KR20180116039A - Synbiotic composition comprising yacon extract and probiotic lactic acid bacteria - Google Patents
Synbiotic composition comprising yacon extract and probiotic lactic acid bacteria Download PDFInfo
- Publication number
- KR20180116039A KR20180116039A KR1020170048723A KR20170048723A KR20180116039A KR 20180116039 A KR20180116039 A KR 20180116039A KR 1020170048723 A KR1020170048723 A KR 1020170048723A KR 20170048723 A KR20170048723 A KR 20170048723A KR 20180116039 A KR20180116039 A KR 20180116039A
- Authority
- KR
- South Korea
- Prior art keywords
- yacon
- extract
- bifidobacterium
- composition
- present
- Prior art date
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Abstract
Description
본 발명은 프리바이오틱 야콘 추출물 및 프로바이오틱 유산균을 포함하는 신바이오틱 조성물에 관한 것이다.The present invention relates to a synbiotic composition comprising a prebiotic yacon extract and a probiotic lactic acid bacterium.
장내미생물은 인체의 장내에서 상호공생 또는 길항 관계를 유지하면서 섭취된 음식물과 소화관으로부터 분리되는 생체성분을 이용하고 증식 또는 배설되고 있다. 이 미생물은 Bifidobacterium, Lactobacillus, Streptococcus 속과 같은 유익균이 있다. Intestinal microorganisms are multiplying or excreted using biocomponents separated from food and digestive tracts while maintaining mutual symbiosis or antagonism in the intestines of the human body. These microorganisms are beneficial bacteria such as Bifidobacterium, Lactobacillus and Streptococcus.
프로바이오틱스(probiotics)를‘살아있는 미생물로 적당한 양을 섭취하면 건강에 유익한 세균’이라고 정의내렸으며(FAO/WHO. 2001), 프리바이오틱스는 프로바이오틱스의 성장에 필요한 영양소를 의미한다. 프로바이오틱스는 Bifidobacterium 속, Clostridium 속, Lactobacillus 속, Bacillus 속, Saccharomyces 속 등이 있으며, 대표적인 유익균인 비피도박테리움(Bifidobacterium)의 장내에서 유기산과 항생물질을 생산하여 외부로부터의 병원균이나 유해균총의 성장을 억제한다. 이 유산균은 대표적인 프로바이오틱스로서 각종 발효 식품, 장류, 의약품 및 가축의 사료 첨가제 등 다양하게 활용되고 있다. 최근에는 유산균의 영양적인 효과와 더불어 숙주의 장내에서 유산균의 다양한 건강 기능성을 강조한 연구들이 많이 이루어지고 있다.Defining probiotics as 'healthy germs when they consume modest amounts of living microbes' (FAO / WHO, 2001), prebiotics are the nutrients necessary for the growth of probiotics. Probiotics include Bifidobacterium, Clostridium, Lactobacillus, Bacillus, and Saccharomyces, which produce organic acids and antibiotics in the intestines of a representative bifidobacterium, Bifidobacterium, . This lactic acid bacterium is a representative probiotics, and various fermented foods, pastes, medicines and feed additives for livestock are being utilized variously. Recently, there have been many studies emphasizing the nutritional effects of lactic acid bacteria and various health functionalities of lactic acid bacteria in the intestinal tract.
이러한 유산균의 생체 내 활성 증가를 위해 생균에 해당하는 프로바이오틱스(probiotics)와, 이들의 영양원이 되는 프리바이오틱스(prebiotics)를 혼합한 신바이오틱스(synbiotics)에 대한 관심이 증가되고 있다. 그러나, 종래의 신바이오틱스에 사용되는 프리바이오틱스는 숙주의 소화 기관에서 상당한 소화가 진행되어 프로바이오틱스의 영양 공급원으로서 효율적으로 작용하지 못하는 문제가 있었고, 또한 프리바이오틱스의 종 특이성이 부족하여 특정한 유산균의 활성을 증가시키고자 할 때에 효율적이지 못한 문제가 있었다.In order to increase the in vivo activity of such a lactic acid bacterium, there is an increasing interest in synbiotics in which probiotics corresponding to live bacteria and prebiotics as nutrients thereof are mixed. However, the prebiotics used in the conventional synbiotics have a problem that digestion of digestive organs of the host progresses considerably and thus they do not act efficiently as a nutritional supply source of probiotics. Also, since the species specificity of prebiotics is insufficient, There is a problem in that it is not effective in increasing the activity.
한편, 야콘은 남미 안데스 원산으로 고구마와 비슷하게 생긴 괴근(덩이뿌리)에 프락토올리고당 및 항산화 성분과 같은 다양한 기능성 인자를 함유하고 있다. 지금까지 야은 이화학 성분 분석, 아미노산 분석, 프락토올리고당 함량 변화, 항염 활성, 추출물의 항산화 및 항암 활성 등과 같은 약리작용 효능이나 식품학적 가치에 관한 연구가 주로 이루어졌다. 그러나, 야콘 추출물이 프리바이오틱으로 장내 미생물 프로바이오틱에 미치는 영향에 대한 연구와 이를 활용한 제품개발은 미비한 실정이다.On the other hand, yacon originates from the South American Andes, and contains various functional factors such as fructooligosaccharides and antioxidants in rootstock, which is similar to sweet potato. So far, researches on pharmacological effects and pharmacological values such as analysis of physicochemical components, amino acid analysis, changes in fructo-oligosaccharide content, anti-inflammatory activity, antioxidant and anticancer activities of extracts have been conducted. However, studies on the effect of yacon extracts on prebiotic and intestinal microbiological probiotics and the development of products utilizing them have been insufficient.
따라서, 본 발명자들은 야콘 추출물이 프리바이오틱으로서 프로바이오틱스와 함께 신바이오틱 식품에 이용 가능성을 확인하고 본 발명을 완성하였다.Therefore, the inventors of the present invention have confirmed the availability of yacon extract as synbiotic food together with probiotics as a prebiotic and completed the present invention.
본 발명의 목적은 프리바이오틱스로서 야콘 추출물을 포함하고, 프로바이오틱스로서 상기 야콘 추출물을 특이적으로 이용하는 비피도박테리움 속 미생물을 함유하는 신바이오틱스 조성물을 제공하는 것이다.It is an object of the present invention to provide a synbiotics composition containing a yacon extract as a prebiotic and a microorganism belonging to the genus Bifidobacterium which specifically uses the yacon extract as a probiotics.
본 발명의 목적은 프리바이오틱스로서 야콘 추출물을 포함하고, 프로바이오틱스로서 상기 야콘 추출물을 특이적으로 이용하는 비피도박테리움 속 미생물을 함유하는 신바이오틱스 식품 조성물을 제공하는 것이다.An object of the present invention is to provide a synbiotics food composition containing a yacon extract as a prebiotic and a microorganism belonging to the genus Bifidobacterium which specifically uses the yacon extract as a probiotics.
본 발명의 다른 목적은 프리바이오틱스로서 야콘 추출물을 포함하고, 프로바이오틱스로서 상기 야콘 추출물을 특이적으로 이용하는 비피도박테리움 속 미생물을 함유하는 신바이오틱스 사료 첨가제를 제공하는 것이다.Another object of the present invention is to provide a synbiotics feed additive comprising a yacon extract as a prebiotic and a microorganism belonging to the genus Bifidobacterium which specifically uses the yacon extract as a probiotics.
본 발명은 야콘 추출물 및 비피도박테리움 속 미생물을 포함하는 조성물을 제공한다. The present invention provides a composition comprising a yacon extract and a microorganism belonging to the genus Bifidobacterium.
본 발명의 야콘 추출물은 야콘의 괴근으로부터 추출한 추출물을 의미한다. 본 발명의 추출과정은 당업계에 알려진 통상적인 방법으로 수행될 수 있다. 예를 들면, 야콘 괴근에 물 또는 유기용매를 넣고, 환류 추출하여 침적시킨 후 여과포 여과와 원심분리를 통해 잔사와 여액을 분리하고, 분리된 여액을 감압 농축하여 야콘 추출물을 얻을 수 있다. 바람직하게는 상기 야콘 괴근은 추출 전에 동결건조된 것 일 수 있다.The yacon extract of the present invention means an extract extracted from yacon root. The extraction process of the present invention can be carried out by a conventional method known in the art. For example, water or an organic solvent is added to a yacon mass, and the mixture is subjected to reflux extraction, followed by filtration, followed by filtration and centrifugation to separate the residue and the filtrate. The separated filtrate is concentrated under reduced pressure to obtain a yacon extract. Preferably, the yacon troche is lyophilized prior to extraction.
본 발명에 사용 가능한 유기용매는 에탄올, 메탄올, 부탄올, 에테르, 에틸아세테이트, 클로로포름 또는 이들 유기용매와 물의 혼합용매에서 선택될 수 있으며, 바람직하게는 80% 에탄올을 사용할 수 있다. 이때, 추출 온도는 10~80℃가 바람직하며, 추출 시간은 6~24시간이 바람직하다. 상기 추출 온도 및 추출 시간을 벗어나면 추출 효율이 떨어지거나 성분의 변화가 생길 수 있다.The organic solvent which can be used in the present invention may be selected from ethanol, methanol, butanol, ether, ethyl acetate, chloroform or a mixed solvent of these organic solvents and water, preferably 80% ethanol. At this time, the extraction temperature is preferably 10 to 80 ° C, and the extraction time is preferably 6 to 24 hours. If the extraction temperature and the extraction time are exceeded, the extraction efficiency may be deteriorated or the component may be changed.
상기에서 용매를 이용하여 추출물을 얻은 이후 당업계에 알려진 통상적인 방법으로 상온에서 냉침, 가열 및 여과하여 액상물을 얻을 수 있으며, 또는 추가로 용매를 증발, 분무 건조 또는 동결 건조할 수 있다.After obtaining the extract using a solvent, the extract can be obtained by cooling, heating and filtering at room temperature by a conventional method known in the art, or the solvent can be further evaporated, spray dried or freeze-dried.
본 발명의 비피도박테리움 속 미생물은 비피도박테리움 속에 속하는 미생물을 포함할 수 있다. 바람직하게는 Bifidobacterium bifidum BGN4 , Bifidobacterium longum BORI 및 Bifidobacterium lactis로 이루어진 군으로부터 선택되는 하나 이상의 미생물일 수 있다. The microorganism belonging to the genus Bifidobacterium of the present invention may include a microorganism belonging to the genus Bifidobacterium. Preferably, Bifidobacterium bifidum BGN4 , Bifidobacterium longum BORI and Bifidobacterium lactis , < / RTI >
본 발명의 야콘 괴근 추출물은 프리바이오틱스로 작용하여 장내 유익균으로 알려진 미생물 중에서도 비피도박테리움 속 미생물의 증식에 보다 특이적으로 이용 될 수 있다. 따라서 다른 야콘 괴근 추출물과 비피도박테리움 속 미생물을 함께 포함시킴으로써 비피도박테리움 속 미생물 증식효과를 높일 수 있다. The extract of yacon grass root of the present invention acts as a prebiotic and can be used more specifically for the growth of microorganisms of the genus Bifidobacterium among the microorganisms known as beneficial bacteria in the intestines. Therefore, it is possible to increase the microbial growth effect of the genus Bifidobacterium by including other microorganisms such as Bacillus subtilis and Bifidobacterium.
본 발명의 식품 조성물은 상기 야콘 추출물 및 비피도박테리움속 미생물의 혼합비가 중량비로 1 : 2-5 비율로 포함될 수 있다. 바람직하게는, 중량비가 1 : 2-4 비율일 수 있다.In the food composition of the present invention, the mixing ratio of the yacon extract and the Bifidobacterium genus microorganism may be 1: 2-5 by weight. Preferably, the weight ratio can be 1: 2-4.
본 발명은 야콘 추출물 및 비피도박테리움 속 미생물을 포함하는 식품 조성물을 제공한다. The present invention provides a food composition comprising a yacon extract and a microorganism belonging to the genus Bifidobacterium.
일 실시예에서, 본 발명의 식품 조성물은 야콘 추출물 및 비피도박테리움 속 미생물 외에 식품의 종류에 따라 맥아당, 무수결정포도당, 난소화성말토덱스트린, 치커리 추출물 및 비타민 C를 더 포함할 수 있고, 이에 제한되는 것은 아니다.In one embodiment, the food composition of the present invention may further comprise maltose, anhydrous crystalline glucose, indigestible maltodextrin, chicory extract, and vitamin C, depending on the type of food, in addition to the yacon extract and Bifidobacterium sp. But is not limited to.
이러한 본 발명의 상기 식품 조성물은 섭취에 편리한 형태로 다양한 경구 제형을 가질 수 있으며, 예를 들어 과립형 분말제, 액상제, 겔제 또는 고제(膏劑)의 제형을 가질 수 있다. The food composition of the present invention may have various oral formulations in a convenient form for ingestion and may have, for example, a granular powder, a liquid preparation, a gel or a solid preparation.
본 발명의 식품 조성물은 일반 식품이나 건강기능 식품에 포함되어 제공될 수도 있다. 상기 식품의 종류에는 특별한 제한은 없다. 상기 조성물을 첨가할 수 있는 식품의 예로는 드링크제, 육류, 소시지, 빵, 비스킷, 떡, 초콜릿, 캔디류, 스낵류, 과자류, 피자, 라면, 기타 면류, 거류, 아이스크림류를 포함한 낙농 제품, 각종 수프, 음료수, 알코올 음료 및 비타민 복합제 등이 있으며, 통상의 건강식품을 모두 포함한다.The food composition of the present invention may be provided in a general food or a health functional food. There is no particular limitation on the kind of the food. Examples of the foods to which the composition can be added include food products such as drinks, meat, sausage, bread, biscuits, rice cakes, chocolate, candies, snacks, confectionery, pizza, ramen, other noodles, Beverages, alcoholic beverages, and vitamin complexes, all of which include normal health foods.
또한, 본 발명은 야콘 추출물 및 비피도박테리움 속 미생물을 포함하는 조성물을 주원료로 하는 사료 첨가제를 제공한다. 본 발명의 야콘 추출물의 추출방법 및 비피도박테리움 속 미생물에 대해서는 상술한 바와 같다.The present invention also provides a feed additive comprising as a main ingredient a composition comprising a yacon extract and a microorganism belonging to the genus Bifidobacterium. The extraction method of the yacon extract of the present invention and the microorganism belonging to the genus Bifidobacterium are as described above.
본 발명의 상기 사료 첨가제는 가축용 사료에 첨가하여 가축에게 급여함으로써, 가축의 장내 유익 균총 증가, 장내 유해균총(Enterobacteriaceae) 감소 및 증체량이 증진되는 것을 특징으로 하나, 이에 제한되는 것은 아니다. The feed additive of the present invention is characterized in that it is added to a feed for livestock and fed to the livestock to increase the intestinal beneficial microflora, reduce the enterobacteriaceae, and increase the amount of the feed.
본 발명은 프리바이오틱스로서 야콘 추출물을 포함하고, 프로바이오틱스로서 상기 야콘 추출물을 특이적으로 잘 이용하는 비피도박테리움 속의 특정 미생물을 포함하는 신바이오틱스 조성물을 제공함으로써, 이를 섭취한 숙주의 장 내에 특정 유산균이 우위적으로 증식할 수 있도록 도와줄 수 있어 장내 유산균 환경을 조절할 수 있다.The present invention provides a synbiotics composition comprising a specific microorganism belonging to the genus Bifidobacterium that contains a yacon extract as a prebiotic and specifically utilizes the yacon extract as a probiotics, It can help the lactic acid bacteria to proliferate predominantly, thereby controlling the intestinal lactic acid bacteria environment.
도 1은 야콘 괴근 에탄올 추출물에 포함된 당 성분의 종류 및 함량을 나타낸 것입니다.
도 2는 야콘 괴근 추출물이 포함된 배지와 그렇지 않은 배지에서 비피도박테리움 속 미생물 3종의 증식여부를 확인하여 세균수를 측정한 결과를 그래프로 나타낸 것입니다.
도 3은 야콘 괴근 추출물이 포함된 배지와 그렇지 않은 배지에서 락토바실러스 속 미생물 3종의 증식여부를 확인하여 세균수를 측정한 결과를 그래프로 나타낸 것입니다.Fig. 1 shows the kinds and content of sugar components contained in the ethanol extract of yacon mass.
FIG. 2 is a graph showing the results of measuring the number of bacteria by confirming the growth of three species of Bifidobacterium sp. In a medium containing a yacon trout extract and a medium not containing the extract.
FIG. 3 is a graph showing the results of measurement of the number of bacteria by confirming the proliferation of three Lactobacillus sp. Microorganisms in a medium containing yacon trout extract and a medium not containing it.
이하, 본 발명에 대하여 실시예를 통하여 보다 상세히 설명한다. 본 명세서에 기재되지 않은 내용은 본원 발명의 기술 분야 또는 유사 분야에서 통상의 기술자라면 충분히 인식하고 유추할 수 있는 것이므로 그 설명을 생략한다.Hereinafter, the present invention will be described in more detail by way of examples. The contents not described in this specification can be sufficiently recognized and inferred by those of ordinary skill in the technical field or the like of the present invention, and the description thereof will be omitted.
<실시예 1> 야콘 추출물의 제조≪ Example 1 > Preparation of yacon extract
실시예 1-1 야콘의 준비Example 1-1 Preparation of Yacon
야콘을 국내에서 수집하여 강원도 평창군 진부면 시험포장(해발고도 560 m)에 2015년 5월 15일에 정식한 후 10월 15일에 수확하였다. 시료로 사용된 야콘은 농촌진흥청 표준재배법에 준하여 재배되었다. 야콘의 괴근 시료는 분석직전까지 초저온냉동고(-70℃)에 보관하면서 사용하였으며, 샘플은 동결건조기(EYELA FDU-2100, EYELA Corp., Tokyo, Japan)로 5일 동안 건조한 후 믹서기(Mini multi deluxe BL126DK blender, Tefal Co., Kaleo, France)를 사용해 분쇄하였다. 각 시험 항목에 대한 시료의 분석은 생체중으로 표기하였고, 4회 반복 실시하였다.Yacon was collected in Korea and harvested on May 15, 2015 after it was planted on May 15, 2015 at the test packaging (560 m above sea level) of Chungbu-myeon, Pyeongchang-gun, Gangwon Province. Yacon used as a sample was cultivated in accordance with RDA standards. The samples were dried in a freezer (EYELA FDU-2100, EYELA Corp., Tokyo, Japan) for 5 days, and then placed in a mixer (Mini multi deluxe BL126DK blender, Tefal Co., Kaleo, France). Analysis of samples for each test item was expressed as live weight and repeated four times.
실시예 1-2 야콘 에탄올 추출물의 제조Example 1-2 Preparation of Ethanol Extract of Yacon
야콘 괴근 추출물은 동결건조된 야콘 괴근 10 g에 20배의 80% 에탄올를 각각 가하여 교반 추출기(VS-8480, Vision Scientific Co., Ltd., Seoul, Korea)로 25℃에서 24시간 동안 200rpm으로 추출하여 여과지(Whatman No. 2, Bukinghamshire, UK)로 여과한 다음, 회전진공농축기(EYELA CCA-1110, Tokyo, Japan)를 이용하여 용매를 제거한 후 추출물로 사용하였다. 제조된 각각의 추출물은 추출버퍼로 최종 10 mL로 정용한 후 실험에 사용하였다.The extracts of Yacon Grassroot extract were prepared by adding 20-fold 80% ethanol to 10 g of lyophilized yacon roots and extracting them at 200 rpm for 24 hours at 25 ° C with a stirring extractor (VS-8480, Vision Scientific Co., After filtration through a filter paper (Whatman No. 2, Bukinghamshire, UK), the solvent was removed using a rotary vacuum concentrator (EYELA CCA-1110, Tokyo, Japan) and used as an extract. Each extract was diluted to 10 mL with extraction buffer and used in the experiment.
<실시예 2> 비피도박테리움 속 미생물 배양 배지 준비Example 2 Preparation of Bifidobacterium Microorganism Culture Medium
본 발명에 사용될 비피도박테리움 속 미생물의 MRS-Control 배지 조성물로는 Dextrose 20g, Meat Peptone 10g, Beef extract 10g, Yeast extract 5g, Sodium acetate 5g, Disodium phosphate 3g, ammonium citrate 0.5g, tween 80 1g, Magnesium sulfate 0.1g, maganese sulfate 0.05g, 0.05% L-cystrein hydrochloride을 증류수 1,000mL에 녹여 121℃에 15분간 멸균하고 사용하였다As the MRS-Control medium composition of the genus Bifidobacterium, 10 g of Dextrose, 10 g of Meat Peptone, 10 g of Beef extract, 5 g of yeast extract, 5 g of sodium acetate, 3 g of Disodium phosphate, 0.5 g of ammonium citrate, 0.1 g of magnesium sulfate, 0.05 g of maganese sulfate and 0.05% of L-cystrein hydrochloride were dissolved in 1,000 mL of distilled water and sterilized at 121 ° C for 15 minutes
(표 1). MRS-Yacon 배지 조성물로는 상기 1,000mL 대조구(MRS-Control) 배지에 대한 1%의 야콘 추출물을 첨가하여 실험에 사용하기 전 3회 이상 계대 배양하여 활성화 시킨 후 사용하였다(표 1).(Table 1). As the MRS-Yacon medium composition, 1% yacon extract was added to the 1,000 mL control (MRS-Control) medium, and the mixture was subcultured three times or more before use in the experiment (Table 1).
Meat peptone
Beef extract
Yeast extract
Sodium acetate
Disodium phosphate
ammonium citrate
Tween 80
Magnesium sulfate
Manganese sulfate
L-cystein hydrochloride
Dextrose
Meat peptone
Beef extract
Yeast extract
Sodium acetate
Disodium phosphate
ammonium citrate
Tween 80
Magnesium sulfate
Manganese sulfate
L-cysteine hydrochloride
10.0g
10.0g
5.0g
5.0g
3.0g
0.5g
1.0g
0.1g
0.05g
0.05%
20.0 g
10.0 g
10.0 g
5.0g
5.0g
3.0 g
0.5 g
1.0 g
0.1 g
0.05 g
0.05%
Meat peptone
Beef extract
Yeast extract
Sodium acetate
Disodium phosphate
ammonium citrate
Tween 80
Magnesium sulfate
Manganese sulfate
L-cystein hydrochloride
Yacon extractDextrose
Meat peptone
Beef extract
Yeast extract
Sodium acetate
Disodium phosphate
ammonium citrate
Tween 80
Magnesium sulfate
Manganese sulfate
L-cysteine hydrochloride
Yacon extract
10.0g
10.0g
5.0g
5.0g
3.0g
0.5g
1.0g
0.1g
0.05g
0.05%
1%20.0 g
10.0 g
10.0 g
5.0g
5.0g
3.0 g
0.5 g
1.0 g
0.1 g
0.05 g
0.05%
One%
* 각 처리의 배지를 증류수 1,000mL에 녹여 121℃ 15분간 멸균후 사용 * Dissolve each treatment medium in 1,000 mL of distilled water and sterilize at 121 ℃ for 15 minutes.
<< 실시예Example 3> 3> 야콘Yacon 괴근Miniature 추출물의 구성성분 및 함량 Components and content of extract
프락토올리고당 분석은 식품공전(KFDA, 2005)의 방법에 준하여 실시하였다. 생체시료 10 g에 80% 에탄올 50 mL를 가하여 균질기(Ultra-Turrax T25 Basic Homogenizer, IKA Co., Staufen, Germany)로 9,500 rpm으로 1분간 마쇄하고, 여과지(No. 2, Whatman International, Maidstone, UK)로 여과하여 50 mL로 정용하였다. 추출한 전처리 용액을 멤브레인 필터(0.45 μm, Milipore, Bedford, MA, USA)로 여과한 후 분석에 사용하였다. 프락토올리고당의 표준용액 조제(Fructooligosaccharides Set, Wako, Osaka, Japan)를 위해 케스토즈(1-kestose= GF2), 니스토즈(nystose=GF3), 프락토푸란노실니스토즈(1F-fructofuranosylnystose =GF4)를 0.4 g씩 취하여 각 각 20 mL의 메스플라스크에 넣고 증류수로 표선까지 채운것을 표준원액으로 이용하였다. 고속액체크로마토그래피(Waters 2695 HPLC, Waters Co., Milford, MA, USA) 분석조건은 컬럼(Carbohydrate LC-NH2 Φ4.6x250 mm 5 μm, Supercosil, Bellefonte, USA) 온도를 30℃로 유지하였고, 이동상은 아세토니트릴:증류수(acetonitrile:water = 70:30, v/v)를 이용하였으며, 유속은 1.0 mL/min로 하였고, 굴절률 검출기(Waters 2414 Refractive Index Detector, Waters, Milford, MA, USA)를 이용하였다. 프락토올리고당이 10%이 함유되어 있었다.Fructooligosaccharide analysis was carried out according to the method of Food Code (KFDA, 2005). 50 ml of 80% ethanol was added to 10 g of the biological sample and the mixture was ground with a homogenizer (Ultra-Turrax T25 Basic Homogenizer, IKA Co., Staufen, Germany) at 9,500 rpm for 1 minute and filtered paper (No. 2, Whatman International, Maidstone, UK) and diluted to 50 mL. The extracted pretreatment solution was filtered through a membrane filter (0.45 μm, Milipore, Bedford, Mass., USA) and used for analysis. (1-kestose = GF2), nystose (GF3), and 1F-fructofuranosylnystose (GF4) for a standard solution preparation of fructooligosaccharides (Fructooligosaccharides Set, Wako, Osaka, Japan) Were weighed in 0.4 g each, placed in 20 mL volumetric flasks, and filled with distilled water to the markers. The HPLC (Waters 2695 HPLC, Waters Co., Milford, Mass., USA) analysis conditions were as follows: Column (Carbohydrate LC-NH2 Φ 4.6 × 250
유리당 분석은 식품공전(KFDA, 2005)의 방법에 준하여 실시하였다. 생체시료 10 g에 80% 에탄올 50 mL를 가하여 균질기(Ultra-Turrax T25 Basic Homogenizer, IKA Co., Staufen, Germany)로 9,500 rpm으로 1분간 마쇄하고, 50 mL로 정용한 다음, 여과지(No. 2, Whatman International, Maidstone, UK)로 여과한 후, 분석장치(Sugar analyser, Toa Dkk, Tokyo, Japan)로 분석하였다. 분석조건은 용출용매로 0.2M 수산화나트륨(NaOH)를 사용하였으며 당전용 분석 컬럼(PCI-520 Φ4.6 ×150 mm 0.5 μm, Dkk Toa, Tokyo, Japan)을 이용하였다. 표준용액은 글루코스, 프럭토스, 수크로스, 말토스의 표준품 각각 0.2, 0.4, 0.5, 0.5 g을 물 50 mL로 녹여 3차 증류수로 100 mL까지 채운 후, 희석하여 표준용액으로 사용하였다. 전처리된 시료와 표준용액 2 mL를 취하여 멤브레인 필터(0.45 μm, Milipore, Bedford, MA, USA)로 여과한 후 자동주입장치(Auto sampler ICA-200AS, Toa DKK, Tokyo, Japan)를 이용하여 분석하였다. 이때 용출 용매의 유속은 0.7 mL/min로 하였다.Analysis of free sugar was carried out according to the method of Food Code (KFDA, 2005). 50 mL of 80% ethanol was added to 10 g of the biological sample and the mixture was ground for 1 minute at 9,500 rpm with a homogenizer (Ultra-Turrax T25 Basic Homogenizer, IKA Co., Staufen, Germany) , Whatman International, Maidstone, UK) and analyzed with an analyzer (Sugar analyzer, Toa Dkk, Tokyo, Japan). The analytical conditions were as follows: 0.2 M sodium hydroxide (NaOH) was used as an elution solvent, and a dedicated analytical column (PCI-520 Φ 4.6 × 150 mm 0.5 μm, Dkk Toa, Tokyo, Japan) was used. Standard solutions were prepared by dissolving 0.2, 0.4, 0.5, and 0.5 g of glucose, fructose, sucrose, and maltose in 50 mL of water, and diluting them to 100 mL with tertiary distilled water. 2 mL of the pretreated sample and standard solution was filtered through a membrane filter (0.45 μm, Milipore, Bedford, Mass., USA) and analyzed using an automatic injection device (Auto sampler ICA-200AS, Toa DKK, Tokyo, Japan) . At this time, the flow rate of the elution solvent was 0.7 mL / min.
야콘 괴근의 당함량을 분석한 결과, 표준물질 프락토올리고당과 유리당의 표준물질 피크는 다음과 같다(도 1). 야콘 생체 중 100 g 당 프락토올리고당이 7.6 g이 함유되었고, 유리당으로 수크로스가 0.72 g으로 가장 많았고, 프럭토스 0.34 g, 글루코스 0.26 g 순이었다(표 1). 야콘에는 프럭토스, 글루코스, 수크로스, 이눌린 및 프락토올리고당 등이 함유되어 있는데, 이 중 프락토올리고당이 약 약 8% 정도 함유되어 대부분을 차지하였다. As a result of analyzing the sugar content of yacon mass, the standard substance peak of the standard substance fructooligosaccharide and free sugar is as follows (Fig. 1). 7.6 g of fructo-oligosaccharide per 100 g of yacon body was contained, and sucrose was the most as 0.72 g of free sugars, followed by fructose 0.34 g and glucose 0.26 g (Table 1). Yacon contains fructose, glucose, sucrose, inulin, and fructooligosaccharides. Of these, fructooligosaccharides account for about 8% of the total.
(g/100g)Fractooligosaccharides
(g / 100 g)
(g/100g)Glucose
(g / 100 g)
(g/100g)Fructose
(g / 100 g)
(g/100g)Sucrose
(g / 100 g)
zMean±S.E. zMean ± S.E.
<실시예 4> 비피도박테리움 속 미생물 증식 효과에 미치는 야콘 추출물의 영향Example 4 Influence of Yacon Extract on Microbial Growth of Bifidobacterium sp.
본 실험에서 사용한 비피도박테리움 비피둠 BGN4(Bifidobacterium bifidum BGN4), 비피도박테리움 롱검 BORI(Bifidobacterium longum BORI), 비피도박테리움 락티스(Bifidobacterium lactis)는 비피도(주)(Bifido Co, Ltd., Hongcheon, Korea)의 균주를 사용하였다. 상기 1000ml MRS 배지에 대한 2% 비피도박테리움 속 미생물 3종을 상기 실시예 2에서 제조한 각각의 MRS 배지에 접종하여 37℃에 18시간 배양한 후 4℃에 냉장 보관하였다. 이들 균주는 0.05% L-cysteine hydrochloride을 넣은 성분을 증류수 1,000ML에 녹여 121℃에 15분간 멸균하고 사용하였다. 장내 비피더스의 생육곡선은 균주를 MRS-Control 배지 및 MRS-Yacon 배지에 접종하여 시간별 생균수를 측정하였는데 최종 12시간까지(0, 4, 8, 10, 12시간)으로 2일간 생균수 값을 측정하면서 관찰하였다. 야콘 추출물을 이용한 배지에 이용하여 총균수 측정방법은 비피도(주)의 방법에 따라 측정하였다. 즉, MRS 배지를 사용하여 발현한 총균수를 카운트하여 평균치±표준편차를 나타내었다(표 3 및 도 2). The Bifidobacterium Bifidobacterium BGN4 ( Bifidobacterium bifidum BGN4), Bifidobacterium ronggeom BORI (Bifidobacterium longum BORI ), Bifidobacterium lactis ) was used as a strain of Bifido Co, Ltd., Hongcheon, Korea. Three of the 2% Bifidobacterium sp. Microorganisms on the 1000 ml MRS medium were inoculated into each of the MRS media prepared in Example 2, incubated at 37 ° C for 18 hours, and then refrigerated at 4 ° C. These strains were dissolved in 1,000 mL of distilled water and sterilized at 121 ° C for 15 minutes in 0.05% L-cysteine hydrochloride. The growth curve of intestinal bifidus was obtained by inoculating the strains into the MRS-Control medium and the MRS-Yacon medium and measuring the number of viable cells per hour. The viable cell count was measured for 2 days at the end of 12 hours (0, 4, 8, 10, Respectively. The total number of bacteria was measured by using the method of Bifido Co., Ltd. in the medium using a yacon extract. That is, the total number of bacteria expressed using the MRS medium was counted, and the average value ± standard deviation was shown (Table 3 and FIG. 2).
배양
시간(h)
culture
Time (h)
그 결과, 야콘 괴근 추출물이 포함된 배지에 접종한 상기 비피도박테리움 속 미생물 3종의 증식이 대조군과 비교하여 향상됨을 확인할 수 있었다. As a result, it was confirmed that the proliferation of the three microorganisms of the genus Bifidobacterium inoculated into the medium containing the extract of yacon trout was improved compared to the control.
< 비교예 1> 락토바실러스 속 미생물 증식 효과에 미치는 야콘 추출물의 영향 본 발명자들은 락토바실러스 속 미생물 3종(Lactobacillus acidophilus, Lactobacillus plantarum , Lactobacillus fermentum)에 대하여 실시예 2 및 실시예4와 동일하게 미생물 증식 여부를 측정하였다(표 4 및 도 3). <Comparative Example 1> Lactobacillus effect of yacon extract on the Bacillus spp proliferative effect The present inventors same microbial growth and Examples 2 and 4 with respect to the Lactobacillus spp three kinds (Lactobacillus acidophilus, Lactobacillus plantarum, Lactobacillus fermentum) (Table 4 and Figure 3).
시간(h)culture
Time (h)
그 결과, 야콘 괴근 추출물이 포함된 배지에 접종한 상기 락토바실러스 속 미생물 3종의 증식이 대조군과 비교하여 유사하거나 시간이 지날수록 증식을 억제하는 것을 확인할 수 있었다. As a result, it was confirmed that the proliferation of the three Lactobacillus sp. Microorganisms inoculated on the medium containing the yacon trout extract was similar to that of the control group or inhibited the proliferation as time passed.
이와 같은 결과로, 야콘 괴근 추출물은 장내 유익균 중에서도 비피도박테리움 속 미생물을 특이적으로 증식시키는 효과를 가짐을 확인할 수 있었다. 따라서 야콘 괴근 추출물을 비피도박테리움 속 미생물과 함께 포함함으로써 유산균 증식효과가 우수한 신바이오틱 조성물이 될 수 있다. As a result, it was confirmed that the extract of Yacon Medicinalum had an effect of specifically proliferating microorganisms of the genus Bifidobacterium among the beneficial bacteria in the intestines. Therefore, by including the extract of Yacon grass root together with the microorganism of Bifidobacterium, it can be a synbiotic composition excellent in the effect of proliferating lactic acid bacteria.
Claims (7)
상기 비피도박테리움 속 미생물은 Bifidobacterium bifidum BGN4, Bifidobacterium longum BORI 및 Bifidobacterium lactis으로 이루어진 군으로부터 선택되는 하나 이상의 미생물인 것인 조성물.The method according to claim 1,
The microorganism belonging to the genus Bifidobacterium is Bifidobacterium bifidum BGN4, Bifidobacterium longum BORI, and Bifidobacterium lactis .
상기 야콘 추출물은 동결건조 시킨 야콘 괴근을 에탄올로 추출한 것인 조성물.The method according to claim 1,
Wherein the lyophilized yacon root is extracted with ethanol.
상기 조성물은 야콘 추출물 및 비피도박테리움 속 미생물이 1 : 2~4 비율로 혼합된 것인, 조성물.The method according to claim 1,
Wherein the composition is a mixture of yacon extract and genus Bifidobacterium in a ratio of 1: 2 to 4.
A feed additive comprising the composition of any one of claims 1 to 4.
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