KR102214532B1 - health food composition using pomegranate fermented by lactic acid bacteria and manufacturing method thereof - Google Patents
health food composition using pomegranate fermented by lactic acid bacteria and manufacturing method thereof Download PDFInfo
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- KR102214532B1 KR102214532B1 KR1020190003673A KR20190003673A KR102214532B1 KR 102214532 B1 KR102214532 B1 KR 102214532B1 KR 1020190003673 A KR1020190003673 A KR 1020190003673A KR 20190003673 A KR20190003673 A KR 20190003673A KR 102214532 B1 KR102214532 B1 KR 102214532B1
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- South Korea
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- lactic acid
- acid bacteria
- pomegranate
- fermented
- food composition
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A23V2250/00—Food ingredients
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- A—HUMAN NECESSITIES
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- A—HUMAN NECESSITIES
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- A—HUMAN NECESSITIES
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- A23V2400/11—Lactobacillus
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Abstract
본 발명은 석류 유산균 발효물을 이용한 건강식품 조성물과 이의 제조방법에 관한 것으로서, 더욱 상세하게는 석류를 유산균으로 발효시킨 유산균 발효물을 함유하여 항비만, 항염증 등의 효능이 우수한 건강식품 조성물과 이의 제조방법에 관한 것이다.
본 발명의 석류 유산균 발효물을 이용한 건강식품 조성물은 우유와 석류추출물을 혼합하여 유산균에 의해 발효시킨 유산균 발효물을 함유한다.The present invention relates to a health food composition using a pomegranate lactic acid bacteria fermentation product and a method for manufacturing the same, and more particularly, a health food composition having excellent efficacy such as anti-obesity and anti-inflammatory effects by containing a lactic acid bacteria fermentation product fermented from pomegranate with lactic acid bacteria It relates to a method of manufacturing the same.
The health food composition using the fermented product of pomegranate lactic acid bacteria of the present invention contains a fermented product of lactic acid bacteria fermented by lactic acid bacteria by mixing milk and pomegranate extract.
Description
본 발명은 석류 유산균 발효물을 이용한 건강식품 조성물과 이의 제조방법에 관한 것으로서, 더욱 상세하게는 석류를 유산균으로 발효시킨 유산균 발효물을 함유하여 항비만, 항염증 등의 효능이 우수한 건강식품 조성물과 이의 제조방법에 관한 것이다. The present invention relates to a health food composition using a pomegranate lactic acid bacteria fermentation product and a method for manufacturing the same, and more particularly, a health food composition having excellent efficacy such as anti-obesity and anti-inflammatory properties by containing a lactic acid bacterium fermentation product fermented with pomegranate lactic acid bacteria, and It relates to a method of manufacturing the same.
생활환경이 고속화됨에 따라 그에 따른 식생활이 고칼로리 위주로 인스턴트화되어 비만증, 고혈압을 비롯하여 각종 성인병에 대한 보고가 급증하고 있다. 핵가족화, 개인주의화, 야간활동의 인구증가, 대중소비시대, 소비패턴의 다양화, 편리성 추구경향, 그리고 여성의 사회진출 증가 등의 사회경제적인 환경의 변화로 식생활 구조의 변화를 겪고 있다.As the living environment increases, the resulting diet is instantaneous mainly of high calories, and reports of obesity, hypertension, and various adult diseases are increasing rapidly. The dietary life structure is undergoing changes due to changes in the socioeconomic environment such as nuclear familyization, individualization, population increase in nighttime activities, mass consumption era, diversification of consumption patterns, tendency to pursue convenience, and increase in women's advancement to society.
그리고 이러한 식사 구성에 대한 의식변화와 간편식에 대한 소비자의 요구 증가는 편의성을 추구하는 경향으로 나타나기 시작하였다. 그러나 이러한 현대인의 잘못된 식생활은 영양부족, 영양 과잉 및 잘못된 영양정보에 의한 영양불량을 초래하기 쉽다. 영양부족으로 인한 영양불량은 빈혈 등 여러 질병을 일으키게 되고 영양소의 과잉섭취는 체중 과다 및 비만을 유발하여 오늘날 보건문제로 대두하고 있을 뿐 아니라 동맥경화증, 당뇨병, 고혈압과 같은 만성 퇴행성 질환의 발병을 더욱 가중시키게 된다.In addition, the change of consciousness about the composition of meals and the increasing consumer's demand for convenience food began to appear as a tendency to pursue convenience. However, these modern people's wrong eating habits are liable to lead to malnutrition due to malnutrition, excess nutrition, and incorrect nutritional information. Malnutrition caused by malnutrition leads to various diseases such as anemia, and excessive intake of nutrients leads to overweight and obesity, which is becoming a health problem today, as well as chronic degenerative diseases such as arteriosclerosis, diabetes, and hypertension. Weighted.
특히, 비만으로 인한 동맥경화, 고혈압, 심장질환, 당뇨병, 고지혈증 등 성인병발병의 증가 및 이로 인해 생기는 각종 암, 악성종양, 심장병, 신장병 등의 발병과 수명단축 등의 심각한 상태에 이르고 있다.In particular, the increase in adult diseases such as arteriosclerosis, hypertension, heart disease, diabetes, and hyperlipidemia due to obesity, and various cancers, malignant tumors, heart disease, kidney disease, etc.
또한 식생활 습관이 질병과 매우 밀접한 관계가 있다는 과학적 증거가 속속 드러나면서 식생활을 포함한 생활양식의 변화를 통해 건강을 증진시키고자 하는 소비자들이 빠르게 늘어나고 있다. 이에 각종 질병을 예방하고 건강에 도움을 주기 위한 건강보조식품 및 기능성 식품에 대한 연구가 활발히 진행되고 있다. In addition, as scientific evidence that dietary habits are very closely related to diseases has emerged, consumers are rapidly increasing to improve their health through lifestyle changes including dietary habits. Accordingly, research on health supplementary foods and functional foods to prevent various diseases and help health is being actively conducted.
한편, 석류(pomegranate)는 석류과에 속하는 낙엽활엽 교목으로 아프가니스탄과 서부북부에 자생하던 식물이며, 고대 이집트와 그리스에서 종교의식과 예술, 신화에 사용되었다. On the other hand, pomegranate is a deciduous broad-leaved tree belonging to the pomegranate family, a plant native to Afghanistan and the western north, and was used in religious ceremonies, art, and mythology in ancient Egypt and Greece.
석류는 새콤달콤한 맛을 나타내며 살균, 살충 등의 효과가 있어 과거에는 한약재처럼 사용하거나 껍질이나 씨앗은 버리고 씨앗을 싸고 있는 과육만을 먹는 것으로 생각되어 왔으나, 최근 석류의 효능에 관한 연구 결과가 보고되면서 건강증진 음용수로써의 개발에 많은 관심을 기울이고 있다.Pomegranate has a sweet and sour taste and is effective in sterilization and insecticide. In the past, it was thought to be used like herbal medicine or to discard the skin or seeds and eat only the flesh wrapped in the seeds. However, recent research results on the efficacy of pomegranate have been reported. A lot of attention is paid to the development of it as an enhanced drinking water.
석류의 효능을 살펴보면 열매와 껍질 속에는 고혈압이나 동맥경화를 예방할 수 있는 효과가 있고, 또한 인터넷 등에서도 에스트로겐 함유로 갱년기 장애, 생리 부조, 비만, 각종의 부인병이나 부스럼 등에도 효과가 있으며, 빈혈이나 병후 체력 회복, 건강한 사람들의 영양 보조, 체력 증강 등에 이용되는 과실로 설명하고 있다.Looking at the efficacy of pomegranate, it has the effect of preventing hypertension and arteriosclerosis in the fruit and skin. Also, it is effective in menopausal disorders, menstrual relief, obesity, various women's diseases and swelling, etc. by containing estrogen in the fruit and skin. It is described as a fruit used for physical fitness recovery, nutritional support for healthy people, and physical strength enhancement.
그러나 석류는 천연식물성 생약 성분을 갖고 있기 때문에 열이나 산 등에 쉽게 변질될 우려가 있어 천연 상태를 유지하면서 섭취하기 좋도록 만드는 것이 중요한데, 지금까지의 국내 연구는 초보 단계를 벗어나지 못한 상태이다. However, since pomegranate contains natural plant-based herbal ingredients, it is important to make it easy to consume while maintaining its natural state, as it is likely to be easily deteriorated by heat or acid, and domestic research so far has not escaped the introductory stage.
한편, 유산균 발효는 서양에서는 발효유와 소시지 중심의 낙농제품 제조에 이용되어왔고, 우리나라에서는 삼국시대 초기부터 김치, 장류, 젓갈을 비롯한 농수산식품의 제조에 많이 이용되어 왔다.On the other hand, lactic acid bacteria fermentation has been used in the manufacture of dairy products centered on fermented milk and sausages in the West, and in Korea, it has been widely used in the manufacture of agricultural and fishery foods including kimchi, paste, and salted fish from the beginning of the Three Kingdoms period.
유산균(Lactic acid bacteria)은 발효식품들의 관능특성에 영향을 줄 수 있는 아로마, 향미료, 효소등과 같은 물질들을 생산할 뿐만 아니라 몇몇 유산균에는 발효식품들의 물성, 조직감, 점성제, 당화, 안정성, 수분결합제, 젤화, 농축특성등에 영향을 줄 수 있는 엑소폴리사카라이드(EPS)를 생산하는 것으로 보고되어 있다.Lactic acid bacteria not only produce substances such as aromas, flavors, and enzymes that can affect the organoleptic properties of fermented foods, but also some lactic acid bacteria include the physical properties, texture, viscosity, saccharification, stability, and water binding agents of fermented foods. , It has been reported to produce exopolysaccharides (EPS) that can affect gelation and concentration properties.
엑소폴리사키라이드는 산업적인 유용한 측면의 물리적인 특성뿐만 아니라 인체에 유용한 생리적 특성인 면역증진, 항궤양 활성, 혈중 콜레스테롤 감소 등의 유용한 효과를 나타낸다고 보고되어 왔으며, 일반적으로 안정한 물질로 인식되어 왔다. Exopolysakiride has been reported to exhibit useful effects such as immunity enhancement, anti-ulcer activity, and blood cholesterol reduction, which are useful physiological properties for the human body, as well as the physical properties of industrial useful aspects, and has been generally recognized as a stable substance.
1990년대 후반부터 프로바이오틱스 및 유산균의 기능성을 강조한 제품들이 홍수처럼 출하되고 있으나 기능성을 과장한 제품들이 다수를 이루어 소비자 신뢰도 하락의 원인이 되었다. 최근 천연소재에 프로바이오틱스를 접목한 신바이오틱스의 기능성 및 효능에 관한 연구가 이루어짐에 따라 이에 대한 소비자들의 관심도 증가 추세에 있다. Since the late 1990s, products emphasizing the functionality of probiotics and lactic acid bacteria have been shipped like a flood, but many products with exaggerated functionality have led to a decline in consumer confidence. Recently, as studies on the functionality and efficacy of new biotics that combine probiotics with natural materials have been conducted, the interest of consumers is also increasing.
국내에서 프로바이오틱스는 인체 의약용인 정장제나 유산균 제제, 사료 첨가제로서의 생균제, 식품공전에서 정의한 건강식품의 일종인 유산균 식품 등 세 가지 의미로 사용하고 있다. In Korea, probiotics are used in three meanings: intestinal agents or lactic acid bacteria preparations for human medicine, probiotics as feed additives, and lactic acid bacteria foods, a kind of health food defined in the Food Code.
종래의 유산균 제품은 대부분 발효유 제품 형태로서 다양한 소재 이용을 모색해야하는 실정으로, 시너지 효과를 기대할 수 있는 기타 여러 우수 식품소재와의 결합을 통한 양질의 제품 개발이 필요한 실정이다. Most of the conventional lactic acid bacteria products are in the form of fermented milk products, and it is necessary to seek the use of various materials, and it is necessary to develop high-quality products through combination with other excellent food materials that can expect a synergistic effect.
본 발명은 상술한 문제점을 개선하고자 창출된 것으로서, 건강에 유용한 석류를 유산균으로 발효시켜 만든 새로운 프로바이오틱스를 다양한 제형의 식품으로 제공할 수 있도록 함으로써 항비만, 항염증 등의 효능이 우수하여 건강에 유용한 건강식품 조성물과 이의 제조방법을 제공하는 그 목적이 있다. The present invention was created to improve the above-described problems, and it is useful for health by providing new probiotics made by fermenting pomegranate useful for health with lactic acid bacteria as foods of various formulations, thereby having excellent anti-obesity and anti-inflammatory effects. It is an object of providing a health food composition and a method of manufacturing the same.
상기의 목적을 달성하기 위한 본 발명의 석류 유산균 발효물을 이용한 건강식품 조성물은 우유와 석류추출물을 혼합하여 유산균에 의해 발효시킨 유산균 발효물을 함유한다.The health food composition using the fermented pomegranate lactic acid bacteria of the present invention for achieving the above object contains a fermented product of lactic acid bacteria fermented by lactic acid bacteria by mixing milk and pomegranate extract.
상기 유산균은 락토바실러스 속 균주이다.The lactic acid bacteria is a strain of the genus Lactobacillus.
상기 균주는 락토바실러스 아시도필루스(Lactobacillus acidophilus), 락토바실러스 플란타룸(Lactobacillus plantarum), 락토바실러스 페르멘텀(Lactobaciilus fermentum) 중에서 선택된 어느 하나이다. The strain is any one selected from Lactobacillus acidophilus , Lactobacillus plantarum , and Lactobaciilus fermentum .
또한, 상기 건강식품 조성물은 음료, 과립, 정제, 파우더, 환, 캡슐 중에서 선택된 어느 하나의 제형으로 형성된다. In addition, the health food composition is formed in any one formulation selected from beverages, granules, tablets, powders, pills, and capsules.
또한, 상기 유산균 발효물은 상기 석류 추출물과 함께 가르시니아 캄보지아(Garcinia cambogia) 추출물을 상기 유산균에 의해 발효시킨다.In addition, the lactic acid bacteria fermented product is fermented by the lactic acid bacteria Garcinia cambogia ( Garcinia cambogia ) extract together with the pomegranate extract.
그리고 상기의 목적을 달성하기 위한 본 발명의 석류 유산균 발효물을 이용한 건강식품 조성물의 제조방법은 우유에 석류추출물을 혼합하는 혼합단계와; 상기 혼합단계에서 수득한 혼합물을 멸균시키는 멸균단계와; 상기 멸균단계 후 상기 혼합물에 유산균을 접종하여 배양시켜 유산균 발효물을 수득하는 배양단계;를 포함한다. And a method for preparing a health food composition using the fermented pomegranate lactic acid bacteria of the present invention for achieving the above object comprises: a mixing step of mixing a pomegranate extract with milk; A sterilization step of sterilizing the mixture obtained in the mixing step; After the sterilization step, a culture step of inoculating and culturing the mixture to obtain a fermented product of lactic acid bacteria; includes.
또한, 상기 혼합단계는 상기 우유에 가르시니아 캄보지아(Garcinia cambogia) 추출물을 더 첨가한다. In addition, in the mixing step, Garcinia cambogia extract is further added to the milk.
상술한 바와 같이 본 발명은 건강에 유용한 석류를 유산균으로 발효시켜 만든 새로운 프로바이오틱스를 다양한 제형의 식품으로 제공할 수 있도록 함으로써 항비만, 항염증 등의 효능이 우수한 건강식품 조성물과 이의 제조방법을 제공할 수 있다. As described above, the present invention provides a health food composition excellent in efficacy such as anti-obesity and anti-inflammatory effects and a method of manufacturing the same by allowing new probiotics made by fermenting pomegranate useful for health with lactic acid bacteria to be provided as foods of various formulations. I can.
또한, 기존의 유산균발효유 제품은 액상형태로 이루어진 것들이 대다수라 유통기간 등 보존성에 있어 상대적인 약점을 가지고 있으나 본 발명은 유산균 발효유의 맛은 유지하면서 유통기간을 늘리고, 석류와 같은 천연재료를 유산 발효시켜 다양한 제형으로 가공함으로써 종래의 제품과 차별화된 프로바이오틱스 제품을 제공할 수 있다. In addition, since most of the existing lactic acid bacteria fermented milk products are made in liquid form, they have relatively weak points in preservation such as shelf life, but the present invention extends the shelf life while maintaining the taste of the lactic acid bacteria fermented milk, and lactic acid fermentation of natural ingredients such as pomegranate By processing into various formulations, it is possible to provide probiotic products differentiated from conventional products.
도 1은 실시예 1의 시료를 동결건조시킨 후 수율을 측정한 결과를 나타낸 표이고,
도 2는 실시예 1의 시료의 유리당 분석결과를 나타낸 표이고,
도 3은 실시예 1의 시료에 대한 관능검사 결과를 나타낸 표이고,
도 4는 석류추출물의 농축 정도에 따라 기호도를 평가하기 위해 실시한 관능검사 결과를 나타낸 표이고,
도 5는 실시예 2의 시료에 대한 세포독성을 측정한 결과를 나타낸 그래프이고,
도 6은 실시예 2의 시료에 대한 항염증을 측정한 결과를 나타낸 그래프이고,
도 7은 실시예 2의 시료에 대한 항비만을 측정한 결과를 나타낸 그래프이다. 1 is a table showing the results of measuring the yield after lyophilizing the sample of Example 1,
2 is a table showing the analysis results per glass of the sample of Example 1,
3 is a table showing the sensory test results for the sample of Example 1,
4 is a table showing the results of a sensory test conducted to evaluate the acceptability according to the concentration of pomegranate extract,
5 is a graph showing the results of measuring cytotoxicity to the sample of Example 2,
6 is a graph showing the results of measuring anti-inflammatory for the sample of Example 2,
7 is a graph showing the results of measuring anti-obesity for the sample of Example 2.
이하, 본 발명의 바람직한 실시 예에 따른 석류 유산균 발효물을 이용한 건강식품 조성물과 이의 제조방법에 대하여 구체적으로 설명한다. Hereinafter, a health food composition using a fermented pomegranate lactic acid bacteria according to a preferred embodiment of the present invention and a method of manufacturing the same will be described in detail.
본 발명의 건강식품 조성물은 유산균 발효물을 함유한다. 가령, 본 발명의 건강식품 조성물은 유산균 발효물을 1 내지 99중량%로 함유할 수 있다. 또한, 본 발명의 건강식품 조성물은 유산균 발효물 100중량%로 함유할 수 있음은 물론이다. The health food composition of the present invention contains fermented lactic acid bacteria. For example, the health food composition of the present invention may contain 1 to 99% by weight of fermented lactic acid bacteria. In addition, it goes without saying that the health food composition of the present invention may contain 100% by weight of fermented lactic acid bacteria.
유산균 발효물은 우유와 석류추출물을 혼합한 혼합물을 유산균에 의해 발효시킨 것이다. 유산균 발효물은 액상 또는 분말일 수 있다. Lactic acid fermented product is a mixture of milk and pomegranate extract fermented by lactic acid bacteria. The lactic acid fermentation product may be liquid or powder.
우유로 시중에서 판매되는 통상적인 제품을 이용하거나, 원유를 이용할 수 있다. As milk, conventional products sold on the market may be used, or crude milk may be used.
석류추출물은 석류로부터 추출된다. 석류는 열매를 통째로 이용하거나, 껍질을 제외한 나머지 열매 부위를 이용할 수 있다. Pomegranate extract is extracted from pomegranate. For pomegranate, the whole fruit can be used, or the rest of the fruit can be used excluding the skin.
석류추출물을 추출하기 위해 다양한 추출방법이 적용 가능하다. 가령 석류에 추출용매를 가해 추출할 수 있다. 이때 추출용매로 물, 탄소수 1 내지 4의 저급 알코올, 다가 알코올 또는 이들의 혼합물로부터 선택된 적어도 어느 하나를 이용할 수 있다. Various extraction methods can be applied to extract pomegranate extract. For example, pomegranate can be extracted by adding an extraction solvent. At this time, at least one selected from water, a lower alcohol having 1 to 4 carbon atoms, a polyhydric alcohol, or a mixture thereof may be used as the extraction solvent.
탄소수 1 내지 4의 저급 알코올로 메탄올, 에탄올 등을 이용할 수 있고, 다가 알코올로 부틸렌글리콜 및 프로필렌글리콜, 펜틸렌글리콜 등을 이용할 수 있다. 그리고 혼합물로는 물 및 저급 알코올의 혼합물, 물 및 다가 알코올의 혼합물, 저급 알코올 및 다가 알코올의 혼합물, 또는 물 및 저급알코올 및 다가 알코올의 혼합물을 이용할 수 있다.Methanol, ethanol, etc. can be used as the lower alcohol having 1 to 4 carbon atoms, and butylene glycol, propylene glycol, pentylene glycol, and the like can be used as the polyhydric alcohol. And as the mixture, a mixture of water and a lower alcohol, a mixture of water and a polyhydric alcohol, a mixture of a lower alcohol and a polyhydric alcohol, or a mixture of water and a lower alcohol and a polyhydric alcohol may be used.
추출의 일 예로 추출용매를 석류에 대하여 중량비로 2 내지 20배를 가한 후 20 내지 120℃에서 1 내지 48시간 동안 추출한 후 여과하여 액상의 석류추출물을 얻을 수 있다. 또한, 여과 후 농축하여 농축액의 형태로 석류추출물을 얻을 수 있음은 물론이다. 또한, 농축액을 동결건조 또는 분무건조 방식 등을 통해 분말형태로 얻을 수 있음은 물론이다.As an example of extraction, a liquid pomegranate extract may be obtained by adding an extraction solvent 2 to 20 times by weight to pomegranate, and then extracting at 20 to 120°C for 1 to 48 hours and then filtering. In addition, it is of course possible to obtain a pomegranate extract in the form of a concentrated solution by concentrating after filtration. In addition, of course, the concentrate can be obtained in powder form through freeze drying or spray drying.
유산균으로 락토바실러스 속 균주를 이용한다. 이러한 균주로 락토바실러스 아시도필루스(Lactobacillus acidophilus), 락토바실러스 플란타룸(Lactobacillus plantarum), 락토바실러스 페르멘텀(Lactobaciilus fermentum) 중에서 선택된 어느 하나를 이용할 수 있다. Lactobacillus strains are used as lactic acid bacteria. These strains include Lactobacillus acidophilus , Lactobacillus plantarum , Lactobacillus fermentum ( Lactobaciilus). fermentum ) can be used.
위와 같이 석류 추출물을 유산균으로 발효시킨 유산균 발효물은 석류를 이용한 새로운 프로바이오틱스 제품을 제공할 수 있다. 이는 종래의 프로바이오틱스에 비해 기능성이 우수하여 차별화된 프로바이오틱스 제품을 제공한다. Lactic acid fermented product obtained by fermenting pomegranate extract with lactic acid bacteria as described above can provide a new probiotic product using pomegranate. This provides a differentiated probiotic product with superior functionality compared to conventional probiotics.
석류 추출물을 유산균으로 발효시킨 유산균 발효물은 항비만, 항염증 등의 효능이 우수하여 건강에 매우 유용한다. Lactic acid fermented products obtained by fermenting pomegranate extract with lactic acid bacteria are very useful for health as they have excellent anti-obesity and anti-inflammatory effects.
본 발명의 건강식품 조성물은 유산균 발효물 외에 공지의 식품첨가물이 첨가될 수 있다.In addition to fermented lactic acid bacteria, known food additives may be added to the health food composition of the present invention.
공지의 식품첨가물의 예로 단당류, 이당류, 다당류, 당알콜 등의 당류와, 타우마틴, 스테비아 추출물, 사카린, 아스파르탐 등의 향미제와, 영양제, 비타민, 식용 전해질, 풍미제, 착색제, 증진제(예, 치즈, 초콜릿 등), 펙트산, 알긴산, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알콜, 탄산화제, 정제수 등이 이용될 수 있다.Examples of known food additives include sugars such as monosaccharides, disaccharides, polysaccharides, and sugar alcohols, flavoring agents such as taumatin, stevia extract, saccharin, and aspartame, nutrients, vitamins, edible electrolytes, flavoring agents, coloring agents, enhancers (e.g. , Cheese, chocolate, etc.), pectic acid, alginic acid, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents, purified water, and the like.
'식품 조성물'이란, 영양소를 한 가지 또는 그 이상 함유하고 있는 천연물 또는 가공품을 의미하며, 바람직하게는 어느 정도의 가공 공정을 거쳐 직접 먹을 수 있는 상태가 된 것을 의미하며, 통상적인 의미로서, 식품, 식품첨가제, 기능성 식품 및 음료를 모두 포함하는 것을 말한다.The term'food composition' refers to a natural product or processed product containing one or more nutrients, and preferably refers to a state that can be eaten directly through a certain amount of processing process, and as a general meaning, food , Food additives, functional foods and beverages.
그리고 '건강식품 조성물'이란 식품에 물리적, 생화학적, 생물공학적 수법 등을 이용하여 해당 식품의 기능을 특정 목적에 작용, 발현하도록 부가가치를 부여한 식품군이나 식품 조성이 갖는 생체방어리듬조절, 질병방지와 회복 등에 관한 체내조절기능을 생체에 대하여 충분히 발현하도록 설계하여 가공한 식품을 의미한다.In addition,'health food composition' refers to a food group or food composition that has added value to act and express the function of the food for a specific purpose by using physical, biochemical, and biotechnological techniques. It refers to food that is designed and processed to sufficiently express the body's regulating function for recovery, etc.
본 발명에 따른 건강식품 조성물로는 예를 들어, 각종 식품류, 음료, 껌, 차, 비타민 복합제, 기능성 식품 등이 있다. 나아가 본 발명은 여러 가지 영양제, 비타민, 광물(전해질), 합성 풍미제 및 천연 풍미제 등의 풍미제, 착색제 및 충진제(치즈, 초콜릿 등), 펙트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알코올, 탄산음료에 사용되는 탄산화제 등을 함유할 수 있으며, 상기 성분들은 독립적으로 또는 조합하여 사용할 수 있다.The health food composition according to the present invention includes, for example, various foods, beverages, gum, tea, vitamin complexes, and functional foods. Furthermore, the present invention provides various nutrients, vitamins, minerals (electrolytes), synthetic flavors and flavoring agents such as natural flavors, colorants and fillers (cheese, chocolate, etc.), pectic acid and salts thereof, alginic acid and salts thereof, organic acids, It may contain a protective colloid thickener, a pH adjuster, a stabilizer, a preservative, a glycerin, an alcohol, a carbonating agent used in carbonated beverages, and the like, and the components may be used independently or in combination.
또한, 건강식품 조성물에는 식품학적으로 허용 가능한 식품 보조 첨가제를 포함할수 있으며, 기능성 식품의 제조에 통상적으로 사용되는 적절한 담체, 부형제 및 희석제를 더욱 포함할 수 있다.In addition, the health food composition may include food additives acceptable for food, and may further include suitable carriers, excipients, and diluents commonly used in the manufacture of functional foods.
본 발명의 건강식품 조성물은 다양한 형태로 제형이 가능하므로 그 형태는 특별히 제한되지 않는다. 가령, 건강식품 조성물은 음료, 과립, 정제, 파우더, 환, 캡슐 중에서 선택된 어느 하나의 제형으로 형성될 수 있다. 이러한 음료, 과립, 정제, 파우더, 환, 캡슐 제형을 갖는 건강식품 조성물은 휴대가 간편하고 언제 어디서나 수시로 섭취하기가 용이하다.Since the health food composition of the present invention can be formulated in various forms, the form is not particularly limited. For example, the health food composition may be formed in any one formulation selected from beverages, granules, tablets, powders, pills, and capsules. Health food compositions having such beverages, granules, tablets, powders, pills, and capsules are easy to carry and easy to take anytime, anywhere.
한편, 본 발명의 건강식품 조성물은 다른 예로 석류추출물과 함께 가르시니아 캄보지아 추출물을 유산균에 의해 발효시킨 유산균 발효물을 함유할 수 있다. On the other hand, the health food composition of the present invention may contain a fermented product obtained by fermenting garcinia cambogia extract with lactic acid bacteria together with pomegranate extract as another example.
가르시니아 캄보지아(Garcinia cambogia)는 클루시아과(Clusiaceae)의 식물로서 열매의 껍질 부위를 사용하며, 껍질에는 탄수화물이 지방으로 저장되는 것을 억제하는 기능이 있는 하이드록시시트르산(Hydroxycitric acid, HCA)이 풍부하게 함유되어 있는 것으로 알려져 있다. Garcinia cambogia is a plant of the Clusiaceae family, which uses the skin part of the fruit, and the skin contains abundant hydroxycitric acid (HCA), which has the function of inhibiting the storage of carbohydrates as fat. It is known to be.
가르시니아 캄보지아 추출물은 탄수화물이 지방으로 합성되는 것을 억제함으로써 석류 추출물과 함께 시너지 효과를 가져 항염증 및 항비만 기능성을 더욱 향상시킬 수 있다. Garcinia cambogia extract can further improve anti-inflammatory and anti-obesity functions by having a synergistic effect with pomegranate extract by inhibiting the synthesis of carbohydrates into fat.
가르시니아 캄보지아 추출물은 가르시니아 캄보지아 열매의 껍질로부터 추출한다. 추출방법은 상술한 석류추출물의 추출방법과 동일하므로 구체적인 설명은 생략한다. Garcinia cambogia extract is extracted from the skin of the fruit of Garcinia cambogia. Since the extraction method is the same as the extraction method of the pomegranate extract described above, a detailed description will be omitted.
이하, 상술한 본 발명의 기능성 식품의 제조방법에 대하여 살펴본다.Hereinafter, a method for manufacturing the functional food of the present invention described above will be described.
본 발명의 기능성 식품의 제조방법은 크게 우유에 석류추출물을 혼합하는 혼합단계, 혼합단계에서 수득한 혼합물을 멸균시키는 멸균단계와, 멸균단계 후 혼합물에 유산균을 접종하여 배양시켜 유산균 발효물을 수득하는 배양단계를 포함한다. 각 단계별로 상세하게 설명한다. The manufacturing method of the functional food of the present invention largely includes a mixing step of mixing pomegranate extract with milk, a sterilization step of sterilizing the mixture obtained in the mixing step, and inoculation of lactic acid bacteria into the mixture after the sterilization step and cultivation to obtain a fermented product of lactic acid bacteria. It includes a culture step. It will be described in detail for each step.
1. 혼합단계1. Mixing step
먼저, 석류추출물을 준비한다. First, prepare pomegranate extract.
석류추출물은 상술한 바와 같이 석류에 추출용매를 가해 추출할 수 있다. 추출용매로 물, 탄소수 1 내지 4의 저급 알코올, 다가 알코올 또는 이들의 혼합물로부터 선택된 적어도 어느 하나를 이용할 수 있다. Pomegranate extract can be extracted by adding an extraction solvent to pomegranate as described above. As the extraction solvent, at least one selected from water, a lower alcohol having 1 to 4 carbon atoms, a polyhydric alcohol, or a mixture thereof may be used.
추출의 일 예로 추출용매를 석류 열매에 대하여 중량비로 2 내지 20배를 가한 후 20 내지 120℃에서 1 내지 48시간 동안 추출한 후 여과하여 액상의 석류추출물을 얻을 수 있다. 또한, 여과 후 농축하여 농축액의 형태로 석류추출물을 얻을 수 있음은 물론이다. As an example of extraction, a liquid pomegranate extract may be obtained by adding an extraction solvent 2 to 20 times by weight to the pomegranate fruit and then extracting at 20 to 120°C for 1 to 48 hours and then filtering. In addition, it is of course possible to obtain a pomegranate extract in the form of a concentrated solution by concentrating after filtration.
준비된 석류추출물은 우유와 혼합하여 혼합물을 얻는다. The prepared pomegranate extract is mixed with milk to obtain a mixture.
혼합물의 일 예로 우유 100중량부에 대하여 석류추출물 10 내지 100중량부를 혼합하여 얻을 수 있다. An example of a mixture may be obtained by mixing 10 to 100 parts by weight of a pomegranate extract with respect to 100 parts by weight of milk.
또한, 혼합물의 다른 예로 우유에 석류추출물, 가르시니아 캄보지아 추출물을 혼합하여 얻을 수 있다. 가령, 우유 100중량부에 대하여 석류추출물 10 내지 100중량부와, 가르시니아 캄보지아 추출물 10 내지 100중량부를 혼합하여 얻을 수 있다. 가르시니아 캄보지아 추출물은 가르시니아 캄보지아 열매의 껍질로부터 추출하고, 추출방법은 상술한 석류추출물의 추출방법과 동일하다. In addition, as another example of the mixture, it can be obtained by mixing pomegranate extract and garcinia cambogia extract with milk. For example, it can be obtained by mixing 10 to 100 parts by weight of pomegranate extract and 10 to 100 parts by weight of garcinia cambogia extract based on 100 parts by weight of milk. The garcinia cambogia extract is extracted from the peel of the garcinia cambogia fruit, and the extraction method is the same as that of the pomegranate extract described above.
또한, 혼합물의 다른 예로 당을 더 첨가할 수 있다. 가령, 우유 100중량부에 대하여 석류추출물 10 내지 100중량부와, 당 0.1 내지 5중량부를 혼합하여 얻을 수 있다. 또한, 우유 100중량부에 대하여 석류추출물 10 내지 100중량부와, 가르시니아 캄보지아 추출물 10 내지 100중량부, 당 0.1 내지 5중량부를 혼합하여 얻을 수 있다.In addition, as another example of the mixture, sugar may be further added. For example, it can be obtained by mixing 10 to 100 parts by weight of pomegranate extract and 0.1 to 5 parts by weight per 100 parts by weight of milk. In addition, it can be obtained by mixing 10 to 100 parts by weight of pomegranate extract, 10 to 100 parts by weight of garcinia cambogia extract, and 0.1 to 5 parts by weight per 100 parts by weight of milk.
당으로 글루코스(glucose)와 같은 단당류, 락토오스(lactose) 또는 수크로스(sucrose)와 같은 이당류 등을 이용할 수 있다. As sugar, monosaccharides such as glucose, disaccharides such as lactose or sucrose, and the like may be used.
2. 멸균단계2. Sterilization step
다음으로, 혼합단계에서 수득한 혼합물을 멸균시킨다.Next, the mixture obtained in the mixing step is sterilized.
이를 위해 혼합물을 오토클레이브(autoclave)에 투입하여 121℃, 1.2기압에서 10 내지 20분간 멸균처리한다. 멸균된 혼합물은 무균상 내에서 방랭시킨다.To this end, the mixture is put into an autoclave and sterilized for 10 to 20 minutes at 121°C and 1.2 atm. The sterilized mixture is allowed to cool in a sterile bed.
3. 배양단계3. Culture step
다음으로, 멸균돤 혼합물에 유산균을 접종하여 배양시켜 유산균 발효물을 수득한다. Next, lactic acid bacteria are inoculated and cultured in the sterilized mixture to obtain a fermented product of lactic acid bacteria.
유산균으로 락토바실러스 아시도필루스(Lactobacillus acidophilus), 락토바실러스 플란타룸(Lactobacillus plantarum), 락토바실러스 페르멘텀(Lactobaciilus fermentum) 중에서 선택된 어느 하나를 이용한다. Lactobacillus acidophilus as lactic acid bacteria ( Lactobacillus acidophilus ), Lactobacillus plantarum ( Lactobacillus plantarum ), Lactobacillus fermentum ( Lactobaciilus fermentum ) is used any one selected from.
유산균을 MRS 배지에 접종하여 37℃의 배양기에서 24시간 액체 배양하여 유산균 배양액을 얻는다. 그리고 멸균된 혼합물 100중량부에 대하여 유산균 배양액 0.5 내지 5중량부를 접종한 후 저온배양기를 이용해 37℃의 배양기에서 96시간 배양함으로써 최종적으로 유산균 발효물을 얻을 수 있다. Lactobacillus is inoculated in MRS medium and liquid cultured in an incubator at 37° C. for 24 hours to obtain a lactic acid bacteria culture solution. And after inoculating 0.5 to 5 parts by weight of the lactic acid bacteria culture solution based on 100 parts by weight of the sterilized mixture By incubating for 96 hours in an incubator at 37°C using a low-temperature incubator, a fermented product of lactic acid bacteria can be finally obtained.
이와 같이 수득한 유산균 발효물은 액상 그 자체로 본 발명의 건강식품 조성물이 될 수 있다. 또한, 유산균 발효물을 동결건조하여 분말 형태로 만들 수 있다. The fermented product of lactic acid bacteria thus obtained may be a health food composition of the present invention as a liquid. In addition, the fermented product of lactic acid bacteria can be freeze-dried to form a powder.
또한, 유산균 발효물에 식품학적으로 허용 가능한 식품 보조 첨가제를 첨가하거나, 적절한 담체, 부형제 및 희석제 등을 첨가하여 제형화시킬 수 있다. 이러한 제형으로 음료, 과립, 정제, 파우더, 환, 캡슐 중에서 선택된 어느 하나를 들 수 있다. In addition, the lactic acid bacteria fermented product may be formulated by adding food additives acceptable for food, or by adding an appropriate carrier, excipient, and diluent. Such formulations may include any one selected from beverages, granules, tablets, powders, pills, and capsules.
이하, 하기의 실시예 및 실험을 통하여 본 발명에 대해 설명하고자 한다. 다만, 하기의 실시 예는 본 발명을 구체적으로 설명하기 위한 것으로, 본 발명의 범위를 하기의 실시 예로 한정하는 것은 아니다.Hereinafter, the present invention will be described through the following examples and experiments. However, the following examples are intended to specifically illustrate the present invention, and are not intended to limit the scope of the present invention to the following examples.
(실시예 1)(Example 1)
석류 열매에 중량비로 10배의 물을 가하여 80℃에서 5시간씩 2회 반복하여 추출한 후 10brix로 농축하여 석류 추출물을 얻었다. Pomegranate fruit was extracted with 10 times the weight of water at 80° C. for 5 hours each, and then concentrated to 10 brix to obtain a pomegranate extract.
그리고 우유에 석류추출물과 글루코스를 첨가하여 수득한 혼합물을 오토클레이브를 이용하여 121℃, 1.2기압에서 15분간 멸균처리한 후 무균상 내에서 방랭시켰다. Then, the mixture obtained by adding pomegranate extract and glucose to milk was sterilized for 15 minutes at 121°C and 1.2 atm using an autoclave, and then left to cool in a sterile bed.
그리고 멸균된 혼합물 490ml에 유산균 배양액을 10ml를 접종한 후 저온배양기를 이용해 37℃의 배양기에서 96시간 배양하여 유산균 발효물을 제조하였다. And after inoculating 10ml of lactic acid bacteria culture solution to 490ml of sterilized mixture The lactic acid bacteria fermented product was prepared by incubating for 96 hours in an incubator at 37°C using a low-temperature incubator.
상기 유산균 배양액은 락토바실러스 아시도필루스(Lactobacillus acidophilus) KCCM 41619, 락토바실러스 플란타룸(Lactobacillus plantarum) KCCM 12116, 락토바실러스 페르멘텀(Lactobaciilus fermentum) KCCM 35469 각각을 MRS 배지에 접종하여 37℃의 배양기에서 24시간 액체 배양하여 얻었다. The lactic acid bacteria culture medium is Lactobacillus acidophilus ( Lactobacillus acidophilus ) KCCM 41619, Lactobacillus plantarum ( Lactobacillus plantarum ) KCCM 12116, Lactobacillus fermentum ( Lactobaciilus fermentum ) KCCM 35469 was obtained by inoculating each of the MRS medium and performing liquid culture for 24 hours in an incubator at 37°C.
혼합물의 조성과 발효시 사용된 균주에 따라 하기 표 1에 나타난 것처럼 유산균 발효물은 총 9종류로 제조하였다. As shown in Table 1 below, according to the composition of the mixture and the strain used during fermentation, a total of 9 types of fermented lactic acid bacteria were prepared.
<유산균 발효물의 수율><Yield of fermented lactic acid bacteria>
실시예 1의 시료를 동결건조시킨 후 수율을 측정한 결과를 도 1에 나타내었다. The results of measuring the yield after lyophilizing the sample of Example 1 are shown in FIG. 1.
도 1을 참조하면, 석류추출물을 30% 첨가해 L. fermentum으로 배양한 유산균 발효물이 수율 13.9%로 가장 높게 나타났으며, 나머지 시료도 이와 큰 차이를 나타내지 않았다. 따라서 석류추출물의 첨가량이 유산균 발효물의 수율에는 큰 영향을 미치지 않는 것으로 보인다. Referring to Figure 1, the lactic acid bacteria fermented product cultured with L. fermentum by adding 30% pomegranate extract showed the highest yield at 13.9%, and the remaining samples also did not show a significant difference. Therefore, the amount of pomegranate extract added does not seem to have a significant effect on the yield of fermented lactic acid bacteria.
<유산균 발효물의 유리당 분석><Analysis of free sugar of fermented lactic acid bacteria>
유리당 분석은 Wilson 등의 방법에 따라 분석하였다. 실시예 1의 시료를 원심분리(3,000rpm, 30min)하여 상징액을 취한 후 여과지(Whatman No.2)로 여과하고, Sepak C18으로 정제시킨 다음 0.45㎛ membrane filter(Millipore Co., U.S.A)로 여과한 여액을 HPLC(High Performance Liquid Chromatography)를 이용하여 분석하였다. 유리당 분석결과를 도 2에 나타내었다. Free sugar analysis was analyzed according to the method of Wilson et al. The sample of Example 1 was centrifuged (3,000 rpm, 30 min) to take a supernatant, filtered through a filter paper (Whatman No. 2), purified with Sepak C18, and then filtered through a 0.45 μm membrane filter (Millipore Co., USA). The filtrate was analyzed using High Performance Liquid Chromatography (HPLC). The results of the analysis of free sugar are shown in FIG. 2.
도 2를 참조하면, L. acidophilus 균주로 발효시킨 유산균 발효물이 유리당 총 함량이 가장 높은 것으로 나타났다. 따라서 L. acidophilus 균주를 이용하는 경우 가장 발효가 잘 될 것으로 보인다. Referring to FIG. 2, it was found that the lactic acid bacteria fermented product fermented with L. acidophilus strain had the highest total free sugar content. Therefore, it seems that fermentation will be the best if L. acidophilus strain is used.
<관능검사><Sensory test>
실시예 1의 시료를 대상으로 관능검사를 실시하였다. 관능검사는 훈련된 패널 20명을 패널(panel)로 구성하여 색, 맛, 향, 전체적인 기호도를 7단계 평가법으로 실시하였다. A sensory test was performed on the sample of Example 1. The sensory test consisted of 20 trained panelists in a panel, and color, taste, aroma, and overall acceptability were evaluated in a seven-step method.
채점 기준은 매우 좋음 7, 좋음 6, 약간 좋음 5, 보통 4, 약간 거부감 3, 거부감 2, 강한 거부감 1로 하였고, 2시간 간격으로 시료의 번호를 바꾸어 같은 panel로 3회 반복하며 각 반복 시 가장 높은 점수와 가장 낮은 점수를 제외하고 평균 득점을 구했다. The scoring criteria were very good 7, good 6, slightly good 5, average 4, slightly rejection 3, rejection 2, strong rejection 1, and change the number of samples every 2 hours and repeat 3 times with the same panel. The average score was calculated by excluding the high score and the lowest score.
검사결과를 SPSS 통계분석 프로그램(SPSS, ver. 16.0, USA)을 이용해 각 실험군간 평균치와 표준편차를 계산하고 통계적 유의성은 유의성 p<0.05 수준으로 얻어진 결과는 one-way ANOVA 중 Duncan’multiple range test로 검증하였다. The test results are calculated using the SPSS statistical analysis program (SPSS, ver. 16.0, USA), and the statistical significance is the significance p<0.05, and the results obtained are the Duncan' multiple range test among the one-way ANOVA. Verified as.
관능검사 결과를 도 3에 나타내었다. The sensory test results are shown in FIG. 3.
관능검사결과, 석류추출물 30% 첨가 L. fermenterim 시료가 향, 색, 맛, 종합기호도 항목에서 가장 낮은 기호도를 나타냈다. 사용한 유산균 종류와 관계없이 석류추출물 30% 첨가한 시료들이 석류 특유의 향과 맛에 의해 낮은 결과를 보였고, 석류추출물 10%, 20% 첨가한 시료들에서는 석류추출물을 첨가하지 않은 대조구에 비해 크게 낮지 않은 결과를 나타내었다. 개인별 선호도에 의한 차이는 보이지만, 석류의 함량과 기호도를 고려시 석류추출물 20% 첨가 L. acidophilus 유산균 발효물이 개발제품으로 적절할 것으로 판단된다. As a result of sensory evaluation, the L. fermenterim sample with 30% pomegranate extract showed the lowest degree of preference in terms of flavor, color, taste, and overall preference. Regardless of the type of lactic acid bacteria used, samples with 30% pomegranate extract showed lower results due to the peculiar aroma and taste of pomegranate, and samples with 10% and 20% pomegranate extract were not significantly lower than the control without pomegranate extract. Showed no results. Although there are differences according to individual preferences, considering the content and preference of pomegranate, L. acidophilus lactic acid fermented product with 20% pomegranate extract is considered to be appropriate as a developed product.
석류추출물의 농축 정도에 따라 기호도를 평가하기 위해 관능검사를 추가로 실시하였다. A sensory test was additionally performed to evaluate the acceptability according to the concentration of pomegranate extract.
우유에 농축정도(1, 3, 5, 10brix)를 달리한 석류추출물을 20%를 첨가한 혼합물에 L. acidophilus 배양액을 첨가하여 발효시킨 유산균 발효물를 시료로 하여 관능검사를 실시하였고, 그 결과를 도 4에 나타내었다. A sensory test was conducted using the lactic acid bacteria fermented product fermented by adding L. acidophilus culture solution to a mixture containing 20% of pomegranate extract with different concentration levels (1, 3, 5, 10 brix) to milk. It is shown in Figure 4.
도 4를 참조하면, 농축정도에 따른 기호도는 5brix > 3brix >1brix > 10brix 순으로 나타났다. 개인별 선호도에 의한 차이는 보이지만, 기호도에서 우수한 5brix이 석류추출물이 개발제품으로 적절할 것으로 판단된다. Referring to FIG. 4, the degree of preference according to the concentration was shown in the order of 5 brix> 3 brix> 1 brix> 10 brix. Although there are differences due to individual preferences, 5brix pomegranate extract, which is excellent in preference, is considered to be appropriate as a developed product.
(실시예 2)(Example 2)
석류 열매에 중량비로 10배의 물을 가하여 80℃에서 5시간씩 2회 반복하여 추출한 후 5brix로 농축하여 석류 추출물을 얻었다. 그리고 가르시니아 캄보지아 열매 껍질에 중량비로 10배의 물을 가하여 80℃에서 6시간 동안 추출한 후 2brix로 농축하여 가르시니아 캄보지아 추출물을 얻었다.Pomegranate fruit was extracted by repeating extraction twice at 80° C. for 5 hours by adding 10 times water by weight to the pomegranate fruit, and then concentrated to 5 brix to obtain a pomegranate extract. Then, 10 times the weight of water was added to the peel of the fruit of Garcinia cambogia, extracted at 80° C. for 6 hours, and concentrated to 2brix to obtain a garcinia cambogia extract.
그리고 우유에 석류 추출물, 가르시니아 캄보지아 추출물, 글루코스를 첨가한 혼합물을 오토클레이브를 이용하여 121℃, 1.2기압에서 15분간 멸균처리한 후 무균상 내에서 방랭시켰다. 그리고 멸균된 혼합물 490ml에 유산균 배양액을 10mL를 접종한 후 저온배양기를 이용해 37℃의 배양기에서 96시간 배양하여 유산균 발효물을 제조하였다. And the mixture of pomegranate extract, garcinia cambogia extract, and glucose added to milk was sterilized for 15 minutes at 121°C and 1.2 atm using an autoclave, and then allowed to cool in a sterile bed. And after inoculating 10 mL of lactic acid bacteria culture solution into 490 mL of sterilized mixture The lactic acid bacteria fermented product was prepared by incubating for 96 hours in an incubator at 37°C using a low-temperature incubator.
상기 유산균 배양액은 락토바실러스 아시도필루스(Lactobacillus acidophilus) KCCM 41619를 MRS 배지에 접종하여 37℃의 배양기에서 24시간 액체 배양하여 얻었다.The lactic acid bacteria culture solution was obtained by inoculating Lactobacillus acidophilus KCCM 41619 in MRS medium and culturing liquid in an incubator at 37° C. for 24 hours.
혼합물의 조성에 따라 하기 표 2에 나타난 것처럼 유산균 발효물은 총 5종류로 제조하였다. According to the composition of the mixture, as shown in Table 2 below, lactic acid fermentation products were prepared in a total of 5 types.
구분
division
실시예 2의 시료를 대상으로 세포독성 실험, 항염증 실험, 항비만 실험을 하였다. 그리고 비교예로서 실시예 2에 사용된 5brix 석류 추출물을 이용하였다. The sample of Example 2 was subjected to cytotoxicity, anti-inflammatory, and anti-obesity experiments. And as a comparative example, 5brix pomegranate extract used in Example 2 was used.
<세포독성 실험><Cytotoxicity test>
Raw 264.7을 96 well plate에 1 ×105 cell/mL의 농도로 배양하여 실험에 사용하였다. 시료를 각각 10, 50, 100, 500㎍/mL의 농도로 24시간 처리한 후, 배지를 제거하고 새로운 배지에 10uL MTS를 첨가하여 4시간 동안 반응시킨 후 540nm로 흡광도를 측정하였다. 각 시료의 세포 생존율은 시료 무처리군을 100%로 하여 상대적으로 계산하였다. Raw 264.7 was cultured in a 96 well plate at a concentration of 1 × 10 5 cells/mL and used in the experiment. Samples were treated at concentrations of 10, 50, 100, and 500 µg/mL, respectively, for 24 hours, the medium was removed, and 10 µL MTS was added to the new medium to react for 4 hours, and the absorbance was measured at 540 nm. The cell viability of each sample was calculated relative to the sample untreated group as 100%.
세포독성을 측정한 결과를 도 5에 나타내었다. The results of measuring cytotoxicity are shown in FIG. 5.
도 5를 참조하면, 석류추출물과 A, B, C, D, E의 세포생존율을 측정한 결과 가장 고농도인 500㎍/mL의 농도에서 각각 96.2, 95.1, 97.3, 96.2, 98.1, 97.0%의 세포생존율을 나타내 시료 모두에서 독성을 보이지 않았다.5, as a result of measuring the cell viability of pomegranate extract and A, B, C, D, E, 96.2, 95.1, 97.3, 96.2, 98.1, and 97.0% cells at the highest concentration of 500 μg/mL, respectively. It showed a survival rate and did not show toxicity in all of the samples.
<항염증 실험><Anti-inflammatory experiment>
Raw 264.7을 1 ×105 cell/mL 농도로 96-well plate에 분주한 후 각각의 시료를 농도별로 처리하여 24시간 배양하였다. 세포배양 상등액 100μL와 Griess 시약(A reagent: 1% sulfanilamide, B reagent: 0.1% naphthylethylendiamine in 25% phosphoric acid) 각 50μL를 혼합하여 96-well plate에서 10분간 반응시킨 후 microplate reader를 이용하여 540nm에서 흡광도를 측정하였다. NO2- 표준곡선은 NaNO2를 농도별로 조제하여 사용하였다.Raw 264.7 was dispensed into a 96-well plate at a concentration of 1 × 10 5 cells/mL, and each sample was treated by concentration and cultured for 24 hours. 100 μL of cell culture supernatant and 50 μL of Griess reagent (A reagent: 1% sulfanilamide, B reagent: 0.1% naphthylethylendiamine in 25% phosphoric acid) were mixed, reacted for 10 minutes in a 96-well plate, and absorbance at 540 nm using a microplate reader. Was measured. NO 2 -The standard curve was used by preparing NaNO 2 by concentration.
항염증 실험 결과를 도 6에 나타내었다. 6 shows the results of the anti-inflammatory experiment.
도 6을 참조하면, 석류추출물과 A, B, C, D, E의 NO 생성율을 측정한 결과, LPS로 염증을 유도한 처리구를 100%로 하였을 때 각각의 NO 생성율은 500㎍/mL의 농도에서 각각 98.1, 98.2, 96.2, 97.3, 91.2, 95.3%로 나타나 D의 시료에서 가장 높은 NO생성 억제율을 보였다.Referring to FIG. 6, as a result of measuring the NO production rate of pomegranate extract and A, B, C, D, E, when the treatment group inducing inflammation with LPS was 100%, each NO production rate was 500 µg/mL. 98.1, 98.2, 96.2, 97.3, 91.2, and 95.3% were respectively shown in D sample, showing the highest inhibition rate of NO production.
<항비만 실험><Anti-obesity experiment>
ㅇ 3T3-L1 cell의 지방세포 분화억제 확인ㅇ Confirmation of inhibition of adipocyte differentiation of 3T3-L1 cell
12 well plate에 3T3-L1 cell를 1 ×104 cell/mL로 seeding하여 2일 간격으로 배지를 교환하고 4일간 배양하였다. 지방세포로 분화를 일으키기 위해 differentiation medium(MDI: 0.5 mM IBMX, 1 mg/mL Insulin, 0.25 μM dexamethasone)을 처리하였다. 각 시료의 분화를 억제 효과를 확인하기 위해서 각각 10, 50, 100, 500μg/mL으로 동시 처리하였다. 48시간 배양을 한 후, insulin medium(1mg/mL Insulin)으로 교체하고 48시간을 더 배양을 하였다. 배양이 끝나면, Oil red O 염색하여 지방세포로 분화 억제하는 것을 확인하였다. 3T3-L1 cells were seeded in a 12 well plate at 1 × 10 4 cells/mL, the medium was exchanged every 2 days, and cultured for 4 days. In order to induce differentiation into adipocytes, a differentiation medium (MDI: 0.5 mM IBMX, 1 mg/mL Insulin, 0.25 μM dexamethasone) was treated. Each sample was simultaneously treated at 10, 50, 100, and 500 μg/mL in order to confirm the inhibitory effect on differentiation. After 48 hours of incubation, insulin medium (1mg/mL Insulin) was replaced and cultured for an additional 48 hours. After the culture was completed, it was confirmed that the differentiation into adipocytes was inhibited by staining with Oil red O.
ㅇ Oil red O stainingㅇ Oil red O staining
각 시료가 지방전구세포에 미치는 영향을 확인하기 위해 3T3-L1 cell line을 분화를 유도하여 Oil red O 염색하여 확인하였다. 분화된 3T3-L1 세포에 추출물을 처리하여 48시간 동안 배양한 후, PBS로 3회 세척한 뒤 4% paraform aldehyde로 4℃에서 30분간 고정하였다. 고정된 세포는 PBS로 3회 세척한 후 4℃에서 1시간 동안 0.5% Oil red O 염색시약(in 60% isopropanol and 40% water 녹은 후 0.45μm filter로 여과한 용액)으로 염색한 뒤 PBS로 3회 세척하였다. 염색된 지방은 DMSO에 용해한 후 500nm에서 흡광도를 측정하였다. 지방함량은 대조군의 흡광도 평균값에 대해 백분율로 계산하였다. In order to confirm the effect of each sample on adipocytes, the 3T3-L1 cell line was identified by inducing differentiation and staining with Oil red O. The differentiated 3T3-L1 cells were treated with the extract and cultured for 48 hours, washed three times with PBS, and then fixed at 4°C for 30 minutes with 4% paraform aldehyde. The fixed cells were washed 3 times with PBS, stained with 0.5%Oil red O staining reagent (solution dissolved in 60% isopropanol and 40% water and filtered with 0.45μm filter) for 1 hour at 4℃ and then 3 with PBS. Washed twice. The dyed fat was dissolved in DMSO and absorbance was measured at 500 nm. Fat content was calculated as a percentage of the average absorbance value of the control group.
항비만 실험 결과를 도 7에 나타내었다. Figure 7 shows the results of the anti-obesity experiment.
도 7을 참조하면, 석류추출물과 A, B, C, D, E의 lipid accumulation을 측정한 결과, 지방세포로의 분화를 유도한 처리구(MDI: 0.5 mM IBMX, 1mg/mL Insulin, 0.25μM dexamethasone)의 흡광도를 100%로 하였을 때 lipid accumulation은 각각 100.0, 99.1, 98.2, 98.5, 87.6, 91.8%로 나타나, D의 시료에서 가장 높은 항비만 효과를 보였다.Referring to FIG. 7, as a result of measuring lipid accumulation of pomegranate extract and A, B, C, D, and E, treatments that induced differentiation into adipocytes (MDI: 0.5 mM IBMX, 1 mg/mL Insulin, 0.25 μM dexamethasone) When the absorbance of was 100%, lipid accumulation was 100.0, 99.1, 98.2, 98.5, 87.6, and 91.8%, respectively, showing the highest anti-obesity effect in D sample.
이상, 본 발명은 일 실시 예를 참고로 설명되었으나 이는 예시적인 것에 불과하며, 당해 기술분야에서 통상의 지식을 가진 자라면 이로부터 다양한 변형 및 균등한 실시 예가 가능하다는 점을 이해할 것이다. 따라서 본 발명의 진정한 보호 범위는 첨부된 청구범위에 의해서만 정해져야 할 것이다.In the above, the present invention has been described with reference to an exemplary embodiment, but this is only exemplary, and those of ordinary skill in the art will understand that various modifications and equivalent embodiments are possible therefrom. Therefore, the true scope of protection of the present invention should be determined only by the appended claims.
Claims (7)
상기 유산균은 락토바실러스 아시도필루스(Lactobacillus acidophilus)이고,
상기 유산균 발효물은 항염증 및 항비만 활성을 갖는 것을 특징으로 하는 석류 유산균 발효물을 이용한 건강식품 조성물. Contains a lactic acid bacteria fermented product fermented by lactic acid bacteria by mixing 49% by weight of milk, 10% by weight of pomegranate extract, 40% by weight of Garcinia cambogia extract and 1% by weight of glucose,
The lactic acid bacteria is Lactobacillus acidophilus ,
The lactic acid bacteria fermented product is a health food composition using a pomegranate lactic acid bacteria fermented product, characterized in that it has anti-inflammatory and anti-obesity activity.
상기 혼합단계에서 수득한 혼합물을 멸균시키는 멸균단계와;
상기 멸균단계 후 상기 혼합물에 유산균을 접종하여 배양시켜 유산균 발효물을 수득하는 배양단계;를 포함하고,
상기 유산균은 락토바실러스 아시도필루스(Lactobacillus acidophilus)이고,
상기 유산균 발효물은 항염증 및 항비만 활성을 갖는 것을 특징으로 하는 석류 유산균 발효물을 이용한 건강식품 조성물의 제조방법.
A mixing step of mixing 49% by weight of milk, 10% by weight of pomegranate extract, 40% by weight of Garcinia cambogia extract, and 1% by weight of glucose;
A sterilization step of sterilizing the mixture obtained in the mixing step;
After the sterilization step, a culture step of inoculating and culturing the mixture to obtain a fermented lactic acid bacteria; includes
The lactic acid bacteria is Lactobacillus acidophilus ,
The lactic acid bacteria fermented product is a method for producing a health food composition using a pomegranate lactic acid bacteria fermented product, characterized in that it has anti-inflammatory and anti-obesity activity.
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| KR102514071B1 (en) | 2022-08-01 | 2023-03-28 | (주)스마트푸릇 | Fermented pomegranate composition for improving edema or obesity, containing high content of ellagic acid and gallic acid with increased absorption into the body and its use having skin whitening, wrinkle improvement or antioxidant function, and method for manufacturing the same by using EFC method |
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| KR102514074B1 (en) * | 2022-08-10 | 2023-03-27 | (주)스마트푸릇 | Fermented pomegranate composition for improving vascular health, prostate or sexual function, containing high content of ellagic acid and gallic acid with increased absorption into the body and its use having skin whitening, wrinkle improvement or antioxidant function, and method for manufacturing the same by using EFC method |
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| KR102514071B1 (en) | 2022-08-01 | 2023-03-28 | (주)스마트푸릇 | Fermented pomegranate composition for improving edema or obesity, containing high content of ellagic acid and gallic acid with increased absorption into the body and its use having skin whitening, wrinkle improvement or antioxidant function, and method for manufacturing the same by using EFC method |
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