KR100711930B1 - Ginseng polysaccharides composition having the use of stabilization and propagation of lactic acid bacteria - Google Patents
Ginseng polysaccharides composition having the use of stabilization and propagation of lactic acid bacteria Download PDFInfo
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- KR100711930B1 KR100711930B1 KR1020050068790A KR20050068790A KR100711930B1 KR 100711930 B1 KR100711930 B1 KR 100711930B1 KR 1020050068790 A KR1020050068790 A KR 1020050068790A KR 20050068790 A KR20050068790 A KR 20050068790A KR 100711930 B1 KR100711930 B1 KR 100711930B1
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- KR
- South Korea
- Prior art keywords
- ginseng
- polysaccharide
- lactic acid
- extraction
- acid bacteria
- Prior art date
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Abstract
본 발명은 인삼 다당체를 함유하는 조성물에 관한 것이다. 상기 조성물은 유산균 동결건조 보호효과, 유산균 보관 안정화 효과, 유산균 증식 효과 및 대장암 등의 원인효소의 생산억제효과가 우수하다. The present invention relates to a composition containing ginseng polysaccharides. The composition is excellent in the lactic acid bacteria freeze-drying protective effect, lactic acid bacteria storage stabilization effect, lactic acid bacteria growth effect and the inhibitory effect of the production of causative enzymes such as colon cancer.
인삼 다당체, 유산균 동결건조 보호효과, 유산균 보관 안정화 효과, 유산균 증식 효과, 대장암 등의 원인효소의 생산억제효과 Ginseng Polysaccharide, Lactobacillus Lyophilization Protection Effect, Lactic Acid Bacteria Storage Stabilization Effect, Lactic Acid Bacteria Growth Effect, Inhibitory Effect of Causative Enzymes Production of Colorectal Cancer
Description
도 1은 세파덱스 G-75 컬럼에서의 홍삼다당체 분획의 용출패턴을 나타낸 것이다. Figure 1 shows the elution pattern of the red ginseng polysaccharide fraction in the Sephadex G-75 column.
본 발명은 활성성분으로서 인삼 다당체를 함유하는 조성물에 관한 것이다. 또한, 본 발명은 유산균 동결건조 보호효과, 유산균 보관 안정화 효과, 유산균 증식 효과 및 대장암 등의 원인효소의 생산억제효과를 지닌 조성물에 관한 것이다. The present invention relates to a composition containing ginseng polysaccharide as an active ingredient. In addition, the present invention relates to a composition having a lactic acid bacteria freeze-drying protective effect, lactic acid bacteria storage stabilization effect, lactic acid bacteria growth effect and the production inhibitory effect of causative enzymes such as colon cancer.
인삼은 식물 분류학상 오가과 인삼속에 속하는 다년생 숙근초로서 지구상에 약 11 종이 알려져 있으며, 대표적인 종의 예로는 아시아 극동 지역 (북위 33-48°: 한국, 북만주, 러시아 일부)에 자생하며 약효가 매우 우수한 고려 인삼 (Panax ginseng C.A.Meyer); 미국, 캐나다에서 자생 및 재배하고 있는 미국삼 (Panax quinquefolium L.); 중국 운남성 동남부로부터 광서성 서남부 지역에서 야생 또는 재배하고 있는 전칠삼 (Panax notoginseng F.H.Chen); 및 일본, 중국 서남부, 네팔 에 이르기까지 분포되어 있는 죽절삼 (Panax japonicus C.A.Meyer) 등이 있다.Ginseng is a perennial root of the genus Ogasus ginseng by plant taxonomy. About 11 species of ginseng are known on the earth. For example, Ginseng is native to the Far East of Asia (33-48 ° north: Korea, Northern Manchuria, and Russia) and has very good efficacy. Korean ginseng (Panax ginseng CAMeyer); American ginseng (Panax quinquefolium L.) growing and growing in the United States and Canada; Panax notoginseng F.H.Chen, wild or cultivated in southwestern Guangxi province from southeastern Yunnan Province of China; And Panax japonicus C.A.Meyer, which is distributed in Japan, southwest China, and Nepal.
인삼은 신농본초경에 상품으로 수재되어 있을 뿐만 아니라 예로부터 귀중한 보약으로 사용되어 오고 있다. 지금까지 많은 약리 실험을 통해 인삼은 스트레스에 대한 생체의 비특이적 저항성을 강화시키고 항산성 작용을 갖고 있음이 밝혀져 있다. 그 외에 고혈압의 개선, 인슐린 작용증강, 알록산 (ALLOXAN) 당뇨 마우스에서의 혈당강하효과, 흰쥐의 간 RNA 합성, 단백질 합성, 당 및 지질대사 촉진효과, 항암효과 등이 있음이 밝혀져 있다. Ginseng is not only stored as a commodity in the new agricultural plant, but has been used as a valuable medicine since ancient times. Many pharmacological experiments to date have revealed that ginseng strengthens the nonspecific resistance of the body to stress and has an acidic action. In addition, it has been shown to improve hypertension, enhance insulin action, hypoglycemic effect in alloxan diabetic mice, liver RNA synthesis in rats, protein synthesis, sugar and lipid metabolism promoting effect, and anticancer effect.
상기 인삼은 주로 한국, 중국, 일본 등의 아시아 국가에서 생약의 형태로 정신 의학적 질병, 신경계의 질병 및 당뇨병 등의 여러 가지 질병에 대해 사용되어 왔으며, 상기 인삼의 주요 성분인 사포닌 (saponin)은 강장, 강정, 진정조형 및 항고혈압 등에 효과를 보이는 것으로 알려져 있다. The ginseng has been used for various diseases such as psychiatric diseases, nervous system diseases and diabetes in the form of herbal medicine mainly in Asian countries such as Korea, China, Japan, and saponin, which is a major component of the ginseng, is tonic It is known to have effects on stiffness, sedation, and antihypertension.
현재 인삼의 사용은, 재배하여 채취한 상태 그대로의 수삼을 상온에서 건조시킨 백삼, 수삼을 98-100℃에서 가열 처리하여 제조되는 홍삼, 또는 120-180℃에서 가열 처리하여 제조되는 열처리 인삼 (선삼, 흑삼 등)의 형태로 사용되고 있다. 인삼의 성분은 인삼 특유의 성분인 다당체, 인삼사포닌 (진세노시드; ginsenoside)과 타 식물에도 존재하는 폴리아세틸렌, 핵산염기배당체 (뉴클레오시드), 알카로이드, 페놀성 화합물, 단백질 및 아미노산, 향기성분 등이 밝혀져 있다. 인삼의 다당체 등을 다시 산성, 중성 및 알카리성 다당체로 분류하고, 인삼의 특이성분인 진세노시드는 34종 (백삼 22 종, 홍삼 30 종) 분리하였으며, 그 외에도 최근에 화합물 K (20-O-β-D-글루코피라노실-20(S)-프로토파낙사디올)와 같은 인삼 사포닌 대사체들이 분리되어 보고되고 있다. 현재 알려진 바에 따르면, 인삼의 약리 효능 중 항암효과, 항당뇨효과, 면역증강작용 등은 인삼의 다당체와 사포닌에 기인하는 것으로 밝혀졌다. Currently, the use of ginseng is white ginseng, dried ginseng as it is grown and collected at room temperature, red ginseng prepared by heating at 98-100 ° C, or heat-treated ginseng prepared by heating at 120-180 ° C. , Black ginseng, etc.). The components of ginseng are polysaccharides, ginsenosides, polyacetylene, nucleic acid base glycosides (nucleosides), alkaloids, phenolic compounds, proteins, amino acids, fragrances that are also present in ginseng. Etc. are revealed. Ginseng polysaccharides were further classified into acidic, neutral and alkaline polysaccharides, and ginsenosides, which are specific to ginseng, were isolated from 34 species (22 white ginseng and 30 red ginseng), and recently, compound K (20-O-β). Ginseng saponin metabolites such as -D-glucopyranosyl-20 (S) -protopanaxadiol) have been reported separately. As currently known, the anti-cancer effect, anti-diabetic effect, and immuno-enhancing effect among the pharmacological effects of ginseng were found to be due to the polysaccharide and saponin of ginseng.
사람의 장내에는 약 300 종류, 100 조 개 이상의 세균이 서식하고 있으며, 사람의 건강에 상당한 영향을 주고 있다. 장내에 서식하는 장내세균은 사람의 건강을 지켜주는 유용균과 나쁜 영향을 주는 유해균이 있으며 이들 양자의 균형에 의하여 건강 상태가 조절되고 있다. 유용균의 대표적인 것이 유산균인 락토바실러스 속 (Lactobacillus sp.), 비피도박테리움 속 (Bifidobacterium sp.) 균주이며, 유해균의 대표적인 균주로는 대장균, 식중독균, 포도구균이며 이들은 장내의 부패를 촉진하여 노화가 빨리 일어나게 하고 발암물질을 생산한다. About 300 kinds of bacteria and more than 100 trillion bacteria live in the intestines of humans, and have a significant impact on human health. Intestinal bacteria living in the intestine are useful bacteria that protect human health and harmful bacteria that have a bad effect, and the health condition is controlled by the balance of both. Representative strains of useful bacteria are Lactobacillus sp. And Bifidobacterium sp., Which are lactic acid bacteria, and representative strains of harmful bacteria are Escherichia coli, food poisoning bacteria and Staphylococcus aureus. Get up quickly and produce carcinogens.
유산균은 포도당 또는 유당과 같은 탄수화물을 분해 이용하여 유산을 많이 만드는 세균 (박테리아)으로서 단백질을 분해하지만 부패시키는 능력이 없으며 인체에 해로운 물질들을 생성하지 않으며 유익한 작용을 하는 세균을 말한다. 유산균은 락토바실러스 속, 비피도박테리움 속 균주 이외에도 스트렙토코커스 속 (Streptococcus sp.), 페디오코커스 속 (Pediococcus sp.), 루코노스톡 속 (Leuconostoc sp.) 등이 있다. 유산균은 다른 균주들과 달리 단백질을 분해하여 암모니아, 아민, 황화수소 등의 독성물질을 만들지 않는다. 따라서, 유산균이 아무리 장에 많이 증식하여도 독성물질을 만들지 않기 때문에 몸에 유익한 작용만을 하는 균으로 생각되고 있다. Lactobacillus is a bacterium (bacteria) that breaks down carbohydrates such as glucose or lactose and makes a lot of lactic acid. It is a bacterium that does not decompose proteins but does not decay, does not produce harmful substances, and has a beneficial effect. The lactic acid bacteria include Streptococcus sp., Pediococcus sp., And Leuconostoc sp. In addition to Lactobacillus and Bifidobacterium strains. Unlike other strains, lactic acid bacteria do not break down proteins to produce toxic substances such as ammonia, amines and hydrogen sulfide. Therefore, no matter how much lactic acid bacteria multiply in the intestine, it does not make a toxic substance, so it is thought to be a bacterium having a beneficial effect on the body.
이러한 유산균의 생리활성은 첫째로, 유기산을 생산하여 장내의 pH를 저하시 킴으로써 장내 유해세균의 증식을 억제하며 정상적인 장내균총을 유지시켜 준다. 둘째로, β-갈락토시다아제를 생산하여 유당소화 불량증이 있는 사람에게 유리하게 작용한다. 셋째로, 장내의 pH를 저하시켜 발암 (carcinogenesis)에 직접적인 영향을 미치고 유산, 초산은 부패균의 성장을 억제하여 N-니트로소 화합물, 페놀성 산물 등의 발암물질을 감소시켜 항암활성을 나타낸다. 넷째로, 비피도박테리아는 히드록시메틸글루타레이트를 생산하여 HMG-CoA 환원효소를 억제하는 작용을 나타내므로 콜레스테롤 저하효과를 나타낸다. 다섯째로, 비타민 B1, B2, B6, B12, 니코틴산, 엽산 등을 합성한다. 여섯째로, 설사의 원인균을 억제하고 장내균총을 정상화함으로써 설사를 방지한다. The physiological activity of these lactic acid bacteria firstly, by reducing the pH of the intestine by producing an organic acid inhibits the growth of harmful bacteria in the intestine and maintains the normal intestinal flora. Secondly, it produces β-galactosidase, which favors people with lactose intolerance. Third, lowering the pH of the intestine has a direct effect on carcinogenesis (carcinogenesis), lactic acid, acetic acid inhibits the growth of decaying bacteria to reduce the carcinogens such as N-nitroso compounds, phenolic products exhibit anticancer activity. Fourthly, Bifidobacteria produces hydroxymethylglutarate and thus exhibits the effect of inhibiting HMG-CoA reductase and thus lowers cholesterol. Fifth, vitamins B 1 , B 2 , B 6 , B 12 , nicotinic acid and folic acid are synthesized. Sixth, diarrhea is prevented by suppressing the causative organism of diarrhea and normalizing the intestinal flora.
장내세균은 섭취하는 음식물, 생활환경, 스트레스에 의해 영향을 받아 유해균과 유익균의 균형을 상실할 수 있다. 예를 들면, 육류의 섭취가 많은 서양인들은 장내 유익균의 감소를 가져오게 되나, 채소류의 섭취가 많은 동양인들은 유익균의 감소가 적어 서양인보다 대장암 발생이 낮다. Enterobacteriaceae may be affected by the foods they eat, their living environment and stress, resulting in a loss of balance between harmful and beneficial bacteria. For example, Westerners who consume a lot of meat will have a lower intestinal beneficial bacterium, but Asians who consume more vegetables will have a lower incidence of colorectal cancer than Westerners due to a lower number of beneficial bacteria.
장내세균총 중 유익균인 유산균을 우세균으로 유지시켜 주기 위한 방법으로는 두 가지 방법이 시도되고 있다. 첫째는, 비피더스균을 경구적으로 섭취하는 방법이다. 이 방법의 단점은 장내의 비피더스균은 숙주 (즉, 장내세균이 살고 있는 사람이나 동물) 고유의 특이성을 나타냄으로써 경구적으로 섭취한 비피더스균은 대장 내에서 정착하지 못하고 배설되는 것으로 나타났다. 둘째는, 비피더스균에게 선택적으로 이용되는 물질, 즉 비피더스 인자 (bifidogenic factor)를 경구 적으로 섭취하는 방법으로 우리가 음식물을 통해서 비피더스 인자를 섭취한다면 어렵지 않게 유산균을 증식시킬 수 있을 것이다. 유산균 증식인자란 유산균에게 선택적으로 이용되며 소화관에서 흡수되지 않는 물질로 올리고당이 유효하다. 즉, 소화효소에 의해서 분해되지 않고 대장까지 도달하며 대장에 서식하고 있는 유산균을 비롯한 장내 유용균에게 이용되므로 유해세균 또는 병원성 세균의 증식을 억제하는 물질을 말한다. Two methods have been attempted to maintain lactic acid bacteria, which are beneficial bacteria in the intestinal flora, as dominant bacteria. First, the method of orally ingesting bifidus bacteria. Disadvantage of this method is that the intestinal bifidus bacillus exhibits the specificity of the host (ie, the person or animal in which the intestinal bacterium lives), so that the orally ingested bifidus bacterium cannot be excreted in the large intestine. Secondly, by orally ingesting bifidogenic factor, a substance selectively used by bifidus bacteria, if we ingest bifidus factor through food, we can easily grow lactic acid bacteria. Lactobacillus growth factors are selectively used by lactic acid bacteria and oligosaccharides are effective substances that are not absorbed in the digestive tract. In other words, it is a substance that inhibits the growth of harmful bacteria or pathogenic bacteria because it is used by intestinal useful bacteria including lactic acid bacteria in the colon and reaches the colon without digestion by digestive enzymes.
유산균 증식인자의 종류는, 첫째, 아미노 당 (amino sugar)이며 아미노 당을 함유하는 비피더스 인자에는 β-에틸-N-아세틸-D-글루코사민, N-아세틸글루코사민 등이 있다. 둘째, 락토오스, 이눌린 등과 같이 천연 올리고당이 있다. 셋째, 푸룩토올리고당 (fructooligosaccharide)과 같이 합성 올리고당이 있다. The lactic acid bacteria growth factor is, firstly, an amino sugar, and the bifidus factor containing the amino sugar includes β-ethyl-N-acetyl-D-glucosamine, N-acetylglucosamine and the like. Second, there are natural oligosaccharides such as lactose and inulin. Third, there are synthetic oligosaccharides, such as fructooligosaccharides.
유산균들은 유기산을 생성하는 등의 반응을 촉매하여 부패반응을 일으키지 않고 유해균의 증식억제, 면역증강 등의 생리활성을 나타낸다. 그러므로 유산균 생균제제들이 임상에서 응용되고 있다. 그러므로 유산균제제들을 사용하기 위해서는 유산균을 배양하고 제품으로서 생산하기 위해 동결건조하고 보관하는 과정을 거쳐야 한다. 그러나 이 과정에서 유산균은 쉽게 치사 (사멸)한다. 특히, 비피도박테리움 속, 락토바실러스 속 균와 클로스트리디움 부티리쿰 (Clostridium butyricum) 균주는 혐기성 균주로 다른 유산균에 비해 더욱 안정성이 낮아 빠르게 치사한다. 그러므로 다양한 안정화제 (보호물질)들이 개발되고 있다. Lactic acid bacteria catalyze reactions such as the generation of organic acids and do not cause rot reactions, and exhibit physiological activities such as inhibition of proliferation of harmful bacteria and immune enhancement. Therefore, lactic acid bacteria probiotics have been applied in clinical practice. Therefore, in order to use lactic acid bacteria preparations, they must be lyophilized and stored for cultivation and production as a product. However, in this process, lactic acid bacteria are easily killed. In particular, Bifidobacterium spp., Lactobacillus spp. And Clostridium butyricum strains are anaerobic strains, which are more stable than other lactic acid bacteria and are rapidly killed. Therefore, various stabilizers (protectors) have been developed.
유산균제제를 만드는 과정에서 가장 중요한 과정은 동결건조이며, 이 동결건 조과정은 동결과 건조로 구별할 수 있다. 전자는 균체를 상해 (손상)되지 않도록 하면서 동결하는 공정이고, 후자는 동결한 시료중의 용매 (물)를 진공 하에서 승화시켜서 건조하는 공정이다. 이 과정들을 진행하는 과정에서 유산균 세포내 얼음의 결정형태, 염류의 농도, 불완전 탈수, 완전 탈수, 프리라디칼 축척, 침투압쇼크 등이 유산균을 치사시키는 원인이 된다. 그러므로 생유산균을 장기간 보존하기 위해서는 안정하게 유지시키기 위해서는 많은 요인에 주의를 기울여야 한다. 특히 이러한 원인을 제거할 수 있는 보호물질 (보호제)은 유산균제제의 안정성에 중요하다. The most important process in the production of lactic acid bacteria is lyophilization, which can be divided into freeze and dry. The former is a step of freezing the cells without causing injury (damage), and the latter is a step of subliming and drying the solvent (water) in the frozen sample under vacuum. In the course of these processes, crystalline forms of ice in lactic acid bacteria, salt concentrations, incomplete dehydration, complete dehydration, free radical accumulation, and infiltration pressure shock are the causes of lethal lactic acid bacteria. Therefore, attention should be paid to a number of factors in order to maintain stability for long-term preservation of live lactobacillus. In particular, protective substances (protectants) that can eliminate these causes are important for the stability of lactic acid bacteria preparations.
유산균의 동결과정에서 냉각속도의 조절이 쉽지 않아 세포내외의 동결과정이 차이로 인해서 유산균이 상해를 입기 쉽다. 그러므로 이 과정을 조절하기 위해 사용하는 보호물질로는 포도당, 설탕, 유당, 탈지유, 알부민, 폴리에틸렌글리콜 등이 사용된다. In the freezing process of lactic acid bacteria, the cooling rate is not easy to control, so the lactic acid bacteria are easily injured due to the difference between freezing process inside and outside the cells. Therefore, the protective substances used to control this process include glucose, sugar, lactose, skim milk, albumin and polyethylene glycol.
동결한 생유산균의 건조과정에서 얼지 않고 남아 있는 결합수의 건조가 유산균의 손상에 가장 큰 원인이다. 그러므로 이 공정의 유산균의 상해를 억제하기 위해서 포도당, 설탕, 아미노산 (예를 들면, 글루탐산, 아스파라긴산, 구연산, 알기닌, 리신) 등이 사용되고 있다. Drying of the bound water that remains unfrozen during the drying of frozen Lactobacillus bacteria is the main cause of damage to lactic acid bacteria. Therefore, glucose, sugar, amino acids (for example, glutamic acid, aspartic acid, citric acid, arginine, lysine) and the like are used to suppress the injury of lactic acid bacteria in this step.
동결건조한 생유산균은 아주 건조한 상태이므로 이 과정에서 오래 보관하면 쉽게 흡습하고 쉽게 균주가 사멸하게 된다. 그러므로 이 보관과정에서의 유산균의 상해를 억제하기 위해서 포도당, 글루코사민 등이 사용된다. 그 외에도 이러한 목적으로 젤라틴, 카르복실메틸셀룰로스, 이노시톨, 솔비톨 등이 이용된다.Lyophilized biolactic acid bacteria are very dry, so if stored for a long time during this process, they are easily hygroscopic and easily killed. Therefore, glucose and glucosamine are used to suppress the injury of lactic acid bacteria during this storage process. In addition to this purpose gelatin, carboxymethyl cellulose, inositol, sorbitol and the like are used.
한국특허등록공보 제144130호에서는 특정한 구조를 지닌 인삼 다당체를 개시하고 있으나, 이 인삼 다당체가 면역증강효과, 항암효과, 암예방효과 및 방사선 방어효과를 가진다는 것만 개시되어 있으며, 한국특허공개공보 제2001-50186호에서는 특정한 구조를 지닌 인삼 다당체를 개시하고 있으나, 이 인삼 다당체가 조혈촉진작용, 골수방어 작용, 암세포 살해 면역세포 생성효과, 암예방효과 및 방사선 민감효과를 가진 것만 검증하였을 뿐, 본 발명의 유산균 동결건조 보호효과, 유산균 보관 안정화 효과, 유산균 증식 효과 및 대장암 등의 원인효소의 생산억제효과는 전혀 언급이 없다. Korean Patent Registration No. 144130 discloses a ginseng polysaccharide having a specific structure, but only discloses that the ginseng polysaccharide has an immune enhancing effect, an anticancer effect, a cancer prevention effect, and a radiation protective effect. 2001-50186 discloses ginseng polysaccharides having a specific structure, but only the ginseng polysaccharides have been verified to have hematopoietic promotion, bone marrow defense, cancer cell killing immune cell production effect, cancer prevention effect and radiation sensitivity effect. Lactobacillus freeze-drying protection effect of the invention, lactic acid bacteria storage stabilization effect, lactic acid bacteria growth effect and the inhibitory effect of the production of causative enzymes such as colon cancer are not mentioned at all.
본 발명의 목적은 인삼 다당체를 함유하는 조성물을 제공하는 것이다. 본 발명의 또 다른 목적은 유산균 동결건조 보호효과, 유산균 보관 안정화 효과, 유산균 증식 효과 및 대장암 등의 원인효소의 생산억제효과를 지닌 인삼 다당체를 함유하는 조성물을 제공하는 것이다. 본 발명의 이러한 용도는 상기 공지된 용도와는 전혀 상이한 신규한 것이다. It is an object of the present invention to provide a composition containing ginseng polysaccharides. Still another object of the present invention is to provide a composition containing ginseng polysaccharide having lactic acid bacteria freeze-drying protection effect, lactic acid bacteria storage stabilization effect, lactic acid bacteria growth effect and production inhibitory effect of causative enzymes such as colon cancer. This use of the invention is a novel one completely different from the above known use.
본 발명은 인삼, 홍삼 또는 고온처리 인삼 다당체 중 어느 하나 이상을 조성물의 총중량을 기준으로 1-99.9 중량% 함유하는 것을 특징으로 하는 조성물을 제공한다. The present invention provides a composition comprising at least one of ginseng, red ginseng or hot-treated ginseng polysaccharide 1-99.9% by weight based on the total weight of the composition.
본 발명의 인삼은 고려인삼, 미국삼, 화기삼, 베트남 인삼, 히말라야인삼, 전칠삼 등을 모두 포함하고, 바람직하게는 고려인삼이다. The ginseng of the present invention includes all of Korean ginseng, American ginseng, Hwagi ginseng, Vietnamese ginseng, Himalayan ginseng, Jeonchisam and the like, and preferably Korean ginseng.
또한, 본 발명은 상기 다당체가 글루코오스 또는 갈락토오스를 포함하는 6탄당 40-50%, 갈락투론산 또는 람노오스를 포함하는 산성 다당 30-50%, 또는 히스티딘, 아르기닌, 아스파르트산, 트레오닌, 세린 또는 글루탐산을 포함하는 단백질 3-6% 를 함유하는 것인 조성물을 제공한다. In addition, the present invention is the polysaccharide 40-50% hexasaccharides containing glucose or galactose, acidic polysaccharide 30-50% including galacturonic acid or rhamnose, or histidine, arginine, aspartic acid, threonine, serine or glutamic acid It provides a composition containing 3-6% protein comprising a.
인삼, 홍삼 및 고온처리 인삼의 다당체는 전분과 펙틴상 성분이 70-80% 함유되어있다. 이 중 20-30% 전분을 제외하고 파낙산 A-U 등 20 종 이상의 다당체 성분이 분리되었다 (한국인삼연초연구원, 천일인쇄사, 대전, p75, 1994; Konno 등, Planta Med., 50, 434, 1984). 이 다당들은 대체적으로 분자량이 1 만 달톤 이상으로 고분자이며, 구성당류는 갈락투론산, 람노오스 등의 산성 다당류가 30-50%, 글루코오스, 갈락토오스 등의 6 탄당이 40-50% 함유되어 있으며 (이성동 등, 고려인삼학회지, 14, 10, 1990), 그 외에 히스티딘, 아르기닌, 아스파르트산, 트레오닌, 세린, 글루탐산 등의 아미노산으로 구성된 단백질을 3-6% 함유하고 있다. Polysaccharides of ginseng, red ginseng and high-temperature ginseng contain 70-80% of starch and pectin-like components. Except for 20-30% starch, more than 20 polysaccharides such as Panaxan AU were isolated (Korea Ginseng and Tobacco Research Institute, Cheonil Printing Co., Daejeon, p75, 1994; Konno et al., Planta Med., 50, 434, 1984) . These polysaccharides are generally polymers with molecular weights of 10,000 Daltons or more, and constituent sugars contain 30-50% of acidic polysaccharides such as galacturonic acid and rhamnose and 40-50% of hexasaccharides such as glucose and galactose ( The Korean Ginseng Society, 14, 10, 1990), and others contain 3-6% of proteins composed of amino acids such as histidine, arginine, aspartic acid, threonine, serine, and glutamic acid.
또한, 본 발명은 하기 공정을 거쳐 정제된 다당체를 함유하는 조성물도 제공한다. The present invention also provides a composition containing the polysaccharide purified through the following process.
(a) 인삼 (수삼 또는 백삼), 홍삼 (95-100℃에서 쪄서 건조한 삼) 또는 고온처리 인삼 (100℃ 이상에서 찌고 건조한 삼) 양 (무게)의 약 1 내지 20 배, 바람직하게는 약 5 배 내지 10 배 부피 (ℓ)의 에탄올 (50-100%) 등의 저급 알코올로 4 내지 100℃, 바람직하게는 20 내지 50℃ 의 추출 온도에서 약 0.5 시간 내지 2 일, 바람직하게는 1 시간 내지 1 일 동안 추출, 환류냉각 추출 또는 초음파 추출 등의 추출방법을 1 회 내지 5 회, 바람직하게는 2 회 내지 3 회 반복하여 추출하는 제 1 단계;(a) about 1 to 20 times the amount (weight) of ginseng (water ginseng or white ginseng), red ginseng (ginseng dried at 95-100 ° C.) or hot-treated ginseng (ginseng dried at 100 ° C. or higher), preferably about 5 times Lower alcohols, such as fold to 10 times volume (L) of ethanol (50-100%), at an extraction temperature of 4 to 100 ° C., preferably 20 to 50 ° C., about 0.5 to 2 days, preferably 1 hour to A first step of extracting the extraction method such as extraction, reflux cooling extraction or ultrasonic extraction for one day by repeating the extraction once to five times, preferably two to three times;
(b) 상기 추출액 중 상징액을 제거하고 남은 잔사를 약 1 내지 20 배, 바람직하게는 10 배의 증류수로 4 내지 150℃, 바람직하게는 25-60℃에서 1 시간 내지 24 시간, 바람직하게는 5 시간 추출, 환류 추출 또는 초음파 추출 등의 추출방법을 1 회 내지 5 회, 바람직하게는 2 회 내지 3 회 반복하여 수득된 추출액을 여과하는 제 2 단계;(b) the residue remaining after removing the supernatant in the extract is about 1 to 20 times, preferably 10 times distilled water at 4 to 150 ℃, preferably 25 to 60 ℃ 1 to 24 hours, preferably 5 A second step of filtering the extract obtained by repeating an extraction method such as time extraction, reflux extraction or ultrasonic extraction once to five times, preferably two to three times;
(c) 상기 여과액을 흐르는 물에서 1 일 내지 10 일, 바람직하게는 5 일 내지 7 일간 투석하는 제 3 단계;(c) a third step of dialysis in the water flowing through the filtrate for 1 to 10 days, preferably 5 to 7 days;
(d) 상기 투석액을 증류수, 저급 알코올 또는 이들의 약 1 : 0.1 내지 1 : 10 의 부피비를 갖는 혼합용매로 침전시키는 제 4 단계;(d) a fourth step of precipitating the dialysate with distilled water, lower alcohol or a mixed solvent having a volume ratio of about 1: 0.1 to 1:10;
(e) (d)의 침전액을 물에 현탁하고 다시 원심분리하여 상층액을 수득하는 제 5 단계;(e) a fifth step of suspending the precipitate of (d) in water and centrifuging again to obtain a supernatant;
인삼, 홍삼 또는 고온 처리 인삼으로부터 다당체를 제조하는 방법은 (a), (b), (c), (d), (e) 공정을 순서대로 진행한다 (분리 방법 1). 경우에 따라서는 (c), (d)와 (e) 전부 또는 일부 공정 단계는 생략할 수도 있다 (분리 방법 2). The method for producing a polysaccharide from ginseng, red ginseng, or high-temperature treated ginseng proceeds in the order of (a), (b), (c), (d), and (e) (separation method 1). In some cases, all or part of the steps (c), (d) and (e) may be omitted (separation method 2).
또한, 본 발명은 유산균 동결건조 보호효과, 유산균 보관 안정화 효과, 유산균 증식 효과 또는 대장암 등의 원인효소의 생산억제효과가 우수한 조성물을 제공한다. In addition, the present invention provides a composition excellent in lactic acid bacteria freeze-drying protective effect, lactic acid bacteria storage stabilization effect, lactic acid bacteria growth effect or production inhibitory effect of causative enzymes such as colon cancer.
본 발명의 조성물은 통상의 방법에 따른 적절한 담체, 부형제 및 희석제를 추가로 포함할 수 있다. The compositions of the present invention may further comprise suitable carriers, excipients and diluents according to conventional methods.
본 발명의 조성물에 포함될 수 있는 담체, 부형제 및 희석제로는 락토오스, 덱스트로오스, 수크로오스, 소르비톨, 만니톨, 자일리톨, 에리스리톨, 말티톨, 전분, 아카시아 고무, 알지네이트, 젤라틴, 칼슘 포스페이트, 칼슘 실리케이트, 셀룰로오스, 메틸 셀룰로오스, 미정질 셀룰로오스, 폴리비닐 피롤리돈, 물, 메틸히드록시 벤조에이트, 프로필히드록시 벤조에이트, 탈크, 마그네슘 스테아레이트 및 광물유를 들 수 있다. Carriers, excipients and diluents that may be included in the compositions of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, Methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxy benzoate, propylhydroxy benzoate, talc, magnesium stearate and mineral oil.
본 발명의 조성물은, 각각 통상의 방법에 따라 산제, 과립제, 정제, 캡슐제, 현탁액, 에멀젼, 시럽, 에어로졸 등의 경구형 제형, 외용제, 좌제 및 멸균 주사용액의 형태로 제형화하여 사용될 수 있다. The compositions of the present invention may be used in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, oral formulations, external preparations, suppositories, and sterile injectable solutions, respectively, according to conventional methods. .
본 발명의 조성물의 사용량은 환자의 나이, 성별, 체중에 따라 달라질 수 있으나, 0.1 내지 500 mg/kg의 양을 하루 1 회 내지 수회 투여할 수 있다. 또한, 그 투여량은 투여경로, 질병의 정도, 성별, 체중, 나이 등에 따라서 증감될 수 있다. 따라서, 상기 투여량은 어떠한 면으로든 본 발명의 범위를 한정하는 것은 아니다. The amount of the composition of the present invention may vary depending on the age, sex, and weight of the patient, but the amount of 0.1 to 500 mg / kg may be administered once to several times a day. In addition, the dosage may be increased or decreased depending on the route of administration, degree of disease, sex, weight, age, and the like. Therefore, the above dosage does not limit the scope of the present invention in any aspect.
본 발명의 조성물은 상기와 같은 제형으로 유산균 동결건조 보호효과, 유산균 보관 안정화 효과, 유산균 증식 효과 또는 대장암 등의 원인효소의 생산억제효과를 갖는 약제, 식품 및 음료 등에 다양하게 이용될 수 있다. 본 인삼 다당체 조성물을 첨가할 수 있는 식품으로는, 예를 들면 각종 유산균제제 등 건강식품, 갱년기 및 노인성 식품, 미용식품 등의 각종 식품류, 음료, 껌, 차, 비타민 복합제 등을 포함한다. 상기 조성물은 독성 및 부작용이 거의 없으므로 예방 목적으로 장기간 복용시에도 안심하고 사용할 수 있는 약제이다. 상기 식품에 첨가될 수 있는 조성물의 양은 일반적으로 전체 식품 중량의 0.01 내지 15 중량%로 가할 수 있으며, 식품 건강 음료 조성물은 100 ml을 기준으로 0.02 ~ 10 g, 바람직하게는 0.3 ~ 1 g의 비율로 가할 수 있다. The composition of the present invention can be used in various formulations, such as lactic acid bacteria freeze-drying protective effect, lactic acid bacteria storage stabilization effect, lactic acid bacteria growth effect or a drug having a production inhibitory effect of causative enzymes such as colon cancer. Examples of the food to which the ginseng polysaccharide composition can be added include various foods such as health foods such as various lactic acid bacteria, menopausal and senile foods, beauty foods, beverages, gums, teas, vitamin complexes, and the like. Since the composition has little toxicity and side effects, it is a drug that can be used safely even for long-term administration for the purpose of prevention. The amount of the composition that can be added to the food can generally be added in 0.01 to 15% by weight of the total food weight, the food health beverage composition is a ratio of 0.02 to 10 g, preferably 0.3 to 1 g based on 100 ml Can be added.
본 발명의 건강 음료 조성물의 액체 성분에는 특별한 제한점은 없으며, 통상의 음료와 같이 여러 가지 향미제 또는 천연 탄수화물 등을 추가 성분으로 포함할 수 있다. 상기 천연 탄수화물의 예들로 모노사카라이드, 예를 들면 포도당, 과당 등; 디사카라이드, 예를 들면 말토오스, 수크로오스 등; 및 폴리사카라이드, 예를 들면 덱스트린, 시클로덱스트린 등과 같은 통상적인 당, 및 자일리톨, 소르비톨, 에리스리톨 등의 당알코올을 포함한다. 상술한 것 이외의 향미제로서 천연 향미제 (타우마틴, 스테비아 추출물 (예를 들면 레바우디오시드 A, 글리시르히진 등) 및 합성 향미제 (사카린, 아스파르탐 등) 를 유리하게 사용할 수 있다. 상기 천연 탄수화물의 비율은 본 발명의 조성물 100 ml 당 일반적으로 약 1 ~ 20 g, 바람직하게는 약 5 ~ 12 g 이다. The liquid component of the health beverage composition of the present invention is not particularly limited, and may include various flavors or natural carbohydrates, etc. as additional ingredients, as in general beverages. Examples of the natural carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose and the like; And conventional sugars such as polysaccharides such as dextrin, cyclodextrin and the like, and sugar alcohols such as xylitol, sorbitol, erythritol and the like. As flavoring agents other than those mentioned above, natural flavoring agents (tauumatin, stevia extract (e.g., Rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. The proportion of said natural carbohydrate is generally about 1-20 g, preferably about 5-12 g per 100 ml of the composition of the present invention.
상기 외에 본 발명의 조성물은 여러 가지 영양제, 비타민, 광물 (전해질), 합성 풍미제 및 천연 풍미제 등의 풍미제, 착색제 및 증진제 (치즈, 초콜렛 등), 펙트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알코올, 탄산 음료에 사용되는 탄산화제 등을 포함할 수 있다. 그 밖에 본 발명의 조성물은 천연 과일 쥬스 및 과일 쥬스 음료 및 야채 음료의 제조를 위한 과육을 포함할 수 있다. 이러한 성분은 독립적으 로 또는 조합하여 사용할 수 있다. 이러한 첨가제의 비율은 그렇게 중요하진 않지만, 본 발명의 조성물 100 중량% 당 약 20 중량% 이하의 범위에서 선택되는 것이 일반적이다. In addition to the above, the composition of the present invention includes various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, colorants and enhancers (such as cheese and chocolate), pectic acid and salts thereof, alginic acid and salts thereof. , Organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, and the like. In addition, the composition of the present invention may include a natural fruit juice and a pulp for the production of fruit juice drinks and vegetable drinks. These components can be used independently or in combination. The proportion of such additives is not so critical, but is generally selected in the range of about 20% by weight or less per 100% by weight of the composition of the present invention.
하기에서는 실시예 및 실험예를 통하여 본 발명을 더욱 구체적으로 설명한다. 그러나, 이들 실시예 및 실험예는 본 발명에 대한 이해를 돕기 위한 예시의 목적으로만 제공된 것일 뿐 본 발명의 범주 및 범위가 하기 실시예에 의해 제한되는 것은 아니다. Hereinafter, the present invention will be described in more detail with reference to Examples and Experimental Examples. However, these Examples and Experimental Examples are provided only for the purpose of illustration for better understanding of the present invention, and the scope and scope of the present invention are not limited by the following Examples.
실시예 1Example 1
인삼 중량 (g)에 1-20 배 (바람직하게는 10 배)의 에탄올을 가하여 실온에서 냉침추출을 2회 반복하여 추출액과 잔사물을 수득하였다. 나머지 잔사물을 각각 60, 100 또는 120℃에서 1-5 시간 (바람직하게는 3 시간) 열처리하고, 이에 1-20 배 (바람직하게는 10 배)의 증류수를 가한 후, 80℃에서 2 시간동안 추출하여 여과하여 얻은 추출액을 흐르는 물에서 7 일간 투석하였다. 이를 통해 수득한 투석액을 50% 에탄올로 침전시켜 침전액을 증류수에 현탁시킨 다음, 그대로 또는 다시 원심분리하여 얻은 상징액을 동결건조하여 인삼 다당체로 사용하였다.1-20 times (preferably 10 times) of ethanol was added to ginseng weight (g), and cold extraction was repeated twice at room temperature to obtain an extract and a residue. The remaining residue was heat treated at 60, 100 or 120 ° C. for 1-5 hours (preferably 3 hours), and then 1-20 times (preferably 10 times) of distilled water was added thereto, followed by 2 hours at 80 ° C. The extract obtained by extraction and filtration was dialyzed in running water for 7 days. The dialysis solution thus obtained was precipitated with 50% ethanol, and the precipitate was suspended in distilled water. The supernatant obtained by centrifugation as it was or again was lyophilized and used as a ginseng polysaccharide.
실시예 2 Example 2
수삼을 98℃에서 1-5 시간 (바람직하게는 3 시간) 동안 처리하고 60℃에서 1-24 시간 (바람직하게는 5 시간) 건조한 홍삼에 1-20 배 (바람직하게는 10 배)의 70% 에탄올을 가하여 실온에서 냉침추출을 2 회 반복하여 추출하고 남은 잔사물에 1-20 배 (바람직하게는 10 배)의 증류수를 가한 후, 80℃에서 2 시간 동안 추출하여 여과하여 얻은 추출액을 50% 에탄올로 침전시켜 침전액을 증류수에 현탁시킨 다음, 그대로 또는 다시 원심분리하여 얻은 상징액을 동결건조하여 홍삼 다당체로 사용하였다. The fresh ginseng is treated for 1-5 hours (preferably 3 hours) at 98 ° C and 70% of 1-20 times (preferably 10 times) to 1-24 hours (preferably 5 hours) dry red ginseng at 60 ° C. After extracting twice by cold extraction at room temperature with cold ethanol, 1-20 times (preferably 10 times) of distilled water was added to the remaining residue, and the resultant was extracted by filtration for 2 hours at 80 ° C and filtered. After precipitation with ethanol, the precipitate was suspended in distilled water, and the supernatant obtained by centrifugation as it was or lyophilized was used as red ginseng polysaccharide.
실시예 3 Example 3
세절한 인삼을 120℃ 감압 하에서 1-5 시간 (바람직하게는 2 시간) 동안 처리하고 60℃에서 5 시간 건조한 고온 처리 인삼에 1-20 배 (바람직하게는 10 배)의 70% 에탄올을 가하여 실온에서 냉침추출을 2 회 반복하여 추출하고 남은 잔사물에 1-20 배 (바람직하게는 10 배)의 증류수를 가한 후, 80℃에서 2 시간 동안 추출하여 여과하여 얻은 추출액을 50% 에탄올로 침전시켜 침전액을 증류수에 현탁시킨 다음, 그대로 또는 다시 원심분리하여 얻은 상징액을 동결건조하여 고온처리 인삼 다당체로 사용하였다.Fine ginseng is treated for 1-5 hours (preferably 2 hours) at 120 ° C. under reduced pressure, and 1-20 times (preferably 10 times) of 70% ethanol is added to the high temperature dried ginseng dried at 60 ° C. for 5 hours at room temperature. The extract was extracted twice by cold extraction at, and 1-20 times (preferably 10 times) of distilled water was added to the remaining residue, followed by extraction at 80 ° C. for 2 hours, and the filtrate was precipitated with 50% ethanol. The precipitate was suspended in distilled water, and then the supernatant obtained by centrifugation as it was or lyophilized was used as a high-temperature ginseng polysaccharide.
실시예 4 Example 4
수삼을 98℃에서 3 시간 동안 처리하고 60℃에서 5 시간 건조한 홍삼에 1-20 배 (바람직하게는 10 배)의 70% 에탄올을 가하여 실온에서 냉침추출을 2 회 반복하여 추출하고 남은 잔사물에 1-20배 (바람직하게는 15 배)의 증류수를 가한 후, 4℃에서 15 시간 동안 추출하여 여과하여 얻은 추출액을 원심분리하여 얻은 상징액을 동결건조하여 홍삼 다당체로 사용하였다.The fresh ginseng was treated for 3 hours at 98 ° C, and 1-20 times (preferably 10 times) of 70% ethanol was added to the red ginseng dried at 60 ° C for 5 hours, followed by cold extraction twice at room temperature. After adding 1-20 times (preferably 15 times) of distilled water, the supernatant obtained by centrifugation of the extract obtained by extracting and filtering at 4 ° C. for 15 hours was used as red ginseng polysaccharide.
분리한 인삼, 홍삼, 고온처리인삼 다당체의 당함량 측정Determination of Sugar Content of Isolated Ginseng, Red Ginseng, and High-Temperature Ginseng Polysaccharides
이 인삼들로부터 분리한 다당류에 대한 진산의 당 함량은 안트론-황산법으로 측정하였다. 시료 1 mg/㎖의 농도로 분리 인삼 다당류을 증류수에 용해시킨 용액과 이를 2 배씩 계열희석하여 만든 용액을 0.25 ㎖에 4℃에 보관했던 안트론-황산 시약을 3 ㎖ 씩 얼음 위에서 진탕하고 10 분간 반응시켜 620 nm에서 함량을 측정하였다. 이 때 표준품으로는 글루코오스을 사용하여 검량선으로부터 작성하였다.The sugar content of jinsan to polysaccharides isolated from these ginsengs was measured by the Antron-Sulfate method. A solution of separated ginseng polysaccharides in distilled water and a double dilution series solution at a concentration of 1 mg / mL of the sample was shaken on ice with 3 ml of anthrone-sulfuric acid reagent, which was stored at 0.25 ° C at 4 ° C, for 10 minutes. The content was measured at 620 nm. At this time, glucose was used as a standard product and was prepared from a calibration curve.
분리한 인삼, 홍삼, 고온처리인삼 다당체의 산성 다당 함량Acidic Polysaccharide Contents of Isolated Ginseng, Red Ginseng, and High-Temperature Ginseng Polysaccharides
분리한 다당체를 우론산 (uronic acid) 함량을 측정하는 방법으로 측정하였다. 다당체 시료 10 mg/㎖의 농도로 분리 인삼 다당류을 증류수에 용해시킨 용액과 이를 2 배씩 계열희석하여 만든 용액을 각각 0.4 ml, 0.0125 M 황산-붕산나트륨을 넣어 잘 진탕하고 100℃에서 5 분간 가열한 후, 얼음 위에서 0.15% m-히드록시비페닐 용액 0.04 ㎖을 넣고 잘 진탕하여 다시 15 분간 강온하여 실온에서 식히고, 520 nm에서 흡광도를 측정하여 함량을 계산하였다. 표준품으로는 갈락투론산을 사용하여 검량선을 작성하였다.The isolated polysaccharide was measured by measuring the uronic acid content. Separate the solution of ginseng polysaccharides in distilled water and double dilution of the solution of polysaccharide samples at a concentration of 10 mg / mL, and shake them well with 0.4 ml and 0.0125 M sodium sulfate-borate, respectively, and heat them at 100 ° C for 5 minutes. , 0.04 ml of 0.15% m-hydroxybiphenyl solution was added to ice, shaken well, cooled for another 15 minutes, cooled to room temperature, and absorbance was measured at 520 nm. A calibration curve was prepared using galacturonic acid as a standard product.
홍삼다당체 용액을 세파덱스 컬럼 (Sephadex G-75, 1.6×50 cm, Pharmacia사)을 사용하여 그 용출패턴을 확인하였다. 컬럼은 증류수로 용출하였고 유속은 10 ㎖/h 로 하였다. 용출된 분획은 0.5 ㎖씩 분획하였다. 분자량 마커 (marker)는 알돌라제, 소혈청 알부민, 리보뉴클레아제 A를 280 nm에서 측정하였고 컬럼에서 용출된 다당체들은 카르바졸 반응 (carbazole reaction)에 의해 발색한 후 540 nm에서 흡광도를 측정하였다. 홍삼 다당체의 용출양상은 도 1과 같았으 며, 홍삼다당체의 분자량은 1-10 만달톤 정도였으며 가장 많은 것은 분자량 4-7 만 달톤 정도인 것이였다. 인삼 다당체와 고온처리 인삼 다당체도 비슷한 양상을 보였다.The red ginseng polysaccharide solution was identified using a Sephadex column (Sephadex G-75, 1.6 × 50 cm, Pharmacia) to confirm its elution pattern. The column was eluted with distilled water and the flow rate was 10 mL / h. The eluted fractions were fractionated by 0.5 ml. Molecular weight markers measured aldolase, bovine serum albumin, ribonuclease A at 280 nm, and the polysaccharides eluted from the column were absorbed at 540 nm after color development by carbazole reaction. . The elution pattern of the red ginseng polysaccharide was as shown in Figure 1, the molecular weight of the red ginseng polysaccharide was about 1-10 million daltons, the most was about 4-70,000 daltons. Ginseng polysaccharides and high-temperature treated ginseng polysaccharides showed similar results.
위의 실험법으로 측정한 결과, 당, 산성 다당 및 단백 함량은 하기 표 1와 같다.As a result of the measurement by the above experimental method, sugar, acidic polysaccharide and protein content are shown in Table 1 below.
유산균 안정성 효과Lactic acid bacteria stability effect
실험예 1Experimental Example 1
유산균인 비피도박테리움 브레브 (Bifidobacterium breve) K-110 (KCCM 10097), 락토바실러스 아시도필러스 (Lactobacillus acidophilus) (KCTC 3168), 스트렙토코커스 패시움 (Streptococcus faecium) (KCTC 3077) 균주를 TSB (Tryptic Soy Broth) 배지나 합성배지 (식용 효모 추출물, 비타민 C, 덱스트로스 함유)에서 배양하여 얻은 균을 원심분리하여 집균하고 생리식염수로 2 회 세척한다. 이 균주를 10% 함유 스킴밀크, 포도당, 인삼 (백삼) 다당체, 홍삼다당체 또는 고온처리인삼 다당체를 첨가하여 현탁하고, 액화질소로 10 분간 급냉시킨 후 동결건조기 (일본, Eyler Co.)로 건조시켜 균수를 측정하고, 이어서 4℃와 25℃에서 보관하면서 균주의 안정성을 측정하였다. 한편, 본 발명의 유산균은 특정 유산균에 한정되지 않는다 (KCCM (Korean Culture Center of Microorganisms) 한국미생물 보존센터; KCTC (Korean Collection for Type Cultures) 한국생명공학원 유전자은행).The strain LB Bifidobacterium breve K-110 (KCCM 10097), Lactobacillus acidophilus (KCTC 3168), Streptococcus faecium (KCTC 3077) (Tryptic Soy Broth) Bacteria obtained by culturing in medium or synthetic medium (containing edible yeast extract, vitamin C, dextrose) are collected by centrifugation and washed twice with physiological saline. This strain was suspended by adding 10% of skim milk, glucose, ginseng (white ginseng) polysaccharide, red ginseng polysaccharide, or high-temperature treated ginseng polysaccharide, quenched with liquefied nitrogen for 10 minutes, and dried by lyophilizer (Japan, Eyler Co.). The number of bacteria was measured, and then the stability of the strain was measured while storing at 4 ℃ and 25 ℃. On the other hand, lactic acid bacteria of the present invention is not limited to specific lactic acid bacteria (KCCM (Korean Culture Center of Microorganisms) Korea Microorganism Conservation Center; KCTC (Korean Collection for Type Cultures) Korea Institute of Biotechnology Gene Bank).
생존율 (%) = (살아남은 균수/처음 사용한 균수) x 100Survival rate (%) = (number of cells survived / number of cells used first) x 100
유당은 많은 사람, 특히 한국인을 포함한 동양인에게는 유당 불내증을 일으키는 당류인 점을 감안한다면 인삼다당류는 동결건조보호제로서 상당히 우수하다. Considering that lactose is a sugar that causes lactose intolerance for many people, especially Asians including Koreans, ginseng polysaccharides are excellent as lyophilized protective agents.
각기 다른 유산균안정화 보호제를 첨가하여 만든 유산균을 4℃에서 보관하면서 생균수를 측정하였을 때, 현재 좋은 유산균 안정화제인 유당보다 인삼다당류, 홍삼다당류, 고온처리 인삼 다당류 등은 아주 우수한 안정화효과를 보였다.Lactobacillus prepared by adding different lactic acid bacterium stabilizing agents was stored at 4 ℃, and ginseng polysaccharides, red ginseng polysaccharides, and high-temperature treated ginseng polysaccharides showed better stabilizing effects than current lactose stabilizers.
각기 다른 유산균안정화 보호제와 동결건조보호제를 동시에 첨가하여 만든 유산균을 4℃에서 보관하면서 생균수를 측정하였을 때 역시 유당의 안정화제 보다 더 우수한 효과를 보여 유산균 안정화에 유당보다 인삼다당류, 홍삼다당류, 고온처리인삼 다당류가 더 크게 기여하고 있음을 알 수 있었다. Lactobacillus prepared by adding different lactic acid stabilizer and lyophilized protector at the same time was measured at 4 ° C and the number of viable cells was also better than lactose stabilizer. Ginseng polysaccharides, red ginseng polysaccharide, The treated ginseng polysaccharides were found to contribute more significantly.
각기 다른 유산균안정화 보호제를 첨가하여 만든 유산균을 25℃에서 보관하면서 생균수를 측정하였을 때 인삼다당류, 홍삼다당류, 고온처리인삼 다당류들은 현재 좋은 유산균 안정화제인 유당과 비슷하거나 우수한 안정화효과를 보였다.Ginseng polysaccharides, red ginseng polysaccharides, and high-temperature treated ginseng polysaccharides showed similar or superior stabilization effects to lactose, a good lactic acid bacteria stabilizer, when the number of live cells was stored at 25 ° C.
유산균 증식효과Lactobacillus multiplication effect
실험예 2Experimental Example 2
GAM 배지에 추출한 인삼추출 다당체, 홍삼추출 다당체를 각각 0.25, 0.5%가 되게 가하고 고압증기멸균하였으며 유산균증식인자로 알려진 락툴로스 (lactulose)를 대조물질로 이용하여 시료와 동일한 방법으로 배지를 제조하고, 비피도박테리움 브레브 K-110 (KCCM 10097), 비피도박테리움 롱검 (Bifidobacterium longum) H-1 (KCCM 10493), 락토바실러스 아시도필러스, 락토바실러스 메센테로이데스 (Lactobacillus mesenteroides) (경희대 약대 미생물학 교실)를 이식하여 pH와 유산균수를 측정하였다. Ginseng extract polysaccharides and red ginseng extract polysaccharides were added to the GAM medium to 0.25, 0.5%, respectively, and autoclave sterilized and lactulose (lactulose), known as a lactic acid bacteria growth factor, was used as a control material. Bifidobacterium Breb K-110 (KCCM 10097), Bifidobacterium longum H-1 (KCCM 10493), Lactobacillus mesenteroides ( Lactobacillus mesenteroides ) (Kyung Hee University Pharmacy Microbiology Class) were transplanted to measure pH and the number of lactic acid bacteria.
인삼, 홍삼, 고온처리 인삼의 다당류의 유산균 증식 효과를 측정한 결과 우수한 유산균 증식효과를 보였다. Lactic acid bacteria proliferation effect of polysaccharides of ginseng, red ginseng, and high-temperature treated ginseng was excellent.
유해균의 성장억제효과Growth inhibitory effect of harmful bacteria
실험예 3: 장내세균총의 세균유해효소에 미치는 효과Experimental Example 3: Effect on Enterobacteriaceae
사람의 장내세균총을 105 개가 되도록 희석하여 5 ㎖의 GAM (-덱스트로스) 배지에 인삼 다당체 또는 홍삼 다당체를 첨가하여 만든 배양액에 이식하고, 24 시간 배양한 후, 균주가 생산한 대장암 및 간장해의 원인효소인 β-글루쿠로니다제와 트립토판나아제의 효소활성을 측정하였다. Dilute human enterobacteriaceae to 10 5 and transplant it into culture medium made by adding ginseng polysaccharide or red ginseng polysaccharide to 5 ml of GAM (-dextrose) medium, and cultured for 24 hours, and then colon cancer and soy sauce produced by the strain Enzyme activity of β-glucuronidase and tryptophanase, the causative enzymes of the sun, was measured.
β-글루쿠로니다아제 효소활성은 0.1M 인산 완충액 (pH 7.0) 0.38 ㎖에 10 mM p-니트로페닐-β-D-글루쿠로니드 0.02 ㎖, 효소액 0.1 ㎖를 가하여 37℃ 에서 1 시간 반응시키고 0.5 N NaOH 0.5 ㎖를 첨가하여 반응을 종료시키고, 증류수 1 ㎖를 가한 다음, 원심분리 (2000×g, 20 분)하고, 상등액에 대해 405 nm에서 흡광도를 측정하였다. β-glucuronidase enzyme activity was reacted for 1 hour at 37 ° C by adding 0.02 ml of 10 mM p-nitrophenyl-β-D-glucuronide and 0.1 ml of enzyme solution to 0.38 ml of 0.1 M phosphate buffer (pH 7.0). The reaction was terminated by adding 0.5 mL of 0.5 N NaOH, 1 mL of distilled water was added, followed by centrifugation (2000 × g, 20 min), and the absorbance was measured at 405 nm for the supernatant.
트립토판나아제 효소활성은 완전반응 혼합물 (0.1 M 비사인 (bicine) pH 8.0), 4% 피리독살 5-포스페이트, 20% 송아지 혈청 알부민) 0.2 ㎖, 0.02 M 트립토판 0.2 ㎖ 및 효소액 0.1 ㎖을 가하여 30 분 반응시키고, 착색제 (p-디메틸아미노벤즈알데히드 14.7 g, 95% 에탄올 948 ㎖, C-H2SO4 52 ㎖) 2 ㎖을 가하여 반응을 종료시킨 후, 원심분리 (2000×g, 20 분)하고 상등액으로 550 ㎚에서 흡광도를 측정하였다.Tryptophanase enzyme activity was obtained by adding 0.2 ml of 0.02 M tryptophan and 0.1 ml of enzyme solution by adding 30 ml of a complete reaction mixture (0.1 M bisine pH 8.0), 4% pyridoxal 5-phosphate, 20% calf serum albumin) After the reaction was completed, 2 ml of a colorant (14.7 g of p-dimethylaminobenzaldehyde, 948 ml of 95% ethanol, 52 ml of CH 2 SO 4 ) was added to terminate the reaction, followed by centrifugation (2000 × g, 20 minutes), followed by a supernatant solution. Absorbance was measured at 550 nm.
하기 표 7에 대장균 HGU-3 (β-글루쿠로니다제 생성균)과 유산균을 혼합배양한 경우의 대장암 및 간장해 유발효소의 저해율을 나타내었다. Table 7 below shows the inhibition rate of colon cancer and hepatic inducing enzymes when E. coli HGU-3 (β-glucuronidase-producing bacteria) and lactic acid bacteria were mixed in culture.
인삼 또는 홍삼의 다당체들은 발암효소로 알려진 장내세균이 생산하는 β-글루쿠로니다아제 및 트립토판나아제의 생산을 크게 억제하는 효과를 보였다. Polysaccharides of ginseng or red ginseng showed an effect of significantly inhibiting the production of β-glucuronidase and tryptophanase produced by enterobacteriaceae known as carcinogen.
상기에서 설명한 바와 같이, 본 발명의 인삼 다당체를 함유하는 조성물은 유산균 동결건조 보호효과, 유산균 보관 안정화 효과, 유산균 증식 효과 또는 대장암 등의 원인효소의 생산억제효과가 우수하다. As described above, the composition containing the ginseng polysaccharide of the present invention is excellent in the lactic acid bacteria lyophilization protective effect, lactic acid bacteria storage stabilization effect, lactic acid bacteria growth effect or production inhibitory effect of causative enzymes such as colon cancer.
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Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
KR102012652B1 (en) | 2018-12-12 | 2019-08-21 | 비타민하우스(주) | Composition for preserving freeze-dried lactic acid bacteria comprising cyclodextrin |
KR20230014932A (en) * | 2021-07-21 | 2023-01-31 | 주식회사 에치와이 | Lactobacillus Paracasei HY7017 With Enhanced Functional Characteristics and Enhanced Immunity Function by Using Red Ginseng as a Nutrient Source, and Use Thereof |
Families Citing this family (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
KR101486536B1 (en) * | 2012-11-19 | 2015-01-26 | 재단법인 금산국제인삼약초연구소 | Microorganism culture medium, freeze drying protectant, method for culture and drying protection by using red ginseng starch |
KR102458980B1 (en) * | 2020-07-22 | 2022-10-25 | 농업회사법인 고려홍삼원 주식회사 | Low molecular weight enzyme-treated Red Ginseng or Panax Ginseng Polysaccharide extracts and prebiotics and probiotics uses thereof |
KR102540263B1 (en) * | 2020-12-24 | 2023-06-07 | 주식회사 한국인삼공사 | Composition For Improving Intestinal Microbial Flora Including Ginseng Extract Or Fraction Thereof |
Citations (1)
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KR0144130B1 (en) * | 1993-12-17 | 1998-07-15 | 신재인 | Ginseng polysaccharide "jinsan" which have immuno potentive effect |
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KR0144130B1 (en) * | 1993-12-17 | 1998-07-15 | 신재인 | Ginseng polysaccharide "jinsan" which have immuno potentive effect |
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
KR102012652B1 (en) | 2018-12-12 | 2019-08-21 | 비타민하우스(주) | Composition for preserving freeze-dried lactic acid bacteria comprising cyclodextrin |
KR20230014932A (en) * | 2021-07-21 | 2023-01-31 | 주식회사 에치와이 | Lactobacillus Paracasei HY7017 With Enhanced Functional Characteristics and Enhanced Immunity Function by Using Red Ginseng as a Nutrient Source, and Use Thereof |
KR102587892B1 (en) | 2021-07-21 | 2023-10-12 | 주식회사 에치와이 | Lactobacillus Paracasei HY7017 With Enhanced Functional Characteristics and Enhanced Immunity Function by Using Red Ginseng as a Nutrient Source, and Use Thereof |
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