KR101179101B1 - A polysaccharide having immune enhancing activity from green algae and method for preparing the same - Google Patents
A polysaccharide having immune enhancing activity from green algae and method for preparing the same Download PDFInfo
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- KR101179101B1 KR101179101B1 KR1020100046073A KR20100046073A KR101179101B1 KR 101179101 B1 KR101179101 B1 KR 101179101B1 KR 1020100046073 A KR1020100046073 A KR 1020100046073A KR 20100046073 A KR20100046073 A KR 20100046073A KR 101179101 B1 KR101179101 B1 KR 101179101B1
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- KR
- South Korea
- Prior art keywords
- polysaccharide
- polysaccharides
- green algae
- supernatant
- immune
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- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
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- A61K36/02—Algae
- A61K36/03—Phaeophycota or phaeophyta (brown algae), e.g. Fucus
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- B01D15/26—Selective adsorption, e.g. chromatography characterised by the separation mechanism
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Abstract
본 발명은 녹조류를 에틸알코올을 이용하여 색소, 지질 및 저분자 단백질을 제거한 후, 증류수와 에틸알코올로 추출하여 다당류를 수득하고, 이를 이온 교환 컬럼을 이용하여 면역 증강 활성을 갖는 다당류를 분리/분획하는 녹조류 유래 다당류의 제조방법 및 이를 통해 제조된 다당류를 면역 증강 활성을 가지는 약제 및 건강기능식품 원료의 개발에 관한 것으로, 본 발명에 따른 면역증강활성을 갖는 녹조류 유래 다당류는 마우스 대식세포인 Raw 264.7 세포에서 면역관련 사이토카인인 NO, TNF-α, IL-1β, PGE2등의 물질을 분비하여 면역 활성을 증강시켜주며, 식품산업에서는 면역증강용 기능성 식품으로, 면역기능이 약한 사람의 면역 활성 증강을 위한 보조 식품으로 사용이 가능한, 식품 소재 개발 산업에 있어 매우 우수한 발명인 것으로 사료된다.The present invention removes pigments, lipids and low molecular weight proteins using ethyl alcohol, and then extracts them with distilled water and ethyl alcohol to obtain polysaccharides, and isolates / fractionates polysaccharides having immune enhancing activity using ion exchange columns. The present invention relates to a method for preparing green algae-derived polysaccharide and to a pharmaceutical and health functional food raw material having immune enhancing activity. Enhances immune activity by secreting immune-related cytokines such as NO, TNF-α, IL-1β, and PGE 2. In the food industry, it is a functional food for enhancing immunity. It is considered to be a very good invention in the food material development industry, which can be used as a supplement for food.
Description
본 발명은 면역 증강 활성을 갖는 녹조류 유래 다당류 및 그 제조방법에 관한 것으로, 보다 구체적으로는 녹조류를 에틸알코올을 이용하여 색소, 지질 및 저분자 단백질을 제거한 후, 증류수와 에틸알코올로 추출하여 다당류를 수득하고, 이를 이온 교환 컬럼을 이용하여 면역 증강 활성을 갖는 다당류를 분리/분획하는 녹조류 유래 다당류의 제조방법 및 이를 통해 제조된 다당류를 면역 증강 활성을 가지는 약제 및 건강기능식품 원료로 개발하는 것이다.
The present invention relates to a green algae-derived polysaccharide having an immune enhancing activity and a method of preparing the same, and more specifically, to remove green algae using ethyl alcohol to remove pigments, lipids and low molecular proteins, and then extract distilled water and ethyl alcohol to obtain polysaccharides. In addition, the method of preparing a green algae-derived polysaccharide which separates / fractionates a polysaccharide having an immune enhancing activity using an ion exchange column, and the polysaccharide prepared as a pharmaceutical and health functional food raw material having an immune enhancing activity.
해조류는 통상 바다에서 자생 또는 양식되는 거대조류를 통틀어 이르는 말로 서식하는 수심과 조체의 빛깔에 따라 녹조류, 갈조류, 홍조류로 나뉜다. 그 중, 녹조류는 일반적으로 만조와 간조의 사이 또는 해안 지역에 많이 서식 하는데, 특히 우리나라에서는 서해안에 많이 서식하고 있다. 녹조류에는 미네랄, 비타민, 단백질, 식이섬유 등이 풍부하게 함유되어 있으며, 매생이, 감태 (가시파래), 파래 등의 녹조류는 예부터 우리 선조들이 즐겨 먹는 먹거리 중의 하나였다. 그러나 이렇게 영양이 풍부한 녹조류는 현재까지 연구가 많이 되어 있지 않으며, 또한 산업적으로 크게 이용을 하지 않고 있는 실정이다. 녹조류의 세포벽에는 “울반” 이라 불리는 수용성 다당류가 존재 하는데, 현재까지 알려진 생리활성으로 항암, 항바이러스, 항혈액응고 등이 있다. 그러나, 아직까지 울반의 면역 증강에 관한 연구는 거의 없는 실정이다. The algae are usually horses that grow wild or grow in the sea, and are divided into green algae, brown algae, and red algae according to the depth and color of the algae. Among them, green algae generally inhabit between the high tide and the low tide or coastal areas, especially in the west coast in Korea. Green algae are rich in minerals, vitamins, proteins, and fiber, and green algae, such as falcon, Ecklonia cava, and green, have been one of the foods our ancestors enjoyed since ancient times. However, this nutrient-rich green algae has not been studied so far and has not been widely used industrially. There are water-soluble polysaccharides called “ulban” on the cell wall of green algae, and the known physiological activities include anticancer, antiviral, and anticoagulant. However, there are few studies on the immune enhancement of Ulban.
현재까지 해조류 유래 다당류에 대한 특허를 살펴보면, 대한민국 등록특허 제2002-2245980호에서는 ‘김으로부터 고지혈증 개선효과가 있는 포피란을 추출 및 정제하는 방법’에서는 김(pophyra yezoensis)으로부터 포피란 (porphyran)을 추출 및 정제하는 방법과, 종래 기술의 단점인 낮은 제품수율, 인체 건강상 유해한 맹독성의 세칠피리디니움 크로라이드(cetylpyridinium chloride) 사용, 다량의 에탄올 소비 등을 개선하는 방안이 제시 되었으며, 대한민국 특허출원 제 10-2007-0049465호는 ‘콜레스테롤 대사 개선 및 비만억제 기능성을 동시에 보유한 천연복합 농축액 및 그 제조방법’은 미역, 김, 다시마, 톳 등에서 다당류를 추출하고, 갈조류에서 퓨코스테롤을 추출한 후, 마늘 추출물인 알리신과 혼합하여 복합 농축액을 만들었다. Looking at the patents on algae-derived polysaccharides so far, Korean Patent No. 2002-2245980 discloses porphyran from laver (pophyra yezoensis) in the method of extracting and purifying porphyrans with antihyperlipidemic effect from laver. A method for extracting and purifying, low product yields, the use of cetylpyridinium chloride, and the consumption of a large amount of ethanol, which are disadvantages of the prior art, which are harmful to human health, have been proposed. No. 10-2007-0049465 refers to 'a natural complex concentrate having both improved cholesterol metabolism and anti-obesity functionality, and a method for preparing the same', extracts polysaccharides from seaweed, seaweed, kelp, and sesame, and extracts fucosterol from brown algae, A mixed concentrate was prepared by mixing garlic extract with allicin.
또한 녹조류의 면역증강 활성에 대하여, 대한민국 등록특허 제10-0617495호 ‘한국산 파래 유래의 면역증강용 추출물, 그 추출방법 및 이를 함유하는 약제’에 한국산 녹조류에 증류수를 가하여 교반한 후 이를 원심분리하여 상등액을 취하는 수용성 성분의 추출방법 또는 상기 상등액에 황산암모늄을 가하여 단백질 성분을 침전시킨 후, 크로마토그래피법에 의해 렉틴 분획 물질을 분리하는 렉틴 성분의 추출방법에 의해 수득한 녹조류 추출물이 정상세포에 대한 세포독성 없이 면역력을 효과적으로 활성화시킬 수 있어 면역증강용 약제로 효과적으로 사용할 수 있음이 개시되어 있다.In addition, the immuno-enhancing activity of the green algae, Korea Patent No. 10-0617495 No. 10, 'Immuno-enrichment extract derived from Korean green seaweed, the extraction method and the drug containing the same' was added by stirring distilled water to the Korean green algae and centrifuged it. The green algae extract obtained by the extraction method of the water-soluble component taking the supernatant or the extraction of the lectin component which separates the lectin fraction material by chromatography by adding ammonium sulfate to the supernatant and separating the lectin fraction material by chromatography It is disclosed that it can effectively activate the immunity without cytotoxicity can be effectively used as an immunopotentiating agent.
또한 최 등의 연구(최용석, 구재근; 가시파래 (Enteromorpha prolifera)로부터 산성다당의 당구조 특성; 한국어업기술학회 2001년도 춘계 수산관련학회 공동학술대회발표요지집; 2001. 01.01) 에 녹조류 중 갈파래가 황산기를 함유한 다당을 다량 함유하고 있으며, 이 산성 다당은 항종양성, 항 바이러스성, 면역활성증강, 항혈액 응고작용 등이 보고되어 있으며, 국내에서도 홑파래에서 추출한 당단백질이 Sarcoma-180 cell에 대한 항암효과 및 면역활성이 있음이 개시되어 있다. In addition, Choi et al. (Cho Yong-seok Choi, Jae-geun Koo; Characterization of sugar structure of acidic polysaccharides from Enteromorpha prolifera; Journal of Korean Society of Fisheries and Technology, Spring Conference of Korean Fisheries Association 2001; 01.01, 2001) It contains a large amount of polysaccharides, and this acid polysaccharide has been reported to have antitumor, antiviral, immune activity enhancement, and anticoagulant activity. It is disclosed that there is an anticancer effect and immune activity.
하지만 녹조류로부터 유기용매를 순차적으로 처리하여 다당류를 제조하고, 이들의 면역증강 활성에 대한 연구는 수행된 바 없다.
특히, 상기 어느 문헌들에서도 건조한 감태 원료에 직접 에탄올을 첨가하여 얻은 펠렛에 다시 아세톤을 처리하여 지질 및 단백질을 제거한 후 분자량이 큰 다당류 물질만을 수득하는 방법은 암시 조차 되어 있지 않다. However, polysaccharides are prepared by sequentially treating organic solvents from green algae, and their immuno-enhancing activity has not been studied.
In particular, none of the above documents suggests that the pellet obtained by directly adding ethanol to the dried Ecklonia cava is treated with acetone again to remove lipids and proteins, thereby obtaining only a high molecular weight polysaccharide material.
이에 본 발명자들은 우리나라에서 많이 생산이 되나 산업적으로 거의 이용하지 않고 있는 녹조류를 산업화에 이용하기 위해 예의 연구를 거듭한 결과, 특히 감태(가시파래) 유래 다당류가 면역 증강 활성을 가지며, 이를 이온 교환 컬럼 크로마토그래피로 정제 하였을 경우 활성이 크게 증가되는 분획물을 수득할 수 있었다. 따라서 정제된 다당류 분획물을 이용하여 면역증강용 기능성 식품의 원료개발이 가능 할 것으로 밝혀내어 본 발명을 완성하게 되었다.
Accordingly, the present inventors have intensively researched to use green algae, which are produced in Korea but are rarely used industrially, for industrialization. In particular, polysaccharides derived from E. coli (thorn) have an immune enhancing activity, and this is an ion exchange column. When purified by chromatography it was possible to obtain a fraction that the activity is greatly increased. Therefore, using the purified polysaccharide fraction, it was found that the development of raw materials of functional foods for immuno-enhancement was completed to complete the present invention.
따라서 본 발명의 목적은 면역 증강 활성을 갖는 녹조류 유래 다당류의 제조 방법을 제공하는 것이다.It is therefore an object of the present invention to provide a method for producing green algae-derived polysaccharides having immune enhancing activity.
또한 본 발명의 다른 목적은 상기 면역 증강 활성을 갖는 녹조류 유래 다당류를 유효성분으로 함유하는 면역 증강 활성 건강기능식품을 제공하는 것이다.
In addition, another object of the present invention is to provide an immune enhancing active dietary supplement containing the green algae-derived polysaccharide having the immune enhancing activity as an active ingredient.
상기와 같은 본 발명의 목적은 대표적인 녹조류로 가시파래 (Enteromorpha prolifera)를 선별하여, 이로부터 면역증강활성을 가지는 다당류를 제조한 후, 제조된 다당류의 산화질소(NO), 종양 괴사 인자-α (TNF-α), 인터류킨 1β (IL-1β) 및 프로스타글란딘 E2 (PGE2)등 면역 관련 사이토카인의 분비 효과를 평가한 결과, 본 발명에 따른 녹조류 유래 다당류가 상기의 면역증진 관련 사이토카인을 분비하는데 결정적으로 작용함을 확인하였고, 상기에서 수득한 다당류를 이온 교환 컬럼 크로마토그래피를 이용하여 정제 할 경우, 면역증강활성이 크게 증가함을 확인함으로써 달성되었다.
The purpose of the present invention as described above is to select the thorny seaweed ( Enteromorpha prolifera ) as a representative green algae, to prepare a polysaccharide having immuno-enhancing activity therefrom, nitric oxide (NO), tumor necrosis factor-α of the polysaccharide prepared As a result of evaluating the secretion effects of immune-related cytokines such as TNF-α), interleukin 1β (IL-1β), and prostaglandin E 2 (PGE 2 ), the green algae-derived polysaccharides according to the present invention secrete the above-described immune-promoting cytokines. It was confirmed that it acts decisively, and when the polysaccharide obtained above was purified by ion exchange column chromatography, it was achieved by confirming that the immuno-enhancing activity is greatly increased.
본 발명은 면역증강 활성을 갖는 녹조류 유래 다당류의 제조방법을 제공한다.The present invention provides a method for producing green algae-derived polysaccharides having immuno-enhancing activity.
본 발명에 따른 방법은 건조된 녹조류를 에틸알코올로 교반추출 후 원심분리로 펠렛을 획득하여 색소, 지질 및 저분자 단백질을 제거하는 단계; 상기 수득한 펠렛을 물과 에틸알코올을 이용하여 다당류를 수득하는 단계; 수득된 다당류를 이온 교환 컬럼 크로마토그래피를 이용하여 정제하는 단계로 이루어진다.The method according to the present invention comprises the steps of extracting the dried green algae with ethyl alcohol to obtain pellets by centrifugation to remove pigments, lipids and low molecular weight proteins; Obtaining the polysaccharide using the obtained pellet using water and ethyl alcohol; The obtained polysaccharide consists of a step of purifying using ion exchange column chromatography.
본 발명에 따른 방법은 보다 구체적으로 건조 녹조류에 에틸알코올을 첨가하고, 교반하면서 가열한 후, 원심분리하여 펠렛을 수득하는 단계; 수득된 펠렛을 아세톤을 사용하여 펠렛에 남아있는 색소, 지질 및 저분자 단백질을 제거 하는 단계; 상기 색소, 지질 및 저분자 단백질이 제거된 시료를 원심분리하여 최종 펠렛을 수득한 후, 건조하여 분말화하는 단계; 상기에서 수득된 분말에 증류수를 첨가하고, 교반하면서 가열 한 후, 원심분리하여 상등액을 수득하는 단계; 수득된 상등액을 농축하고, 농축된 상등액에 에틸알코올을 첨가한 후, 원심분리하여 상등액을 수득하는 단계; 수득된 상등액에 에틸알코올을 첨가하여 침전물을 수득하고 이를 여과, 세척하여 다당류를 수득하는 단계; 및 상기 수득된 다당류를 이온 교환 컬럼 크로마토그래피로 정제하는 단계로 이루어진다.The method according to the present invention more specifically comprises adding ethyl alcohol to the dried alga, heating with stirring, and then centrifuging to obtain pellets; Removing the remaining pigments, lipids and low molecular weight proteins from the pellets using acetone; Centrifuging the sample from which the pigments, lipids and low molecular weight proteins have been removed to obtain final pellets, and then drying and powdering; Adding distilled water to the powder obtained above, heating with stirring, and centrifuging to obtain a supernatant; Concentrating the obtained supernatant, adding ethyl alcohol to the concentrated supernatant, and centrifuging to obtain a supernatant; Adding ethyl alcohol to the obtained supernatant to obtain a precipitate, which is filtered and washed to obtain a polysaccharide; And purifying the obtained polysaccharide by ion exchange column chromatography.
또한 본 발명은 상기 방법으로 제조된 면역증강 활성을 갖는 녹조류 유래 다당류를 유효성분으로 함유하는 면역증강 활성 약제학적 조성물을 제공한다.In another aspect, the present invention provides an immuno-enhancing active pharmaceutical composition containing a green algae-derived polysaccharide having an immuno-enhancing activity prepared by the above method as an active ingredient.
또한 본 발명은 상기 방법으로 제조된 면역증강 활성을 갖는 녹조류 유래 다당류를 유효성분으로 함유하는 면역증강 활성 건강기능식품 조성물을 제공한다.In another aspect, the present invention provides an immuno-enhanced health functional food composition containing a green algae-derived polysaccharide having an immuno-enhancing activity as an active ingredient prepared by the above method.
본 발명에 있어서, 상기 녹조류라 함은 파래, 클로렐라, 청각, 매생이, 해캄을 포함하나, 이에 제한되는 것은 아니며, 당업계에 알려진 다른 녹조류일 수도 있다.
In the present invention, the green algae include green, chlorella, auditory, falconus, but is not limited thereto, and may be other green algae known in the art.
본 발명에 있어서, 유효성분이라 함은 내재된 약리작용에 의해 그 의약품의 효능효과를 직접 또는 간접적으로 발현한다고 기대되는 물질 또는 물질군(약리학적 활성성분 등이 밝혀지지 않은 생약 등을 포함한다)으로서 주성분을 포함하는 것을 의미한다.In the present invention, the term "active ingredient" refers to a substance or a group of substances (including medicinal herbs whose pharmacologically active ingredients are not known) which are expected to express directly or indirectly the efficacy effect of the medicine by inherent pharmacological action. It means containing a main component as.
본 발명에 있어서, "건강기능식품"은 건강기능식품에 관한 법률 제6727호에 따른 인체에 유용한 기능성을 가진 원료나 성분을 사용하여 제조 및 가공한 식품을 의미하며, "기능성"이라 함은 인체의 구조 및 기능에 대하여 영양소를 조절하거나 생리학적 작용 등과 같은 보건 용도에 유용한 효과를 얻을 목적으로 섭취하는 것을 의미한다.In the present invention, "health functional food" means a food manufactured and processed using raw materials or ingredients having functional properties useful for the human body according to the Health Functional Food Act No. 6767, and "functional" means a human body It means the ingestion for the purpose of obtaining a useful effect in health use such as nutrient control or physiological action on the structure and function of.
본 발명의 녹조류 유래 다당류를 포함하는 약제학적 조성물은 약학적 조성물의 제조에 통상적으로 사용하는 적절한 담체, 부형제 및 희석제를 더 포함할 수 있다.Pharmaceutical compositions comprising the green algae-derived polysaccharide of the present invention may further comprise suitable carriers, excipients and diluents commonly used in the manufacture of pharmaceutical compositions.
본 발명의 녹조류 유래 다당류를 포함하는 약제학적 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 pharmaceutical composition comprising the green algae-derived polysaccharide of the present invention may be prepared in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, oral formulations, external preparations, suppositories, and sterile injectable solutions, respectively. It can be formulated and used in the form.
본 발명의 녹조류 유래 다당류를 포함하는 약제학적 조성물에 포함될 수 있는 담체, 부형제 및 희석제로는 락토즈, 덱스트로즈, 수크로스, 솔비톨, 만니톨, 자일리톨, 에리스리톨, 말티톨, 전분, 아카시아 고무, 알지네이트, 젤라틴, 칼슘 포스페이트, 칼슘 실리케이트, 셀룰로즈, 메틸셀룰로즈, 미정질 셀룰로스, 폴리비닐 피롤리돈, 물, 메틸히드록시벤조에이트, 프로필히드록시벤조에이트, 탈크, 마그네슘 스테아레이트 및 광물유를 들 수 있다. 제제화할 경우에는 보통 사용하는 충진제, 증량제, 결합제, 습윤제, 붕해제, 계면활성제 등의 희석제 또는 부형제를 사용하여 조제된다. 경구투여를 위한 고형제제에는 정제, 환제, 산제, 과립제, 캡슐제 등이 포함되며, 이러한 고형제제는 상기 추출액에 적어도 하나 이상의 부형제 예를 들면, 전분, 칼슘카보네이트(calciumcarbonate), 수크로스(sucrose) 또는 락토오스(lactose), 젤라틴 등을 섞어 조제된다. 또한 단순한 부형제 이외에 마그네슘 스테아레이트, 탈크 같은 윤활제들도 사용된다. 경구를 위한 액상 제제로는 현탁제, 내용액제, 유제, 시럽제 등이 해당되는데 흔히 사용되는 단순희석제인 물, 리퀴드 파라핀 이외에 여러 가지 부형제, 예를 들면 습윤제, 감미제, 방향제, 보존제 등이 포함될 수 있다. 비경구 투여를 위한 제제에는 멸균된 수용액, 비수성용제, 현탁제, 유제, 동결건조 제제, 좌제가 포함된다. 비수성용제, 현탁제로는 프로필렌글리콜(propylene glycol), 폴리에틸렌 글리콜, 올리브 오일과 같은 식물성 기름, 에틸올레이트와 같은 주사 가능한 에스테르 등이 사용될 수 있다. 좌제의 기제로는 위텝솔(witepsol), 마크로골, 트윈(tween) 61, 카카오지, 라우린지, 글리세로제라틴 등이 사용될 수 있다.Carriers, excipients and diluents which may be included in the pharmaceutical composition comprising the green algae-derived polysaccharide of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, Gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. In the case of formulation, a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, or a surfactant is usually used. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and such solid preparations include at least one excipient such as starch, calcium carbonate, sucrose in the extract. Or lactose, gelatin, or the like is mixed. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Oral liquid preparations include suspensions, solvents, emulsions, and syrups, and may include various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. . Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.
본 발명에 따른 녹조류 유래 다당류의 바람직한 투여량은 환자의 상태 및 체중, 질병의 정도, 약물형태, 투여경로 및 기간에 따라 다르지만, 당업자에 의해 적절하게 선택될 수 있다. 그러나 바람직한 효과를 위해서, 본 발명의 녹조류 유래 다당류는 1일 0.0001 내지 100 mg/kg으로, 바람직하게는 0.001 내지 100 mg/kg으로 투여하는 것이 좋다. 투여는 하루에 한번 투여할 수도 있고, 수회 나누어 투여할 수도 있다. 상기 투여량은 어떠한 면으로든 본 발명의 범위를 한정하는 것은 아니다.Preferred dosages of the green algae-derived polysaccharides according to the present invention vary depending on the condition and weight of the patient, the extent of the disease, the form of the drug, the route of administration and the duration, and may be appropriately selected by those skilled in the art. However, for the desired effect, the green algae-derived polysaccharide of the present invention is preferably administered at 0.0001 to 100 mg / kg, preferably at 0.001 to 100 mg / kg. The administration may be carried out once a day or divided into several times. The dose is not intended to limit the scope of the invention in any way.
본 발명에 따른 녹조류 유래 다당류는 쥐, 생쥐, 가축, 인간 등의 포유동물에 다양한 경로로 투여될 수 있다. 투여의 모든 방식이 예상될 수 있는데, 예를 들면, 경구, 직장 또는 정맥, 근육, 피하, 자궁내 경막 또는 뇌혈관내(intracerebroventricular) 주사 및 피부에 직접 투여될 수 있다. Green algae-derived polysaccharides according to the present invention can be administered to mammals such as mice, mice, livestock, humans, and the like by various routes. All modes of administration can be expected, for example, oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dural or intracerebroventricular injections and directly to the skin.
또한, 본 발명에 따른 녹조류 유래 다당류는 면역증강 활성을 목적으로 식품 또는 음료에 첨가될 수 있다. 녹조류 유래 다당류를 첨가할 수 있는 식품으로는, 예를 들어, 각종 식품류, 음료, 껌, 차, 비타민 복합제, 건강 기능성 식품류, 분말, 과립, 정제, 캡슐 또는 음료 등이 있다. 이 때, 식품 또는 음료 중의 상기 녹조류 유래 다당류의 양은 전체 식품 중량의 0.01 내지 15 중량%로 가할 수 있으며, 건강 음료 조성물은 100 ㎖를 기준으로 0.02 내지 5 g, 바람직하게는 0.3 내지 1 g의 비율로 가할 수 있다.In addition, the green algae-derived polysaccharide according to the present invention may be added to food or beverage for the purpose of immuno-enhancing activity. Examples of foods to which green algae-derived polysaccharides can be added include various foods, beverages, gums, teas, vitamin complexes, health functional foods, powders, granules, tablets, capsules, and beverages. At this time, the amount of the green algae-derived polysaccharide in the food or beverage may be added at 0.01 to 15% by weight of the total food weight, the health beverage composition is based on 100 ml of 0.02 to 5 g, preferably 0.3 to 1 g Can be added.
본 발명의 식품 조성물은 지시된 비율로 필수 성분으로서 녹조류 유래 다당류를 함유하는 외에는 다른 성분에는 특별한 제한이 없으며 통상의 음료와 같이 여러 가지 향미제 또는 천연 탄수화물 등을 추가 성분으로서 함유할 수 있다. 상술한 천연 탄수화물의 예는 모노사카라이드, 예를 들어, 포도당, 과당 등; 디사카라이드, 예를 들어 말토스, 슈크로스 등; 및 폴리사카라이드, 예를 들어 덱스트린, 시클로덱스트린 등과 같은 통상적인 당, 및 자일리톨, 소르비톨, 에리트리톨 등의 당알콜이다. 상술한 것 이외의 향미제로서 천연 향미제(타우마틴, 스테비아 추출물(예를 들어 레바우디오시드 A, 글리시르히진 등) 및 합성 향미제(사카린, 아스파르탐 등)를 유리하게 사용할 수 있다. 상기 천연 탄수화물의 비율은 본 발명의 조성물 100 ㎖당 일반적으로 약 1 내지 20g, 바람직하게는 약 5 내지 12g이다. The food composition of the present invention is not particularly limited to other ingredients except for containing green algae-derived polysaccharides as essential ingredients in the ratios indicated, and may contain various flavors or natural carbohydrates as additional ingredients, such as ordinary drinks. Examples of the above-mentioned 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 sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents other than those described 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 carbohydrates 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 녹조류 유래 다당류는 여러 가지 영양제, 비타민, 광물(전해질), 합성 풍미제 및 천연 풍미제 등의 풍미제, 착색제 및 중진제(치즈, 초콜릿 등), 펙트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알콜, 탄산음료에 사용되는 탄산화제 등을 함유할 수 있다. 그밖에 본 발명의 녹조류 유래 다당류는 천연 과일 주스 및 과일 주스 음료 및 야채 음료의 제조를 위한 과육을 함유할 수 있다. 이러한 성분은 독립적으로 또는 조합하여 사용할 수 있다. 이러한 첨가제의 비율은 그렇게 중요하진 않지만 본 발명의 녹조류 유래 다당류는 100 중량부 당 0 내지 약 20 중량부의 범위에서 선택되는 것이 일반적이다.
In addition to the above-mentioned green algae-derived polysaccharide of the present invention, various nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavoring agents, such as colorants and neutralizing agents (cheese, chocolate, etc.), pectic acid and salts thereof, alginic acid And salts thereof, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, and the like. In addition, the green algae-derived polysaccharide of the present invention may contain a fruit flesh for producing natural fruit juice and fruit juice beverage and vegetable beverage. These components may be used independently or in combination. The proportion of such additives is not so critical, but the green algae-derived polysaccharides of the present invention are generally selected in the range of 0 to about 20 parts by weight per 100 parts by weight.
본 발명에서는 해조류로서 녹조류로 가시파래를 원료로 사용하여 면역증강 활성을 갖는 다당류를 제조하였다. 우선 상기 선택된 녹조류를 건조하고, 건조된 시료를 80% 에틸알코올로 상온에서 12시간 동안 교반 추출을 수행하고, 원심분리하여 펠렛을 수득한다. 수득된 펠렛을 80% 에틸알코올을 이용하여 상온에서 12시간동안 교반 추출 후, 원심분리하여 펠렛을 수득한다. 수득된 펠렛은 아세톤을 이용하여 색소, 지질 및 저분자 단백질을 제거 후, 상온에서 건조하여 건조된 녹조류 분말을 획득한다. In the present invention, polysaccharides having immuno-enhancing activity were prepared using green seaweed as a raw material as algae as a seaweed. First, the selected green alga is dried, and the dried sample is subjected to stirring extraction at room temperature for 12 hours with 80% ethyl alcohol, and centrifuged to obtain pellets. The obtained pellets were extracted by stirring for 12 hours at room temperature using 80% ethyl alcohol, followed by centrifugation to obtain pellets. The obtained pellet is acetone to remove the pigment, lipids and low molecular weight protein, and then dried at room temperature to obtain a dried green algae powder.
상기에서 수득한 녹조류 분말을 증류수를 첨가하여 60℃에서 2시간 교반 추출 후, 원심분리를 이용하여 상등액을 수득한다. 펠렛은 다시 증류수를 첨가하여 60℃에서 2시간 교반 추출 후, 원심분리를 이용하여 상등액을 수득한다. 상기의 2회에서 수득된 상등액을 감압 농축기를 이용하여 농축 후, 에틸알코올을 첨가하여 다당류를 침전 시켰고, filtration을 이용하여 다당류를 획득 하였다. 획득된 다당류는 Ion exchange chromatography를 이용하여 3개의 fraction으로 분리 하였고, 분리하기 전의 다당류와 분리한 후 다당류를 이용하여 NO, TNF-α, IL-1β, PGE2등의 면역관련 사이토카인 분비 능력을 평가하였다. 이와 같은 녹조류 면역관련 사이토카인의 분비능을 관찰한 결과, 본 발명에 따른 녹조류 유래 다당류는 NO, TNF-α, IL-1β, PGE2등의 면역관련 사이토카인을 분비함으로써 면역 증강활성이 우수한 것으로 나타났다.
After distilled water was added to the green algae powder obtained above, the mixture was stirred for 2 hours at 60 ° C., and a supernatant was obtained by centrifugation. The pellet was again extracted by stirring with distilled water at 60 ° C. for 2 hours, and then obtained a supernatant by centrifugation. The supernatant obtained in the above two times was concentrated using a vacuum concentrator, and then ethyl alcohol was added to precipitate polysaccharides, and polysaccharides were obtained by filtration. The obtained polysaccharides were separated into three fractions by ion exchange chromatography. The polysaccharides were separated from the polysaccharides before separation, and the polysaccharides were used to secrete immune-related cytokines such as NO, TNF-α, IL-1β, and PGE 2 . Evaluated. As a result of observing the secretion ability of the green algae immune-related cytokine, the green algae-derived polysaccharide according to the present invention was shown to have excellent immune enhancing activity by secreting immune-related cytokines such as NO, TNF-α, IL-1β, PGE 2 .
상기한 바와 같이, 본 발명에 따른 면역증강활성을 갖는 녹조류 유래 다당류는 마우스 대식세포인 Raw 264.7 세포에서 면역관련 사이토카인인 NO, TNF-α, IL-1β, PGE2등의 물질을 분비하여 면역 활성을 증강시켜주며, 식품산업에서는 면역증강용 기능성 식품으로, 면역기능이 약한 사람의 면역 활성 증강을 위한 보조 식품으로 사용이 가능한, 식품 소재 개발 산업에 있어 매우 우수한 발명인 것으로 사료된다.
As described above, the green algae-derived polysaccharide having immuno-stimulating activity according to the present invention secretes immune-related cytokines NO, TNF-α, IL-1β, and PGE 2 from raw 264.7 cells, which are mouse macrophages. It is believed to be an excellent invention in the food material development industry, which enhances the activity, and is used as a functional food for enhancing immune function in the food industry, and can be used as a supplement for enhancing immune activity of a person with weak immune function.
도 1은 가시파래 유래 다당류와 분리된 다당류의 NO 분비능 측정 결과를 나타낸 그래프이다
도 2는 가시파래 유래 다당류와 분리된 다당류의 TNF-α 분비능 측정 결과를 나타낸 그래프이다
도 3은 가시파래 유래 다당류와 분리된 다당류의 IL 1β 분비능 측정 결과를 나타낸 그래프이다
도 4는 가시파래 유래 다당류와 분리된 다당류의 PGE2 분비능 측정 결과를 나타낸 그래프이다1 is a graph showing the measurement results of NO secretion of the polysaccharide separated from the visible blue-derived polysaccharides
Figure 2 is a graph showing the measurement results of TNF-α secretion of the polysaccharide separated from the visible polysaccharide derived polysaccharides
Figure 3 is a graph showing the measurement results of IL 1β secretion of the polysaccharide separated from the spinach-derived polysaccharides
Figure 4 is a graph showing the results of measuring the PGE2 secretion of polysaccharides separated from thorn-derived polysaccharides
이하 본 발명의 바람직한 실시 형태를 실시예를 참고로 하여 보다 상세하게 설명한다. 하지만 본 발명의 범위가 이러한 실시예에 한정되는 것은 아니다.
Hereinafter, preferred embodiments of the present invention will be described in more detail with reference to Examples. However, the scope of the present invention is not limited to these examples.
실시예 1: 녹조류에서 다당류 추출Example 1 Polysaccharide Extraction from Green Algae
본 실시예에서는 녹조류 가시파래로부터 다당류를 추출하였다. In the present embodiment, polysaccharides were extracted from green algae barley.
우선 가시파래를 60℃에서 2일간 열풍건조하였다. 건조된 해조류 50g에 80% 에틸알코올 500mL를 첨가하여 상온에서 12시간 동안 교반하면서 가열한 후, 원심분리하여 펠렛을 수득하였다. 이와 같은 과정을 2회 반복하고, 수득된 펠렛을 아세톤을 사용하여 펠렛에 남아있는 색소, 지질 및 저분자 단백질을 제거 하였다. 상기 색소, 지질 및 저분자 단백질이 제거된 시료를 원심분리하여 최종 펠렛을 수득한 후, 상온에서 18시간 건조하여 분말화 하였다. First, the visible seaweed was hot-air dried at 60 ° C. for 2 days. 500 g of 80% ethyl alcohol was added to 50 g of dried seaweed, and the mixture was heated with stirring for 12 hours at room temperature, followed by centrifugation to obtain pellets. This process was repeated twice, and the resulting pellets were removed using acetone to remove pigments, lipids and small molecule proteins remaining in the pellets. Centrifuged to remove the pigment, lipid and low molecular protein samples to obtain a final pellet, and then dried at room temperature for 18 hours to powder.
상기에서 수득된 분말 20g에 400mL의 증류수를 첨가하여 65℃에서 2시간동안 교반 가열 한 후, 원심분리하여 상등액을 수득하였다. 이와 같은 과정을 2회 반복하고, 수득된 상등액은 감압농축기를 이용하여 100mL로 농축하였다. 농축된 상등액에 총 알코올의 함량이 30%(v/v)가 되게 에틸알코올을 첨가한 후, 4℃에서 4시간 동안 보관하고, 원심분리 후 상등액을 수득하였다. 수득된 상등액에 총 알코올의 함량이 80%(v/v)가 되게 에틸알코올을 첨가한 후, 4℃에서 18시간 보관하였다. 알코올에 의해 침전된 침전물을 0.4μm 여과지로 여과하여 다당류를 수득하고, 이를 수세하여 최종적으로 가시파래 유래 다당류를 수득하였다. 이와 같이 수득한 다당류를 조(crude) 다당류라 명하였다.
To 20 g of the powder obtained above, 400 mL of distilled water was added and stirred and heated at 65 ° C. for 2 hours, followed by centrifugation to obtain a supernatant. This process was repeated twice, and the obtained supernatant was concentrated to 100 mL using a vacuum concentrator. Ethyl alcohol was added to the concentrated supernatant so that the total alcohol content was 30% (v / v), and then stored at 4 ° C. for 4 hours to obtain a supernatant after centrifugation. Ethyl alcohol was added to the obtained supernatant so that the total alcohol content was 80% (v / v), and then stored at 4 ° C. for 18 hours. The precipitate precipitated by alcohol was filtered through 0.4 μm filter paper to obtain a polysaccharide, which was washed with water to finally obtain a spiny blue derived polysaccharide. The polysaccharide thus obtained was named crude polysaccharide.
실시예 2: 이온 교환 Example 2: Ion Exchange 컬럼column 크로마토그래피를이용한Chromatography 다당류의 정제 Purification of Polysaccharides
본 실시예에서는 이온 교환 컬럼 크로마토그래피(DEAE Sepharose Fast Flow column)를 이용하여 상기 실시예 1에서 수득한 가시파래 유래 조 다당류를 정제하였다.In this example, the crude polysaccharide derived from spiny blue obtained in Example 1 was purified using ion exchange column chromatography (DEAE Sepharose Fast Flow column).
우선 상기 실시예 1에서 수득된 조 다당류 200mg을 10mL 증류수에 용해 후, 극초단파(microwave bomb)를 이용하여 더 작은 입자로 용해시켰다. 극초단파에 의해 가열되어 뜨거운 다당류를 이온 교환 컬럼 크로마토그래피에 주입한 후, 증류수를 2시간 동안 흘려주었다. 4시간 동안 용리한 용액에서 수득한 분획물을 F1분획물이라 명하고, 용매를 0.5 M NaCl 용액으로 바꾸어 준 후, 다시 4시간 후 동한 용리한 용액에서 수득한 분획물을 F2 분획물이라 하였으며, 1.0 M NaCl 용액으로 4시간 동안 용리한 용액에서 수득한 분획물을 F3 분획물이라 하였다. 각 용매에서 수득한 다당류를 Mw 8000의 투석막(dialysis membrane)에 주입하여, 일반수에서 2일간, 증류수에서 1일간 투석한 후 동결건조하여 최종 정제된 F1, F2, F3 다당류를 수득하였다.
First, 200 mg of the crude polysaccharide obtained in Example 1 was dissolved in 10 mL of distilled water, and then dissolved into smaller particles by using a microwave bomb. After heating by microwave, hot polysaccharides were injected into ion exchange column chromatography, and distilled water was flowed for 2 hours. The fraction obtained from the solution eluted for 4 hours was called F1 fraction, the solvent was changed to 0.5 M NaCl solution, and again after 4 hours, the fraction obtained from the same eluted solution was called F2 fraction, and 1.0 M NaCl solution. The fraction obtained from the solution eluted for 4 hours was called F3 fraction. Polysaccharides obtained from each solvent were injected into a dialysis membrane of Mw 8000, dialyzed in distilled water for 2 days in normal water, and then lyophilized to obtain final purified F1, F2, and F3 polysaccharides.
실험예Experimental Example 1 : 가시파래 유래 다당류의 1: of polysaccharides NONO 생성능Generation 평가 evaluation
본 실험예에서는 상기 실시예 1과 2에서 각각 수득한 가시파래 유래 다당류의 NO의 생성능을 측정하였다. 마우스 대식세포는 105 cell/well를 사용하였으며, 세포 배양 24 시간 후, 다당류를 6.25, 12.5, 25 ug/mL 농도로 배양된 마우스 대식세포에 첨가하여 다시 24시간 동안 배양한 후, 상등액을 회수 하였다. 세포에서 분비되는 NO의 양은 회수된 상등액과 그리스 시약(Griess reagent)을 동량으로 첨가하여 플레이트 리더기 540 nm를 이용하여 측정하였다. 평가 결과를 도 1에 나타내었다.In the present experimental example, the NO production ability of the polysaccharide derived from prickly pear obtained in Examples 1 and 2 was measured. Mouse macrophages were used for 10 5 cell / well, and after 24 hours of cell culture, polysaccharides were added to mouse macrophages incubated at 6.25, 12.5, and 25 ug / mL concentrations, followed by culturing for 24 hours, and then the supernatant was recovered. It was. The amount of NO secreted from the cells was measured using a plate reader 540 nm with the same amount of the recovered supernatant and Greases reagent. The evaluation results are shown in FIG. 1.
도 1에 나타낸 바와 같이, 가시파래에서 수득된 다당류 및 이를 이온 교환 컬럼 크로마토그래피로 정제한 다당류는 마우스 대식세포인 Raw 264.7 세포를 자극하여, NO의 분비를 유도하였다. 특히 F2 분획의 경우, 6.25 ug/mL 농도에서도 비교 대조군으로 사용한 LPS(lipopolysaccharide) 10ug/mL와 유사한 NO의 분비량을 나타내었다.
As shown in FIG. 1, polysaccharides obtained from spiny blue and polysaccharides purified by ion exchange column chromatography stimulated Raw 264.7 cells, which are mouse macrophages, to induce NO secretion. Particularly, in the case of the F2 fraction, the secretion amount of NO was similar to that of 10ug / mL of LPS (lipopolysaccharide) used as a comparative control even at a concentration of 6.25 ug / mL.
실험예Experimental Example 2 : 가시파래 유래 다당류의 2: of polysaccharides TNFTNF -α 및 -α and ILIL -1β -1β 생성능Generation 평가 evaluation
본 실험예에서는 상기 실시예 1과 2에서 각각 수득한 가시파래 유래 다당류의 TNF-α 및 IL-1β의 생성능을 측정하였다. 마우스 대식세포는 105 cell/well를 사용하였으며, 세포 배양 24 시간 후, 다당류를 6.25, 12.5, 25 ug/mL 농도로 배양된 마우스 대식세포에 첨가하여 다시 24시간 동안 배양한 후, 상등액을 회수 하였다. 회수된 상등액은 TNF-α 및 IL-1β의 사이토카인을 측정하는 키트를 사용하였으며, 각 사이토카인의 분비 흡광도는 플레이트 리더기 450nm를 이용하여 측정하였다. 평가 결과를 도 2와 도 3에 나타내었다.In this experimental example, the production capacity of TNF-α and IL-1β of the polysaccharide derived from prickly pear obtained in Examples 1 and 2, respectively, was measured. Mouse macrophages were used for 10 5 cell / well, and after 24 hours of cell culture, polysaccharides were added to mouse macrophages incubated at 6.25, 12.5, and 25 ug / mL concentrations, followed by culturing for 24 hours, and then the supernatant was recovered. It was. The collected supernatant was used to measure the cytokines of TNF-α and IL-1β, and the secretion absorbance of each cytokine was measured using a plate reader 450nm. The evaluation results are shown in FIGS. 2 and 3.
도 2와 도 3에 나타낸 바와 같이, 가시파래에서 수득된 다당류 및 이를 이온 교환 컬럼 크로마토그래피로 정제한 다당류는 마우스 대식세포인 Raw 264.7 세포를 자극하여, TNF-α 및 IL-1β의 분비를 유도하였다. 특히 TNF-α의 경우 F1 및 F2 분획이 6.25 ug/mL 농도에서도 비교 대조군으로 사용한 LPS 10 ug/mL와 유사한 분비량을 나타내었다. 한편, IL-1β는 세 종류의 분획 중에서 F2가 가장 많은 IL-1β의 분비량을 나타내었으며, 특히, 6.25 ug/mL에서 비교 대조군으로 사용한 LPS 10 ug/mL와 유사한 분비량을 나타내었다.
As shown in Figures 2 and 3, polysaccharides obtained from spiny blue and polysaccharides purified by ion exchange column chromatography stimulate raw 264.7 cells, which are mouse macrophages, to induce secretion of TNF-α and IL-1β. It was. In particular, in the case of TNF-α, the F1 and F2 fractions showed a similar secretion amount to
실험예Experimental Example 3 : 가시파래 유래 다당류의 3: spiny-derived polysaccharide PGEPGE 22 생성능Generation 평가 evaluation
본 실험예에서는 상기 실시예 1과 2에서 각각 수득한 가시파래 유래 다당류의 PGE2의 생성능을 측정하였다. 마우스 대식세포는 105 cell/well를 사용하였으며, 세포 배양 24 시간 후, 다당류를 6.25, 12.5, 25 ug/mL 농도로 배양된 마우스 대식세포에 첨가하여 다시 24시간 동안 배양한 후, 상등액을 회수하였다. 회수된 상등액은 PGE2 사이토카인을 측정하는 키트를 사용하였으며, 사이토카인의 분비 흡광도는 플레이트 리더기 450nm를 이용하여 측정하였다. 평가 결과를 도 4에 나타내었다. In this Experimental Example, the production capacity of PGE 2 of the spiny blue derived polysaccharides obtained in Examples 1 and 2, respectively, was measured. Mouse macrophages were used for 10 5 cell / well, and after 24 hours of cell culture, polysaccharides were added to mouse macrophages incubated at 6.25, 12.5, and 25 ug / mL concentrations, followed by culturing for 24 hours, and then the supernatant was recovered. It was. The recovered supernatant was used as a kit for measuring PGE 2 cytokines, and the secretion absorbance of cytokines was measured using a plate reader 450 nm. The evaluation results are shown in FIG. 4.
도 4에 나타낸 바와 같이, 가시파래에서 수득된 다당류 및 이를 이온 교환 컬럼 크로마토그래피로 정제한 다당류는 마우스 대식세포인 Raw 264.7 세포를 자극하여, PGE2의 분비를 유도하였다. F1 분획은 12.5 ug/mL 농도에서 비교 대조군으로 사용한 LPS 10 ug/mL와 유사한 분비량을 나타내었다. 또한 F2 분획은 그보다 낮은 농도인 6.25 ug/mL 농도에서 비교 대조군으로 사용한 LPS 10ug/mL와 유사한 분비량을 나타내었다. As shown in FIG. 4, polysaccharides obtained from spiny blue and polysaccharides purified by ion exchange column chromatography stimulated Raw 264.7 cells, which are mouse macrophages, to induce the secretion of PGE 2 . The F1 fraction showed a secretion similar to 10 ug / mL LPS used as a comparative control at a concentration of 12.5 ug / mL. In addition, the F2 fraction showed a secretion similar to that of LPS 10ug / mL used as a comparative control at a lower concentration of 6.25 ug / mL.
Claims (5)
수득된 펠렛을 아세톤을 사용하여 펠렛에 남아있는 색소, 지질 및 저분자 단백질을 제거 하는 단계;
상기 색소, 지질 및 저분자 단백질이 제거된 시료를 원심분리하여 최종 펠렛을 수득한 후, 건조하여 분말화하는 단계;
상기에서 수득된 분말에 증류수를 첨가하고, 교반하면서 가열 한 후, 원심분리하여 상등액을 수득하는 단계;
수득된 상등액을 농축하고, 농축된 상등액에 에틸알코올을 30%(v/v)가 되게 첨가한 후, 원심분리하여 상등액을 수득하는 단계;
수득된 상등액에 에틸알코올을 80%(v/v)가 되게 첨가하여 침전물을 수득하고 이를 여과, 세척하여 다당류를 수득하는 단계; 및
상기 수득된 다당류를 이온 교환 컬럼 크로마토그래피로 정제하는 단계
로 이루어진 감태 유래 다당류의 제조 방법.
Adding 80% ethyl alcohol to 10% (v / w) in the dried Ecklonia cava, heating with stirring, and centrifuging to obtain pellets;
Removing the remaining pigments, lipids and low molecular weight proteins from the pellets using acetone;
Centrifuging the sample from which the pigments, lipids and low molecular weight proteins have been removed to obtain final pellets, and then drying and powdering;
Adding distilled water to the powder obtained above, heating with stirring, and centrifuging to obtain a supernatant;
Concentrating the obtained supernatant, adding ethyl alcohol to 30% (v / v) to the concentrated supernatant, and centrifuging to obtain a supernatant;
Adding ethyl alcohol to 80% (v / v) to the obtained supernatant to obtain a precipitate, which was filtered and washed to obtain a polysaccharide; And
Purifying the obtained polysaccharide by ion exchange column chromatography
Method for producing Ecklonia derived polysaccharide.
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