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KR101471033B1 - Novel Strains of Weissella sp. F22 with Anti-Obesity Ability - Google Patents

Novel Strains of Weissella sp. F22 with Anti-Obesity Ability Download PDF

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KR101471033B1
KR101471033B1 KR1020130125614A KR20130125614A KR101471033B1 KR 101471033 B1 KR101471033 B1 KR 101471033B1 KR 1020130125614 A KR1020130125614 A KR 1020130125614A KR 20130125614 A KR20130125614 A KR 20130125614A KR 101471033 B1 KR101471033 B1 KR 101471033B1
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weissella
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임상동
김기성
최인욱
하상근
조성아
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한국식품연구원
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    • A23V2200/00Function of food ingredients
    • A23V2200/30Foods, ingredients or supplements having a functional effect on health
    • A23V2200/332Promoters of weight control and weight loss
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    • C12R2001/00Microorganisms ; Processes using microorganisms
    • C12R2001/01Bacteria or Actinomycetales ; using bacteria or Actinomycetales

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Abstract

The present invention relates to strains of Weissella sp. F22 having anti-obesity ability. The strains of the present invention have excellent anti-obesity ability, show growth inhibition activity against food poisoning-inducing virus, and have bile tolerance and strong acid resistance. Also, the strains show acid production fermentation characteristic, thereby being used as strains for fermentation starter to produce fermented milk products.

Description

비만억제능이 우수한 신규한 웨이젤라 에스피 F22 균주{Novel Strains of Weissella sp. F22 with Anti-Obesity Ability} Novel Strains of Weissella sp. ≪ RTI ID = 0.0 > F22 with Anti-Obesity Ability}

본 발명은 신규한 웨이젤라 에스피(Weissella sp.)F22 균주에 관한 것으로, 더욱 상세하게는 발효유 제조에 적합한 비만억제능이 있는 신규한 웨이젤라 에스피(Weissella sp.)F22에 관한 것이다.
The present invention relates to a novel Weissella sp. Strain F22, and more particularly to a novel Weissella sp. F22 which has an ability to inhibit obesity suitable for producing fermented milk.

비만은 당질이나 지질 등의 에너지원이 생체 조직, 특히 피하 지방 조직이나 장기 주변 조직의 지방세포에 중성지방으로서 비정상적으로 축적된 결과, 체중이 골격계 혹은 생리 기능의 한계를 넘어서 증가한 상태를 의미한다. 특히, 장간막 등의 내장 주위에 축적되는 내장 지방조직은 당질대사나 지질대사의 이상을 발생시키는 원인으로 알려져 있고, 심혈관질환, 동맥경화증, 고혈압증, 당뇨병, 지방간 등의 생활 습관병으로 진전하는 경우도 있다. 비만의 치료방법은 크게 식이-운동요법, 수술 요법 및 약물 요법이 있다. 식이-운동 요법은 저칼로리-저지방 섭취와 산소를 소비하는 육체의 활동을 통한 치료 방법인데, 이는 인내심을 가지고 반복적, 지속적으로 수행되어야 하기 때문에 대중적인 효과를 보기는 어려운 것으로 인식되고 있다. 수술요법은 외과적 수술을 통해 체지방을 물리적으로 제거하는 방법으로서 단기간에 효과를 볼 수 있는 장점이 있지만, 수술을 해야 하는 점, 효과의 지속성이 없다는 점, 비용이 많이 든다는 점 등 때문에 제한적으로 활용되고 있다. 약물 요법은 식욕 감소를 유발하거나 지방 흡수를 억제하는 약제들을 이용해서 비만을 치료 또는 예방하고자 하는 방법이다. 최근에는 비만치료제 개발을 위한 많은 연구가 진행되고 있으며, 그 중에 하나가 췌장 지방분해효소 저해제(pancreatic lipase inhibitor)이다. 췌장 지방분해효소(pancreatic lipase)는 트리글리세라이드(triglyceride)를 2-모노아실글리세롤(2-monoacylglycerol)과 지방산(fatty acid)으로 분해하는 핵심 효소로 작용한다[Bitou, N., M. Nimomiya, T. Tsjita and H. Okuda. 1999. Screening of lipase inhibitors from marine algae. Lipids 34, 441-445]. 대표적인 췌장 지방분해소 저해제는 Streptomyces toxitricini로부터 유래된 lipstatin의 유도체인 tetrahydrolipstatin (Orlistat)으로서 섭취된 지방의 약 30%를 저해할 정도로 효능이 우수한 것으로 알려져 있으며[Drent, M. L., et al., 1995. Orlistat (RO 18-0647), a lipase inhibitor, in the treatment of human obesity: a multiple dose study. Int. J. Obes. 19, 221 -226.], 현재 의약품으로도 시판중이다. 그러나 이와 같은 효능에도 불구하고 tetrahydrolipstatin은 위장장애, 과민증, 담즙분비장애, 지용성 비타민 흡수억제등의 부작용이 있는 것으로 알려져 있다. 따라서 최근에는 부작용이 없는 식품 및 천연물로부터 췌장 지방분해효소 저해제의 개발을 위한 연구가 진행되고 있다. Obesity means that the energy source such as carbohydrate or lipid is abnormally accumulated as a triglyceride in living tissue, especially subcutaneous adipose tissue or adipose tissue of organ organs, and the body weight is increased beyond the limit of skeletal system or physiological function. In particular, visceral adipose tissue accumulated around the intestines of the mesentery and the like is known to cause the abnormalities of carbohydrate metabolism and lipid metabolism, and there are cases in which lifestyle-related diseases such as cardiovascular disease, arteriosclerosis, hypertension, diabetes, . Treatment methods for obesity are diet - exercise, surgery and drug therapy. Dietary-exercise therapy is a method of treating low-calorie-low-fat meals and oxygen-consuming physical activity, which is perceived to be difficult to see because of the need to be patient, repetitive, and continuous. Surgery is a method of physically removing body fat through surgical operations, which is advantageous in that it can be effective in a short period of time. However, it is limited due to the necessity of surgery, lack of persistence of effect, and cost . Pharmacotherapy is a method to treat or prevent obesity by using drugs that induce appetite reduction or inhibit fat absorption. Recently, many studies for the development of obesity treatment drugs have been carried out. One of them is pancreatic lipase inhibitor. Pancreatic lipase acts as a key enzyme that breaks down triglyceride into 2-monoacylglycerol and fatty acid [Bitou, N., M. Nimomiya, T. Tsjita and H. Okuda. 1999. Screening of lipase inhibitors from marine algae. Lipids 34, 441-445]. Representative pancreatic lipolytic inhibitors are tetrahydrolipstatin (Orlistat), a derivative of lipstatin derived from Streptomyces toxitricini , which is known to be effective in inhibiting about 30% of the fat ingested [Drent, ML, et al., 1995. Orlistat RO 18-0647), a lipase inhibitor, in the treatment of human obesity: a multiple dose study. Int. J. Obes. 19, 221 -226.], And is currently also being marketed as a pharmaceutical product. Despite these benefits, however, tetrahydrolipstatin has been known to have side effects such as gastrointestinal disorders, hypersensitivity, biliary secretion, and lipid-soluble vitamin absorption inhibition. Recently, studies for the development of pancreatic lipolytic enzyme inhibitors from foods and natural products without side effects have been carried out.

유산균은 통상 소비하는 당에 대하여 약 50% 이상의 유산을 대사산물로 생성하는 세균을 의미하는 것으로, 글루코오스로부터 다량의 유산을 생성하면서 식품이나, 사람과 동물의 장내에서 인체에 해로운 물질, 예컨대, 인돌(indole), 스카톨(skatole), 페놀(phenol), 아민(amine), 암모니아과 같은 물질을 생성하지 않고 부패를 방지하는 등의 유익한 작용을 하는 세균을 개념적으로 총칭하는 의미로 사용된다. 유산균은 오래 전부터 발효유, 유산균 음료, 발효 버터 등의 유제품 등의 식품을 제조할 때에 사용되고 있으나, 유산균 자체도 정장효과를 비롯하여, 여러 가지 약리작용을 가지기 때문에, 건강식품이나 의약품 등의 소재로도 이용되고 있다. 대한민국 공개특허 제10-2010-0109661호에는 혈중콜레스테롤 강하와 비만억제 효능을 갖는 신규의 락토바실러스 커베터스 균주가 개시되어 있고, 대한민국 공개특허 제10-2012-0128260호에는 비피도박테리움의 혼합 유산균을 포함하는 비만 및 비만관련질환의 치료 또는 예방용 조성물이 개시되어 있으며, 대한민국 공개특허 제10-2010-0010015호에는 혈중 콜레스테롤 저하 및 항비만 활성을 갖는 락토바실러스존소니 균주가 개시되어 있다.
Lactic acid bacteria refers to a bacterium that produces about 50% or more of lactic acid as a metabolite in a sugar that is commonly consumed. It is a food or a substance harmful to human body in human and animal intestines, such as an indole is used to conceptually generically refer to bacteria that do not produce substances such as indole, skatole, phenol, amine, and ammonia and prevent decay. Lactic acid bacteria have long been used in the manufacture of foods such as fermented milk, lactic acid bacteria beverages, fermented butter, and other dairy products, but lactic acid bacteria themselves have various pharmacological effects as well as a dressing effect and are therefore also used as materials for health foods and medicines . Korean Patent Laid-Open No. 10-2010-0109661 discloses a novel Lactobacillus carvettus strain having a blood cholesterol lowering effect and an obesity-inhibiting effect, and Korean Patent Publication No. 10-2012-0128260 discloses a novel Lactobacillus species Discloses a composition for treating or preventing obesity and obesity-related diseases including lactic acid bacteria, and Korean Patent Laid-Open No. 10-2010-0010015 discloses a lactobacillus Johnsonia strain having a blood cholesterol lowering and anti-obesity activity.

본 명세서 전체에 걸쳐 다수의 논문 및 특허문헌이 참조되고 그 인용이 표시되어 있다. 인용된 논문 및 특허문헌의 개시 내용은 그 전체로서 본 명세서에 참조로 삽입되어 본 발명이 속하는 기술 분야의 수준 및 본 발명의 내용이 보다 명확하게 설명된다.
Numerous papers and patent documents are referenced and cited throughout this specification. The disclosures of the cited papers and patent documents are incorporated herein by reference in their entirety to better understand the state of the art to which the present invention pertains and the content of the present invention.

대한민국 공개특허 제10-2010-0109661호Korean Patent Publication No. 10-2010-0109661 대한민국 공개특허 제10-2012-0128260호Korean Patent Publication No. 10-2012-0128260 대한민국 공개특허 제10-2010-0010015호Korean Patent Publication No. 10-2010-0010015

본 발명자들은 비만억제능이 뛰어난 젖산균을 발굴하기 위해 연구 노력하였다. 그 결과, 분변으로부터 비만억제능이 매우 우수한 웨이젤라 에스피(Weissella sp.)F22 균주를 분리 및 동정하는데 성공하였으며, 이 균주의 발효제품에 대한 응용가능성을 실험적으로 확인하여 본 발명을 완성하였다. The present inventors have sought to find lactic acid bacteria having excellent ability to inhibit obesity. As a result, we succeeded in isolating and identifying Weissella sp. F22 strain having excellent ability to inhibit obesity from feces. The applicability of this strain to a fermentation product was confirmed experimentally and the present invention was completed.

따라서, 본 발명의 목적은 비만억제능이 우수한 신규 젖산균주를 제공하는데 있다. Accordingly, an object of the present invention is to provide a novel lactic acid bacteria strain having excellent ability to inhibit obesity.

본 발명의 다른 목적은 상기 균주를 포함하는 발효 출발 물질 조성물을 제공하는데 있다. Another object of the present invention is to provide a fermentation starting material composition comprising the strain.

본 발명의 또 다른 목적은 상기 발효 출발 물질 조성물을 발효시켜 제조한 발효물을 제공하는데 있다. It is still another object of the present invention to provide a fermented product prepared by fermenting the fermentation starting material composition.

본 발명의 또 다른 목적은 상기 발효물을 포함하는 식품 조성물을 제공하는데 있다. It is still another object of the present invention to provide a food composition comprising the fermented product.

본 발명의 목적 및 장점은 하기의 발명의 상세한 설명, 청구의 범위 및 도면에 의해 보다 명확하게 된다.
The objects and advantages of the present invention will become more apparent from the following detailed description of the invention, claims and drawings.

본 발명의 일 양태에 따르면, 본 발명은 비만억제능이 우수한 웨이젤라 에스피(Weissella sp.)F22 (수탁번호: KACC 91867P) 균주를 제공한다. According to one aspect of the present invention, there is provided a strain of Weissella sp. F22 (accession number: KACC 91867P) excellent in the ability to inhibit obesity.

본 발명의 웨이젤라 에스피(Weissella sp.)F22 균주는 젖산균으로서 분변에서 분리하였으며, 후술하는 실시예에 기재된 방법에 따라 스크리닝 및 동정하였다. 본 발명의 미생물의 동정은 미생물의 형태학적, 생리학적 및 생화학적 특성을 기초로 하여 이루어졌다. The Weissella sp. F22 strain of the present invention was isolated from feces as lactic acid bacteria and screened and identified according to the method described in the following Examples. The identification of the microorganism of the present invention is based on the morphological, physiological and biochemical characteristics of the microorganism.

미생물 분리 및 동정Microbial Isolation and Identification

본 발명의 신규 미생물은 성인분변에서 분리되었으며, 펩톤(peptone) 희석액으로 희석하여 NaN3를 첨가한 변형 MRS배지(하기 실시예 1의 표 1 참조)에 0.1 ㎖ 씩 평면도말법으로 접종한 후 37℃에서 48시간 배양하고 각 균락을 변형 MRS배지에서 순수 분리한 다음 노란색으로 변한 균락을 잠정적 젖산균으로 선발하였다. 선발된 균주는 전기 변형 MRS 배지에 도말한 후, 호기 배양하여 순수분리 하였다. 순수 분리된 균주를 동정한 결과, 균주는 그람 양성의 간균이며, 산소유무와 상관없이 잘 생장하고 카탈라아제와 운동성에 대해서는 음성으로 확인되었다. 또한, 15℃에서는 생장하지 않았고 45℃에서 생장하였으며, 글루코오스(glucose)로부터 가스와 알기닌으로부터 암모니아를 생성하지 않았다. 49종의 당발효 시험을 실시하였을 때 글루코스 등 24종으로부터 산을 생성하였다. (하기 실시예의 표 3 참조)The novel microorganism of the present invention was isolated from adult feces and was inoculated with modified MRS medium (see Table 1 of Example 1 below) diluted with peptone diluent and NaN 3 in a planar state by 0.1 ml, For 48 hours. Each strain was purified from the transformed MRS medium and then transformed into yellow as a potential lactic acid bacterium. The selected strains were plated on the electrotransformed MRS medium and cultured aerobically to isolate pure strains. As a result of the identification of pure isolates, the strain was Gram positive bacillus and showed good growth regardless of presence or absence of oxygen and negative for catalase and motility. In addition, it did not grow at 15 ° C, grew at 45 ° C, and did not produce ammonia from gas and arginine from glucose. When 49 sugar fermentation tests were carried out, acid was produced from 24 kinds such as glucose. (See Table 3 in the following examples)

16S rRNA 유전자 염기서열을 이용하여 얻어진 결과(도 1) 웨이젤라 에스피(Weissella sp.)인 것으로 최종 동정하였다. 본 발명의 성인분변에서 분리한 비만억제능력이 우수한 균주를 웨이젤라 에스피(Weissella sp.) F22로 명명하고, 2013년 10월 1일 농업생명공학연구원 한국농업미생물자원센터(KACC)에 수탁번호 KACC91867P로 기탁하였다. The result obtained using the 16S rRNA gene sequence (Fig. 1) was finally identified as Weissella sp. Weissella sp. F22 was named as a strain having excellent ability to inhibit obesity in adult feces of the present invention and designated as Accession No. KACC91867P (National Institute of Agricultural Biotechnology, KACC) ≪ / RTI >

미생물의 생장Microbial growth

생균수와 pH를 측정하여 본 발명 젖산균의 생장을 온도에 따라 시험한 결과, 생장최적온도는 40℃이었다. The viable cell count and pH were measured, and the growth of the lactic acid bacteria of the present invention was tested according to the temperature. As a result, the optimum growth temperature was 40 캜.

항생제 내성Antibiotic resistance

항생제 내성시험을 위해 항생제 존재하에서 생장여부를 관찰하여 항생제의 최저억제농도(MIC, minimal inhibitory concentration) 값을 측정한 결과, 폴리마이신, 스트렙토마이신 및 반코마이신에 대해 내성이 있는 반면 페니실린과 바시트라신에 감수성이 높았다(하기 실시예 표 4 참조). The antimicrobial resistance test showed that the antimicrobial activity of the antibiotics in the presence of antibiotics was not affected by the antimicrobial resistance to polymyxin, streptomycin and vancomycin, (See Table 4 in the following Examples).

효소 활성Enzyme activity

분리한 본 발명 젖산균에 대한 효소활성시험을 수행한 결과, 웨이젤라 에스피(Weissella sp.) F22는 산 포스파타제에서 효소 활성이 높게 나타났다. As a result of the enzyme activity test on the isolated lactic acid bacteria of the present invention, Weissella sp. F22 showed high enzyme activity in acid phosphatase.

Benzopyrene을 발암성 물질로 전환시키는 발암효소인 β-glucuronidase의 경우에는 효소활성이 없는 것으로 나타나 안전성을 확인할 수 있었다. In the case of β-glucuronidase, a carcinogenic enzyme that converts benzopyrene to a carcinogenic substance, there is no enzyme activity and its safety can be confirmed.

담즙내성 및 pH 내성Biliary and pH tolerance

본 발명 젖산균을 MRS 배지에서의 성장을 시험하여 담즙에 대한 내성을 측정한 결과, 7 시간 배양 후 담즙을 첨가하지 않을 때와 첨가할 때와는 0.25 log의 차이로 약간 억제를 받기는 하나 담즙에 대한 충분한 내성을 보유하는 것으로 확인되었다. 또한, pH 내성을 시험한 결과, 대조구 pH인 6.4에 비해 강산인 pH 2에서 조차도 생장에 영향을 받지 않음에 따라 강한 내산성을 가지고 있음을 확인하였다. When the lactic acid bacteria of the present invention were tested for growth resistance in the MRS medium, the tolerance to bile was measured to be slightly inhibited by 0.25 log difference between when the bile was not added and when the bile was added after 7 hours of culture, And that the tolerance was sufficient. As a result of the test for pH resistance, it was confirmed that even at pH 2, which is a strong acid, it is not influenced by growth and has a strong acid resistance, compared with the control pH of 6.4.

항균력Antibacterial activity

항균력 시험에서는 본 발명의 웨이젤라 에스피(Weissella sp.) F22는 스테피로코커스 오레우스(Staphylococcus aureus)에 대해서 87.3%로서 가장 억제력이 높았고, 살모넬라 타이피머리움(Salmonella typhimurium)에 대해 60.0%의 억제력을 나타내었다. 반면 에스케리키아 콜리(Escherichia coli)에 대해 억제력이 없었다.In the antimicrobial activity test, Weissella sp. F22 of the present invention showed the highest inhibitory effect against Staphylococcus aureus (87.3%) and the inhibitory effect against Salmonella typhimurium (60.0%) against Salmonella typhimurium Respectively. On the other hand, there was no inhibition against Escherichia coli .

비만억제활성Obesity inhibitory activity

본 발명의 엔테로코커스 휘칼리스(Enterococcus faecalis) MD366 균주는 우수한 췌장 지방분해효소(pancreatic lipase) 저해활성을 보였으며, 3T3-L1 지방전구세포를 이용한 지방세포분화 활성 실험에서 우수한 항-지방세포분화활성(anti-adipogenic activity)을 나타내었다. The Enterococcus faecalis of the present invention, The MD366 strain showed excellent pancreatic lipase inhibitory activity and showed excellent anti-adipogenic activity in the adipocyte differentiation activity test using 3T3-L1 adipocyte precursor cells.

본 발명의 다른 양태에 따르면, 본 발명은 웨이젤라 에스피(Weissella sp.) F22 (수탁번호: KACC 91867P) 균주가 접종된 발효기질을 포함하는 발효 출발 물질 조성물을 제공한다. According to another aspect of the present invention, there is provided a fermentation starter composition comprising a fermentation substrate inoculated with a strain of Weissella sp. F22 (Accession No .: KACC 91867P).

본 발명의 젖산균은 발효 출발 물질의 스타터(starter) 균주로서 사용가능하다. The lactic acid bacteria of the present invention can be used as a starter strain of a fermentation starting material.

본 발명의 일 구현예에 따르면, 본 발명의 발효 출발 물질 조성물에서 상기 발효기질은 우유이다. According to one embodiment of the present invention, in the fermentation starting material composition of the present invention, the fermentation substrate is milk.

본 발명의 또 다른 양태에 따르면, 본 발명은 상기 발효 출발 물질 조성물을 발효시켜 제조한 발효물로서, 웨이젤라 에스피(Weissella sp.) F22(수탁번호: KACC 91867P) 균주가 포함된 발효물을 제공한다. According to another aspect of the present invention, there is provided a fermentation product prepared by fermenting the above fermented starter composition, comprising a fermentation product containing a strain of Weissella sp. F22 (accession number: KACC 91867P) do.

본 발명의 또 다른 양태에 따르면, 본 발명은 상기 웨이젤라 에스피(Weissella sp.) F22(수탁번호: KACC 91867P) 균주가 포함된 발효물을 포함하는 식품 조성물을 제공한다.
According to yet another aspect of the present invention, the present invention provides a food composition comprising a fermentation product comprising the strain Weissella sp. F22 (Accession No. KACC 91867P).

본 발명은 비만억제능이 뛰어난 웨이젤라 에스피(Weissella sp.) F22 균주에 관한 것이다. 본 발명의 균주는 비만억제능이 우수하고, 식중독 유발균에 대해 생장억제 활성이 있으며, 담즙내성과 강한 내산성을 가진다. 또한, 탈지유에 산 생성 발효특성을 나타내므로, 발효유제품 제조를 위한 발효스타터 균주로 사용할 수 있다.
The present invention relates to a strain of Weissella sp. F22 which is excellent in the ability to inhibit obesity. The strain of the present invention is excellent in the ability to inhibit obesity, has a growth inhibitory activity against food poisoning-causing bacteria, and has bile resistance and strong acid resistance. Further, since the skim milk exhibits acid-producing fermentation characteristics, it can be used as a fermentation starter strain for producing fermented milk products.

도 1은 성인분변에서 분리된 F22 균주의 계통수를 나타낸 결과이다.
도 2는 3T3-L1 지방전구세포를 사용하여 본 발명의 젖산균 웨이젤라 에스피(Weissella sp.) F22 균주의 지방세포분화 억제활성을 측정한 결과이다.
도 3은 본 발명의 젖산균 웨이젤라 에스피(Weissella sp.) F22에 대해 10% 탈지분유를 사용하여 34℃, 37℃ 및 40℃에서의 24시간 동안 생장률을 측정한 결과이다.
도 4는 본 발명의 젖산균 웨이젤라 에스피(Weissella sp.) F22에 대해 10% 탈지분유를 사용하여 34℃, 37℃ 및 40℃에서 24시간 동안 배지에서의 pH 변화를 측정한 결과이다.
도 5는 본 발명의 젖산균 웨이젤라 에스피(Weissella sp.) F22의 담즙내성을 측정한 결과이다.
도 6은 본 발명의 젖산균 웨이젤라 에스피(Weissella sp.) F22의 탈지유에서의 발효 특성을 측정한 결과이다.
Fig. 1 shows the results of the phylogenetic tree of F22 isolates isolated from adult feces.
FIG. 2 shows the results of measuring the inhibitory activity of the lactic acid bacterium Weissella sp. F22 according to the present invention on adipocyte differentiation using 3T3-L1 adipose precursor cells.
FIG. 3 shows the results of measurement of growth rate at 34 ° C., 37 ° C. and 40 ° C. for 24 hours using 10% skimmed milk powder of the lactic acid bacterium Weissella sp. F22 of the present invention.
Fig. 4 shows the results of measurement of changes in pH in the medium for 24 hours at 34 ° C, 37 ° C and 40 ° C using 10% skim milk powder for the lactic acid bacterium Weissella sp. F22 of the present invention.
Fig. 5 shows the result of measuring the bile resistance of the lactic acid bacterium Weissella sp. F22 of the present invention.
6 shows the results of measurement of fermentation characteristics of the lactic acid bacterium Weissella sp. F22 of the present invention in skimmed milk.

이하, 실시예를 통하여 본 발명을 더욱 상세히 설명하고자 한다. 이들 실시예는 오로지 본 발명을 보다 구체적으로 설명하기 위한 것으로, 본 발명의 요지에 따라 본 발명의 범위가 이들 실시예에 의해 제한되지 않는다는 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 embodiments are only for describing the present invention in more detail and that the scope of the present invention is not limited by these embodiments in accordance with the gist of the present invention .

실시예 Example

실시예 1: 미생물의 분리 Example 1: Isolation of microorganisms

시료로 사용된 원유와 성인분변을 수거하여 브롬크레졸 퍼플(Bromcresol purple)과 소듐아자이드(sodium azide)를 첨가한 변형 MRS 배지(표 1 참조)에 0.3 ㎖씩 평면도말법으로 접종한 후 37℃에서 48시간 배양하였다. 그런 다음, 각각의 균락을 상기한 변형 MRS 배지에서 순수분리하고, 노란색으로 변한 균락을 잠정적 젖산균으로 선발하였다. 선발된 균주는 상기 변형 MRS 배지에 3회 백금이로 도말한 후 호기배양 하여 순수분리 하였다. 하기 표 1은 미생물 분리에 사용된 변형 MRS 배지의 조성을 나타낸다. Crude milk and adult fecal samples used as samples were inoculated into modified MRS media (see Table 1) supplemented with Bromcresol purple and sodium azide (see Table 1) in a flatness condition of 0.3 ml, And cultured for 48 hours. Then, each of the herbicides was separated pure from the above-mentioned modified MRS medium, and the yellow-transformed herb was selected as a potential lactic acid bacterium. The selected strains were plated on the modified MRS medium three times and then cultured aerobically to isolate pure strains. Table 1 below shows the composition of the modified MRS medium used for microbial isolation.

성분  ingredient 성분비(g/l) Component Ratio (g / l) 프로테오스 펩톤 #3 Proteose Peptone # 3 10.0 10.0 쇠고기 추출물Beef extract 10.0  10.0 효모 추출물Yeast extract 5.05.0 락토오스 Lactose 20.0 20.0 트윈 80 Twin 80 1.01.0 암모늄 시트레이트Ammonium citrate 2.02.0 소디움 아세테이트Sodium acetate 5.05.0 마그네슘 설페이트Magnesium sulfate 0.10.1 망간 설페이트Manganese sulfate 0.05  0.05 디포타슘 포스페이트 Dipotassium phosphate 2.02.0 소듐 아자이드Sodium azide 0.250.25 브롬크레졸 퍼플 Bromocresol purple 0.040.04

실시예 2: 비만억제능 젖산균주 선발 Example 2: Obesity inhibitory ability of lactic acid bacteria

원유에서 분리한 1,272개 균주와 분변에서 분리한 269개 균주 중 돼지 췌장지방분해효소(porcine pancreatic lipase)를 이용하여 지방분해효소(lipase) 활성저해 정도를 측정하여 지방분해 억제활성(anti-lipase activity)이 65% 이상인 4개 균주를 먼저 선별하였다. 상기 선별된 4개의 균주를 10% 탈지분유(skim milk) 용액에 1% 접종하고 37℃에서 배양한 후 얻어진 배양액을 동결건조한 다음 3T3-L1 세포주에 100㎍/ml 농도를 첨가하여 이 세포의 지방세포로의 분화 및 지방축적(adipogenesis)을 유도하는지 여부를 측정하는 지방분화 유도활성(adipogenic activity) 실험을 실시하였다. 이 실험의 결과는 다음 표 2와 도 2에 나타내었다. Among 1,272 strains isolated from crude oil and 269 strains isolated from feces, porcine pancreatic lipase was used to measure lipase activity inhibition, and anti-lipase activity ) Were 65% or more. The selected four strains were inoculated 1% in 10% skim milk solution, cultured at 37 ° C, lyophilized, and then added to a 3T3-L1 cell line at a concentration of 100 μg / ml. Adipogenic activity experiments were conducted to determine whether the cells induced differentiation and lipid accumulation (adipogenesis) of the captive. The results of this experiment are shown in Table 2 and FIG.

Figure 112013095097794-pat00001
Figure 112013095097794-pat00001

상기 표 2에서 보는 바와 같이 분리된 젖산균 F22 균주는 지방분해억제율이 68.03%이고, 지방분화율이 87.02%로서 다른 균주에 비해 가장 비만억제 능력이 우수한 균주임을 확인하였고 이 균주를 선발하였다.
As shown in Table 2, it was confirmed that the isolated strain of lactic acid bacteria F22 was 68.03% in lipolysis inhibition rate and 87.02% in lipolysis rate.

실시예 3: 미생물의 동정 Example 3: Identification of microorganisms

미생물의 분류학적 동정은 Hammes 등[The prokaryotes, 1563-1578, 2nd Edition, Springer-Verlag Co.(1992)]의 방법에 따라 수행하였으며, 그 결과는 표 3에 나타내었다. 이들 균주는 모두 그람 양성, 간균이었으며, 산소유무와 상관없이 잘 생장하였고, 카탈라아제와 운동성은 음성이었다. 또한, 15℃에서는 생장하지 않고 45℃에서 생장하였으며 포도당(glucose)으로부터 가스(gas)와 알기닌(arginine)으로부터 암모니아를 생성하지 않았다. 49종의 당발효 시험을 실시하였을 때 글루코스 등 24종으로부터 산을 생성하였다. 16S rRNA 유전자 염기서열을 이용하여 얻어진 결과(도 1) 웨이젤라 에스피(Weissella sp.) F22로 판명되었다. 상기 과정을 통해 성인분변에서 분리한 비만억제능력이 우수한 균주를 웨이젤라 에스피(Weissella sp.) F22로 명명하고, 2013년 10월 1일 농업생명공학연구원 한국농업미생물자원센터(KACC)에 수탁번호 KACC 91867P로 기탁하였다. The taxonomic identification of microorganisms was performed according to the method of Hammes et al. [The prokaryotes, 1563-1578, 2nd Edition, Springer-Verlag Co. (1992)]. All of these strains were Gram - positive, Bacillus, well grown regardless of presence or absence of oxygen, and catalase and motility were negative. In addition, it did not grow at 15 ℃ and grew at 45 ℃, and did not produce ammonia from gas and arginine from glucose. When 49 sugar fermentation tests were carried out, acid was produced from 24 kinds such as glucose. The result obtained using the 16S rRNA gene sequence (Fig. 1) was found to be Weissella sp. F22. Weissella sp. F22 was named as an isolate of obesity-inhibiting ability isolated from adult feces by the above procedure and was deposited on October 1, 2013 by KACC (Korea Research Institute of Bioscience and Biotechnology) KACC 91867P.

아래 표 3은 본 발명 신규 젖산균 웨이젤라 에스피(Weissella sp.) F22의 생리적 및 생화학적 특성을 정리하여 나타내었다. Table 3 below summarizes the physiological and biochemical characteristics of the novel lactic acid bacterium Weissella sp. F22 of the present invention.

그램 반응
세포 형태
포자 형성
운동성
호기적 성장
혐기적 성장
카탈라제 반응
15℃에서 성장
45℃에서 성장
포도당으로 부터 가스생성
알기닌으로 부터 암모니아 생성
Gram reaction
Cell morphology
Spore formation
motility
Aerobic growth
Anaerobic growth
Catalase reaction
Grow at 15 ℃
Grow at 45 ° C
Gas production from glucose
Ammonia production from arginine
+
rod
-
-
+
+
-
-
+
-
-
+
rod
-
-
+
+
-
-
+
-
-
당이용 능력 Party ability 글리세롤
에리티톨
D-아라비노스
L-아라비노스
리보스
글루코스
프락토스
만노스
소르보스
람노스
둘시톨
이노시톨
만니톨
솔비톨
α-메칠-D-만노사이드
α-메칠-D-글루코사이드
N 아세칠 글루코스아민
아미그달린
아르부틴
에스쿨린
살리신
셀로비오스
말토스
락토스
멜리비오스
Glycerol
Erythritol
D-arabinose
L-arabinose
Ribos
Glucose
Fructose
Mannos
Sorbos
Rams North
Dissytol
Inositol
Mannitol
Sorbitol
alpha -methyl-D-mannose
alpha -methyl-D-glucoside
N-acetylglucosamine
Amigalline
Arbutin
Esculin
Salincin
Cellobiose
Maltose
Lactose
Melibiose
+
-
-
+
+
+
+
+
-
-
-
-
+
+
+
-
+
+
+
+
+
+
+
+
+
+
-
-
+
+
+
+
+
-
-
-
-
+
+
+
-
+
+
+
+
+
+
+
+
+
D-사이로스
L-사이로스
아도니톨
β-메칠-D-사이로사이드
갈락토스
수크로스
트레할로스
이눌린
멜레지토스
라피노스
스타치
글리코겐
실리톨
겐티오비오스
D-투라노스
D-라이소스
D-타가토스
D-후코스
L-후코스
D-아라비톨
L-아라비톨
글루코네이트
2-케토-글루코네이트
5-케토-글루코네이트
D-Cirrus
L-Cirrus
Adonitol
beta -methyl-D-thiazole
Galactose
Sucrose
Trehalose
Inulin
Melitos
Raffinos
Starch
Glycogen
Silica
Gentiobios
D-Turanos
D-lysos
D-tagatose
D-Fucose
L-Fucose
D-arabitol
L-arabitol
Gluconate
2-keto-gluconate
5-keto-gluconate
-
-
-
-
+
+
+
-
-
-
-
-
-
+
+
-
+
-
-
-
-
-
-
-
-
-
-
-
+
+
+
-
-
-
-
-
-
+
+
-
+
-
-
-
-
-
-
-

실시예 4: 미생물의 생장시험 Example 4: Microbial growth test

젖산균의 생장은 생균수 및 pH를 측정하여 시험하였다. 생균수는 10% 탈지분유 150 ㎖에 젖산균을 1 ㎖용 피펫으로 1 방울 접종한 후 34℃, 37℃, 40℃에서 3시간 간격으로 24시간 까지 배양한 각 시료를 0.1% 펩톤용액에 희석하여 BCP plate count agar 평판에서 부어 굳힌 후 35℃에서 48시간 배양하여 계수하였고, 온도 및 시간별로 pH 변화를 측정하였다. 측정 결과는 도 3 및 도 4에 나타내었다. 도 3과 도 4에서 볼 수 있는 바와 같이 젖산균 생장 최적온도는 40℃이었다.
The growth of lactic acid bacteria was measured by measuring viable cell count and pH. The number of viable cells was inoculated to 150 ml of 10% skim milk powder, and 1 ml of a lactic acid bacterium was inoculated with 1 ml of a pipette. Each sample was incubated at 34 ° C, 37 ° C, and 40 ° C for 3 hours at intervals of 3 hours, BCP plate count agar was poured on a plate, hardened, incubated at 35 ° C for 48 hours and counted by pH and temperature. The measurement results are shown in FIG. 3 and FIG. As can be seen from Figs. 3 and 4, the optimum temperature for lactic acid bacteria growth was 40 占 폚.

실시예 5: 항생제 내성시험 Example 5: Antibiotic resistance test

항생제 내성시험은 트립틱 소이 브로스(tryptic soy broth, Difco, USA)를 사용하여 2배 희석방법에 의해 생장여부를 관찰하여 최저억제농도(MIC) 값을 정하였으며, 측정한 결과는 표 4에 나타내었다. 아래 표 4에는 웨이젤라 에스피(Weissella sp.) F22의 항생제 감수성을 측정한 결과가 나타나 있다. The antimicrobial resistance test was carried out by using a tryptic soy broth (Difco, USA), and the growth inhibition was examined by a 2-fold dilution method to determine the lowest inhibitory concentration (MIC) . Table 4 below shows the results of measuring antibiotic susceptibility of Weissella sp. F22.

항생제Antibiotic 최저억제농도(㎍/㎖)The lowest inhibitory concentration (占 퐂 / ml) 아미노글리코사이드계Aminoglycoside system 아미카신 Amikacin 320320 겐타마이신Gentamicin 2020 가나마이신 Kanamycin 400400 네오마이신Neomycin 400400 스트렙토마이신*Streptomycin * 16001600 β-락탐계beta -lactam system 페니실린-G*Penicillin-G * 55 메치실린 Methacillin 8080 옥사실린Oxacillin 3030 암피실린Ampicillin 320320 그램-양성 스펙트럼Gram-positive spectrum 바시트라신*Bashtrasin * 3.753.75 리팜피신Rifampicin 3030 노보바이오신Novobaiosin 3030 린코마이신Lincomycin 800800 그램-음성 스펙트럼Gram-voice spectrum 폴리마이신B*Polymycin B * 24002400 광범위 스펙트럼Broad spectrum 클로람페니콜 Chloramphenicol 4040 반코마이신*Vancomycin * 16001600

* : units/㎖
*: units / ml

상기 표 4에서 볼 수 있는 바와 같이 웨이젤라 에스피(Weissella sp.) F22는 타 계열의 항생제에 비해 폴리마이신, 스트렙토마이신 및 반코마이신에 대해 내성이 있는 반면 페니실린과 바시트라신에 감수성이 높았다.
As can be seen in Table 4 above, Weissella sp. F22 is more resistant to polymyxin, streptomycin and vancomycin than penicillin and bacitracin, but is more susceptible to penicillin and bacitracin than other antibiotics.

실시예 6: 효소활성시험 Example 6: Enzyme activity test

효소활성시험은 MRS 액체배지에서 37℃, 18시간 동안 배양한 균주를 생리식염수로 희석하여 105 - 106 cfu/㎖ 수준의 시료를 조제한 후, API ZYM 키트(API bioMerieux, France)를 이용하여 37℃에서 5시간 동안 배양한 다음 효소반응을 시켰다. 효소활성은 표준색상표를 비교하여 0-5의 수치로 표시한 결과는 표 5에 나타내었다. 아래 표 5는 웨이젤라 에스피(Weissella sp.) F22의 효소활성을 비교한 결과이다. The enzyme activity test was performed by diluting a strain cultured in MRS liquid medium at 37 ° C for 18 hours with physiological saline to prepare a sample of 10 5 to 10 6 cfu / ml, and then using API ZYM kit (API bioMerieux, France) Incubated at 37 ° C for 5 hours, and then subjected to enzyme reaction. The enzyme activity is shown in Table 5 as the value of 0-5 compared with the standard color trademark. Table 5 below shows the results of comparing enzyme activities of Weissella sp. F22.

효 소enzyme 효소활성Enzyme activity 알칼라인 포스파타제
에스테라제(C4)
에스테라제 리파제(C8)
리파제(C14)
류신 아릴아미다제
발린 아릴아미다제
시스틴 아릴아미다제
트립신
카이모트립신
산 포스파타제
나프톨-AS-BI-포스포하이드로라제
α-갈락토시다제
β-갈락토시다제
β-글루쿠로니다제
α-글루코시다제
β-글루코시다제
N-아세칠-β-글루코스아민니다제
α-만노시다제
β-푸코시다제
Alkaline phosphatase
Esterase (C4)
Esterase lipase (C8)
Lipase (C14)
Leucine arylamidase
Valine arylamidase
Cysteine aryl amidase
Trypsin
Chymotrypsin
Acid phosphatase
Naphthol-AS-BI-phosphohydrolase
? -galactosidase
? -galactosidase
β-glucuronidase
alpha -glucosidase
? -glucosidase
N-acetyl- beta -glucosamine amine
? -mannosidase
? -fucosidase
1
1
0
1
2
2
0
2
0
4
2
2
1
0
1
1
0
0
1
One
One
0
One
2
2
0
2
0
4
2
2
One
0
One
One
0
0
One

*: 0에서 5까지 단계별로 표준색이 명시되어 있으며, 0은 음성이고 5는 최대의 강도를 나타낸다. 1은 5 나노몰(nanomoles), 2는 10 나노몰, 3은 20 나노몰, 4는 30 나노몰, 5는 40 나노몰 이상의 효소활성을 나타낸다.
*: Standard color is specified in steps from 0 to 5, 0 is negative and 5 is maximum intensity. 1 shows 5 nano moles, 2 has 10 nano moles, 3 has 20 nano moles, 4 has 30 nano moles, and 5 has an enzyme activity of 40 nano moles or more.

상기 표 5에서 볼 수 있는 바와 같이, 웨이젤라 에스피(Weissella sp.) F22는 산 포스파타제에서 효소 활성이 높게 나타났다. Benzopyrene을 발암성 물질로 전환시키는 발암효소인 β-glucuronidase의 경우에는 효소활성이 없는 것으로 나타나 안전성을 확인할 수 있었다.
As shown in Table 5 above, Weissella sp. F22 showed high enzyme activity in acid phosphatase. In the case of β-glucuronidase, a carcinogenic enzyme that converts benzopyrene to a carcinogenic substance, there is no enzyme activity and its safety can be confirmed.

실시예 7: 담즙내성시험 Example 7: Bile tolerance test

담즙내성시험은 길리랜드와 월커의 방법(Gilliland & Walker, J. Dairy Sci., 73, 905, 1990)에 따라 측정하였으며, 측정 결과는 도 5에 나타내었다. 도 5에서 볼 수 있는 바와 같이 7 시간 배양 후 담즙을 첨가하지 않을 때와 첨가할 때와는 0.25 log의 차이로 약간 억제를 받기는 하나 담즙에 대한 내성이 있는 것으로 나타났다.
The bile tolerance test was performed according to the method of Gilliland and Walker (Gilliland & Walker, J. Dairy Sci., 73, 905, 1990), and the measurement results are shown in FIG. As can be seen from FIG. 5, after the incubation for 7 hours, the difference was 0.25 log between the addition and the absence of bile.

실시예 8: pH 내성시험 Example 8: pH resistance test

pH 내성 시험은 클라크 등의 방법(Clark et al., Cultured Dairy Products J., 28(4), 11, 1993)에 따라 측정하였으며, 측정 결과는 표 6에 나타내었다. 아래 표 6에는 염산용액에서 3 시간 후의 웨이젤라 에스피(Weissella sp.) F22 내성을 비교한 결과를 나타내었다. The pH tolerance test was carried out according to the method of Clark et al. (Clark et al., Cultured Dairy Products J., 28 (4), 11, 1993), and the measurement results are shown in Table 6. Table 6 below shows the results of comparing Weissella sp. F22 resistance after 3 hours in a hydrochloric acid solution.

배양시간(hr)Culture time (hr) pH 2pH 2 pH 3pH 3 pH 4pH 4 pH 6.4pH 6.4 00 7.377.37 7.407.40 7.417.41 7.397.39 1One 7.417.41 7.477.47 7.507.50 7.467.46 22 7.507.50 7.557.55 7.567.56 7.587.58 33 7.487.48 7.537.53 7.557.55 7.547.54

단위: CFU per ㎖ (Log number)
Unit: CFU per ㎖ (Log number)

상기 표 6에서 볼 수 있는 바와 같이 본 발명의 균주는 대조구 pH인 6.4에 비해 강산인 pH 2에서 조차도 생장에 영향을 받지 않음에 따라 매우 강한 내산성을 가짐을 확인할 수 있었다.
As can be seen from Table 6, the strain of the present invention was not affected by growth even at pH 2, which is a strong acid, compared to the control pH of 6.4.

실시예 9: 항균력 시험 Example 9: Antimicrobial activity test

항균력 시험은 길리랜드와 스펙(Gilliland & Speck, J. Food Prot., 40(12), 820, 1977)에 따라 측정하였으며, 측정 결과는 아래 표 7에 나타내었다. 아래 표 7은 MRS 배지에서 웨이젤라 에스피(Weissella sp.) F22 식중독균 억제(*)활성을 측정한 결과이다. The antimicrobial activity test was performed according to Gilliland & Speck, J. Food Prot., 40 (12), 820, 1977, and the measurement results are shown in Table 7 below. Table 7 below shows the results of measuring the activity of Weissella sp. F22 food poisoning bacteria (*) in the MRS medium.

식중독균Food poisoning bacteria 생 장Growth 식중독균a Food poisoning bacteria a Weissella sp. F22*,a Weissella sp. F22 *, a 억제율(%) % Inhibition CFU/㎖CFU / ml pHpH CFU/㎖CFU / ml pHpH Escherichia coli Escherichia coli 1.2×108 1.2 × 10 8 6.466.46 1.8×108 1.8 x 10 8 5.545.54 -- Salmonella typhimurium Salmonella typhimurium 2.0×106 2.0 × 10 6 6.486.48 8.0×105 8.0 × 10 5 5.505.50 60.060.0 Staphyloccous aureus Staphylococcus aureus 1.5×108 1.5 × 10 8 6.546.54 1.9×107 1.9 × 10 7 5.405.40 87.387.3

* Weissella sp. F22의 초기 수 : 3.3 X 106 CFU/㎖ * Weissella sp. Initial number of F22: 3.3 X 10 < 6 > CFU / ml

a 37℃에서 6시간 배양 후 측정
a Measurement after incubation at 37 ° C for 6 hours

상기 표 7에서 볼 수 있는 바와 같이, 본 발명의 균주 웨이젤라 에스피(Weissella sp.) F22는 스테피로코커스 오레우스(Staphylococcus aureus)에 대해서 87.3%로서 가장 억제력이 높았고, 살모넬라 타이피머리움(Salmonella typhimurium)에 대해 60.0%의 억제력을 나타내었다. 반면 에스케리키아 콜리(Escherichia coli)에 대해 억제력이 없었다.
As can be seen in Table 7, the most inhibitory effect was high as 87.3% with respect to the strain-way gelatinase SP (Weissella sp.) F22 is stearyl Pyrococcus fluorescein mouse (Staphylococcus aureus) of the invention, Salmonella tie blood head Titanium (Salmonella typhimurium ) was inhibited by 60.0%. On the other hand, there was no inhibition against Escherichia coli .

실시예 10: 웨이젤라 에스피(Weissella sp.) F22 균주의 제품적용을 위한 pH 변화측정 Example 10: Measurement of pH change for product application of Weissella sp. Strain F22

본 발명의 균주가 발효유 스타터로서 적합한지 여부를 확인하기 위하여 웨이젤라 에스피(Weissella sp.) F22를 탈지유에 배양하면서 시험에 사용하였다. 탈지유에 웨이젤라 에스피(Weissella sp.) F22를 각각 1.0%씩 접종하고 37℃에서 3시간 별로 24시간까지 배양한 후의 pH 변화를 측정하였으며, 측정결과는 도 6에 나타내었다. 도 6에서 볼 수 있는 바와 같이 탈지유에서 발효가 이루어졌으며, 따라서, 본 발명의 젖산균은 발효유의 스타터 균주로 적합한 것으로 나타났다.
To confirm whether the strain of the present invention is suitable as a fermented milk starter, Weissella sp. F22 was used for the test while culturing in skimmed milk. The skim milk was inoculated with 1.0% each of Weissella sp. F22 and cultured at 37 ° C for 3 hours for 24 hours. The pH change was measured. The measurement results are shown in FIG. As can be seen from FIG. 6, fermentation was carried out in skimmed milk. Thus, the lactic acid bacteria of the present invention were found to be suitable as starter strains of fermented milk.

이상으로 본 발명의 특정한 부분을 상세히 기술하였는 바, 당업계의 통상의 지식을 가진 자에게 있어서 이러한 구체적인 기술은 단지 바람직한 구현 예일 뿐이며, 이에 본 발명의 범위가 제한되는 것이 아닌 점은 명백하다. 따라서, 본 발명의 실질적인 범위는 첨부된 청구항과 그의 등가물에 의하여 정의된다고 할 것이다.
While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the same is by way of illustration and example only and is not to be construed as limiting the scope of the present invention. Accordingly, the actual scope of the present invention will be defined by the appended claims and their equivalents.

농업생명공학연구원Agricultural Biotechnology Research Institute KACC91867PKACC91867P 2013100120131001

Claims (5)

비만억제능이 우수한 웨이젤라 에스피(Weissella sp.) F22(수탁번호: KACC 91867P) 균주.
Weissella sp. F22 (accession number: KACC 91867P) which is excellent in the ability to inhibit obesity.
제 1 항에 있어서, 상기 웨이젤라 에스피(Weissella sp.) F22 (수탁번호: KACC 91867P) 균주는 췌장 지방분해효소(pancreatic lipase) 억제활성 및 항-지방세포분화활성(anti-adipogenic activity)을 갖는 것을 특징으로 하는 균주.
The strain according to claim 1, wherein the strain Weissella sp. F22 (accession number: KACC 91867P) has a pancreatic lipase inhibitory activity and an anti-adipogenic activity Lt; / RTI >
비만억제능이 우수한 웨이젤라 에스피(Weissella sp.) F22(수탁번호: KACC 91867P) 균주가 접종된 발효기질을 포함하는 발효 출발 물질 조성물.
A fermentation starting material composition comprising a fermentation substrate inoculated with a strain of Weissella sp. F22 (accession number: KACC 91867P) having excellent inhibitory activity against obesity.
제 3 항의 발효 출발 물질 조성물을 발효시켜 제조한 발효물로서, 웨이젤라 에스피(Weissella sp.) F22(수탁번호: KACC 91867P) 균주가 포함된 발효물.
A fermentation product obtained by fermenting the fermentation starting material composition of claim 3, wherein the fermentation product is a strain of Weissella sp. F22 (accession number: KACC 91867P).
제 4 항의 발효물을 포함하는 식품 조성물. A food composition comprising the fermented product of claim 4.
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Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20190068068A (en) 2017-12-08 2019-06-18 한국식품연구원 Pediococcus acidilactici AO22 strain with Anti-Obesity Ability and Composition for treatment or improvement or preventing of obesity comprising the same
KR20190068075A (en) 2017-12-08 2019-06-18 한국식품연구원 Lactobacillus pentosus AO21-1 strain with Anti-Obesity Ability and Composition for treatment or improvement or preventing of obesity comprising the same
KR20190068078A (en) 2017-12-08 2019-06-18 한국식품연구원 Lactobacillus paracasei AO356 strain with Anti-Obesity Ability and Composition for treatment or improvement or preventing of obesity comprising the same
KR20240110171A (en) 2023-01-05 2024-07-15 국립공주대학교 산학협력단 A composition containing a Lactobacillus reuteri KCL018strain having an ability to inhibit fat accumulation in fat cells, its culture medium or cell extract
KR20240110172A (en) 2023-01-05 2024-07-15 국립공주대학교 산학협력단 A composition containing a Lactobacillus pentosus KCL032 strain having an ability to inhibit fat accumulation in fat cells, its culture medium or cell extract

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20090113478A (en) * 2008-04-28 2009-11-02 주식회사 엔유씨전자 Probiotic Lactobacillus plantarum having anti-obesity and brain function improvement activity
KR20100010015A (en) * 2008-07-21 2010-01-29 신현길 Lactobacillus johnsonii hfi 108 having blood cholesterol level lowering and anti-obesity activity
KR20100109661A (en) * 2009-04-01 2010-10-11 주식회사한국야쿠르트 Lactobacillus curvatus hy7601 having inhibitory activity against blood cholestrol and obesity, and product containing thereof as an effective factor
KR20120128260A (en) * 2011-05-17 2012-11-27 삼육대학교산학협력단 Phamaceutical or food composition for treating or preventing obesity disease comprising a mixture of lactic acid bacteria

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20090113478A (en) * 2008-04-28 2009-11-02 주식회사 엔유씨전자 Probiotic Lactobacillus plantarum having anti-obesity and brain function improvement activity
KR20100010015A (en) * 2008-07-21 2010-01-29 신현길 Lactobacillus johnsonii hfi 108 having blood cholesterol level lowering and anti-obesity activity
KR20100109661A (en) * 2009-04-01 2010-10-11 주식회사한국야쿠르트 Lactobacillus curvatus hy7601 having inhibitory activity against blood cholestrol and obesity, and product containing thereof as an effective factor
KR20120128260A (en) * 2011-05-17 2012-11-27 삼육대학교산학협력단 Phamaceutical or food composition for treating or preventing obesity disease comprising a mixture of lactic acid bacteria

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20190068068A (en) 2017-12-08 2019-06-18 한국식품연구원 Pediococcus acidilactici AO22 strain with Anti-Obesity Ability and Composition for treatment or improvement or preventing of obesity comprising the same
KR20190068075A (en) 2017-12-08 2019-06-18 한국식품연구원 Lactobacillus pentosus AO21-1 strain with Anti-Obesity Ability and Composition for treatment or improvement or preventing of obesity comprising the same
KR20190068078A (en) 2017-12-08 2019-06-18 한국식품연구원 Lactobacillus paracasei AO356 strain with Anti-Obesity Ability and Composition for treatment or improvement or preventing of obesity comprising the same
KR102077720B1 (en) 2017-12-08 2020-02-14 한국식품연구원 Pediococcus acidilactici AO22 strain with Anti-Obesity Ability and Composition for treatment or improvement or preventing of obesity comprising the same
KR102136718B1 (en) 2017-12-08 2020-08-14 한국식품연구원 Lactobacillus pentosus AO21-1 strain with Anti-Obesity Ability and Composition for treatment or improvement or preventing of obesity comprising the same
KR20240110171A (en) 2023-01-05 2024-07-15 국립공주대학교 산학협력단 A composition containing a Lactobacillus reuteri KCL018strain having an ability to inhibit fat accumulation in fat cells, its culture medium or cell extract
KR20240110172A (en) 2023-01-05 2024-07-15 국립공주대학교 산학협력단 A composition containing a Lactobacillus pentosus KCL032 strain having an ability to inhibit fat accumulation in fat cells, its culture medium or cell extract

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