KR101909927B1 - Probiotics Comprising Yeast From Kefir - Google Patents
Probiotics Comprising Yeast From Kefir Download PDFInfo
- Publication number
- KR101909927B1 KR101909927B1 KR1020170031377A KR20170031377A KR101909927B1 KR 101909927 B1 KR101909927 B1 KR 101909927B1 KR 1020170031377 A KR1020170031377 A KR 1020170031377A KR 20170031377 A KR20170031377 A KR 20170031377A KR 101909927 B1 KR101909927 B1 KR 101909927B1
- Authority
- KR
- South Korea
- Prior art keywords
- kefir
- grape seed
- yeast
- present
- derived
- Prior art date
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Abstract
본 발명은 케피어 유래 효모를 포함하는 장 기능 개선용 프로바이오틱스에 관한 것이다. 본 발명에 따른 케피어 유래 효모를 포함하는 신규한 프로바이오틱스는 장 부착능이 우수하여 장내 유해균의 성장을 억제할 수 있다. 또한, 케피어 유래 효모를 포도씨 분말 또는 추출물과 함께 사용함으로써 현저히 향상된 항산화 활성을 발휘할 수 있다. 본 발명에 다른 케피어 유래 효모는 장 기능 개선용 건강기능식품으로 활용될 수 있으며, 장내 유해균의 생육을 억제 및 항산화를 위한 약학 조성물로서도 활용될 수 있다.The present invention relates to probiotics for improving bowel function comprising kefir-derived yeast. The novel probiotics containing the kefir derived yeast according to the present invention are excellent in the ability to adhere to the intestines and can inhibit the growth of harmful bacteria in the intestines. In addition, the kefir-derived yeast can be used in combination with the grape seed powder or the extract to exhibit remarkably improved antioxidative activity. The kefir-derived yeast according to the present invention can be used as a health functional food for improving bowel function, and can be used as a pharmaceutical composition for inhibiting the growth of enteric bacteria in the intestines and for antioxidation.
Description
본 발명은 케피어 유래 효모를 포함하는 장 기능 개선용 프로바이오틱스에 관한 것이다.The present invention relates to probiotics for improving bowel function comprising kefir-derived yeast.
케피어(Kefir)는 코카서스 산간 지방에서 유래한 발효유의 일종으로, 티벳지방 승려들이 건강을 위해 음용하던 것이 대중화된 음료이다. 케피어의 구성영양소로는 단백질 및 다당류가 포함되어 있고 비타민 A, 비타민 B1, 비타민 B2, 비타민 B6, 비타민 D, 비타민 K2, 엽산, 니코틴산, 칼슘, 철분, 요오드 등을 고루 포함하고 있다. 이러한 케피어는 유산균과 효모 복합체인 케피어 그레인으로 불리우는 균종(케피어 과립균이라고도 한다)을 개시제로 하여 우유를 발효시켜 얻어진 요구르트상의 음료이다. Kefir is a kind of fermented milk derived from the mountains of the Caucasus. It is a popular beverage that Tibetan monks drink for health. Kepir's constituent nutrients include proteins and polysaccharides and include vitamin A, vitamin B1, vitamin B2, vitamin B6, vitamin D, vitamin K2, folic acid, nicotinic acid, calcium, iron and iodine. Such a kefir is a drink of yoghurt obtained by fermenting milk with an initiator (also referred to as Kefir granule bacterium) called kefir grain, which is a combination of lactic acid bacteria and yeast.
케피어 그레인에 의하여 발효된 음료인 케피어는 인체에 유익한 유산균을 포함하고 있어 장 활동을 원활히 하여 변비를 억제하고 위장 장애를 개선하는데 도움이 되며 면역체계의 강화에 효능이 있다고 알려져 있으나, 케피어 유래의 효모가 갖는 프로바이오틱 기능성에 대해서는 아직 밝혀진 바가 없다. Kefir, a fermented beverage that is fermented by kefir grain, is known to contain beneficial lactic acid bacteria to facilitate intestinal activity, to inhibit constipation, to improve gastrointestinal disorders, and to enhance the immune system. Kefir The probiotic functionality of the resulting yeast has not yet been elucidated.
프로바이오틱스(probiotics)는 건강에 도움이 되는 장내 세균총 즉, 숙주의 건강에 이로움을 제공하는 살아있는 미생물 즉 생균류를 의미한다. 일반적으로 프로바이오틱스는 요거트 등의 발효식품의 일부 또는 식이보충제로서 소비된다. 프로바이오틱스로 알려진 미생물에는 락트산 박테리아(lactic acid bacteria; LAB), 비피도박테리아(bifidobacteria), 바실러스 등이 있다.Probiotics refers to healthy microbes that help the health of a host, that is, live microbes, that are beneficial to the health of the host. Generally, probiotics are consumed as part of fermented foods such as yogurt or as dietary supplements. Microorganisms known as probiotics include lactic acid bacteria (LAB), bifidobacteria, and bacillus.
프로바이오틱스와 유사한 개념으로, 프리바이오틱스(prebiotics)는 장내 유익한 박테리아의 생장을 돕는 난소화성 성분으로서 프로바이오틱스(probiotics)의 영양원이 되어 장내 환경을 개선하는 데 도움을 주는 물질을 말한다. 어떠한 식품 성분이, 대장 내 유용 미생물에 의해 이용되어 미생물의 생육이나 활성을 촉진함으로써 숙주 건강에 좋은 효과를 나타내게 하는 프리바이오틱스로 활용되기 위해서는, 위장관의 상부에서 소화 또는 흡수되지 않아야 하고 대장 내 미생물 중 유용 세균을 선택적으로 활성화시키고 병원균 등의 유해균은 억제할 수 있어야 한다.In a similar concept to probiotics, prebiotics are substances that help to improve the intestinal environment by becoming a nutrient source of probiotics, an indigestible ingredient that helps the growth of beneficial bacteria in the intestines. In order for any food ingredient to be utilized as a prebiotic to be used by the beneficial microorganism in the intestine to promote the growth or activity of the microorganism and thereby exhibit a beneficial effect on host health, it must not be digested or absorbed from the upper part of the gastrointestinal tract, It is necessary to selectively activate the bacterium and to inhibit harmful bacteria such as pathogens.
한편, 포도에는 식물에서 발견되는 항산화물질인 폴리페놀(polyphenol) 계열의 레스베라트롤(resveratrol)이 많이 함유되어 있으며, 레스베라트롤은 항균, 항암, 항바이러스, 항알레르기, 및 항염증 감소효과를 지니며 강력한 항산화 활성의 기능이 있는 물질로 포도부산물인 과피, 씨, 송이 가지에 다량 함유되어 있으므로 포도부산물을 그대로 폐기할 경우 농산물에 함유된 유용한 성분을 손실하게 되는 것은 물론이고, 환경오염의 문제가 되므로 포도부산물의 효용가치를 높이는 연구가 활발하게 진행되고 있다. On the other hand, grapes contain a lot of polyphenol-based resveratrol, which is found in plants. Resveratrol has antioxidant, anti-cancer, anti-virus, antiallergic and anti-inflammatory effects, It is a substance with active function, and it is contained in large amount in grape by-products, perilla, seed and pine branches. Therefore, when grape by-products are discarded as they are, useful components contained in agricultural products are lost, Researches are being actively carried out to increase the utility value of.
본 발명자들은 케피어 유래 효모의 프로바이오틱스로서의 활성을 확인하고, 포도씨 분말이 케피어 유래 효모에 대한 프리바이오틱스로서 작용할 수 있다는 것을 확인함으로써, 본 발명을 완성하였다.The present inventors have confirmed the activity of kefir-derived yeasts as probiotics and have completed the present invention by confirming that the grape seed powder can act as a prebiotic for kefir-derived yeast.
이와 같은 과제를 해결하기 위한 본 발명의 목적은 케피어 유래 효모를 포함하는 장 기능 개선용 프로바이오틱스를 제공하는 것이다. It is an object of the present invention to solve the above problems and provide a probiotic for improving bowel function comprising kefir-derived yeast.
본 발명의 다른 목적은 케피어 유래 효모, 및 포도씨 분말 또는 추출물을 포함하는 장 기능 개선용 프로바이오틱스를 제공하는 것이다. Another object of the present invention is to provide a probiotic for improving bowel function comprising kefir-derived yeast and a grape seed powder or extract.
본 발명의 또 다른 목적은 케피어 유래 효모를 포함하는 장 기능 개선용 건강기능식품 및 약학 조성물을 제공하는 것이다.Another object of the present invention is to provide a health functional food and a pharmaceutical composition for improving intestinal function comprising kefir-derived yeast.
상기 목적을 달성하기 위해, 본 발명은 케피어(Kefir) 유래 효모를 포함하는, 장 기능 개선용 프로바이오틱스(probiotics)를 제공한다. In order to achieve the above object, the present invention provides probiotics for improving bowel function, including Kefir derived yeast.
본 발명에 있어서, 상기 케피어 유래 효모는 사카로마이세스(Saccharomyces) 속 또는 클루베로마이세스(Kluyveromyces) 속인 것을 특징으로 할 수 있으며, 보다 바람직하게는 상기 케피어 유래 효모는 사카로마이세스 세레비지애(Saccharomyces cerevisiae) 또는 클루베로마이세스 막시아누스(Kluyveromyces marxianus)인 것을 특징으로 할 수 있다. In the present invention, the kefir-derived yeast may be characterized by being a genus of Saccharomyces or Kluyveromyces . More preferably, the kefir-derived yeast may be Saccharomyces cerevisiae Saccharomyces cerevisiae or Kluyveromyces < RTI ID = 0.0 > marxianus ).
또한, 본 발명은 상기 케피어 유래 효모가 포도씨 분말 또는 포도씨 추출물을 함유하는 배지에서 배양된 것을 특징으로 하는 프로바이오틱스를 제공한다. Also, the present invention provides a probiotic wherein the kefir-derived yeast is cultured in a medium containing grape seed powder or grape seed extract.
본 발명에서, 상기 포도씨 분말은 와인 제조 부산물인 것을 특징으로 할 수 있다. In the present invention, the grape seed powder may be a byproduct of wine production.
본 발명에서, 상기 포도씨 추출물은 포도씨 분말을 물 또는 알코올을 이용하여 추출한 추출물인 것을 특징으로 할 수 있다. In the present invention, the grape seed extract may be an extract obtained by extracting grape seed powder with water or alcohol.
또한, 본 발명은 케피어 유래 효모, 및 포도씨 분말 또는 포도씨 추출물을 포함하는 프로바이오틱스 조성물을 제공한다. The present invention also provides a probiotic composition comprising kefir-derived yeast and a grape seed powder or a grape seed extract.
또한, 본 발명은 케피어 유래 효모를 유효성분으로 포함하는 장 기능 개선용 건강기능식품을 제공한다. The present invention also provides a health functional food for improving intestinal function comprising kefir-derived yeast as an active ingredient.
바람직하게는, 상기 건강기능식품은 포도씨 분말 또는 포도씨 추출물을 추가로 포함할 수 있다. Preferably, the health functional food may further comprise grape seed powder or grape seed extract.
또한, 본 발명은 케피어 유래 효모를 유효성분으로 포함하는 장내 유해균 생육 억제 및 항산화용 약학 조성물을 제공한다.The present invention also provides a pharmaceutical composition for inhibiting growth of intestinal noxious bacteria and antioxidation comprising kefir-derived yeast as an active ingredient.
바람직하게는, 상기 약학 조성물은 포도씨 분말 또는 포도씨 추출물을 추가로 포함할 수 있다.Preferably, the pharmaceutical composition may further comprise grape seed powder or grape seed extract.
본 발명에 따른 케피어 유래 효모를 포함하는 신규한 프로바이오틱스는 장 부착능이 우수하여 장내 유해균의 성장을 억제할 수 있다. 또한, 케피어 유래 효모를 포도씨 분말 또는 추출물과 함께 사용함으로써 현저히 향상된 항산화 활성을 발휘할 수 있으며, 와인 제조시 발생되는 부산물을 프리바이오틱스로 활용할 수 있기 때문에, 친환경적이고 경제적이다. The novel probiotics containing the kefir derived yeast according to the present invention are excellent in the ability to adhere to the intestines and can inhibit the growth of harmful bacteria in the intestines. In addition, the kefir-derived yeast can be used in combination with the grape seed powder or the extract to exhibit a remarkably improved antioxidant activity, and the byproduct generated in the wine production can be utilized as a prebiotic, thereby being environmentally friendly and economical.
본 발명에 다른 케피어 유래 효모는 장 기능 개선용 건강기능식품으로 활용될 수 있으며, 장내 유해균의 생육을 억제하고 항산화를 위한 약학 조성물로서도 활용될 수 있다.The kefir-derived yeast according to the present invention can be utilized as a health functional food for improving bowel function, inhibiting the growth of enteric bacteria in the intestines and also being used as a pharmaceutical composition for antioxidation.
도 1은 본 발명의 일 실시예에 따른 케피어 유래 효모균의 장 부착능 측정 결과를 나타낸다.
도 2(a)는 본 발명의 일 실시예에 따른 케피어 유래 효모균의 그대로의 세포(intact cells)의 DPPH 라디칼 소거능을 측정한 결과를 나타낸다.
도 2(b)는 본 발명의 일 실시예에 따른 케피어 유래 효모균의 무세포 추출액의 DPPH 라디칼 소거능을 측정한 결과를 나타낸다.
도 3(a)는 본 발명의 일 실시예에 따른 케피어 유래 효모균 및/또는 포도씨 분말의 DPPH 라디칼 소거능을 측정한 결과를 나타낸다.
도 3(b)는 본 발명의 일 실시예에 따른 케피어 유래 효모균 및/또는 포도씨 물 추출물의 DPPH 라디칼 소거능을 측정한 결과를 나타낸다.
도 3(c)은 본 발명의 일 실시예에 따른 케피어 유래 효모균 및/또는 포도씨 80% 메탄올 추출물의 DPPH 라디칼 소거능을 측정한 결과를 나타낸다.FIG. 1 shows the result of measuring the adherence of kefir-derived yeasts according to an embodiment of the present invention.
FIG. 2 (a) shows the results of measuring DPPH radical scavenging activity of intact cells of Kefir-derived yeast strain according to an embodiment of the present invention.
FIG. 2 (b) shows the results of measurement of the DPPH radical scavenging ability of the cell-free extract of Kefir-derived yeast strain according to an embodiment of the present invention.
FIG. 3 (a) shows the results of measurement of DPPH radical scavenging ability of kefir-derived yeast and / or grape seed powder according to an embodiment of the present invention.
FIG. 3 (b) shows the DPPH radical scavenging activity of kefir-derived yeast and / or grape seed extract according to an embodiment of the present invention.
FIG. 3 (c) shows the DPPH radical scavenging activity of kefir-derived yeast and / or 80% methanol extract of grapevine according to an embodiment of the present invention.
다른 식으로 정의되지 않는 한, 본 명세서에서 사용된 모든 기술적 및 과학적 용어들은 본 발명이 속하는 기술 분야에서 숙련된 전문가에 의해서 통상적으로 이해되는 것과 동일한 의미를 갖는다. 일반적으로, 본 명세서에서 사용된 명명법은 본 기술 분야에서 잘 알려져있고 통상적으로 사용되는 것이다.Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In general, the nomenclature used herein is well known and commonly used in the art.
본 발명의 일 구현 양태는, 케피어 유래 효모를 포함하는 장 기능 개선용 프로바이오틱스에 관한 것이다. One embodiment of the present invention relates to probiotics for improving bowel function comprising kefir derived yeast.
본 발명에서 “프로바이오틱스(probiotics)”란, 섭취되어 장에 도달하였을 때에 장내 환경에 유익한 작용을 하는 균주를 의미한다. 일반적으로 프로바이오틱스로는 유산균이 많이 사용되지만, 효모가 프로바이오틱스로서 사용되는 경우 유산균과는 다른 이점들이 존재한다. 예를 들어, 보통 프로바이오틱스는 열에 매우 민감하여 30℃만 넘어가도 기능을 잃기 시작하는 반면, 효모는 37℃에서 가장 활발히 활동을 하기 때문에 온도에 민감하지 않다는 장점이 있다. 또한, 일반적인 프로바이오틱스는 항생제에 의해 무력화될 수 있지만 효모의 경우 항생제에 영향을 받지 않는다. 마지막으로, 일반적인 프로바이오틱스는 위산과 담즙에 약하기 때문에 위산과 담즙에 버티기 위하여 다양한 방법을 적용해야 하지만 효모의 경우 위산과 담즙에 상관없이 생존한다는 특징이 있다. In the present invention, " probiotics " means a strain that is beneficial to the intestinal environment when it is ingested and reaches the intestines. Generally, lactic acid bacteria are widely used as probiotics. However, when yeast is used as a probiotic, there are advantages other than lactic acid bacteria. For example, probiotics are very sensitive to heat and begin to lose their function when they reach 30 ° C, while yeast is most susceptible to temperature because it is most active at 37 ° C. In addition, common probiotics can be neutralized by antibiotics, but yeast is not affected by antibiotics. Finally, since common probiotics are weak against gastric acid and bile, various methods must be applied in order to stay in stomach acid and bile, but yeast is characterized in that it survives regardless of gastric acid and bile.
프로바이오틱스로서 실질적인 기능을 나타내기 위해서는 장내 생존력도 중요하지만, 무엇보다 장 상피세포에 부착하여 균총을 형성하는 능력이 중요한데, 콜로니를 형성하여 기타 병원성 미생물이 자리잡지 못하도록 막아주기 위해서는 장 부착능력이 필수요소이다. Although it is important to have viability in the intestines in order to exhibit practical functions as probiotics, it is important that the ability to adhere to intestinal epithelial cells to form microorganisms is important. In order to prevent colonization of other pathogenic microorganisms, to be.
또한, 항산화 기능은 체내에서 일어나는 산화반응을 억제시키거나 그 속도를 느리게 하는 것인데, 체내에서 일어나는 산화 반응은 노화의 유발과 세포의 변이를 유발하여 암세포를 생성한다. 따라서, 항산화 작용은 이런 체내의 부정적인 반응을 억제하기 위한 예방 방편 중 하나로서 활용되고 있다. 구체적으로, 체내에서는 세포 호흡과 더불어 에너지에 해당하는 ATP생성이 주기능인 세포 내의 미토콘드리아가 호흡의 부산물인 자유 라디칼(free radical)을 생성하는데, 이는 불안정하여 지질, 핵산, 단백질 등의 다른 물질과 급격하고 무차별적으로 반응한다. 따라서 항산화를 통하여 이러한 자유 라디칼의 생성을 막을 수 있다면 간접적으로 노화와 암을 예방할 수 있다. In addition, the antioxidant function inhibits or slows down the oxidation reaction occurring in the body. Oxidation reaction in the body induces aging and cell mutation to generate cancer cells. Therefore, the antioxidant activity is utilized as one of the preventive measures to suppress the negative reaction in the body. Specifically, in the body, in addition to cell respiration, mitochondria within the cell, which is the main function of ATP production, generate free radicals, which are byproducts of respiration. This is unstable and can be caused by other substances such as lipids, nucleic acids, And react indiscriminately. Therefore, if antioxidants can prevent the generation of these free radicals, they can prevent aging and cancer indirectly.
본 발명에서는, 케피어로부터 동정한 효모가 프로바이오틱스 활성, 특히, 우수한 장 부착능 및 항산화 효능을 나타낸다는 것을 확인하였다. 즉, 본 발명은 케피어 유래 효모를 포함하는 신규한 장 기능 개선용 프로바이오틱스에 관한 것이다. In the present invention, it was confirmed that the yeast identified from Kappy exhibited probiotic activity, in particular, excellent intestinal adherence and antioxidant efficacy. That is, the present invention relates to a novel long-range function probiotics including kefir-derived yeast.
본 발명에서, 상기 케피어 유래 효모는 사카로마이세스(Saccharomyces) 속 또는 클루베로마이세스(Kluyveromyces) 속이 바람직하다. 더욱 구체적으로는, 상기 케피어 유래 효모는 사카로마이세스 세레비지애(Saccharomyces cerevisiae) 또는 클루베로마이세스 막시아누스(Kluyveromyces marxianus)인 것이 바람직하다. 본 발명의 일 실시예에서는, 사카로마이세스 세레비지애(ATCC6037) 및 클루베로마이세스 막시아누스(ATCC34440), 클루베로마이세스 막시아누스(KU140723-01), 클루베로마이세스 막시아누스(KU140723-02), 클루베로마이세스 막시아누스(KU140721-04) 및 클루베로마이세스 막시아누스(KU140723-05)가 종래 장 부착능이 우수한 것으로 알려진 락토바실러스 애시도필러스(Lactobacillus acidophilus)에 비하여 더 뛰어난 장 부착능을 나타낼 뿐만 아니라, 항산화 활성도 우수하다는 것을 확인하였다. In the present invention, the kefir-derived yeast is preferably Saccharomyces or Kluyveromyces . More specifically, the kefir-derived yeast is Saccharomyces cerevisiae cerevisiae or Kluyveromyces < RTI ID = 0.0 > marxianus ). In one embodiment of the present invention, there is provided a method for screening a compound of the present invention, which comprises the steps of: Saccharomyces cerevisiae (ATCC 6037) and Clube mariesces membrane cyanosis (ATCC 34440), Clube marine proteoglycans (KU140723-01) (KU140723-02), Clube marcesces cyanosis (KU140721-04) and Clube marcesces cyanosis (KU140723-05) have been found to be effective against Lactobacillus acidophilus It was confirmed that it exhibited more excellent adherence ability as well as antioxidative activity.
상기 케피어 유래 효모는 통상적으로 사용되는 방법에 의하여 케피어로부터 분리할 수 있다. The kefir-derived yeast can be isolated from kefir by a commonly used method.
본 발명의 바람직한 구현 양태에 있어서, 상기 케피어 유래 효모는 포도씨 분말 또는 추출물을 함유하는 배지에서 배양된 것일 수 있다. 본 발명자들은 포도씨 분말 또는 추출물이 케피어 유래 효모의 프로바이오틱스 활성에 유익한 영향을 미치는 프리바이오틱스(prebiotics)로서 작용한 다는 것을 확인하였다. In a preferred embodiment of the present invention, the kefir-derived yeast may be cultured in a medium containing grape seed powder or extract. The present inventors have confirmed that grape seed powder or extract acts as prebiotics which have a beneficial effect on the probiotic activity of yeast derived from Kepir.
본 발명에서 "프리바이오틱스(Prebiotics)"란, 장내 유익한 박테리아의 생장을 돕는 난소화성 성분으로서 프로바이오틱스(probiotics)의 영양원이 되어 장내 환경을 개선하는 데 도움을 주는 물질을 말한다.In the present invention, "Prebiotics" refers to a substance that helps to improve the intestinal environment as a nutrient source of probiotics as an indigestible ingredient that helps the growth of beneficial bacteria in the intestines.
본 발명의 일 실시예에서는 케피어 유래 효모를 포도씨 분말 또는 추출물을 함유하는 배지에서 배양하는 경우 항산화능이 월등히 향상될 수 있다는 것을 확인하였다. 특히, 포도씨 분말을 프리바이오틱스로서 사용하는 경우 케피어 유래 효모의 항산화능이 포도씨 분말 자체가 갖는 항산화능과 케피어 유래 효모의 항산화능의 단순 합산보다 더욱 월등히 향상되었다. In one embodiment of the present invention, it was confirmed that when the yeast derived from Kefir was cultured in a medium containing grape seed powder or extract, the antioxidant ability could be greatly improved. Especially, when the grape seed powder was used as the prebiotic, the antioxidant ability of the kefir derived yeast was much improved than the simple addition of the antioxidant ability of the grape seed powder itself and the antioxidant ability of the kefir - derived yeast.
본 발명에 이용되는 포도의 품종, 산지 또는 수확시기 등에는 특별한 제한은 없다. 본 발명의 일 실시예에서는 샤르도네(Chardonnay) 포도를 이용하였으나, 이에 제한되지 않는다.There are no particular restrictions on the type, area or harvest date of the grapes used in the present invention. In one embodiment of the present invention, Chardonnay grapes were used, but the present invention is not limited thereto.
본 발명의 포도씨 분말은 포도를 재료로 하는 음료나 주류의 제조과정에서 발생되는 부산물로부터 생성될 수 있다. 바람직하게는, 상기 포도씨 분말은 포도주 또는 와인(wine)의 제조과정에서 발생되는 부산물로부터 생성될 수 있다.The grape seed powder of the present invention can be produced from beverages made from grapes or from by-products generated in the course of producing liquors. Preferably, the grape seed powder may be produced from by-products generated during the manufacturing of wine or wine.
여기서, "부산물"은 포도를 원료로 하여 파쇄 및 발효 공정을 거친 후 증류 공정에서 생성되는 와인 또는 포도주 증류액을 제외한 나머지 생성물을 총칭한다.Here, "by-product" is generically referred to as wine or wine distillate produced in a distillation process after crushing and fermentation processes using grapes as raw materials.
통상적으로, 와인 또는 포도주 제조시 발생되는 부산물들은 폐기되고 있다. 그러나 이러한 폐기물은 원료 포도의 40% 정도에 이르러 경제적으로 큰 손실이 아닐 수 없다. 본 발명에 따르면, 와인 제조시 발생되는 부산물을 프리바이오틱스로 활용할 수 있기 때문에, 환경적, 경제적으로도 큰 이점이 있다.Typically, by-products that occur during the manufacture of wine or wine are being discarded. However, these wastes amount to as much as 40% of raw grapes and are economically disastrous. According to the present invention, byproducts generated in the production of wines can be utilized as prebiotics, there is a great advantage in terms of environment and economy.
본 발명의 일 실시예에 따른 "포도씨 분말"(Grape seed flour, GSF)은 와인을 생산하고 남은 부산물로부터 포도씨를 분리, 건조한 뒤, "냉각프레스", "열프레스" 기법 등을 이용하여 포도씨로부터 오일을 추출한 다음, 가루로 분쇄함으로써 생성될 수 있다. 포도씨 분말의 분리, 건조, 압축, 분쇄 방법은 종래 알려진 기술들에 의해 다양한 형태로 수행될 수 있다.Grape seed flour (GSF) according to one embodiment of the present invention separates and dries grape seeds from the remaining byproducts that produce wine, and then extracts grape seed from the grape seed using a "cold press" Extracting the oil, and then pulverizing it into a powder. The method of separating, drying, compressing and pulverizing the grape seed powder can be carried out in various forms by known techniques.
포도씨 분말의 입자 크기는 분쇄 환경에 따라 다양하게 조절할 수 있으며, 더 작은 입자를 가진 분말을 생성하기 위해 체로 거르는 작업을 추가할 수 있다. 포도씨 분말의 입자 크기는 식품, 의약품 등 활용 용도와 목적에 맞게 조절 가능하다.Grain size of the grape seed powder can be varied according to the grinding environment, and sieving operation can be added to produce a powder with smaller particles. The particle size of the grape seed powder can be adjusted to suit the application purpose and purpose such as food, medicines.
본 발명에서, "포도씨 추출물"이란 포도씨 또는 포도씨 분말을 다양한 추출 용매를 처리하여 수득되는 것으로서, 추출 용매로는 물, 알코올(바람직하게는, 메탄올, 에탄올) 등이 이용될 수 있다. 구체적으로, 포도씨 분말을 초음파 처리 후 물 또는 80% 메탄올로 추출하여 얻어진 추출물을 사용할 수 있다.In the present invention, "grape seed extract" is obtained by treating grape seed or grape seed powder with various extraction solvents, and water, alcohol (preferably, methanol, ethanol) and the like can be used as an extraction solvent. Specifically, an extract obtained by sonicating the grape seed powder and extracting it with water or 80% methanol can be used.
본 발명에서, 상기 배지는 효모균을 배양할 수 있는 배지라면 특별히 제한되지 않는다. 예를 들어, MRS Broth 배지를 사용하여 케피어 유래 효모균을 배양할 수 있다. In the present invention, the medium is not particularly limited as long as the medium is capable of culturing yeast. For example, kefir-derived yeast can be cultured using MRS broth medium.
상기 포도씨 분말 또는 추출물은 배지 10mL에 대하여 1 내지 50mg 포함될 수 있으며, 바람직하게는 3 내지 20mg 포함될 수 있고, 가장 바람직하게는 5 내지 10mg 포함될 수 있다.The grape seed powder or extract may be contained in an amount of 1 to 50 mg, preferably 3 to 20 mg, and most preferably 5 to 10 mg per 10 mL of the medium.
본 발명에서, 상기 케피어 유래 효모는 포도씨 분말 또는 추출물을 함유하거나 함유하지 않는 배지 10mL에 대하여 1.0 x 106 내지 1.0 x 108CFU를 사용할 수 있으며, 약 1.0 x 107CFU를 사용하는 것이 바람직하다.In the present invention, the kefir-derived yeast may be used in an amount of 1.0 x 10 6 to 1.0 x 10 8 CFU per 10 mL of medium containing or not containing grape seed powder or extract, preferably about 1.0 x 10 7 CFU Do.
본 발명의 다른 구현 양태는, 케피어 유래 효모, 및 포도씨 분말 또는 추출물을 포함하는 프로바이오틱스 조성물에 관한 것이다. Another embodiment of the present invention relates to a probiotic composition comprising a kefir-derived yeast, and a grape seed powder or extract.
본 발명의 프로바이오틱스 조성물은 케피어 유래 효모, 포도씨 분말 또는 추출물 및 허용가능한 담체를 포함할 수 있다. The probiotic composition of the present invention may comprise kefir-derived yeast, grape seed powder or extract, and acceptable carriers.
케피어 유래 효모를 포도씨 분말 또는 추출물과 함께 사용하는 경우, 포도씨 분말 또는 추출물이 프리바이오틱스로서 작용하여 케피어 유래 효모의 항산화능을 현저히 향상시킬 수 있다. 따라서, 우수한 장 부착능과 항산화 활성을 발휘하는 프로바이오틱스 조성물을 제공할 수 있다. When the kefir-derived yeast is used together with the grape seed powder or the extract, the grape seed powder or the extract acts as a prebiotic, so that the antioxidative ability of the kefir-derived yeast can be remarkably improved. Therefore, it is possible to provide a probiotic composition exhibiting excellent intestinal adhesiveness and antioxidative activity.
본 발명의 프로바이오틱스 조성물은 케피어 유래 효모 1.0 x 107CFU에 대하여 포도씨 분말 또는 추출물을 1 내지 50mg 포함할 수 있으며, 바람직하게는 3 내지 20mg 포함할 수 있고, 가장 바람직하게는 5 내지 10mg 포함할 수 있다.The probiotic composition of the present invention may contain 1 to 50 mg, preferably 3 to 20 mg, and most preferably 5 to 10 mg of the grape seed powder or extract against 1.0 x 10 7 CFU of kefir-derived yeast .
상기 프로바이오틱스 조성물은 장 기능 개선을 위한 다양한 용도에서 활용될 수 있다. 예를 들어, 본 발명의 프로바이오틱스 조성물은 정장 작용을 가지므로 '정장용' 조성물로서 사용 가능하다. '정장용'이란 사람이나 동물의 장내 세균총의 이상 발효에 의하여 야기되는 제반증상을 치료 및/또는 개선하는 용도를 말한다. 상기 증상으로는 이에 한정되지는 않으나, 예를 들어, 설사, 위장염, 염증성 장질환, 신경성 장염 증후군, 소장 미생물 과성장증 등이 포함된다.The probiotic composition can be utilized in various applications for improving bowel function. For example, the probiotic compositions of the present invention have a dressing action and thus can be used as a 'dressing' composition. "Formal use" refers to the use of treating and / or ameliorating symptoms caused by abnormal fermentation of intestinal flora in a person or animal. Such symptoms include, but are not limited to, diarrhea, gastroenteritis, inflammatory bowel disease, neurodegenerative inflammatory syndromes, small intestine microorganisms and growth conditions.
상기 프로바이오틱스 조성물은 당업계에 공지된 방법에 따라 다양한 제형과 방법으로 제조 및 투여될 수 있다. 예를 들어, 본 발명의 효모균 및/또는 포도씨 분말 또는 추출물을 포함하는 조성물은 식품/의약 분야에서 통상적으로 사용되는 담체와 혼합하여 건강기능식품 또는 약학 조성물로서 활용될 수 있다.The probiotic composition can be prepared and administered in a variety of formulations and methods according to methods known in the art. For example, the composition comprising the yeast and / or the grape seed powder or extract of the present invention may be used as a health functional food or a pharmaceutical composition by mixing with a carrier commonly used in the field of food / medicine.
본 발명의 또 다른 구현 양태는, 케피어 유래 효모를 유효성분으로 포함하는 장 기능 개선용 건강기능식품에 관한 것이다. Another embodiment of the present invention relates to a health functional food for improving bowel function comprising kefir-derived yeast as an active ingredient.
본 발명의 건강기능식품은 포도씨 분말 또는 추출물을 추가로 포함할 수 있다. The health functional food of the present invention may further comprise a grape seed powder or an extract.
상기 건강기능식품은 발효식품인 것이 바람직하지만, 이에 제한되지는 않는다. 예를 들어서, 건강기능식품은 음료, 껌, 차, 비타민 복합제, 건강보조 식품류일 수 있으며, 더욱 자세히는, 유제품, 제과물, 조미료, 음료 및 드링크제, 스낵, 캔디류, 젤리류, 아이스크림 및 냉동용 디저트, 아침 곡물류, 영양바, 스낵 바 초콜렛 제품, 가공 식품, 곡물 제품 및 파스타, 스프, 소스 및 드레싱, 과자 제품, 오일 및 지방 제품, 유제품 음료(dairy drink) 및 우유 음료, 차, 두유 및 콩 유제품(soy dairy-like product), 냉동식품, 조리 음식 및 대체 음식, 육류 제품, 치즈, 요구르트, 빵 및 롤빵, 케이크, 쿠키 및 크래커로 이루어진 군에서 선택된 어느 하나일 수 있다. The health functional food is preferably a fermented food, but is not limited thereto. For example, the health functional food may be a beverage, a gum, a tea, a vitamin complex, a health supplement food, and more specifically, a dairy product, a confectionery, a seasoning, a beverage and a drink, a snack, a candy, a jelly, Dairy, breakfast cereals, nutrition bars, snack bars Chocolate products, processed foods, cereal products and pasta, soups, sauces and dressings, confectionery products, oil and fat products, dairy drinks and milk drinks, tea, soy milk and beans And may be any one selected from the group consisting of dairy dairy-like products, frozen foods, cooked foods and alternate foods, meat products, cheese, yogurt, bread and buns, cakes, cookies and crackers.
또한, 상기 건강기능식품은 케피어 유래 효모 및 포도씨 분말 또는 추출물을 포함하는 캡슐, 정제, 분말, 액상 현탁액, 환제 및 과립제 등의 제형으로 제조될 수 있다.The health functional food may be prepared in the form of capsules, tablets, powders, liquid suspensions, pills and granules containing kefir-derived yeast and grape seed powder or extract.
또한, 상기 건강기능식품은 유효성분으로서 케피어 유래 효모 및 포도씨 분말 또는 추출물 뿐만 아니라, 식품 제조 시에 통상적으로 첨가되는 성분을 추가로 포함할 수 있다.The health functional food may further contain, as an active ingredient, not only kefir-derived yeast and grape seed powder or extract, but also ingredients normally added in the manufacture of food.
본 발명의 건강기능식품은 여러 가지 영양제, 비타민, 광물(전해질), 합성 풍미제 및 천연 풍미제 등의 풍미제, 착색제 및 중진제(치즈, 초콜릿 등), 펙트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알코올, 탄산 음료에 사용되는 탄산화제 등을 함유할 수 있으며, 과일주스나 야채 음료 제조를 위한 과육을 함유할 수 있다.The health functional food of the present invention can be used as a nutritive, a vitamin, a mineral (electrolyte), a flavoring agent such as a synthetic flavor agent and a natural flavor agent, a coloring agent and an aging agent (cheese, chocolate etc.), a pectic acid and its salt, Salt, organic acid, protective colloid thickener, pH adjusting agent, stabilizer, preservative, glycerin, alcohol, carbonating agent used in carbonated beverage, and may contain fruit pulp for producing fruit juice or vegetable beverage.
본 발명에 따른 건강기능식품은 장 내에 상피세포에 부착되어 유해균의 생육을 저해하고, 항산화 활성을 발휘하여 암 예방과 노화 방지에 도움을 줄 수 있다.The health functional food according to the present invention adheres to the epithelial cells in the intestines to inhibit the growth of harmful microorganisms and to exhibit antioxidant activity, thus being able to prevent cancer and prevent aging.
본 발명의 또 다른 구현 양태는, 케피어 유래 효모를 유효성분으로 포함하는 장내 유해균 억제용 약학 조성물에 관한 것이다. Another embodiment of the present invention relates to a pharmaceutical composition for inhibiting intestinal harmful bacteria comprising kefir-derived yeast as an active ingredient.
본 발명에서, 상기 약학 조성물은 포도씨 분말 또는 추출물을 추가로 포함할 수 있다. In the present invention, the pharmaceutical composition may further include a grape seed powder or an extract.
본 발명에 따른 약학 조성물은 장 내에 상피세포에 부착되어 유해균의 생육을 저해하고, 항산화 활성을 발휘하여 장 질환의 예방 또는 장 내 건강 개선의 효과가 있다.The pharmaceutical composition according to the present invention has an effect of preventing intestinal diseases or improving intestinal health by inhibiting the growth of noxious bacteria by adhering to epithelial cells in intestines and exhibiting antioxidant activity.
상기 약학 조성물은 케피어 유래 효모를 유효성분으로서 함유하며, 통상의 방법에 따른 적절한 담체, 부형제 또는 희석제를 추가로 포함할 수 있다.The pharmaceutical composition contains kefir-derived yeast as an active ingredient, and may further comprise an appropriate carrier, excipient or diluent according to a conventional method.
상기 담체, 부형제 및 희석제로는 락토즈, 덱스트로즈, 수크로스, 솔비톨, 만니톨, 자일리톨, 에리스리톨, 말티톨, 전분, 아카시아 고무, 알지네이트, 젤라틴, 칼슘 포스페이트, 칼슘 실리케이트, 셀룰로즈, 메틸 셀룰로즈, 미정질 셀룰로스, 폴리비닐 피롤리돈, 물, 메틸히드록시벤조에이트, 프로필히드록시벤조에이트, 탈크, 마그네슘 스테아레이트 및 광물유를 들 수 있다.Examples of the carrier, excipient and diluent include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, Cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.
또한, 본 발명의 약학 조성물은 다양한 경구 또는 비경구 투여 형태로 제형화될 수 있다.In addition, the pharmaceutical composition of the present invention may be formulated into various oral or parenteral administration forms.
경구 투여용 제형으로는 예를 들면 정제, 환제, 경질, 연질 캅셀제, 액제, 현탁제, 유화제, 시럽제, 과립제 등이 있는데, 이들 제형은 유효성분 이외에 희석제(예: 락토즈, 덱스트로즈, 수크로즈, 만니톨, 솔비톨, 셀룰로즈 및/또는 글리신), 활택제(예: 실리카, 탈크, 스테아르산 및 그의 마그네슘 또는 칼슘염 및/ 또는 폴리에틸렌 글리콜)를 추가로 포함할 수 있다. 또한, 상기 정제는 마그네슘 알루미늄 실리케이트, 전분 페이스트, 젤라틴, 트라가칸스, 메틸셀룰로즈, 나트륨 카복시메틸셀룰로즈 및/또는 폴리비닐피롤리딘과 같은 결합제를 함유할 수 있으며, 경우에 따라 전분, 한천, 알긴산 또는 그의 나트륨 염과 같은 붕해제 또는 비등 혼합물 및/또는 흡수제, 착색제, 향미제 및 감미제를 함유할 수 있다. 상기 제형은 통상적인 혼합, 과립화 또는 코팅 방법에 의해 제조될 수 있다.Examples of formulations for oral administration include tablets, pills, hard, soft capsules, liquids, suspensions, emulsions, syrups, granules and the like. These formulations may contain, in addition to the active ingredient, a diluent such as lactose, dextrose, (For example, silica, talc, stearic acid and magnesium or calcium salts thereof and / or polyethylene glycol), in addition to the active ingredient (s). The tablets may also contain binders such as magnesium aluminum silicate, starch paste, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose and / or polyvinylpyrrolidine, optionally mixed with starch, agar, alginic acid Or a disintegrating or boiling mixture such as its sodium salt and / or an absorbent, a colorant, a flavoring agent and a sweetening agent. The formulations may be prepared by conventional mixing, granulating or coating methods.
또한, 비경구 투여용 제형의 대표적인 것은 주사용 제제이며, 주사용 제제의 용매로서 물, 링거액, 등장성 생리 식염수 또는 현탁액을 들 수 있다. 상기 주사용 제제의 멸균 고정 오일은 용매 또는 현탁 매질로서 사용할 수 있으며 모노-, 디-글리세라이드를 포함하여 어떠한 무자극성 고정오일도 이러한 목적으로 사용될 수 있다. 또한, 상기 주사용 제제는 올레산과 같은 지방산을 사용할 수 있다.Representative examples of formulations for parenteral administration include injectable preparations, and water, Ringer's solution, isotonic saline or suspensions may be mentioned as a solvent for the injectable preparation. The sterile, fixed oils of the injectable preparations may be used as a solvent or suspending medium, and any non-irritating fixed oils, including mono-, di-glycerides, may be used for this purpose. The injectable preparation may be a fatty acid such as oleic acid.
본 발명의 약학 조성물의 투여량은 환자의 연령, 성별, 체중, 질환의 중증도에 따라 달라질 수 있다. 실제로 투여되는 본 발명의 약학 조성물의 양 및 투여 횟수는 치료하고자 하는 증상의 종류, 투여 경로, 성별, 건강 상태, 식이, 개체의 연령 및 체중, 및 질환의 중증도와 같은 다양한 인자에 의해 적절하게 결정될 수 있다.The dosage of the pharmaceutical composition of the present invention may vary depending on the age, sex, weight, and severity of the disease of the patient. The amount and the administration frequency of the pharmaceutical composition of the present invention to be actually administered is appropriately determined by various factors such as the kind of symptom to be treated, route of administration, sex, health condition, diet, age and weight of the individual, .
실시예Example
이하, 실시예를 통하여 본 발명을 더욱 상세히 설명하고자 한다. 이들 실시예는 오로지 본 발명을 예시하기 위한 것으로, 본 발명의 범위가 이들 실시예에 의해 제한되는 것으로 해석되지는 않는다는 것은 당업계에서 통상의 지식을 가진 자에게 있어서 자명할 것이다.Hereinafter, the present invention will be described in more detail with reference to Examples. It will be apparent to those skilled in the art that these embodiments are for illustrative purposes only and that the scope of the present invention is not construed as being limited by these examples.
실시예Example 1: One: 케피어Kefir 유래 효모의 장 Derived yeast cell 부착능Attachment 측정 Measure
케피어 유래 효모의 장 부착능을 측정하기 위하여, 장 상피세포 Caco-2 세포를 24 well plate에 융합성(confluent)이 95 내지 99%의 범위가 될 때까지 배양하였다. In order to measure the adherence ability of the kefir-derived yeast, the intestinal epithelial cell Caco-2 cells were cultured in a 24-well plate until confluent was in the range of 95 to 99%.
케피어 유래 효모균으로서, 사카로마이세스 세레비지애(ATCC6037), 클루베로마이세스 막시아누스(ATCC34440), 클루베로마이세스 막시아누스(KU140723-01), 클루베로마이세스 막시아누스(KU140723-02), 클루베로마이세스 막시아누스(KU140721-04) 및 클루베로마이세스 막시아누스(KU140723-05)를 각 well에 106 CFU/ml의 농도로 첨가하고, 5% CO2 존재 하에 37℃에서 1시간 동안 배양하였다. As the kefir-derived yeast, there are Saccharomyces cerevisiae (ATCC 6037), Clube Veromyces membranaceus (ATCC34440), Clube Veromyces membranaceus (KU140723-01), Clube Veromyces membriana (KU140723 (KU140721-04) and Clube Mymeseth membrane cyanosis (KU140723-05) were added to each well at a concentration of 10 6 CFU / ml, and incubated in the presence of 5% CO 2 Followed by incubation at 37 DEG C for 1 hour.
또한, 양성 대조군으로서, 락토바실러스 애시도필러스(Lactobacillus acidophilus)를 동일 조건으로 배양하였다. In addition, as a positive control, Lactobacillus acidophilus was cultured under the same conditions.
배양 후, PBS(phosphate buffered saline)로 3번 세척하여 부착되지 않은 시험균을 제거하고, caco-2 세포에 부착된 시험균의 생균수를 측정하기 위하여 2mL Triton X-100(1% v/v)를 가하여 시험균을 회수하였다. 용해물은 MRS agar에 희석시켜 접종하여 도말하였다. After incubation, the cells were washed 3 times with PBS (phosphate buffered saline) to remove unattached test bacteria. To measure the viable cell count of the test bacteria attached to caco-2 cells, 2 mL Triton X-100 (1% v / v ) Was added to recover the test bacteria. The lysates were inoculated with MRS agar diluted.
시험균의 장 부착능을 세포의 수로 측정하여 도 1에 나타내었다. The ability of the test bacteria to bind to the intestines was measured by the number of cells and is shown in Fig.
실험 결과, 양성 대조군인 락토바실러스 애시도필러스에 비하여, 케피어 유래 효모의 균주들이 5 내지 25% 가량 부착능이 더 높은 것을 확인할 수 있다. 특히, 클루베로마이세스 막시아누스(KU140723-02) 균의 장 부착능이 가능 우수하였다. As a result of the experiment, it can be confirmed that the strains of the yeast strains derived from the kefir are higher by about 5 to 25% than those of the lactobacillus acidophilus strain, which is a positive control. Particularly, the ability of the Adhesion of Clube marasmalum cyanosis (KU140723-02) was excellent.
즉, 케피어 유래 효모균들이 프로바이오틱스 기능성인 장 부착성능이 매우 우수하며, 유해균의 생육을 억제할 수 있다는 것을 확인할 수 있다. In other words, it can be confirmed that the yeast-derived yeasts of Kepir are excellent in the ability to bind the probiotic functional fungus and inhibit the growth of harmful bacteria.
실시예Example 2: 2: 케피어Kefir 유래 효모의 항산화 능력 측정 Antioxidant capacity of yeast
케피어 유래 효모균들의 그대로의 세포(intact cells)와 무세포 추출액 (intracellular cell-free extracts)의 항산화 능력을 DPPH 라디칼 소거능 측정을 이용하여 측정하였다. The antioxidant capacity of intact cells and intracellular cell-free extracts of kefir-derived yeasts was measured by DPPH radical scavenging assay.
실시예 1에서 사용된 것과 동일한 케피어 유래 효모균주들을 3차 계대하여 활성이 높은 균주를 사용하였다. The strains with the highest activity against the tertiary system were used for the yeast strains derived from the same kefir as those used in Example 1.
균주를 PBS 완충용액으로 3회 세척한 후, 10ml의 PBS로 재현탁하여, 그대로의 세포(intact cells) 샘플을 준비하였다. The strain was washed three times with PBS buffer, and resuspended in 10 ml of PBS to prepare a sample of intact cells.
또한, 무세포 추출액 준비를 위하여, 세포 펠렛을 멸균 3차 증류수로 2회 세척한 후 재현탁하여 25분 동안 얼음조에서 5초 간격, 15초 작동으로 초음파처리(ultrasonication)를 실시하였다. 분쇄된 균체를 3,000rpm에서 10분간 원심분리 후, 그의 상등액으로부터 무세포 추출액을 얻었다. In order to prepare cell-free extracts, the cell pellet was washed twice with sterilized distilled water three times, resuspended, and ultrasonicated for 25 minutes in an ice bath at intervals of 5 seconds and for 15 seconds. The pulverized cells were centrifuged at 3,000 rpm for 10 minutes, and a cell-free extract was obtained from the supernatant.
DPPH(α,α-diphenyl-β-picrylhydrazyl)을 0.2 mM 농도로 메탄올에 녹인 용액에 상기 샘플들을 0.1 mL로 주입하고, 30분간 37℃에서 암반응시켰다. 반응된 용액을 원심분리하여 이의 상등액 0.15 mL을 새 플레이트에 옮겼다. DPPH (α, α-diphenyl-β-picrylhydrazyl) was dissolved in methanol at a concentration of 0.2 mM, and the samples were incubated at 37 ° C. for 30 minutes. The reacted solution was centrifuged and 0.15 mL of its supernatant was transferred to a new plate.
DPPH 라디칼 소거능(DPPH radical scavenging activity)은 분광광도계를 이용하여 흡광도 517nm에서 측정하였다. Control group은 PBS, 양성 대조군은 아스코르브산(ascorbic acid)을 이용하였다. The DPPH radical scavenging activity was measured at 517 nm using a spectrophotometer. Control group was PBS, and positive control group was ascorbic acid.
DPPH 라디칼 소거능은 아래와 같은 식에 의하여 계산하였다. DPPH radical scavenging activity was calculated by the following equation.
DPPH 라디칼 소거능(%) = 
(Ac: control의 흡광도, As: Sample의 흡광도, Asb: Sample blank의 흡광도)(Ac: control absorbance, As: absorbance of sample, Asb: absorbance of sample blank)
도 2(a) 및 (b)에서 확인 가능한 바와 같이, 케피어 유래 효모균의 세포 그대로와 무세포 추출액 모두 유사한 패턴으로 항산화 활성을 나타내었다. As can be seen from Figs. 2 (a) and 2 (b), both the cell-free extract and the cell-free extract of Kefir-derived yeast showed antioxidative activity in a similar pattern.
실시예Example 3: 3: 케피어Kefir 유래 효모균과 포도씨 분말/추출물의 항산화 능력 측정 Antioxidant capacity measurement of derived yeast and grape seed powder / extract
3-1. 포도씨 분말 및 추출물 제조3-1. Manufacture of grape seed powder and extract
샤르도네(Chardonnay) 포도를 이용하여 와인을 만들고 남은 부산물을 냉압착(cold-pressed)하여, 포도씨에서 기름을 제거하였다. 그 다음, 남은 생성물을 열풍 건조하고, air blow를 이용하여 껍질 부분을 제거하였다. 열풍 건조 후 남은 생성물을 100mesh(150micron) 사이즈를 가진 체를 이용하여 걸러 고운 입자의 포도씨 분말(Grape seed flour, GSF)을 제조하였다.Chardonnay grapes were used to make wine, and the remaining by-products were cold-pressed to remove grease from the grape seeds. Then, the remaining product was hot-air dried and the skin was removed by air blow. Grape seed flour (GSF) was prepared by filtering the remaining product after hot air drying using a sieve having a size of 100 mesh (150 micron).
한편, 포도씨 추출물(Grape seed extracts, GSE)로는, i) 포도씨 분말을 초음파 처리 후 물로 추출한 추출물(Sonicated grape seed water extracts, SGWE)과, ii) 포도씨 분말을 초음파 처리 후 80% 메탄올로 추출한 추출물(Sonicated grape seed MeOH 80% extracts, SGME)을 준비하였다.Grape seed extracts (GSE) were prepared by i) sonicated grape seed water extracts (SGWE) and ii) grape seed powder was extracted with 80% methanol after ultrasonication Sonicated
3-2. 항산화 능력 측정3-2. Antioxidant capacity measurement
크루이베로마이세스 막시아누스(KU140723-02)(YST)를 2차 계대를 하였다. And the second passage was made of Crude Veromycetes membrane cyanus (KU140723-02) (YST).
MRS broth 10mL이 담긴 튜브에 포도씨 분말(GSF), 포도씨 분말 물 추출물(SGWE) 및 포도씨 분말 80% 메탄올 추출물(SGME)을 각각 5mg와 10mg의 함량으로 첨가하고, 5분 동안 초음파 처리(50초 작동, 10초 간격, 5회 반복) 한 액체 배지에 상기 2차 계대한 균주를 첨가하여 3차 계대를 하였다.To the tube containing 10 mL of MRS broth was added 5 mg and 10 mg of the grape seed powder (GSF), the grape seed powder water extract (SGWE) and the 80% methanol extract of the grape seed (SGME), respectively, and sonicated for 5 minutes , Every 10 seconds, repeated 5 times), and the third strain was added to the liquid medium.
3차 계대 배양한 활성이 높은 실험 균주 배양액을 PBS로 3회 세척한 후, 각 시료와 DPPH를 0.1 mL씩 혼합하여 30분간 37℃에서 암반응을 시켰다. The third culture was washed three times with PBS and 0.1 mL of DPPH was added to each well. The reaction mixture was incubated at 37 ° C for 30 minutes.
반응된 용액을 원심분리를 하여 반응물의 포도씨를 걸러내고, 이의 상등액 0.15 mL을 취해 96 well 플레이트에 옮겨 실시예 2와 동일한 방법으로 DPPH 라디칼 소거능을 측정하였다. 실험결과를 도 3 및 표 1에 나타내었다. The reaction solution was centrifuged to remove the grape seeds of the reaction product, and 0.15 mL of the supernatant was transferred to a 96-well plate. The DPPH radical scavenging activity was measured in the same manner as in Example 2. The results of the experiment are shown in FIG. 3 and Table 1.
No.
Sample
상기 표에서, 예측값은 크루이베로마이세스 막시아누스(KU140723-02)(YST)의 DPPH 라디칼 소거능 측정결과와 포도씨 분말 또는 추출물의 DPPH 라디칼 소거능 측정결과를 단순 합산한 뒤 2로 나눈 값을 나타낸다. In the above table, the predicted value is obtained by simply summing the results of measurement of DPPH radical scavenging ability of Cruyveromyces membrane cyanosis (KU140723-02) (YST) and measurement of DPPH radical scavenging ability of grape seed powder or extract and dividing by 2.
도 3(a)에서, 5mg 포도씨 분말을 첨가한 배지에서 배양한 효모균은 DPPH 라디칼 소거능이 22.58%에서 87.94%로 약 3.89배 가량 증가하였으며, 10mg 포도씨 분말을 첨가한 배지에서 배양한 효모균은 라디칼 소거능은 88.29%로 약 3.91배 가량 증가하였다. 이와 같은 증가는 표 1에서 확인 가능한 바와 같이, 단순히 효모균 자체의 라디칼 소거능과 포도씨 분말의 라디칼 소거능을 평균한 결과(예측값)보다 더 유의미하게 향상된 결과를 나타낸다. 즉, 포도씨 분말이 효모균의 항산화능을 현저히 향상시킬 수 있다는 것을 확인할 수 있다. In FIG. 3 (a), the yeast cultured in the medium supplemented with 5 mg of grape seed powder showed a 3.89-fold increase in DPPH radical scavenging ability from 22.58% to 87.94%. The yeast cultured in the medium supplemented with 10 mg of grape seed powder showed radical scavenging ability Was 88.29%, which was about 3.91 times higher than that of the control group. As can be seen in Table 1, such an increase shows a significantly improved result than the result (predicted value) obtained by simply averaging the radical scavenging ability of the yeast itself and the radical scavenging ability of the grape seed powder. That is, it can be confirmed that the grape seed powder can remarkably improve the antioxidant ability of yeast.
도 3(b) 및 도 3(c)에서도 마찬가지로, 효모균을 포도씨 추출물과 함께 배양한 경우 라디칼 소거능이 약 4배 가량 상승한 것을 확인할 수 있으며, 이 결과 또한 효모균과 포도씨 추출물의 라디칼 소거능을 단순 합산한 결과보다 유의미하게 향상된 값을 나타내었다. 3 (b) and 3 (c), when the yeast cells were cultured together with the grape seed extract, the radical scavenging ability was increased about 4 times. As a result, the radical scavenging ability of yeast and grape seed extract The results were significantly better than the results.
따라서, 포도씨 분말 또는 추출물이 프리바이오틱스로서 작용하여 프로바이오틱스인 케피어 유래 효모균의 항산화 활성을 크게 증폭시킨다는 것을 확인하였다. Therefore, it was confirmed that grape seed powder or extract acts as a prebiotics, which greatly amplifies the antioxidant activity of yeast-derived yeasts, which are probiotics.
이상으로 본 발명의 내용의 특정부분을 상세히 기술하였는 바, 당업계의 통상의 지식을 가진 자에게 있어서, 이러한 구체적 기술은 단지 바람직한 실시양태일 뿐이며, 이에 의해 본 발명의 범위가 제한되는 것이 아닌 점은 명백할 것이다. 따라서, 본 발명의 실질적인 범위는 첨부된 청구항들과 그것들의 등가물에 의하여 정의된다고 할 것이다.While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it will be understood by those skilled in the art that various changes and modifications may be made without departing from the spirit and scope of the invention as defined in the appended claims. It will be obvious. Accordingly, the actual scope of the present invention will be defined by the appended claims and their equivalents.
Claims (12)
A probiotics for improving bowel function comprising yeast derived from Kefir cultivated in a medium containing grape seed powder or grape seed extract.
상기 케피어 유래 효모가 사카로마이세스(Saccharomyces) 속 또는 클루베로마이세스(Kluyveromyces) 속인 것을 특징으로 하는, 프로바이오틱스.
The method according to claim 1,
Characterized in that the kefir derived yeast is of the genus Saccharomyces or Kluyveromyces .
상기 케피어 유래 효모가 사카로마이세스 세레비지애(Saccharomyces cerevisiae) 또는 클루베로마이세스 막시아누스(Kluyveromyces marxianus)인 것을 특징으로 하는, 프로바이오틱스.
The method according to claim 1,
Wherein the kefir derived yeast is selected from the group consisting of Saccharomyces cerevisiae or Kluyveromyces < RTI ID = 0.0 > marxianus ). < / RTI >
상기 포도씨 분말이 와인 제조 부산물인 것을 특징으로 하는, 프로바이오틱스.
The method according to claim 1,
Wherein the grape seed powder is a byproduct of wine production.
상기 포도씨 추출물은 포도씨 분말을 물 또는 알코올을 이용하여 추출한 추출물인 것을 특징으로 하는, 프로바이오틱스.
The method according to claim 1,
Wherein the grape seed extract is an extract obtained by extracting grape seed powder with water or alcohol.
Derived yeast cultured in a medium containing grape seed powder or grape seed extract, and a gravid seed powder or a grape seed extract.
A health functional food for improving intestinal function comprising kefir derived yeast cultured in a medium containing grape seed powder or grape seed extract as an active ingredient.
상기 건강기능식품이 발효식품인 것을 특징으로 하는, 건강기능식품.
9. The method of claim 8,
Wherein the health functional food is a fermented food.
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