured at 37 ° C for 48 hours, and the number of viable bacteria was counted by counting the number of colonies produced, and the number of survival bacteria relative to the number of viable cells in the solvent control was confirmed.
또한, 내담즙성을 확인하기 위해서 락토바실러스 플란타룸 Ln4 균주를 0.3% 옥스갈(oxgall)의 배지에 접종하여 24 시간 동안 정치배양하였다. 배양 후, 균을 수득하여 동일한 방법으로 상대적인 생존 균수를 확인하였다.Lactobacillus plantarum Ln4 strain was inoculated on a medium containing 0.3% oxgall and cultured for 24 hours in order to confirm the biliary properties. After culturing, bacteria were obtained and the relative viable counts were confirmed in the same manner.
그 결과, 하기 [표 1]에서 나타난 바와 같이 락토바실러스 플란타룸 Ln4 균주는 인공 위액 배지에서도 3 시간 동안 99% 이상의 생존률을 유지하였기 때문에 우수한 내산성을 나타내는 것으로 확인하였다(표 1). 또한, 내담즙성에 대해서도 24 시간 동안 80% 이상의 균수를 유지하여, 우수한 내담즙성을 나타내는 것으로 확인하였다(표 1).As a result, as shown in the following Table 1, it was confirmed that Lactobacillus flutarium Ln4 showed excellent acid resistance because it maintained 99% or more survival rate in artificial gastric juice medium for 3 hours (Table 1). In addition, it was confirmed that the biliary properties were maintained at 80% or more for 24 hours, indicating excellent biliary properties (Table 1).
<2-2> <2-2> 락토바실러스Lactobacillus 플란타룸Flora Room Ln4의Ln4 효소 enzyme 생산능Production capacity 확인 Confirm
전통발효식품으로부터 분리된 락토바실러스 플란타룸 Ln4의 특징을 확인하기 위하여, 락토바실러스 플란타룸 Ln4 균주가 발암의 원인이 되는 β-글루쿠로니다제(β-glucuronidase) 등 다양한 효소를 생산할 수 있는지의 여부를 확인하였다.To confirm the characteristics of Lactobacillus plantarum Ln4 isolated from traditional fermented foods, Lactobacillus plantarum Ln4 strain can produce various enzymes such as β-glucuronidase, which causes carcinogenesis. .
구체적으로, 락토바실러스 플란타룸 Ln4 균주를 대수 증식기까지 배양한 다음, 10,000×g에서 10 분 동안 원심분리하여 상층액의 배지를 제거하였다. 배지를 제거한 후 수득한 균주를 PBS 완충용액으로 세척(washig)한 후, API ZYM 키트의 기질을 세척한 균주와 혼합하여 4 시간 동안 배양하였다. 그런 다음, 상기 키트의 용액 A 및 B를 각각 첨가하고 높은 광원에서 5 분간 반응시킨 후, 발색 정도에 따라 효소 생산 여부를 확인하였다.Specifically, the Lactobacillus plantarum Ln4 strain was cultured until the logarithmic growth phase, and then the supernatant was removed by centrifugation at 10,000 xg for 10 minutes. After the medium was removed, the obtained strain was washed with PBS buffer, and the substrate of the API ZYM kit was mixed with the washed strain and cultured for 4 hours. Then, the solutions A and B of the kit were added, respectively, and reacted for 5 minutes at a high light source. Then, enzyme production was checked according to the degree of color development.
그 결과, 하기 [표 2]에 나타난 바와 같이 락토바실러스 플란타룸 Ln4은 발암의 원인이 되는 β-글루쿠로니다제를 생산하지 않는 것을 확인하였다(표 2).As a result, it was confirmed that Lactobacillus plantarum Ln4 did not produce? -Glucuronidase, which is a cause of cancer, as shown in Table 2 below (Table 2).
(상기 표에서 0은 0 내지 5 nmol의 효소를 생산하는 것을 나타내고; 1은 5 내지 10 nmol의 효소를 생산하는 것을 나타내며; 2은 10 내지 20 nmol의 효소를 생산하는 것을 나타내고; 3은 20 내지 30 nmol의 효소를 생산하는 것을 나타내며; 4는 30 내지 40 nmol의 효소를 생산하는 것을 나타내고; 및 5는 40 nmol 이상의 효소를 생산하는 것을 나타낸다.)(In the above table 0 means producing 0-5 nmol of enzyme; 1 means producing 5-10 nmol of enzyme; 2 means producing 10-20 nmol of enzyme; 3 is 20- 30 nmol of enzyme; 4 represents 30 to 40 nmol of enzyme; and 5 represents 40 nmol of enzyme.
<2-3> <2-3> 락토바실러스Lactobacillus 플란타룸Flora Room Ln4의Ln4 세포 독성 확인 Cytotoxicity check
전통발효식품으로부터 분리된 락토바실러스 플란타룸 Ln4의 특징을 확인하기 위하여, 락토바실러스 플란타룸 Ln4 균주의 세포 독성을 확인하였다.To confirm the characteristics of Lactobacillus plantarum Ln4 isolated from traditional fermented foods, the cytotoxicity of Lactobacillus plantarum Ln4 was confirmed.
구체적으로, 정상세포인 MRC-5 세포를 96웰 플레이트에 접종한 다음, 24 시간 동안 배양하여 MRC-5 세포를 2×105 세포/㎖ 농도로 준비하였다. 그리고 락토바실러스 플란타룸 Ln4 균주를 대수 증식기까지 배양한 다음, 10,000×g에서 10 분 동안 원심분리하여 상층액의 배지를 제거하였다. 배지를 제거한 후 수득한 균주를 PBS 완충용액으로 세척한 후, 상기 준비한 MRC-5 배양 플레이트에 수득한 락토바실러스 플란타룸 Ln4 균주를 접종하여 44 시간 동안 배양하였다. 배양 후, PBS 완충용액으로 3 번 반복 세척한 다음, MTT 시약 50 ㎕를 처리하여 4 시간 동안 반응시켰다. 반응 종결 후에는 DMSO 50 ㎕를 처리하여 570 ㎚ 파장에서 흡광도를 측정하여 발색 정도에 따라 사멸한 MRC-5 세포 수준을 통해 세포 독성을 확인하였다.Specifically, MRC-5 cells, which are normal cells, were inoculated on a 96-well plate and cultured for 24 hours to prepare MRC-5 cells at a concentration of 2 × 10 5 cells / ml. Then, the Lactobacillus plantarum Ln4 strain was cultured until the logarithmic growth phase, and then the supernatant was removed by centrifugation at 10,000 × g for 10 minutes. After the medium was removed, the obtained strain was washed with PBS buffer, and then the Lactobacillus plantarum Ln4 strain obtained on the MRC-5 culture plate prepared above was inoculated and cultured for 44 hours. After incubation, the cells were washed three times with PBS buffer solution, and treated with 50 μl of MTT reagent, followed by reaction for 4 hours. After completion of the reaction, 50 μl of DMSO was treated to measure the absorbance at a wavelength of 570 nm, and cytotoxicity was confirmed through the level of MRC-5 cells killed according to the degree of color development.
그 결과 하기 [표 3]에서 나타난 바와 같이 락토바실러스 플란타룸 Ln4 균주는 정상 세포와 함께 배양한 후에도 MRC-5 세포의 사멸을 유도하지 않는 것을 확인하여, 락토바실러스 플란타룸 Ln4 균주가 정상 세포에 대하여 세포 독성을 가지지 않는 것을 확인하였다(표 3).As a result, as shown in Table 3, it was confirmed that the Lactobacillus plantarum Ln4 strain did not induce the death of MRC-5 cells even after culturing with the normal cells, (Table 3). ≪ tb > < TABLE >
<2-4> <2-4> 락토바실러스Lactobacillus 플란타룸Flora Room Ln4의Ln4 장 chapter 부착능Attachment 확인 Confirm
전통발효식품으로부터 분리된 락토바실러스 플란타룸 Ln4의 특징을 확인하기 위하여, 락토바실러스 플란타룸 Ln4 균주의 장 부착능을 확인하였다.To confirm the characteristics of Lactobacillus plantarum Ln4 isolated from traditional fermented foods, the ability of Lactobacillus plantarum Ln4 to bind to the intestines was confirmed.
구체적으로, 대장암 세포인 HT-29 세포를 96웰 플레이트에 접종한 다음, 24 시간 동안 배양하여 MRC-5 세포를 1×105 세포/㎖ 농도로 준비하였다. 그리고 락토바실러스 플란타룸 Ln4 균주를 대수 증식기까지 배양한 다음, 10,000×g에서 10 분 동안 원심분리하여 상층액의 배지를 제거하였다. 배지를 제거한 후 수득한 균주를 PBS 완충용액으로 세척한 후, 상기 준비한 HT-29 배양 플레이트에 수득한 락토바실러스 플란타룸 Ln4 균주를 접종하여 2 시간 동안 배양하였다. 배양 후, PBS 완충용액으로 3 번 반복 세척한 다음, 트리톤-X100 1 ㎖을 가하여 10 분 동안 처리하고 원심분리하고, 상등액을 수득하고 lactobacilli MRS 한천 배지에 도말하고 37℃에서 48 시간 동안 배양한 다음 생성된 콜로니 수를 확인하여, 대장암세포주에 부착한 균수와 배지에 남은 균수(생성된 콜로니 수)를 비교하여 장부착능을 확인하였다.Specifically, HTC-29 cells, colon cancer cells, were inoculated into 96-well plates and cultured for 24 hours to prepare MRC-5 cells at a concentration of 1 × 10 5 cells / ml. Then, the Lactobacillus plantarum Ln4 strain was cultured until the logarithmic growth phase, and then the supernatant was removed by centrifugation at 10,000 × g for 10 minutes. After the medium was removed, the obtained strain was washed with PBS buffer, and then the Lactobacillus plantarum Ln4 strain obtained on the HT-29 culture plate prepared above was inoculated and cultured for 2 hours. After culturing, the cells were washed three times with PBS buffer, and then treated with 1 ml of Triton-X100 for 10 minutes. The supernatant was obtained, plated on lactobacilli MRS agar medium and cultured at 37 ° C for 48 hours The number of colonies formed was checked, and the number of colonies attached to the colon cancer cell line and the number of colonies remaining in the medium were compared to confirm the ability of the colon to adhere to the colon.
그 결과, 하기 [표 4]에서 나타난 바와 같이 락토바실러스 플란타룸 Ln4 균주가 대장암세포에 대하여 높은 장 부착능을 가지는 것을 확인하였다(표 4).As a result, it was confirmed that the Lactobacillus plantarum Ln4 strain had a high ability to bind to colon cancer cells as shown in Table 4 below (Table 4).
<2-5> <2-5> 락토바실러스Lactobacillus 플란타룸Flora Room Ln4의Ln4 항생제 내성 확인 Identification of antibiotic resistance
전통발효식품으로부터 분리된 락토바실러스 플란타룸 Ln4의 특징을 확인하기 위하여, 락토바실러스 플란타룸 Ln4 균주가 CLSI(Clinical and Laboratory Standards Institute, 2012) 기준에 사용되는 항생제에 대하여 내성을 나타내는지 여부를 확인하였다.To characterize Lactobacillus plantarum Ln4 isolated from traditional fermented foods, it is necessary to determine whether the Lactobacillus plantarum Ln4 strain is resistant to antibiotics used in CLSI (Clinical and Laboratory Standards Institute, 2012) Respectively.
구체적으로, 락토바실러스 플란타룸 Ln4 균주를 대수 증식기까지 배양한 다음, 10,000×g에서 10 분 동안 원심분리하여 상층액의 배지를 제거하였다. 배지를 제거한 후 수득한 균주를 PBS 완충용액으로 세척한 후, 105 CFU/㎖ 농도로 희석하여 lactobacilli MRS 한천 배지에 도말하여 배지 표면을 락토바실러스 플란타룸 Ln4로 포화될 때까지 배양하였다. 그런 다음, 멸균한 종이 디스크(paper disc)를 50 ㎕ 암피실린(ampicillin), 테트라사이클린(Tetracycline), 제타마이신(Gentamycin), 시프로플록사신(Ciprofloxacin), 가나마이신(Kanamycin), 클로람페니콜(Chloramphenicol), 스트렙토마이신(Streptomycin) 또는 독소시클린(Doxycycline)에 각각 적시고, 이를 상기 배양한 락토바실러스 플란타룸 Ln4 균주 배지 표면에 올려놓았다. 그리고 37℃에서 24 시간 동안 배양하여 생성된 생육저지환(inhibition zone)의 크기에 따라 항생제 저항성 정도를 판별하였다.Specifically, the Lactobacillus plantarum Ln4 strain was cultured until the logarithmic growth phase, and then the supernatant was removed by centrifugation at 10,000 xg for 10 minutes. After the medium was removed, the obtained strains were washed with PBS buffer, diluted to a concentration of 10 5 CFU / ml, plated on lactobacilli MRS agar medium and cultured until the surface of the medium was saturated with Lactobacillus plantarum Ln4. The sterilized paper disc was then incubated with 50 μl of ampicillin, Tetracycline, Gentamycin, Ciprofloxacin, Kanamycin, Chloramphenicol, Streptomycin Streptomycin or Doxycycline, respectively, and these were placed on the surface of the cultured Lactobacillus plantarum Ln4 culture medium. The culture medium was incubated at 37 ° C. for 24 hours to determine the antibiotic resistance according to the size of the inhibition zone.
그 결과, 하기 [표 5]에 나타난 바와 같이 락토바실러스 플란타룸 Ln4은 암피실린, 테트라사이클린, 클로람페니콜 및 독소시클린에 대하여 민감하고, 제타마이신, 시프로플록사신, 가나마이신 및 스트렙토마이신에 대하여 내성을 나타내는 것을 확인하였다(표 5).As a result, as shown in the following Table 5, Lactobacillus plantarum Ln4 was found to be resistant to ampicillin, tetracycline, chloramphenicol and toxocycline, and resistant to zetamycin, ciprofloxacin, kanamycin and streptomycin (Table 5).
(상기 표에서, R는 항생제 내성(resistant)를 나타내고, S는 항생제 민감성(sensitive)을 나타낸다.)(In the above table, R represents antibiotic resistance and S represents antibiotic sensitivity.)
락토바실러스Lactobacillus 플란타룸Flora Room Ln4의Ln4 생균체Live cell 및 And 사균체의Dead cell 제조 Produce
본 발명의 락토바실러스 플란타룸 Ln4의 약학적 효능을 시험관 내(in vitro) 및 생체 내(in vivo)에서 확인하기 위해, 락토바실러스 플란타룸 Ln4의 생균체와 사균체를 제조하였다.In order to confirm the pharmaceutical efficacy of the Lactobacillus plantarum Ln4 of the present invention in vitro and in vivo , Lactobacillus flutarium Ln4 cells and dead cells were prepared.
구체적으로, 상기 <실시예 1>에서 분리 동정한 락토바실러스 플란타룸 Ln4을 lactobacilli MRS 액체 배지에 접종하고 37℃에서 18 시간 동안 배양한 다음, 5,000×g에서 10 분 동안 원심분리하여 균체의 침전물과 배양액의 상등액으로 분리하였다. 그런 다음, 분리한 침전물을 동결건조하여 이를 생균체로서 수득하였고, 생균체를 121℃에서 5 분 동안 열처리하여 사균체를 제조 및 수득하였다.Specifically, Lactobacillus plantarum Ln4 isolated and isolated in Example 1 was inoculated into a lactobacilli MRS liquid medium and cultured at 37 ° C for 18 hours, followed by centrifugation at 5,000 × g for 10 minutes to obtain a precipitate of cells And the culture supernatant. Then, the separated precipitate was lyophilized to obtain a biomass, and the biomass was heat-treated at 121 ° C for 5 minutes to prepare and obtain dead cells.
락토바실러스 플란타룸 Ln4의 항비만 효과 확인Confirmed anti-obesity effect of Lactobacillus plantarum Ln4
<4-1> 락토바실러스 플란타룸 Ln4의 지방축적 억제능 확인<4-1> Confirmation of inhibitory effect of Lactobacillus plantarum Ln4 on fat accumulation
본 발명의 락토바실러스 플란타룸 Ln4가 시험관 내(In vitro) 수준에서 항비만 효과를 나타낼 수 있는지 여부를 확인하고자, 지방 세포에서의 지방 축적 억제 효과를 확인하였다. In order to confirm whether or not the Lactobacillus plantarum Ln4 of the present invention can exhibit an anti-obesity effect at the in vitro level, the fat accumulation inhibitory effect in adipocytes was confirmed.
구체적으로, 지방 전구세포주인 3T3-L1 세포를 DMEM 배지에 접종한 후, 플레이트에서 포화도가 100%가 될 때까지 37℃의 5% CO2 배양기에서 배양하였다. 플레이트가 세포가 가득 차도록 자라면, 5 ㎍/㎖ 인슐린, 115 ㎍/㎖ IBMX 및 10μM 덱사메타손(dexamethasone)을 포함하는 10% FBS/DMEM 분화용 배지를 세포 배지로서 교환하고, 2 일 동안 추가 배양하였다. 이 때, 상기 분화용 배지에는 상기 <실시예 3>에서 제조한 락토바실러스 플란타룸 Ln4 사균체 100 ㎍/㎖를 추가하였다. 2 일 간의 배양이 끝나면, 락토바실러스 플란타룸 Ln4 사균체 100 ㎍/㎖ 및 5 ㎍/㎖ 인슐린을 포함하는 10% FBS/DMEM 배지로 교환하여 4 내지 6 일간 추가 배양하여, 지방전구세포로부터 분화된 지방세포를 수득하였다. 수득한 지방세포는 10% 포르말린 수용액을 가해 고정시키고, Oil Red O 용액을 처리하여 약 15 분 내지 2 시간 동안 반응하여 세포 내 축적된 지방을 염색하였다. 염색 완료 후, 현미경으로 관찰한 다음 이소프로필 알콜로 Oil Red O 용액을 다시 녹여 518 ㎚의 파장에서 흡광도를 측정하였다. 대조군으로는, 분화시키지 않은 미분화 세포 대조군(ND, non-differentiated) 및 사균체를 첨가하지 않은 배지에서 분화시킨 분화 세포 대조군(D, differentiated)을 사용하였으며, ND 대조군에 대비하여 D 대조군 및 실험군의 흡광도 수준의 증가 정도를 하기 [수학식 1]을 사용해 계산하였다.Specifically, 3T3-L1 cells, which are lipogenic precursor cells, were inoculated into a DMEM medium and then cultured in a 5% CO 2 incubator at 37 ° C until the degree of saturation reached 100% on the plate. If the plates were to become full of cells, 10% FBS / DMEM differentiation medium containing 5 μg / ml insulin, 115 μg / ml IBMX and 10 μM dexamethasone was replaced as cell culture medium and further incubated for 2 days . At this time, 100 g / ml of Lactobacillus plantarum Ln4 cells prepared in Example 3 was added to the differentiation medium. After 2 days of culture, the cells were exchanged with 10% FBS / DMEM medium containing 100 μg / ml of Lactobacillus plantarum Ln4 cells and 5 μg / ml insulin, and further cultured for 4 to 6 days to differentiate Fat cells were obtained. The obtained adipocytes were fixed with an aqueous solution of 10% formalin, treated with Oil Red O solution, and reacted for about 15 minutes to 2 hours to stain the accumulated fat in the cells. After completion of dyeing, observation with a microscope was performed, and the oil red O solution was again dissolved in isopropyl alcohol to measure the absorbance at a wavelength of 518 nm. As a control, differentiated cells (D, differentiated) differentiated in a medium not supplemented with ND, non-differentiated and dead cells were used. In contrast to the ND control, D control and experimental group The degree of increase in the absorbance level was calculated using the following equation (1).
[수학식 1][Equation 1]
그 결과, 도 2에서 나타난 바와 같이 분화 세포 대조군(D)와 비교하였을 때, 본 발명의 락토바실러스 플란타룸 Ln4 처리 실험군에서 지방 축적 수준이 감소된 것을 확인하였으며, 현미경 관찰을 통해서도 적색으로 염색된 지방구의 수 및 크기가 유의적으로 감소하는 것을 확인하였다(도 2).As a result, when compared with the differentiated cell control group (D) as shown in FIG. 2, it was confirmed that the level of fat accumulation was reduced in the Lt4 treatment group of Lactobacillus plantarum Ln4 of the present invention, And the number and size of lipids were significantly reduced (FIG. 2).
<4-2> 고지방식이에 의한 당뇨 유도 동물 모델의 제조<4-2> Production of Diabetic Induced Animal Model by High Fat Diet
락토바실러스 플란타룸 Ln4가 대사성질환이 유도된 동물모델에서 유의적인 항비만 효과를 나타낼 수 있는지 여부를 확인하기 위해, 대상 실험 동물인 당뇨 유도 마우스 모델을 제작하였다.To determine whether Lactobacillus plantarum Ln4 can exhibit significant anti-obesity effects in an animal model induced by metabolic disease, a diabetic-induced mouse model was prepared.
구체적으로, 상기 <실시예 3>에서 제조한 락토바실러스 플란타룸 Ln4 생균체를 증류수에 2.5×109 CFU/㎖ 농도로 현탁하여 균주 시료를 준비하였다. 그런 다음, 6 주령의 C57/BL6 마우스에 고지방 사료를 자유 급여하면서 상기 준비한 균주 시료를 매일 0.2 ㎖씩 경구투여하여 매일 5×108 CFU를 투여하였다. 상기 고지방 사료는 전체 사료의 열량(㎉)을 기준으로, 21.0% 단백질, 24.6% 탄수화물 및 54.4% 지방을 포함하는 펠렛 형태를 사용하였다. 총 5 주간 동일한 환경에서 균주 시료 투여 및 고지방 사료 급여한 후의 마우스를 락토바실러스 플란타룸 Ln4 투여 당뇨 유도 실험군(Ln4)로 사용하였다. Ln4 실험군의 결과를 비교하기 위해, 동일한 환경에서 사육하면서 생균체를 투여하면서 정상 사료를 자율급여한 군은 정상 대조군(Control)으로서 사용하였고, 동일한 환경에서 사육하면서 생균체를 투여하지 않고 고지방 사료를 자율급여한 군을 고지방식이 유도군(HFD)으로서 사용하였다. Ln4 실험군, 정상 대조군 및 HFD 유도군은 각각의 체중을 확인하여 고지방식이에 의해 당뇨 및 대사성질환이 유도되었는지 여부를 확인함과 동시에, 락토바실러스 플란타룸 Ln4에 의한 체중 감소 효과가 나타나는지를 확인하였다.Specifically, the Lactobacillus plantarum Ln4 cells prepared in Example 3 were suspended in distilled water at a concentration of 2.5 x 10 9 CFU / ml to prepare a strain sample. Then, the high-fat diets were freely fed to C57 / BL6 mice at 6 weeks of age and 0.2 ml of the prepared strain samples were orally administered at a daily dose of 5 × 10 8 CFU. The high-fat diet used 21.0% protein, 24.6% carbohydrate and 54.4% fat in the form of pellets based on the calories (㎉) of the whole feed. Mice were treated with Lactobacillus plantarum Ln4-administered diabetic rats (Ln4) after the administration of strain samples and high fat diet in the same environment for a total of 5 weeks. In order to compare the results of the Ln4 experimental group, the control group was used as a control group, while the normal group was fed with the live cells while the animals were kept in the same environment. The autonomic group was used as the induction group (HFD). Ln4 test group, normal control group and HFD-induced group were checked for their weight, and it was confirmed whether or not diabetes and metabolic diseases were induced by the high fat diet method, and whether or not the weight loss effect of Lactobacillus plantarum Ln4 was shown Respectively.
그 결과, 도 3에서 나타난 바와 같이 정상 대조군에 비해 사육 1주일 후부터 HFD 유도군의 체중의 증가가 유의적으로 높은 수준임을 확인하였으며, 이러한 HFD 군의 체중 증가에 비해 Ln4 생균제 경구 투여군(Ln4)에서 체중 증가 수준이 유의적으로 감소하는 것을 확인하였다(도 3).As a result, as shown in FIG. 3, it was confirmed that the body weight gain of the HFD-induced group was significantly higher than that of the normal control group after one week of feeding. Compared to the weight gain of the HFD group, the Ln4- And the weight gain level was significantly decreased (FIG. 3).
<4-3> 락토바실러스 플란타룸 Ln4의 체지방 감소 효능 분석<4-3> Effect of Lactobacillus plantarum Ln4 on body fat reduction
락토바실러스 플란타룸 Ln4가 대사성질환이 유도된 동물모델에서 유의적인 항비만 효과를 나타낼 수 있는지 여부를 확인하기 위해, 대사성질환을 유도시킨 동물모델에서 체지방 감소 효과를 확인하였다.In order to confirm whether Lactobacillus plantarum Ln4 can exhibit a significant anti-obesity effect in an animal model induced by metabolic diseases, the effect of reducing fat in an animal model induced by metabolic disease was confirmed.
구체적으로, 상기 실시예 <4-2>의 방법으로 5주 동안 대사성질환을 유도한 Ln4 생균제 투여군(Ln4) 및 대조군에서 체중 조절 효능을 분석한 후, 대표적인 내장 지방인 부고환 지방을 분리하여 무게를 측정하였다. 그런 다음, 고지방식이를 투여하기 전의 부고환 지방 무게와 비교하여 5주간의 고지방식이 유도에 따른 지방 무게의 증가 수준을 상대적으로 계산하였다.Specifically, the body weight control efficacy was analyzed in the Ln4-treated group (Ln4) and the control group in which the metabolic diseases were induced for 5 weeks by the method of Example <4-2>, and then the representative visceral fat epididymal fat was isolated and weighed Respectively. Then, the elevation method of 5 weeks compared to the epididymal fat weight before the administration of the high fat diet was relatively calculated the increase level of the fat weight according to the induction.
그 결과, 도 4에서 나타난 바와 같이 정상 식이로 급여한 대조군과 비교하였을 때 고지방식이를 유도한 대조군(HFD)은 부고환지방의 무게가 약 2 배 수준으로 증가하였으나, 실험군(Ln4)에서는 HFD 군에 비해 부고환지방 무게의 증가 수준이 유의적으로 감소하는 것을 확인하였다(도 4).As a result, as shown in FIG. 4, the weight of epididymal fat increased to about 2-fold in the control group (HFD) induced by the high fat diet compared with the control diet fed the normal diet, but in the experimental group (Ln4) (Fig. 4). As shown in Fig.
<110> Konkuk University Industrial Cooperation Corp KOREA FOOD RESEARCH INSTITUTE <120> Novel Lactobacillus plantarum Ln4 strain and compositions for the prevention and treatment of obesity containing the same <130> 1061538 <160> 1 <170> KopatentIn 2.0 <210> 1 <211> 943 <212> RNA <213> Lactobacillus plantarum <400> 1 tgctaataca tgcagtcgaa cgaactctgg tattgattgg tgcttgcatc atgatttaca 60 tttgagtgag tggcgaactg gtgagtaaca cgtgggaaac ctgcccagaa gcgggggata 120 acacctggaa acagatgcta ataccgcata acaacttgga ccgcatggtc cgagcttgaa 180 agatggcttc ggctatcact tttggatggt cccgcggcgt attagctaga tggtggggta 240 acggctcacc atggcaatga tacgtagccg acctgagagg gtaatcggcc acattgggac 300 tgagacacgg cccaaactcc tacgggaggc agcagtaggg aatcttccac aatggacgaa 360 agtctgatgg agcaacgccg cgtgagtgaa gaagggtttc ggctcgtaaa actctgttgt 420 taaagaagaa catatctgag agtaactgtt caggtattga cggtatttaa ccagaaagcc 480 acggctaact acgtgccagc agccgcggta atacgtaggt ggcaagcgtt gtccggattt 540 attgggcgta aagcgagcgc aggcggtttt ttaagtctga tgtgaaagcc ttcggctcaa 600 ccgaagaagt gcatcggaaa ctgggaaact tgagtgcaga agaggacagt ggaactccat 660 gtgtagcggt gaaatgcgta gtaatatgga agaacaccag tggcgaaggc ggctgtctgg 720 tctgtaactg acgctgaggc tcgaaagtat gggtagcaaa caggattaga taccctggta 780 gtccataccg taaacgatga atgctaagtg ttggagggtt tccgcccttc agtgctgcag 840 ctaacgcatt aagcattccg cctgggggag tacggccgcc aggctgaaac tcaaaggaat 900 ttgacggggg cccgcacaag cggtcggagc atgtggtttc att 943 <110> Konkuk University Industrial Cooperation Corp KOREA FOOD RESEARCH INSTITUTE <120> Novel Lactobacillus plantarum Ln4 strain and compositions for the prevention and treatment of obesity containing the same <130> 1061538 <160> 1 <170> Kopatentin 2.0 <210> 1 <211> 943 <212> RNA <213> Lactobacillus plantarum <400> 1 tgctaataca tgcagtcgaa cgaactctgg tattgattgg tgcttgcatc atgatttaca 60 tttgagtgag tggcgaactg gtgagtaaca cgtgggaaac ctgcccagaa gcgggggata 120 acacctggaa acagatgcta ataccgcata acaacttgga ccgcatggtc cgagcttgaa 180 agatggcttc ggctatcact tttggatggt cccgcggcgt attagctaga tggtggggta 240 acggctcacc atggcaatga tacgtagccg acctgagagg gtaatcggcc acattgggac 300 tgagacacgg cccaaactcc tacgggaggc agcagtaggg aatcttccac aatggacgaa 360 agtctgatgg agcaacgccg cgtgagtgaa gaagggtttc ggctcgtaaa actctgttgt 420 taaagaagaa catatctgag agtaactgtt caggtattga cggtatttaa ccagaaagcc 480 acggctaact acgtgccagc agccgcggta atacgtaggt ggcaagcgtt gtccggattt 540 attgggcgta aagcgagcgc aggcggtttt ttaagtctga tgtgaaagcc ttcggctcaa 600 ccgaagaagt gcatcggaaa ctgggaaact tgagtgcaga agaggacagt ggaactccat 660 gtgtagcggt gaaatgcgta gtaatatgga agaacaccag tggcgaaggc ggctgtctgg 720 tctgtaactg acgctgaggc tcgaaagtat gggtagcaaa caggattaga taccctggta 780 gtccataccg taaacgatga atgctaagtg ttggagggtt tccgcccttc agtgctgcag 840 ctaacgcatt aagcattccg cctgggggag tacggccgcc aggctgaaac tcaaaggaat 900 ttgacggggg cccgcacaag cggtcggagc atgtggtttc att 943
Claims (6)
A pharmaceutical composition for the prevention and treatment of obesity comprising Lactobacillus plantarum Ln4 strain deposited with Accession No. KCCM 11897P, a culture thereof or a mixture thereof as an active ingredient.
The pharmaceutical composition for preventing and treating obesity according to claim 1, wherein the strain has the 16S rRNA base sequence set forth in SEQ ID NO: 1.
The pharmaceutical composition for preventing and treating obesity according to claim 1, wherein the strain has acid resistance, biliary properties, intestinal adhesiveness and antibiotic resistance.
The pharmaceutical composition for the prevention and treatment of obesity according to claim 1, wherein the Lactobacillus plantarum Ln4 strain exhibits a fat accumulation inhibitory effect and a body fat reducing effect.
Lactobacillus plantarum Ln4 deposited with accession number KCCM 11897P, a culture solution thereof, or a mixture thereof as an active ingredient.
The health functional food according to claim 5, wherein the Lactobacillus plantarum Ln4 strain is characterized by an effect of inhibiting fat accumulation and reducing body fat.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20200060568A (en) * | 2018-11-21 | 2020-06-01 | 주식회사 발효예스 | Fermented product of red beet and manufacturing method thereof |
| KR102163551B1 (en) * | 2020-04-21 | 2020-10-07 | (주) 에이투젠 | Composition for prevention and treatment of lipid-related metabolic disease comprising lactobacillus plantarum ATG-K2 or lactobacillus plantarum ATG-K6 |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20200060568A (en) * | 2018-11-21 | 2020-06-01 | 주식회사 발효예스 | Fermented product of red beet and manufacturing method thereof |
| KR102163551B1 (en) * | 2020-04-21 | 2020-10-07 | (주) 에이투젠 | Composition for prevention and treatment of lipid-related metabolic disease comprising lactobacillus plantarum ATG-K2 or lactobacillus plantarum ATG-K6 |
| WO2021215627A1 (en) * | 2020-04-21 | 2021-10-28 | (주)에이투젠 | Composition for preventing or treating lipid-related metabolic diseases, comprising lactobacillus plantarum atg-k2 or atg-k6 |
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