KR20060019496A - Fermentation liquor using microorganisms from bamboo, and process for preparing the same - Google Patents
Fermentation liquor using microorganisms from bamboo, and process for preparing the same Download PDFInfo
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- KR20060019496A KR20060019496A KR1020050037758A KR20050037758A KR20060019496A KR 20060019496 A KR20060019496 A KR 20060019496A KR 1020050037758 A KR1020050037758 A KR 1020050037758A KR 20050037758 A KR20050037758 A KR 20050037758A KR 20060019496 A KR20060019496 A KR 20060019496A
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- South Korea
- Prior art keywords
- bamboo
- fermentation broth
- bacteria
- microorganisms
- present
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Abstract
본 발명은 대나무의 껍질, 줄기, 잎, 뿌리 등을 채취하여 이를 발효시켜서 얻은 대나무 발효액, 그의 제조 방법 및 이 발효액의 용도에 관한 것으로서, 더욱 구체적으로는 대나무의 껍질, 잎, 줄기, 죽순 등에 유래하는 자연 서식 미생물을 이용하여 발효액을 제조하는 방법, 이 발효액의 다양한 산업적 용도에 관한 것이다. 본 발명에 따른 발효액은 천연물에서 유래한 것으로 부작용이 없으며 여러 가지 유익한 기능성을 나타내므로 의약 또는 식품 성분 등 다양하게 이용할 수 있다.The present invention relates to a bamboo fermentation broth obtained by extracting bamboo bark, stems, leaves, roots, etc., and to fermenting the same, and more specifically, to a bamboo bark, leaf, stem, bamboo shoot, and the like. The present invention relates to a method for producing a fermentation broth using a natural habitat microorganism, and various industrial uses of the fermentation broth. The fermentation broth according to the present invention is derived from natural products and has no side effects and shows various beneficial functions, and thus can be used in various ways such as medicine or food ingredients.
대나무, 발효액 Bamboo, fermented broth
Description
도 1은 본 발명에 따른 발효액의 비브리오균(Vibrio vulnificus)에 대한 항균효과를 나타내는 사진.1 is a photograph showing the antimicrobial effect of Vibrio vulnificus of the fermentation broth according to the present invention.
도 2는 본 발명에 따른 발효액의 에어로모나스균(Aeromonas hydrophila)에 대한 항균효과를 나타내는 사진.Figure 2 is a photograph showing the antimicrobial effect of the fermentation broth according to the present invention ( Aeromonas hydrophila ).
도 3은 본 발명에 따른 발효액의 대장균(Escherichia coli)에 대한 항균효과를 나타내는 사진.Figure 3 is a photograph showing the antimicrobial effect of Escherichia coli ( Esherichia coli ) of the fermentation broth according to the present invention.
본 발명은 대나무 껍질, 줄기, 잎, 뿌리 등을 채취하여 이를 자연적으로 또는 인위적으로 발효시켜서 얻은 대나무 발효액, 그의 제조 방법 및 이 발효액의 용도에 관한 것으로서, 더욱 구체적으로는 대나무의 껍질, 잎, 줄기, 뿌리, 죽순 등에 유래하는 자연 서식 미생물(예, 세균, 곰팡이)로부터 제조된 발효액, 그의 제조 방법 및 이 발효액의 다양한 산업적 용도에 관한 것이다.The present invention relates to a bamboo fermentation broth obtained by harvesting bamboo bark, stems, leaves, roots, and the like naturally or artificially, and to a method of preparing the fermentation broth, and more specifically, to bark, leaves and stems of bamboo. It relates to a fermentation broth prepared from natural habitat microorganisms (eg, bacteria, fungi) derived from roots, roots, bamboo shoots, and the like, methods for preparing the same, and various industrial uses of the fermentation broth.
대나무는 화본과 식물로 우리나라를 위시한 동남아시아에 주로 분포하고 있 으며, 우리나라에 자생하고 있는 대표적인 종류는 왕대(참대), 맹종죽, 조릿대, 신의대 등을 들 수 있다. 대나무는 예로부터 고혈압, 발한, 중풍 등의 치료를 위한 민간약으로 활용이 되어 왔을 뿐만 아니라 대나무의 방부효과도 민간에 널리 알려져서 김치를 담근 후 대나무 잎으로 덥거나, 동치미에 조릿대 잎을 띄워서 보관기간을 연장하는데 사용하였다. 또한 대나무 성분이 민간 약제 및 건강음료로 사용되고 있는 점을 고려할 때, 식품 보존제로 사용이 가능할 것으로 보인다. 대나무의 항균성에 관한 연구로는 주로 김치의 젖산균에 대한 대나무의 항균효과를 보고했지만 대나무 열수 추출물의 당 Brix에 대한 농도로 표시하여 대나무 종류별 정확한 항균 효과로 보기 어렵다. Bamboo is mainly distributed in Southeast Asia, including Korea as a flower and plant, and the representative varieties that grow wild in Korea include Wangdae (Bangdae), Mengjongjuk, Joridae, and Sinuidae. Bamboo has long been used as a folk medicine for the treatment of high blood pressure, sweating, and stroke, and the preservative effect of bamboo is widely known to the public. Used to extend. In addition, considering that bamboo is used as a folk medicine and health drink, it may be used as a food preservative. The study on the antimicrobial activity of bamboo mainly reported the antimicrobial effect of bamboo against lactic acid bacteria of kimchi, but it is hard to see the exact antimicrobial effect of each type of bamboo by expressing the concentration of sugar Brix of hydrothermal extract of bamboo.
대나무를 이용한 산업적 응용의 구체적인 예로는, 대나무 추출물을 이용하여육류제품을 가공하는 방법이 등록 특허공보 제1999-187719호에 개시되어 있고 죽내피 추출물을 함유한 미백화장료가 특허공개공보 2000-0007943호에 개시되어 있다. As a specific example of industrial application using bamboo, a method of processing meat products using bamboo extract is disclosed in Korean Patent Publication No. 1999-187719, and a whitening cosmetic containing bamboo extract is disclosed in Korean Patent Publication No. 2000-0007943. Is disclosed.
그러나, 대나무의 유리한 특성을 산업적으로 최대한으로 활용할 수 있는 묘안에 대해서 여전히 많은 방안들이 요구되고 있는 실정이다.However, many methods are still required for the seedlings that can make the most of the advantageous properties of bamboo industrially.
본 발명자들은 대나무에서 기능성을 나타내는 물질을 탐색하고자 예의 연구한 결과, 대나무의 껍질, 줄기, 죽순, 뿌리, 잎을 채취하여 여기에 부착된 유익한 세균 및 곰팡이 등의 자연미생물을 발효시켜 본 결과, 이 발효액이 다양한 기능성을 나타냄을 확인하고 본 발명을 완성하기에 이르렀다.The present inventors earnestly researched to search for substances that exhibit functionality in bamboo. As a result of extracting the bark, stems, bamboo shoots, roots and leaves of bamboo and fermenting natural microorganisms such as beneficial bacteria and fungi attached thereto, It was confirmed that the fermentation broth exhibited various functionalities and came to complete the present invention.
따라서, 본 발명의 목적은 대나무의 껍질, 잎, 줄기, 죽순, 뿌리를 채취하여 이로부터 곰팡이 등 미생물을 자연 증식시키거나 또는 상기 재료를 깨끗이 세척한 다음 여기에 세균류, 진균류 또는 이들의 복합균을 첨가하여 미생물을 증식시킨 다음 이를 발효시켜서 얻은 대나무 발효액 및 그의 제조 방법을 제공하는데 있다.Therefore, an object of the present invention is to extract the bark, leaves, stems, bamboo shoots, roots of bamboo to naturally grow microorganisms such as mold or to clean the material and then to remove bacteria, fungi or complexes thereof. The present invention provides a bamboo fermentation broth obtained by adding microorganisms and then fermenting the same, and a method for producing the same.
본 발명의 또 다른 목적은 상기 발효액의 다양한 산업적인 용도, 예들 들면 상기 발효액을 포함한 숙취해소용 음료, 위장관 치료제, 가축 및 어류 양식용 사료첨가제 및 어병억제제를 제공하는데 있다.It is still another object of the present invention to provide various industrial uses of the fermentation broth, for example, hangover-relieving beverages including the fermentation broth, treatment for gastrointestinal tract, feed additives for livestock and fish farming, and fish disease inhibitors.
본 발명은, 일면에 있어서, 대나무의 껍질, 잎, 줄기, 죽순, 뿌리를 채취하여 이로부터 세균, 곰팡이 등의 미생물을 자연 증식시키거나 또는 상기 재료를 깨끗이 세척한 다음 여기에 세균류, 진균류 또는 이들의 복합균을 첨가하여 미생물을 증식시킨 다음 이를 발효시켜서 얻은 대나무 발효액, 및 그의 제조 방법을 제공하는데 있다.The present invention, in one aspect, bamboo bark, leaves, stems, bamboo shoots, roots and the like to naturally grow microorganisms such as bacteria, fungi or the like or to clean the material and then to bacteria, fungi or these It is to provide a bamboo fermentation broth obtained by adding a microorganism and then fermenting the microorganism, and a method for producing the same.
본 발명에 따른 대나무 발효액은Bamboo fermentation broth according to the present invention
a) 대나무의 껍질, 잎, 줄기, 뿌리 또는 죽순을 채취하여 여기에 증자시킨 고두밥을 첨가하여 미생물을 자연 증식시키거나 또는 상기 재료들을 깨끗이 세척한 다음 여기에 고두밥을 첨가하고 세균류, 진균류, 또는 이들의 복합균을 접종하여 미생물을 증식시키는 단계,a) Take bamboo shells, leaves, stems, roots or bamboo shoots and add the added gourd rice to increase the microorganisms naturally or clean the above materials and add the gourd rice to them and add bacteria, fungi, or these Inoculating the complex bacteria to multiply the microorganisms,
b) 상기 고두밥에 당원을 첨가하여 당화시킨 다음 이를 발효시켜 발효원액을 제조하는 단계, 및b) preparing a fermentation stock solution by saccharifying by adding sugar source to the soybean rice, and then fermenting it; and
c) 상기 발효원액을 살균한 냉각 지하수 또는 대나무의 껍질, 잎, 줄기, 뿌 리 또는 죽순을 끓인 물로 희석하여 숙성시켜 발효액을 제조하는 단계에 의해 얻어진다. c) the fermentation broth is obtained by diluting the sterilized ground water or bamboo bark, leaves, stems, roots or bamboo shoots with boiled water to mature.
이하, 상기 발효액의 구체적인 제조 방법에 대해서 설명한다.Hereinafter, the specific manufacturing method of the said fermentation broth is demonstrated.
상기 단계 a)에서 사용되는 대나무는 특별히 한정되지 않고, 예를 들면 조릿대, 솜대, 맹종죽, 왕대, 오죽 등의 껍질, 줄기, 잎, 죽순, 뿌리를 이용할 수 있다. 이러한 재료는 적당한 크기로 절단하여 취급이 용이하도록 전처리하여도 좋다. 상기 대나무 재료는 단독 또는 혼합하여 사용할 수 있고, 이를 약간 경사진 바닥의 비닐 위에 직경 약 50cm의 범위로 분산시켜 배치하고 그 위에 증자시킨 고두밥(예, 맵쌀 밥)을 골고루 얹은 다음 7~20 일 동안 15~60 ℃의 온도에서 미생물을 증식시킨다. 그 후 당액으로서 흑설탕 등을 상기 고두밥에 대하여 약 1:0.1~5 배의 비율로 첨가하여, 상기 미생물을 증식시킨 조건과 동일 또는 유사한 조건하에서 3~15일 동안 발효시켜 자연스럽게 우러나오는 발효원액을 수집한다. 고두밥에 당액을 첨가하여 당화시켜서 당화된 고두밥을 발효시키는 공정에 있어서, 당액의 원료는 흑설탕, 설탕, 키토올리고당, 엿기름, 벌꿀, 포도당, 과당, 유당, 플락토올리고당 등을 사용하여도 되고 이들의 혼합물을 사용하여도 좋다. 이어서, 상기 발효액은 상기 대나무의 껍질, 줄기, 뿌리, 잎, 죽순 등을 수거하여 약 2시간 정도 끓인 후 이를 4 ℃로 냉각시킨 추출수로 일정비로 희석하여 숙성시켜 발효액을 얻는다. 희석배수는 1:10~1000배의 범위 내에서 수행할 수 있으며, 약 100배 정도 희석하는 것이 바람직하다. 숙성 조건은 특별히 한정할 필요는 없으나, 보통 실온 내지 45℃에서 수행하는 것이 좋으며, 숙성 기간은 1주 내지 2개월 정도 수행하는 것이 바람직하 다. 희석액으로서 상기 끓인 대나무 재료 추출수 대신에 비등시킨 다음 4℃로 냉각시킨 지하수를 사용하는 것도 가능하다. 이 발효액을 필터(예, 0.2㎛)로 여과시켜 미생물 등의 고형물질과 이물질을 제거한다. 추가로, 필요에 따라서, 여액을 100℃ 까지 약 2 시간 정도 끓여 잔여 미생물을 살균시킨 다음 자연 냉각시켜 목적하는 용도에 사용할 수 있다. 상기 멸균 과정은 순간멸균, 고압 순간 멸균, 초음파 처리 등 다른 물리적 방법으로 수행하여도 좋다.The bamboo used in the step a) is not particularly limited, and for example, bark, stalks, barley porridge, wangdae, oyster shells, stems, leaves, bamboo shoots, roots can be used. Such materials may be cut to a suitable size and pretreated to facilitate handling. The bamboo material may be used alone or in combination, and it is disposed on a vinyl of a slightly inclined bottom in a range of about 50 cm in diameter and evenly topped with steamed rice (eg, spicy rice) on it for 7 to 20 days. Proliferate microorganisms at a temperature of 15-60 ° C. Thereafter, brown sugar or the like is added at a ratio of about 1: 0.1 to 5 times the sugar gourd rice, and the fermentation stock solution naturally collected by fermentation for 3 to 15 days under the same or similar conditions as the conditions for proliferating the microorganisms. do. In the step of fermenting saccharified gourd rice by adding saccharified sugar solution to the gourd rice, the raw material of the sugar liquid may be brown sugar, sugar, chitooligosaccharide, malt, honey, glucose, fructose, lactose, floctooligosaccharide, or the like. Mixtures may be used. Subsequently, the fermentation broth collects the bark, stems, roots, leaves, bamboo shoots, etc. of the bamboo, boils for about 2 hours, and dilutes the fermentation broth by diluting it with a constant ratio of extracted water cooled to 4 ° C. Dilution multiple times can be carried out in the range of 1:10 to 1000 times, it is preferable to dilute about 100 times. The aging conditions do not need to be particularly limited, but are usually performed at room temperature to 45 ° C., and the aging period is preferably performed for about 1 week to 2 months. It is also possible to use ground water cooled to 4 ° C. after boiling as a diluent instead of the boiled bamboo material extract water. This fermentation broth is filtered through a filter (eg 0.2 μm) to remove solids such as microorganisms and foreign substances. In addition, if necessary, the filtrate may be boiled up to 100 ° C. for about 2 hours to sterilize the remaining microorganisms and then naturally cooled to use for the desired application. The sterilization process may be performed by other physical methods such as instant sterilization, high pressure instant sterilization, and ultrasonic treatment.
본 발명의 바람직한 실시형태에 있어서 상기 미생물을 증식시키는 과정은 자연 상태에 존재하는 미생물을 발효시키는 시스템을 채용한 것을 예로 들어 설명한 것이나, 별법으로 대나무의 재료를 깨끗이 세척한 후 여기에 발효 등의 유익한 기능을 제공하는 것으로 알려진 세균, 진균 등의 각종 미생물 또는 복합 미생물 제제 또는 효모 등을 첨가하여 발효시키는 방식 역시 바람직한 결과를 초래할 수 있다.In the preferred embodiment of the present invention, the process of propagating the microorganism has been described as an example of employing a system for fermenting microorganisms existing in a natural state, but alternatively, after washing the bamboo material cleanly, it is beneficial to fermentation and the like. Fermentation with the addition of various microorganisms or complex microbial agents or yeasts, such as bacteria, fungi, etc., which are known to provide function, may also produce desirable results.
이와 같이 미생물 균주를 인위적으로 첨가하여 발효 과정을 수행하는 경우에는 단독의 미생물 또는 미생물군을 사용하는 것보다는 유익한 기능을 하는 것으로 알려진 다양한 미생물들을 복합적으로 사용하는 것이 더욱 바람직한 결과를 초래할 수 있다.As such, when the microbial strain is artificially added to carry out the fermentation process, it may be more desirable to use a combination of various microorganisms known to have a beneficial function than to use a single microorganism or a group of microorganisms.
상기 발효 과정에 참가할 수 있는 미생물은 발효 등의 유익한 기능을 제공하는 것이라면 특별히 제한되지 않으며, 예를 들면 이들에 제한되지 않고, 세균, 진균류를 포함하며, 바람직하게는, 유산균, 광합성균, 바실러스균, 효모, 방선균 또는 이들의 혼합 미생물을 들 수 있다. The microorganism that can participate in the fermentation process is not particularly limited as long as it provides a beneficial function, such as fermentation, for example, is not limited to these, and includes bacteria, fungi, preferably, lactic acid bacteria, photosynthetic bacteria, Bacillus bacteria Yeast, actinomycetes or mixed microorganisms thereof.
유산균 또는 복합 유산균의 예로서는, 이들에 제한되지 않고, 스트렙토코커 스 락티스(Streptococcus lactis), 락토바실러스 플란타룸(Lactobacillus plantarum), 락토바실루스 브레비스(Lactobacillusbrevis), 락토바실러스 카세이(Lactobacillus casei), 락토바실루스 페르멘툼(Lactobacillus fermentum), 스트렙토코커스 락티스(Streptococcus lactis), 페디오코쿠스 아시드락티시이(Pediococcus acidlacticii), 페디오코쿠스 세레비시에(Pediococcus cerevisiae) 등, FDA 등재목록에 있는 GRAS(Generally Regarded As Safe) 미생물에 포함되는 유산균주를 배양 발효시킬 수 있다. 유산균은 기질의 섬유소 분해능이 뛰어나고, 인체, 동물 및 어류의 면역기능을 강화하는 것으로 알려져 있다. Examples of lactic acid bacteria or complex lactic acid bacteria include, but are not limited to, Streptococcus lactis , Lactobacillus plantarum , Lactobacillus brevis, Lactobacillus brevis, Lactobacillus casei , Lactobacillus Lactobacillus fermentum , Streptococcus lactis , Pediococcus acidlacticii , Pediococcus cerevisiae , etc., GRAS (Generally Regarded As) Safe) Lactic acid bacteria contained in microorganisms can be cultured and fermented. Lactic acid bacteria are known to have excellent fibrinolytic properties of substrates and to enhance immune function in humans, animals and fish.
광합성균으로서는 스테아로 써모필러스균(Bacillus stearothermophilus), 로도슈도모나스 캅술라타(Rhodopseudomonas capsulata)를 들 수 있으며, 바실러스 속의 균의 예로서는 바실러스 섭틸리스(Bacillus subtilis), 바실러스 나토(Bacillus natto), 바실러스 메가테리움(Bacillus megaterium), 바실러스 아밀로리퀴파시엔스(Bacillus amyloliquefaciens) 등을 들 수 있고, 방선균의 예로서는 스트렙토마이세스 그리세우스(Streptomyces griseus), 진균의 예로는 사카로마이세스 세레비시에(Saccharomyces cerevisiae), 칸디다 세이크(Candida sake), 모르티에렐라 엘롱가타(Mortierella elongata), 뮤코 시르시넬로이데스(Mucor circinelloides), 아스퍼길루스 속(Aspergillus sp.), 예컨대 아스퍼길루스 자포니쿠스(Aspergillus japonicus), 아스퍼길루스 오리재(Aspergillus oryzae), 페니실리움 왁스마니(Penicillium waksmani), 트리코데르마 리그노럼(Tricoderma lignorum), 리조푸스 속(Rhizopus sp.), 예컨대 리조푸스 올리고스포루스(Rhizopus oligosporus) 등을 들 수 있다. Examples of photosynthetic bacteria include Bacillus stearothermophilus , Rhodopseudomonas capsulata , and Bacillus subtilis , Bacillus natto , and Bacillus natto . Te Leeum (Bacillus megaterium), Bacillus amyl Lori rake Pacifico Enschede (Bacillus amyloliquefaciens) and the like, and examples of actinomycetes Streptomyces draw three mouse (Streptomyces griseus), examples of fungi Mai Seth serenity Vichy as Saccharomyces (Saccharomyces cerevisiae , Candida sake , Mortierella elongata , Mucor circinelloides , Aspergillus sp., such as Aspergillus japonicus ), Aspergillus peogil loose duck material (Aspergillus oryzae), Penny room mani Solarium wax (Penicillium waksmani), Rico van der Matthew League noreom (Tricoderma lignorum), Rizzo in crispus (Rhizopus sp.), For example, Rizzo crispus up and the like Spokane Ruth (Rhizopus oligosporus).
이러한 미생물 또는 복합 미생물 제제는 단지 설명할 목적으로 예를 들어 설명한 것이며, 유익한 것으로 알려진 다양한 미생물을 제한 없이 본 발명에 적용할 수 있음은 본 발명의 특징에 비추어 당업자에게 자명하다. 미생물의 입수는 자연 상태에 존재하는 균을 사용해도 좋으며, 상업적으로도 입수 가능한 것을 사용해도 좋고, 미생물 기탁기관으로부터 분양 받아서 사용해도 좋다.Such microorganisms or complex microbial preparations have been described by way of example only for purposes of explanation, and it will be apparent to those skilled in the art in view of the features of the invention that various microorganisms known to be beneficial may be applied to the invention without limitation. The microorganisms may be obtained by using microorganisms existing in a natural state, commercially available ones, or may be obtained from a microorganism depositing institution.
대나무 재료를 사용하여 미생물을 자연적으로 증식시키는 경우에 물론 유익한 균종 외에도 잡균의 번식이나 오염 가능성은 전혀 배제할 수는 없으나, 수차례의 시험 결과 어떠한 잡균의 존재도 검출되지 않았으며, 이러한 결과는 대나무의 고유한 항균 작용에 의해 잡균이 서식하지 못하게 되고, 간혹 잡균이 침투하게 되더라도 먼저 증식하여 우세하게 된 유익한 세균이나 진균류가 분비하는 효소나 길항 물질의 작용에 의해 잡균의 증식이 억제되거나 방해되는 것으로 보인다.The natural growth of microorganisms using bamboo materials, of course, in addition to beneficial strains, the possibility of propagation or contamination of various bacteria can not be excluded at all, but several tests have not detected the presence of various bacteria. The inherent antimicrobial action prevents the infestation of bacteria, and even if the bacteria invade, the proliferation of bacteria is inhibited or hindered by the action of enzymes or antagonists secreted by beneficial bacteria or fungi that proliferate first. see.
이와 같이 제조된 발효액은 pH 5 이하의 강산성을 띠고 있으며, 대나무의 재료로부터 유래하는 고유의 항균 작용으로 인하여 잡균이 서식하지 못하므로 인체에 해롭지 않고, 더욱이 상기 발효액에 대해서는 고온 멸균 처리하여도 기능 상에 문제가 발생하지 않으므로 물리적 방식으로 추가 멸균하여 안정성을 확보하는 것이 가능하다. 물론, 멸균처리를 하지 않은 원액에 대해서 독성 실험을 한 결과에 의하더라도 전혀 독성을 나타내지 않은 것으로 밝혀졌다.The fermentation broth prepared in this way has a strong acidity of pH 5 or less, and because of the inherent antibacterial action derived from the material of bamboo, it is not harmful to the human body because it is not harmful to the human body. It is possible to ensure stability by additional sterilization in a physical manner since no problem occurs. Of course, the results of the toxicity test on the unsterilized stock solution did not show any toxicity.
본 발명은, 다른 일면에 있어서, 상기한 바의 방법에 의해 얻어지는 대나무 발효액을 포함하는 위장관 질환의 치료용 제약 조성물 또는 기능성 식품 조성물을 제공한다.In another aspect, the present invention provides a pharmaceutical composition or functional food composition for the treatment of diseases of the gastrointestinal tract, comprising a bamboo fermentation broth obtained by the method as described above.
상기 추출물은 후술하는 시험예로부터 확인할 수 있는 바와 같이 인체의 장을 정화하고, 소화율을 향상시키고 면역력을 강화시킬 뿐만 아니라 장내 유익균총을 증식시키는 기능을 하며 소화기능, 위궤양, 숙취해소, 및 소화기 개선 작용을 나타내므로 위장관 질환의 개선 또는 소화 촉진을 위한 의약 조성물로서 사용될 수 있다. 대상이 되는 질환의 예로는 이들에 제한되지는 않고, 위암, 소화불량, 위염 숙취 또는 위궤양 등의 기타 소화기 장애를 들 수 있다.As the extract can be confirmed from the test examples described below, it cleanses the intestines of the human body, improves digestion rate and enhances immunity, as well as functions to proliferate the intestinal beneficial flora, and improve digestive function, stomach ulcer, hangover, and digestive organs. It can be used as a pharmaceutical composition for improving gastrointestinal diseases or promoting digestion because it exhibits action. Examples of diseases of interest include, but are not limited to, gastrointestinal cancer, dyspepsia, gastritis hangover or other digestive disorders such as gastric ulcer.
본 발명의 의약 조성물은 공지의 의약용 담체와 조합하여 제제화할 수 있다. 일반적으로는, 유효 성분을 약학적으로 허용할 수 있는 액상 또는 고체상의 담체와 배합하고, 또한 필요에 따라 용제, 분산제, 유화제, 완충제, 안정제, 부형제, 결합제, 붕괴제, 활택제 등를 가하고, 정제, 과립제, 산제, 분말제, 캡슐제 등의 고형제, 통상액제, 현탁제, 유제 등의 액제로 할 수 있다. 또한 이것을 사용전에 적당한 담체의 첨가에 의해 액상으로 만들 수 있는 건조품으로 할 수도 있다.The pharmaceutical composition of the present invention can be formulated in combination with a known pharmaceutical carrier. Generally, the active ingredient is blended with a pharmaceutically acceptable liquid or solid carrier, and a solvent, a dispersant, an emulsifier, a buffer, a stabilizer, an excipient, a binder, a disintegrant, a lubricant and the like are added as necessary and purified. And solid solutions such as granules, powders, powders, and capsules, liquids such as ordinary liquids, suspensions, and emulsions. Moreover, it can also be made into the dry goods which can be made into a liquid state by addition of a suitable carrier before use.
본 발명의 제약 조성물은 경구제나, 주사제, 점적용제 등의 비경구제 중의 어떠한 것에 의해서도 투여할 수 있다.The pharmaceutical composition of the present invention can be administered by any of parenteral agents such as oral preparations, injections, and drops.
의약용 담체는, 상기 투여형태 및 제형에 따라서 선택할 수 있고, 경구제의 경우는, 예를 들어 전분, 유당, 백당, 만니톨, 카복시메틸셀룰로스, 콘 스타치, 무기염 등이 이용된다. 또한 경구제의 조제에 있어서는 추가로 결합제, 붕괴제, 계면활성제, 윤택제, 유동성 촉진제, 교미제, 착색제, 향료 등을 배합할 수도 있다.A medical carrier can be selected according to the said dosage form and formulation, and, in the case of an oral preparation, starch, lactose, white sugar, mannitol, carboxymethylcellulose, corn starch, an inorganic salt, etc. are used, for example. Moreover, in preparation of an oral preparation, a binder, a disintegrating agent, surfactant, a lubricating agent, a fluidity promoter, a mating agent, a coloring agent, a fragrance | flavor, etc. can also be mix | blended further.
한편, 비경구제의 경우는 통상의 방법에 따라서 본 발명의 유효성분인 대나무 발효액을 포함하는 조성물을 희석제로서의 주사용 증류수, 생리식염수, 포도당 수용액, 주사용 식물유, 참기름, 낙화생유, 대두유, 옥수수유, 프로필렌글리콜, 폴리에틸렌글리콜 등에 용해 내지 현탁시켜서 필요에 따라 살균제, 안정제, 등장화제, 무통화제 등을 가함으로써 조제된다.On the other hand, in the case of parenteral preparations, the composition comprising the bamboo fermentation broth, the active ingredient of the present invention, according to a conventional method as a diluent, distilled water for injection, saline solution, glucose aqueous solution, vegetable oil for injection, sesame oil, peanut oil, soybean oil, corn oil It is prepared by dissolving or suspending in propylene glycol, polyethylene glycol or the like and adding a fungicide, stabilizer, isotonicity agent, analgesic agent and the like as necessary.
본 발명의 의약 조성물은 제제 형태에 따른 적당한 투여경로로 투여된다. 투여방법은 특히 한정할 필요는 없고, 내용, 외용 및 주사에 의해 투여할 수 있다. 주사제는, 예를 들어 정맥내, 근육내, 피하, 피내 등에 투여할 수 있다.The pharmaceutical composition of the present invention is administered by a suitable route of administration according to the formulation form. The administration method need not be particularly limited and can be administered by internal content, external application, and injection. Injections can be administered, for example, intravenously, intramuscularly, subcutaneously, intradermally or the like.
본 발명의 의약 조성물의 투여량은, 그 제제 형태, 투여 방법, 사용 목적 및 이것에 적용되는 환자의 연령, 체중, 증상에 따라서 적절히 설정되고, 일정하지 않지만 일반적으로는 제제 중에 함유되는 유효성분의 양은 성인 1일당 예컨대 10㎍∼200 ㎎/㎏이다. 물론 투여량은, 각종 조건에 의해서 변동하기 때문에, 상기 투여량보다 적은 양으로 충분한 경우도 있고, 또는 범위를 초과하여 필요한 경우도 있다.The dosage of the pharmaceutical composition of the present invention is appropriately set depending on the form of the preparation, the method of administration, the purpose of use, and the age, weight, and symptoms of the patient to be applied thereto. Amounts are, for example, 10 μg-200 mg / kg per adult. Of course, since the dose varies depending on various conditions, an amount smaller than the dose may be sufficient, or it may be necessary beyond the range.
본 발명은, 추가의 일면에 있어서, 상기 대나무 발효액을 유효성분으로 함유하는 기능성 식품 조성물, 및 음료 조성물을 제공한다.In still another aspect, the present invention provides a functional food composition and a beverage composition containing the bamboo fermentation broth as an active ingredient.
본 발명에 따른 대나무 발효액은 소화기능, 위궤양, 숙취해소, 및 소화기 개선 작용을 나타내는 다양한 약리 활성 및 생리 활성 물질을 포함하므로 이를 복용이 간편한 기능성 식품 또는 음료수 조성물의 유효 성분으로서 유용하게 사용될 수 있으며, 유효 성분으로서 대나무 발효액의 함량은 예컨대 조성물의 전체 중량을 기준으로 0.01~20%의 범위로 설정할 수 있다.Bamboo fermentation broth according to the present invention includes a variety of pharmacological and physiologically active substances exhibiting digestive function, gastric ulceration, hangover relief, and digestive action can be usefully used as an active ingredient of a functional food or beverage composition that is easy to take, The content of the bamboo fermentation broth as an active ingredient can be set, for example, in the range of 0.01 to 20% based on the total weight of the composition.
식품의 보조 성분들, 이들의 배합비 등은 당업계에 통상적으로 사용되는 것들을 특별한 기술적 어려움이 없이 채용할 수 있다.Auxiliary components of the food, their blending ratio and the like can employ those conventionally used in the art without particular technical difficulties.
본 발명에 따른 식품 조성물의 제형은, 예를 들면 정제, 캡슐, 과자류 또는 드링크제 등의 음료수의 형태로 제공될 수 있다. The formulation of the food composition according to the present invention may be provided in the form of a beverage, such as, for example, a tablet, capsule, confectionary or drink.
본 발명에 따른 식품 조성물은 원하는 조성물의 형태에 따라 통상의 보조제, 예를 들면 아스파탐, 사카린, 펙틴, 구아검, 탈지분유 및 올리고당으로 이루어진 군 중에서 선택되는 하나 이상의 성분을 추가해도 좋다.The food composition according to the present invention may add one or more ingredients selected from the group consisting of conventional auxiliaries, such as aspartame, saccharin, pectin, guar gum, skim milk powder and oligosaccharides, depending on the form of the desired composition.
본 발명의 식품 조성물에 있어서 감미료로는 통상적으로 사용되는 아스파탐, 사카린, 말리톨, 솔비톨, 자일리톨을 사용할 수 있다. 이들은 본 발명의 식품 조성물의 전체 중량을 기준으로 약 0.1-20 중량 %로 사용하는 것이 적절하다.As the sweetener in the food composition of the present invention, aspartame, saccharin, malitol, sorbitol, and xylitol commonly used may be used. They are suitably used at about 0.1-20% by weight based on the total weight of the food composition of the present invention.
본 발명에 따른 음료 조성물은 원하는 조성물의 형태에 따라 음료용 첨가제, 예를 들면 감미료, 아스파탐, 사카린, 펙틴, 구아검, 탈지분유, 말리톨, 솔비톨, 자일리톨, 올리고당, 구연산, 비타민 등을 첨가하여 기호도나 미감을 증대시키는 것이 좋다. 이들은 본 발명의 음료 조성물의 전체 중량을 기준으로 약 0.01-20 중량%로 사용하는 것이 적절하다.Beverage composition according to the present invention according to the form of the desired composition for the beverage additives, for example, sweeteners, aspartame, saccharin, pectin, guar gum, skim milk powder, malitol, sorbitol, xylitol, oligosaccharides, citric acid, vitamins, etc. It is good to increase the degree of taste and aesthetics. They are suitably used at about 0.01-20% by weight based on the total weight of the beverage composition of the present invention.
음료수는, 예를 들면, 대나무 발효액을 0.01-20% 중량으로 넣고 구연산을 0.01-2%, 사과산 0.01-2%, 타우린 0.1~2%, 비타민 C 0.01-2%, 비타민 B1 0.01-2%, 바이오틴 0.01-2%, 액상과당 0.01-2% 등을 넣고 단맛은 아스파탐을 0.01-2%를 넣어 기능성 음료수로 제조할 수 있다.Drinking water is, for example, a bamboo fermentation broth in an amount of 0.01-20%, citric acid 0.01-2%, malic acid 0.01-2%, taurine 0.1-2%, vitamin C 0.01-2%, vitamin B1 0.01-2%, Biotin 0.01-2%, liquid fructose 0.01-2% and put the sweetness aspartame 0.01-2% can be prepared as a functional beverage.
본 발명은, 더욱 추가의 일면에 있어서, 상기 대나무 발효액을 유효성분으로 함유하는 가축 및 어류의 사료 첨가제 조성물 및 어병억제제을 제공한다.In still another aspect, the present invention provides a feed additive composition and a fish disease inhibitor of livestock and fish containing the bamboo fermentation broth as an active ingredient.
본 발명의 사료 조성물 및 어병억제제는 어류양식용 또는 가축 사료에 혼합하여 사용하는 것이 편리하며, 사료의 중량을 기준으로 0.01% 내지 10.00%의 비율로 혼합하여 사용하는 것이 바람직하다. 그러나, 배합 비율은 사용 목적에 따라 변화시킬 수 있으며, 예를 들어, 개체의 성장 단계, 면역력, 생리적 상태 등에 따라 다소 변화시킬 수 있음은 자명하다. The feed composition and the fish bottle suppressant of the present invention is conveniently used for mixing fish farming or livestock feed, it is preferable to use a mixture of 0.01% to 10.00% based on the weight of the feed. However, it is apparent that the blending ratio may be changed depending on the purpose of use, and may be slightly changed depending on, for example, the growth stage, the immunity, the physiological state, etc. of the individual.
근래 국민의 소득 수준 향상과 더불어 육류 제품 소비량이 증가하고 있으나, 농산물 수입 개방화에 따라 값이 싸고 질이 낮은 수입품의 대량 유통으로 인하여 수입 농산물과의 경쟁력에서 불리한 위치에 있어 국내 양식 및 축산물 산업은 붕괴 위험에 처해 있는 실정이다. 따라서, 우리 정서에 알맞고 소비자들에게 친근감을 주면서도 기능성이 강화된 고품질 축산 및 어류 양식 제품의 개발이 절실히 요구되고 있다. In recent years, the consumption of meat products has increased along with the improvement of the income level of the people, but the domestic aquaculture and livestock industry has collapsed due to the disadvantage of competitiveness with imported agricultural products due to the mass distribution of cheap and low-quality imported products due to the opening of agricultural imports. The situation is in danger. Therefore, there is an urgent need for the development of high quality livestock and fish farming products that are in line with our sentiments and provide consumers with a friendly and functionally enhanced functionality.
더욱이, 1990년대 이후 보편화된 어류양식은 최근 들어 사료 값의 상승과 많은 질병발생으로 어려움을 겪고 있으며, 이를 극복하기 위한 노력이 계속되고 있다. 아직까지 양식에 사용되고 있는 사료의 대부분은 자연에서 어획되는 전갱이, 고등어 등의 생사료를 그대로 사용하거나 생사료에 배합사료를 혼합한 모이스트 펠렛(MP)을 사용함으로서 사료의 효율성 저하 및 환경오염 등 많은 문제를 안고 있으며, 이러한 문제를 해결할 수 있는 획기적인 사료의 개발은 이루어지고 있지 않은 실정임에 비추어 본 발명에 따른 대나무 발효액은 비브리오균을 포함한 어병의 발생을 근본적으로 차단할 수 있으므로 어병치료제 또는 사료의 첨가제 성분으로 효 율적으로 이용될 수 있다.Moreover, fish farming, which has been common since the 1990s, has recently suffered from rising feed prices and many diseases, and efforts have been made to overcome them. Most of the feeds still used in aquaculture use raw feeds such as horse mackerel and mackerel caught in nature, or use moist pellets (MP) mixed with mixed feeds in raw feeds to reduce feed efficiency and environmental pollution. In view of the fact that the development of a groundbreaking feed to solve this problem has not been made in view of the bamboo fermentation broth according to the present invention can fundamentally block the development of fish disease, including Vibrio bacteria, as an additive component of fish disease treatment or feed It can be used efficiently.
본 발명의 사료 조성물에 포함된 대나무 발효액은 가축 및 어류의 장을 정화하고, 소화율을 향상시켜 성장을 촉진하여 생산성을 증대시키고 어류 및 동물의 장내에서 유익균총을 증식시켜 질병을 예방할 뿐 아니라, 병원균 번식을 억제하고 어류 및 가축의 육질을 개선하는 등의 직접적인 효과 뿐만 아니라 질병의 발생억제에 따른 관리비의 저감이 가능하고 친환경적인 양식 산업을 촉진할 수 있을 것으로 기대된다. Bamboo fermentation broth contained in the feed composition of the present invention purifies the intestines of livestock and fish, improves digestibility, promotes growth to increase productivity, and proliferates beneficial bacteria in the intestines of fish and animals to prevent diseases, as well as pathogens. In addition to the direct effects of suppressing breeding and improving the quality of fish and livestock, it is expected to reduce the management costs associated with the suppression of disease and promote eco-friendly aquaculture industry.
<실시예><Example>
이하, 본 발명은 다음의 대표적인 실시예에 의하여 더욱 구체적으로 설명되나, 본 발명이 이들 실시예에 의해 어떤 식으로든 제한되는 것은 아니다. Hereinafter, the present invention is described in more detail by the following representative examples, but the present invention is not limited in any way by these examples.
실시예 1: 대나무 잎 유래의 미생물을 이용한 발효액의 제조Example 1: Preparation of fermentation broth using microorganisms derived from bamboo leaves
전남북 지역에서 자생하는 솜대(Phyllostachys nigra var. henonis) 밭에 떨어져 있는 대나무 잎 2 kg을 모아서 약간 경사진 바닥의 비닐 위에 직경 50cm의 범위로 분산시켜 배치하고 그 위에 증자시킨 맵쌀 밥 5kg을 골고루 얹은 다음 15일 동안 31 ℃ 전후에서 방치하여 미생물을 증식시켰다. 그 후 당액으로서 흑설탕 5kg을 첨가하여 동일한 조건에서 1주일 동안 발효시켜 우러나온 발효원액 800 ml을 수집하였다. 별도로 상기 대나무 잎을 수거하여 2시간 동안 끓인 후 이를 4℃로 냉각시킨 물로 상기 발효액을 100배 희석하여 이를 실온에서 30일 동안 숙성시켜 발 효액(pH 2.75) 80ℓ를 얻었다. 이 발효액을 0.2㎛의 필터로 여과시켜 고형분과 이물질을 제거하고 여액을 100℃로 2시간 동안 끓여 미생물을 사멸시킨 다음 자연 냉각시켜 사용할 때 까지 밀봉한 다음 실온에 보관하였다.Gather 2 kg of bamboo leaves in the field of Phyllostachys nigra var. The microorganisms were propagated by standing at around 31 ° C. for 15 days. Thereafter, 5 kg of brown sugar was added as a sugar solution, followed by fermentation for one week under the same conditions, to collect 800 ml of fermented stock solution. Separately, the bamboo leaves were collected and boiled for 2 hours, and then the fermentation broth was diluted 100 times with water cooled to 4 ° C. and aged at room temperature for 30 days to obtain 80 L of fermentation broth (pH 2.75). The fermentation broth was filtered through a 0.2 μm filter to remove solids and foreign matter, and the filtrate was boiled at 100 ° C. for 2 hours to kill microorganisms, which was then naturally cooled and sealed until use, and then stored at room temperature.
실시예 2: 발효액의 미생물학적 분석Example 2: Microbiological Analysis of Fermentation Broth
실시예 1에서 제조된 멸균전 상태의 발효액을 멸균 생리 식염수로 단계별 희석하여 Miles & Misra법(참고문헌: Miles, A.A. and Misra S.S.(1938, The estimate of bactericidal power of the blood. J. Hygiene, 38, 732-749)에 따라 각 희석액 25㎕를 Brain heart infusion agar(Difco, BHIA) 평판위에 접종하여 25 ℃에서 2일간 배양한 다음 살아난 균의 집락을 계수하여 산정하였다.The presterilized fermentation broth prepared in Example 1 was diluted stepwise with sterile physiological saline (Miles, AA and Misra SS (1938, The estimate of bactericidal power of the blood.J. Hygiene, 38). , 732-749) was inoculated on a brain heart infusion agar (Difco, BHIA) plate 25 ul incubated for 2 days at 25 ℃ and counted the colonies of the surviving bacteria.
1 ml당 총균수는 3.6 x 104 cfu/ml로서 비교적 적은 숫자의 균이 검출되었다. BHIA에서 자라난 집락 9개를 선택하여 동정한 결과 3종류의 균(BA-01, BA-02, BA-03)이 검출되었다. 이중 BA-01균이 66.7%로 가장 많고, BA-02는 22.2%, BA-03는 11.1%의 비율로 검출되었다. 분리균의 성상은 모두 그람양성균으로 OF에서 포도당을 이용하지 못했으며 산과 gas의 생성도 없었다. 운동성은 양성이었지만 BA-03균주는 운동성이 아주 미약하였다. 한편 catalase는 양성이었지만 oxidase, indole 및 황화수소의 생산은 모두 음성이었고 Rapid STR kit로 검사한 프로필은 각각 30000, 70000 및 61,000이었다.The total number of bacteria per ml was 3.6 x 10 4 cfu / ml, and a relatively small number of bacteria were detected. Three colonies (BA-01, BA-02, BA-03) were detected as a result of selecting nine colonies grown in BHIA. Among them, BA-01 was the most common at 66.7%, BA-02 at 22.2%, and BA-03 at 11.1%. All of the isolates were Gram-positive bacteria that did not use glucose in OF and produced no acid and gas. Mobility was positive, but BA-03 strain was very weak. Catalase was positive, but the production of oxidase, indole and hydrogen sulfide were all negative. Profiles tested with Rapid STR kit were 30000, 70000 and 61,000, respectively.
각 균주의 16S RNA 부분염기서열에 기초한 분자계통학적 분석에 의하면 대나 무 발효액의 배양에 관여한 미생물들은 표준 바실러스 섭틸리스(Bacillus subtilis) 균주와 바실러스 메가테리움(Bacillus megaterium) 균주와 각각 98% 및 96%의 유연결과를 가지는 것으로 계통학적으로 이들 종에 속하는 것으로 확인되었다. 한편, 발효에 관여한 진균류는 Phizopus 속에 속하는 것으로 확인되었고, 이를 2004. 8. 12일자로 대전광역시 소재 한국생명공학연구원 생물자원센터(Korean Collection for Type Cultures, KCTC)에 기탁하고 기탁번호 KCTC 10684BP를 부여 받았다. 이들 세균 및 곰팡이류는 모두 유익한 균주로 확인된 종들이다.Molecular systematic analysis based on the 16S RNA partial base sequence of each strain showed that the microorganisms involved in the culture of the fermentation broth were 98% of the Bacillus subtilis and Bacillus megaterium strains, respectively. And 96% of these species were identified systematically as belonging to these species. On the other hand, fungi involved in fermentation were identified as belonging to the genus Phizopus , which was deposited on August 12, 2004 to the Korean Collection for Type Cultures (KCTC), Daejeon Metropolitan City, and deposited accession number KCTC 10684BP. Granted. These bacteria and fungi are all species identified as beneficial strains.
실시예 3: 미생물을 이용한 발효액의 제조Example 3: Preparation of Fermentation Broth Using Microorganisms
실시예 2에서 확인된 미생물들을 깨끗이 세척한 대나무 잎과 고두밥의 배지에 첨가함으로써 실시예 1의 과정을 반복하여 미생물을 증식시킨 후 대나무 발효액을 제조하였다. By adding the microorganisms identified in Example 2 to the medium of freshly washed bamboo leaves and gourd rice, the process of Example 1 was repeated to multiply microorganisms to prepare a bamboo fermentation broth.
실시예 4: 미생물을 이용한 발효액의 제조Example 4 Preparation of Fermentation Broth Using Microorganisms
*복합 유산균 제품인 불가리스(판매원: 남양유업), 바실러스 섭틸리스(Bacillus subtilis) strain Nara-B2 (대전시 목원대학교 미생물생태자원 연구소 보유), 바실러스 메가테리움(Bacillus megaterium) strain Nara-B3 (대전시 목원대학교 미생물생태자원 연구소 보유) 및 누룩을 전체 용량이 200ml 정도 되게 혼합한 후, 깨끗이 세척한 대나무 잎과 고두밥의 배지에 첨가한 후 실시예 1의 과정을 반복하여 미생물을 증식시킨 후 대나무 발효액을 제조하였다. * Bulgaris (Sales: Namyang Dairy), Bacillus subtilis strain Nara-B2 (owned by the Institute of Microbiological Ecology, Mokwon National University), Bacillus megaterium strain Nara-B3 (Daewon Mokwon University) School microbiological ecological resources research institute) and Nuruk were mixed to a total capacity of about 200ml, and then added to the medium of freshly washed bamboo leaves and gourdap, and repeat the process of Example 1 to produce a microbial fermentation broth It was.
실시예 5: 약학적 제제Example 5: Pharmaceutical Formulations
제형의 형태(캅셀제)Form of Formulation (Capsule)
유효성분(실시예 1의 발효물 동결건조품) 50 mg50 mg of active ingredient (fermented product lyophilized product of Example 1)
락토오스 50 mgLactose 50 mg
전분 15 mgStarch 15 mg
스테아르산 마그네슘 2 mg2 mg magnesium stearate
결정성 셀룰로오스 10 mg10 mg of crystalline cellulose
캅셀들은 통상의 방법에 의해 제조하였으며 상기한 성분들을 원료로서 함유하였다. 상기 제제는 연질캅셀 또는 정제로도 제조할 수 있으며, 정제들은 통상의 장용피(예, 히드록시프로필메틸세룰로오스 프탈레이트), 당 코팅 또는 필름(에틸 세룰로오스)를 필요시 가할 수 있다.Capsules were prepared by conventional methods and contain the above ingredients as raw materials. The formulations may also be prepared as soft capsules or tablets, and tablets may be added if conventional enteric coating (eg hydroxypropylmethylcellulose phthalate), sugar coating or film (ethyl cellulose) is required.
실시예 6: 음료수의 제조Example 6 Preparation of Drinks
실시예 1에서 얻은 대나무 발효액 1 ℓ를 넣고, 여기에 구연산 20g, 아스파탐 17 g 및 비타민 C 30 g을 첨가하한 후 나머지를 정제수로 첨가하여 음료수 10 ℓ를 제조하여 저온 살균하였다. 음료수의 대조군은 동충하초 (종근당건강)을 사용하였다. 결과적으로 본 발명 음료수는 종래의 음료수 제품과 색깔, 풍미, 입안 감촉, 단맛 그리고 종합평가 면에서 차이가 없었다.1 L of the bamboo fermentation broth obtained in Example 1 was added, and 20 g of citric acid, 17 g of aspartame, and 30 g of vitamin C were added thereto, followed by addition of the rest with purified water to prepare 10 L of drinking water and pasteurization. As a control group of drinking water, Cordyceps sinensis (Chong Kun Dang Health) was used. As a result, the beverage of the present invention was not different from conventional beverage products in terms of color, flavor, mouth feel, sweetness, and overall evaluation.
시험예 1: 급성독성시험Test Example 1: Acute Toxicity Test
실시예 1에서 얻은 대나무 발효액을 실험동물에 대한 급성 독성 실험을 수행하고 그 독성유무를 관찰하였다. 실험용 동물은 생후 5-7주된 약 130g의 특이한 병변이 없는 수컷(specific pathogenic free male) sprague-dawely rat 50마리를 사용하였고 본 발명의 발효액과 음성대조군으로서 증류수를 사용하여 시험하였다. 먼저, 상기 래트들을 온도 22±2℃, 상대습도 53±2 % 및 형광등 조명(09:00 점등-18:00 소등)의 명암사이클, 150-300 Lux의 조도 조건을 갖춘 실험실 사육 상자에서 약 1주일 정도의 기간에 걸쳐 순화시킨 다음, 건강한 동물들만을 선택하여 평균체중이 일치하도록 각 군으로 나누고 일일 1회 20ml/kg의 양으로 14일 동안 강제 경구투여한 다음, 일반상태의 변화, 중독증상, 운동성, 외관, 자율신경, 체중변화 및 사망동물의 유무에 관하여 점검하였다.The bamboo fermentation broth obtained in Example 1 was subjected to acute toxicity experiments on the experimental animals and observed for its toxicity. The experimental animals used about 130 g of 5-1 week-old specific pathogenic free male sprague-dawely rats and were tested using the fermentation broth of the present invention and distilled water as negative controls. First, the rats were approximately 1 in a laboratory breeding box with a temperature cycle of 22 ± 2 ° C., a relative humidity of 53 ± 2%, a light and dark cycle of fluorescent lighting (09:00 light-18: 00 light off), and illumination conditions of 150-300 Lux. After a week-long period of time, the animals were selected and divided into groups to ensure that the average body weights were matched and forced oral administration for 20 days at 20ml / kg once daily, followed by changes in general condition and symptoms of intoxication. , Motility, appearance, autonomic nerves, weight change and the presence of dead animals were checked.
실험 결과에 의하면, 실험기간 동안 체중에 있어서 5% 이내의 변화를 보였으나 유의성은 없었고, 시료의 양을 시험동물에 투여가능한 최대량인 kg당 20ml의 최고 농도를 선정하였음에도 사망동물이 관찰되지 않아 개략의 치사량 산출은 불가하였으므로 LD50은 20ml/kg B.W. 이상인 것으로 나타났고, 특이한 일반증상이나 부검시 특이한 병변이 관찰되지 않았으므로 이를 종합적으로 판단해보면 상기 조성물은 독성이 없는 것으로 판명되었다.According to the experimental results, there was no significant change in body weight within 5% of the experimental period, but there was no significance, and even though the highest concentration of 20ml / kg was selected, the dead animals were not observed. LD 50 was found to be 20 ml / kg BW or more, and no specific lesions were observed at the time of autopsy or at the time of autopsy. Thus, the composition was found to be non-toxic.
시험예 2: 발효액의 어병세균에 대한 증식 억제 효과Test Example 2: Effect of Fermentation Broth on the Growth of Fish Bacteria
본 실험에 사용한 균의 유래는 하기 표 1에 나타냈다. 병원균은 BHIA에 25℃에서 1일간 배양한 다음 멸균 생리식염수에 현탁시켜 5 x 108 cell/ml와 5 x 105 cell/ml의 균액을 제조하였다. 0.45㎛ 멤브레인 필터로 여과 멸균한 발효액 2.5 ml과 2배의 농도로 고압 증기 멸균한 Brain Heart Infusion Broth(Difco) 2.5 ml을 혼합한 배양액 속에 각 균액을 0.1 ml 씩 접종한 다음 25℃에 배양하면서 일정 시간 후 각각 0.3 ml을 채취하였다. 채취한 배양액은 멸균생리식염수로 10단계 희석시킨 다음 BHIA에 접종하여 25℃에서 1일간 배양하여 자라난 집락을 계수하였다.The origin of the bacteria used in this experiment is shown in Table 1 below. The pathogen was incubated in BHIA for 1 day at 25 ° C. and then suspended in sterile saline to prepare 5 x 10 8 cell / ml and 5 x 10 5 cell / ml. Inoculate 0.1 ml of each microbial solution in a culture mixture of 2.5 ml of fermentation broth filtered with 0.45 ㎛ membrane filter and 2.5 ml of Brain Heart Infusion Broth (Difco) autoclaved at twice the concentration. 0.3 ml of each was taken after time. The collected culture solution was diluted 10 times with sterile physiological saline, inoculated in BHIA and incubated at 25 ° C. for 1 day to count colonies grown.
어병 세균의 in vitro에서 발육억제 효과는 고농도인 107을 접종한 경우 4균주 모두 접종 3시간 후부터 감소하기 시작하여 24시간 후에는 1/10~1/100으로 감소하였지만 본 발명의 대나무 발효액을 첨가하지 않은 대조군은 100~10,000배로 증가하여 고농도의 경우 어느 정도 균의 증식이 억제됨을 확인하였다. 이러한 발육억제 효과는 균의 종류에 따라 차이가 있었는데 패혈증 유발균인 비브리오균(도 1), 식중독 유발균인 에어로모나스균(도 2), 슈도모나스균(도면으로 나타내지 않음) 보다는 장내세균인 대장균(도 3)에 대한 억제 효과가 약한 것으로 나타났다. 한편, 저농도인 104를 접종하였을 때에는 모든 균이 시간이 경과함에 따라 균의 증식이 억제되어 접종 24시간 후에는 검출되지 않았고, 특히 Vibrio, 와 Aeromonas 균에 대한 억제효과가 뛰어나 접종 3~6시간 후에는 검출되지 않았다. 그러나 대조군에서는 접종 24시간 후에는 접종 농도의 10,000~1,000,000배로 균이 증식되었다. 따라서 본 발효액은 고농도의 어병세균에 대한 발육억제 효과는 약한 반면, 세균의 농도가 저농도인 경우에는 균의 종류에 관계없이 발육을 억제하는 것으로 나타났다.In vitro growth inhibitory effect of fish diseased bacteria was reduced after 3 hours of inoculation when inoculated with high concentration 10 7 and decreased to 1/10 to 1/100 after 24 hours, but the bamboo fermentation broth of the present invention was added. The control group did not increase the growth to 100 ~ 10,000 times confirmed that the growth of the bacteria to some extent at high concentrations. The growth inhibition effect was different depending on the type of bacteria, such as visceral bacteria such as Vibrio bacteria (Figure 1), aerobic bacteria causing aerobic bacteria (Figure 2), Pseudomonas bacteria E. coli (not shown) The inhibitory effect on FIG. 3) was found to be weak. On the other hand, when the low concentration of 10 4 was inoculated, the growth of all the bacteria was inhibited as time passed and was not detected after 24 hours of inoculation. Especially, it was excellent in inhibiting effect against Vibrio, and Aeromonas bacteria. It was not detected later. However, in the control group, bacteria were grown at 10,000 to 1,000,000 times the inoculation concentration after 24 hours of inoculation. Therefore, the fermentation broth showed a weak growth inhibitory effect on high concentrations of fish pathogens, while low concentrations of bacteria inhibited the growth regardless of the type of bacteria.
시험예 3: 생쥐의 혈중 알코올 농도에 미치는 효과시험Test Example 3 Effect Test on Blood Alcohol Concentration in Mice
체중 50g의 웅성 생쥐 20마리를 2군으로 나누고, 대조군은 생쥐에 알코올(40%, 15 ml/kg B.W)을 1시간 간격으로 2회 투여한 다음 1시간 후 동일량의 증류수를 투여하였으며, 다시 1시간이 경과한 후에 채혈하였다. 시험군은 대조군과 동일한 방식으로 알코올을 투여한 다음 1시간 후에 실시예 1에서 제조된 대나무 발효액을 동량으로 투여하였다.Twenty male males weighing 50 g were divided into two groups, and the control group was given alcohol (40%, 15 ml / kg BW) twice to the mice at 1 hour intervals, followed by the same amount of distilled water after 1 hour. Blood was collected after 1 hour. The test group administered the same amount of the bamboo fermentation broth prepared in Example 1 one hour after the alcohol administration in the same manner as the control group.
상기 각 생쥐의 혈액을 채취한 후 혈정을 분리한 다음 알코올 반응 혼합액(glycine buffer reagent, Sigma Co.) 3ml에 혈청 10㎕를 가한 다음 분광광도계를 사용하여 340 nm에서 흡광도를 측정하고 그 결과를 다음의 표 2에 나타냈다.After the blood of each mouse was collected, serum was separated, and then 10 μl of serum was added to 3 ml of alcohol reaction mixture (glycine buffer reagent, Sigma Co.), and then the absorbance was measured at 340 nm using a spectrophotometer. Table 2 shows.
상기 표 2의 결과로부터 알 수 있는 바와 같이, 본 발명의 발효액을 처리한 경우 대조군에 비하여 혈중 알코올 농도가 약 20% 정도 낮게 나타났다. 이와 같은 결과는 상기 대나무 발효액의 숙취해소제로서 가능성을 나타낸다.As can be seen from the results of Table 2, when the fermentation broth of the present invention was treated, the blood alcohol concentration was about 20% lower than that of the control group. Such a result shows the possibility as a hangover releasing agent of the bamboo fermentation broth.
시험예 4: 숙취해소에 대한 임상시험Test Example 4: Clinical Trial for Hangover Relief
상기 실시예 6에서 제조된 음료의 숙취해소에 미치는 효과를 검사하기 위하여 30대∼60대의 평소 주량이 소주 2병 전후인 남,녀 자원자 60명을 선발하여 3개군으로 나누어 다음 표 3에 나타낸 항목에 대하여 검사 방식을 설명하고 A 그룹에 대해서는 알코올 도수가 48%인 양주를 200cc를 30분에 걸쳐서 마시게 한 바로 직후 본 발명의 발효액 200cc를 마시게 하고 혈중 알코올 농도를 매 30분 단위로 측정하였다. B 그룹은 A 그룹과 동일한 방식으로 양주를 마시게 하고 곧바로 동일한 양의 물을 마시게 하였다. C 그룹은 양주를 마시기 전에 동일 양의 발효액을 섭취하게 하였다. 측정 결과를 다음의 표 3에 나타내었다.In order to examine the effect on the hangover of the beverage prepared in Example 6, 60 male and female volunteers in their 30s and 60s, who usually had two bottles of soju, were selected and divided into three groups. The test method was described, and for group A, 200cc of alcoholic beverage having 48% alcohol content was drunk 200cc over 30 minutes, and 200cc of fermentation broth of the present invention was drunk, and blood alcohol concentration was measured every 30 minutes. . Group B was allowed to drink liquor in the same manner as group A and immediately drink the same amount of water. Group C had the same amount of fermented broth before drinking liquor. The measurement results are shown in Table 3 below.
위 결과로부터 알수 있는 바와 같이, 본 발명의 발효액 음료를 마신 경우에는 대조군에 비하여 약 23% 정도(90분 경과 기준) 개선된 알코올 분해 효과를 나타내었다.As can be seen from the above results, when drinking the fermented beverage of the present invention showed about 23% (90 minutes) improved alcohol degradation effect compared to the control.
*시험예 5: 대나무 발효액의 위장질환 개선 효과 Test Example 5: Effect of improving the gastrointestinal disease of bamboo fermentation broth
실험동물은 생후 6-8주된 약 150g의 특이한 병변이 없는 수컷(specific pathogenic free male) 스프라그-다울리 쥐를 한국화학연구소 안정성 연구부로부터 구입하여 실험에 사용하였다. 사료와 물은 자유롭게 먹게 하며, 사육실의 온도는 21~24 ℃ , 상대 습도는 40~60%로 유지하였다. 또한 사육실의 밤과 낮이 12시간마다 반복되도록 조절하였다. Experimental animals were purchased from the Korea Research Institute of Chemical Technology and stability of the 6-8 weeks old, about 150 g of specific pathogenic free male Sprague-Dawley rats. Feed and water were allowed to eat freely, and the temperature in the nursery was maintained at 21 ~ 24 ℃ and relative humidity 40 ~ 60%. In addition, the night and day of the nursery was adjusted to repeat every 12 hours.
랫트는 흉부를 절개한 후 심장의 정맥을 이용해 PBS로 체내의 혈액을 제거한 뒤 4% 파라포름알데히드(Para-formaldehyde)로 관류 고정하였다. 고정된 쥐의 위를 적출하여 microcryocut를 이용하여 3-5㎛두께의 동결절편을 제작하였다. 준비된 위 절편은 PBS로 여러 번 씻어주었다. 30% 과산화 수소로 전처리하여 씻어준 후 vectastain ABC kit의 normal goat serum을 0.5% BSA/PBS에 섞어 1시간 동안 반응시켜 비특이적 단백질의 결합을 막았다. TH 일차(primary) 항체와 0.5% BSA/PBS (1:10000) 희석하여 실온에서 1시간에서 2시간 정도 반응시켰다. 시료를 PBS로 3회 세척한 후, 이차 항체(biotinylated anti-rabbit IgG in 1% BSA/ PBS)를 이용하여 1시간 동안 반응시킨 다음 1% BSA/ PBS로 두 번 씻어냈다. 사용하기 30분 전에 vectastain ABC kit (1:50)를 희석하여 50분 동안 반응시킨 후 PBS로 2번 씻고, 50ml DAB 용액과 0.2M PB 용액에 30% hydrogen peroxide 12㎕를 첨가한 용액으로 5분간 염색시켜 0.1M PB로 5분간 반응을 동시에 정지시킨 다음, 광학 현미경으로 관찰하였다. 그리고 위에서의 세포 계수(counting)는 각각 처리된 4개의 염색된 절편을 선택하여 400에서 전 층을 대상으로 조직절편의 단위면적당 TH가 발현된 세포의 수를 측정하였고 계수 결과는 윌콕슨의 순위합 검정 (Wilcoxson rank sum test)를 이용하였다.The rat was incised to the chest, and then blood was removed from the body with PBS using a vein of the heart, followed by perfusion fixation with 4% para-formaldehyde. The stomach of the immobilized rat was extracted and frozen sections of 3-5 μm thickness were prepared using a microcryocut. The prepared sections were washed several times with PBS. After washing with 30% hydrogen peroxide, normal goat serum of vectastain ABC kit was mixed with 0.5% BSA / PBS for 1 hour to prevent nonspecific protein binding. 0.5% BSA / PBS (1: 10000) was diluted with TH primary antibody and reacted at room temperature for 1 to 2 hours. The sample was washed three times with PBS, and then reacted with a secondary antibody (biotinylated anti-rabbit IgG in 1% BSA / PBS) for 1 hour and then washed twice with 1% BSA / PBS. 30 minutes prior to use, dilute vectastain ABC kit (1:50) for 50 minutes, wash twice with PBS, and add 5 ml of 30% hydrogen peroxide to 50 ml DAB solution and 0.2 M PB solution for 5 minutes. The reaction was stopped at the same time by 0.1 M PB for 5 minutes and then observed under an optical microscope. The cell counts above were selected from each of the four stained sections treated, and the number of TH-expressing cells per unit area of the tissue sections was measured at 400 in all layers, and the counting result was Wilcoxon's rank sum. The Wilcoxson rank sum test was used.
Eisenhofer 등 (Eisenhofer E., Hansson G., Harta S., Hoffman G.. Non-neuronal dopamine in the gastrointestinal system. Clinical and Exp. Pharm. & Phys. Supplement 26:14-22, 1999.)에 따르면 위(Stomach)에서 dopamine이 생성되고 TH activity를 가진다고 보고하였다. 그리고 Elsenhofer 등이 위에서의 도파민(dopamine)은 위장관(gastrointestinal tract)의 보호물질로 작용하며 면역조직화학(Immunohistochemistry)과 웨스턴 블롯팅(western blotting)을 이용하여 위에서의 TH-IR 세포들이 존재함을 보고하였다. According to Eisenhofer et al. (Eisenhofer E., Hansson G., Harta S., Hoffman G. Non-neuronal dopamine in the gastrointestinal system.Clinical and Exp. Pharm. & Phys. Supplement 26: 14-22, 1999.) (Stomach) reported dopamine production and TH activity. Elsenhofer et al reported that dopamine in the stomach acts as a protective agent for the gastrointestinal tract and that TH-IR cells in the stomach are present using immunohistochemistry and western blotting. It was.
실시예 1에 따른 대나무 발효액을 복강에 처리했을 때 위에서의 TH 반응 양상 변화를 관찰하였다. 먼저 대조군을 살펴보면 TH-IR 세포들이 점막근판 (Muscularis mucosae)에서 주로 나타났으며 위선 (Gastric glands)에서는 갈색의 씨앗 모양을 하고 있는 한 두 개의 TH-IR 세포들을 관찰하였다.When the bamboo fermentation broth according to Example 1 was treated in the abdominal cavity, changes in the TH reaction pattern were observed. First, the control group showed that TH-IR cells were mainly found in the muscularis mucosae and gastric glands, one or two TH-IR cells in the shape of brown seeds.
반면에 시료를 처리한 조직들에서는 대조군에 비해 점막근판에서 더 많이 발현됨을 확인하였고 발현영역이 위선까지 더 넓어짐을 확인하였다. 그리고 위 조직에 나타난 TH-IR 세포를 일정한 단위면적에서 cell counting을 하여 비교해 본 결과, 대조군에 비한 실험군들의 TH-IR 세포들의 수가 더 많이 나타났으며 최대 2.5배 이상의 숫적 증가를 확인하였다. On the other hand, the tissues treated with the sample were found to be expressed more in the mucosa near the control group, and the expression region was confirmed to be wider up to the hypocrisy. And as a result of comparing cell counting of TH-IR cells in a certain unit area, the number of TH-IR cells in the experimental group was higher than that of the control group, and the number increased by more than 2.5 times.
시험예 6: 본 발명 제품의 임상 시험Test Example 6: Clinical Trial of the Product of the Present Invention
상기 실시예 6에서 제조된 기능성 음료의 효과를 검사하기 위하여 20대∼70대의 남,녀 자원자 250명을 5개군으로 나누어 다음 표 4에 나타낸 항목에 대하여 검사 방식을 설명하고 익숙할 수 있도록 연습시킨 후 1일 평균 600cc를 물 대용으로 음용하는 방식으로 임상 검사를 실시하였다. In order to examine the effects of the functional beverage prepared in Example 6, 250 male and female volunteers in their 20s to 70s were divided into five groups, and the test methods were described and practiced to be familiar with the items shown in Table 4 below. Afterwards, the clinical test was performed in a manner of drinking an average of 600 cc as a water substitute.
평가 기준은 4 단계로 나누고 점수가 높을수록 좋은 결과를 나타내는 것으로 정하였다. 즉, 1: 차이가 없음, 2: 다소 개선됨, 3: 상당히 개선됨, 4: 현저히 개선됨. 제시된 값은 평균치이다. The evaluation criteria were divided into four stages, and the higher the score was, the better the result was. That is, 1: no difference, 2: somewhat improved, 3: significantly improved, 4: significantly improved. The values presented are averages.
위의 결과로부터 알 수 있는 바와 같이, 본 발명의 기능성 음료 조성물은 생체의 신진 대사를 활성화시키고, 위장관의 상태를 개선시키는 등 여러 가지 기능성을 가짐을 알 수 있다. As can be seen from the above results, it can be seen that the functional beverage composition of the present invention has various functionalities such as activating the metabolism of the living body and improving the state of the gastrointestinal tract.
시험예 7: 돼지에 대한 성장촉진 시험 Test Example 7: Growth promotion test for pigs
생후 1개월된 100두의 랜드레이스(Landrace)종 돼지에 대해서 본 발명의 대나무 발효액의 성장 촉진 기능 여부를 시험하였다. 시험을 개시한지 1 개월 동안은 젖먹이 사료를 공급하고, 그 후 2개월 동안은 육성돈 사료를 공급하였으며, 나머지 5개월까지는 비육돈 사료를 공급하였다. 대조군에 대해서는 일반사료를, 시험군에 대해서는 본 발명의 대나무 발효물을 0.50중량%로 첨가한 사료를 공급하였다. 시험결과를 다음의 표 5에 나타낸다.100 Landrace pigs, one month of age, were tested for growth promoting function of the bamboo fermentation broth of the present invention. Feeding feed was provided for 1 month after the start of the trial, feed for pigs for 2 months thereafter, and feed for feed for up to 5 months. For the control group, the feed was added to the feed and 0.50% by weight of the bamboo fermentation product of the present invention for the test group. The test results are shown in Table 5 below.
위 결과로부터 알수 있는 바와 같이, 성장 촉진 효과를 나타내는 1두당 평균 중량을 보면, 대조군에 있어서는 73.28kg, 대나무 발효액을 첨가한 집단에서는 76.88 kg으로 현저한 차이를 나타내었다.As can be seen from the above results, when looking at the average weight per head showing the growth promoting effect, there was a significant difference of 73.28kg in the control group, 76.88 kg in the bamboo fermented broth.
지금까지 설명된 바와 같이 본 발명에 따른 대나무 발효액은 자연 상태에 존재하는 곰팡이 등의 미생물을 발효시켜 제조한 것이므로 인체에 해가 없고, 다양한 기능성을 나타낸다. 따라서, 본 발명의 대나무 발효액은 잡균의 서식을 억제하는 기능을 함으로써 가축 및 어류 양식과 같이 환경오염의 영향을 크게 받는 양식, 축산 분야에서 사용될 수 있으며, 상기 발효액은 인체의 신진 대사를 활성화시키고, 위장관의 상태를 개선시키는 기능을 나타내므로 숙취해소제 등으로도 사용할 수 있다. As described so far, the bamboo fermentation broth according to the present invention is prepared by fermenting microorganisms such as molds existing in a natural state, and thus does not harm the human body and exhibits various functionalities. Therefore, the bamboo fermentation broth of the present invention can be used in aquaculture, animal husbandry, which is greatly affected by environmental pollution, such as livestock and fish farming by functioning to suppress the culture of various bacteria, the fermentation broth activates the metabolism of the human body, It can be used as a hangover relievers because it shows the function of improving the condition of the gastrointestinal tract.
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| KR101285206B1 (en) * | 2012-01-06 | 2013-07-11 | 담양군 | Fermented beverage using bamboo sprout |
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