KR20210013482A - Method for Extracting Active Ingredient of Herb Medicine and Composition for Enhancing Activity of Macrophage Prepared therby - Google Patents
Method for Extracting Active Ingredient of Herb Medicine and Composition for Enhancing Activity of Macrophage Prepared therby Download PDFInfo
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- KR20210013482A KR20210013482A KR1020190090949A KR20190090949A KR20210013482A KR 20210013482 A KR20210013482 A KR 20210013482A KR 1020190090949 A KR1020190090949 A KR 1020190090949A KR 20190090949 A KR20190090949 A KR 20190090949A KR 20210013482 A KR20210013482 A KR 20210013482A
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Abstract
Description
본 발명은 식물 세포벽 분해효소를 활용한 새로운 식물성 생약의 유효성분 추출방법과 이 방법에 의해 제조되는 선천면역계를 대표하는 대식세포 활성 증진효과를 가진 식물성 생약 조성물에 관한 것이다.The present invention relates to a method for extracting an active ingredient of a new plant herbal medicine using a plant cell wall degrading enzyme, and a plant herbal composition having an effect of enhancing macrophage activity representing an innate immune system produced by this method.
생약(生藥, crude drugs)이란 의약품의 일종으로, 천연으로 산출되는 자연물 중 소정의 생물학적 효능이 알려진 것을 그대로 또는 건조하거나, 썰거나 분말화 하는 정도의 간단한 가공처리를 한 의약품 또는 의약품 소재를 말한다. 생약에는 망초, 석고, 영사 등과 같은 광물성, 사향, 웅담, 봉밀 등과 같은 동물성이 있지만 대부분은 식물성이다. Crude drugs are a kind of pharmaceuticals, and refer to pharmaceuticals or pharmaceutical materials that have been subjected to simple processing such as drying, slicing, or powdering of natural products with known biological efficacy. Herbal medicines have minerals such as forget-me-not, gypsum, and yeongsa, and animals such as musk, ungdam, and bongmil, but most of them are vegetable.
한방에서는 다양한 생약을 건조하여 정해진 성분과 함량에 따라 배합하여 사용하는데, 구체적인 제조방법에 따라 한약을 탕제(湯劑, 전제煎劑라 하기도 함), 침제(浸劑), 분제(粉劑), 환제(丸劑) 및 탄제(炭劑) 등으로 구분한다. 이 중 탕제는 소위 '달임'이라는 방식으로 열수추출하여 얻은 즙(탕약)으로 거의 모든 한약 종류에서 활용되고 있다.In oriental medicine, various herbal medicines are dried and used in combination according to the prescribed ingredients and contents.According to the specific manufacturing method, herbal medicines are also referred to as Tangje (湯劑, Jeonje煎劑), acupuncture (浸劑), powder (粉劑), and pills. It is divided into (丸劑) and tanje (炭劑). Among them, Tangje is a juice obtained by hot water extraction in the so-called'dalim' and is used in almost all types of herbal medicine.
생약으로 활용되는 식물에서 '생물학적 유효성분'은 세포내 또는 세포벽에 존재한다. 즉, 건조된 식물성 생약에서 유효성분은 셀룰로즈, 펙틴, 헤미셀룰로즈 등 거대 다당류로 이루어진 딱딱하고 치밀한 세포벽 내부에 또는 세포벽과 세포벽 사이에 존재한다. 탕약 제조를 위한 '달임(decoction)'이란, 고온의 열수로 건조 생약의 딱딱하고 치밀한 세포벽을 느슨하게 해체하고 붕괴시켜 세포벽으로부터 유효성분을 분리(열수추출)하는 과정이다. 이때 일부 세포벽 성분이 열분해되어 펙틴이나 헤미셀룰로즈 자체 또는 이들이 물리적으로 추출과정 중 수식된 형태로 된 면역활성 다당류들도 추출되어 생약의 유효성 증대에 기여한다. 식물조직의 건조과정 및 고온의 달임 과정에서 단백질이 변성되므로 단백질은 탕약의 유효성분이 아니다.In plants used as herbal medicines,'biologically active ingredients' exist in the cell or in the cell wall. That is, in the dried herbal medicine, the active ingredient exists in the hard and dense cell wall made of macropolysaccharides such as cellulose, pectin, and hemicellulose , or between the cell wall and the cell wall . The'decoction' for the manufacture of hot water is a process of separating the active ingredient from the cell wall (hot water extraction) by loosening and disintegrating the hard and dense cell wall of a dried herbal medicine with high temperature hot water. At this time, some cell wall components are pyrolyzed to extract pectin or hemicellulose itself, or immunoactive polysaccharides in a modified form during the physical extraction process, thereby contributing to the increase of the efficacy of the herbal medicine. Protein is not an active ingredient in bath medicine because the protein is denatured in the process of drying plant tissues and decoction at high temperatures.
식물성 생약을 탕약화하는 공정은 다음과 같은 세 가지 문제를 내재하고 있다. 첫째, 탕약화는 통상 90℃ 이상의 열수에서 3~6시간 처리해야 하기 때문에 시간과 비용이 많이 소비된다. 둘째 한약재의 용량과 달이는 시간에 따라 약효와 약리반응에 차이가 난다는 사실이 알려져 있는데, 약탕기의 크기, 재질, 제어특성 등이 매우 다양하기 때문에 결과물인 '턍약'의 품질 균일성을 유지하기 어렵다. 셋째, 예를 들면 휘발성 유효성분을 가진 생약(계피, 박하, 계지, 곽향, 청호, 세신, 백두구, 초두구, 단향, 목향, 회향, 감국 등)의 경우 고온에서 시간이 경과되면서 상당한 유효성분이 휘발되어 사라진다. 이러한 문제를 해결하기 위하여 휘발성 생약을 달임의 후단에 추가하는 방법이 시도되고 있으나, 이는 공정상 복잡하고 특히 대량생산에서는 적용하기 쉽지 않다.The process of making herbal herbal medicines has three problems: First, the weakening of hot water usually requires a lot of time and cost because it has to be treated in hot water of 90°C or higher for 3 to 6 hours. Second, it is known that the medicinal efficacy and pharmacological response differ depending on the dosage and decoction time of medicinal herbs.Since the size, material, and control characteristics of the medicinal herbs are very diverse, it is difficult to maintain the uniformity of the quality of the resultant'Pyeongyak'. . Third, for example, in the case of herbal medicines with volatile active ingredients (cinnamon, mint, gyeji, gwakhyang, cheongho, sesin, baekdugu, chodugu, danhyang, mokhyang, fennel, gamguk, etc.), considerable active ingredients volatilize over time at high temperatures. Become and disappear. In order to solve this problem, a method of adding a volatile herbal medicine to the rear end of the decoction has been attempted, but this is complicated in the process and is not easy to apply, especially in mass production.
등록특허 10-1774564는 인삼에 아밀라제 및 셀룰라제를 순차 처리하고 에탄올로 침전·분리하는 다당류만을 얻는 방식으로, 면역기능 증강활성을 가진 인삼유래 다당분획물을 제조하는 방법에 관한 것이다. 등록특허 10-1774566은 보리잎을 펙티나제로 처리하고 다당류를 역시 에탄올로 침전·분리하는 방식으로 면역기능 증강활성을 가진 다당분획물을 얻는 방법에 관한 것이다. 등록특허 10-1868507은 그라비올라 분말을 셀룰라제 및 펙티나제로 처리하고 분리한 항산화, 항염 및 주름 개선용 화장료 조성물에 관한 것인데, '다당류'에 대한 언급은 없으나 에탄올로 침전·분리 과정을 거치고 있어 최종 산물이 '다당류'임을 알 수 있다.Registered Patent 10-1774564 relates to a method of preparing a ginseng-derived polysaccharide fraction having immune function enhancing activity by sequentially treating ginseng with amylase and cellulase and obtaining only polysaccharides that precipitate and separate with ethanol. Registered Patent 10-1774566 relates to a method of obtaining a polysaccharide fraction having immune function enhancing activity by treating barley leaves with pectinase and separating and separating polysaccharides with ethanol. Registered Patent 10-1868507 relates to a cosmetic composition for antioxidant, anti-inflammatory, and wrinkle improvement obtained by treating graviola powder with cellulase and pectinase, and separating it. Although there is no mention of'polysaccharide', it is undergoing a precipitation and separation process with ethanol. It can be seen that the final product is'polysaccharide'.
이들 종래기술들은 식물 조직에 분해효소를 처리하여 기능성 있는 다당류를 생성하고 이를 순수분리하는 것인데, 애초에 원료 식물체가 가지고 있는 각종 생리학적 유효성분(혹여 그 구조와 기능이 명백히 밝혀지지 않은 것이더라도)을 효율적으로 추출하는 것에는 관심이 없다.These conventional techniques are to produce functional polysaccharides by treating plant tissues with degrading enzymes, and to purely separate them.In the first place, various physiologically active ingredients (even if their structure and function are not clearly identified) are used. I am not interested in extracting efficiently.
한편, 생약으로부터 유래된 다양한 물질을 이용한 식품이 환경 및 식품에 혼입(잔류)되어 있는 환경호르몬 영향을 상쇄하거나 면역시스템 활성화로 질병에 대한 생체방어 시스템을 보강한다는 연구결과가 밝혀짐에 따라 이들을 이용한 기능성식품 또는 약용식품 산업화가 주목을 받고 있다. 예를 들자면, 생약 자원을 이용한 건강기능성식품, 건강보조식품, 천연색소 및 천연향미료, 차 및 약용주, 조리 중에 첨가하는 식품첨가제로서의 소스 및 향신료 등이 주목받고 있다. 이러한 추세에 맞추어 안전성과 약효가 충분히 알려져 있고 채취 또는 재배가 용이하여 다양한 한방약에 널리 사용되는 몇몇 범용성 있는 생약재들을 소재로 하여 건강기능성식품 또는 건강보조식품으로 활용할 수 있도록 제공하는 것도 시의적절하다 할 것이다.On the other hand, as research results show that foods using various substances derived from herbal medicines compensate for the effects of environmental hormones mixed (residue) in the environment and food, or reinforce the biological defense system against diseases by activating the immune system, Industrialization of functional foods or medicinal foods is drawing attention. For example, health functional foods using herbal resources, health supplements, natural colors and natural flavors, tea and medicinal liquor, sauces and spices as food additives added during cooking are attracting attention. In line with this trend, it is also timely to provide some general-purpose herbal materials widely used in various herbal medicines as materials for their safety and medicinal effects are sufficiently known and easy to collect or cultivate so that they can be used as health functional foods or health supplements. will be.
본 발명은 종래 '달임 방식'보다 온건하고 효율적으로 식물성 생약에서 생물학적 유효성분을 추출할 수 있는 방법을 제공하는 것을 목적으로 한다. It is an object of the present invention to provide a method for extracting biologically active ingredients from herbal medicines more moderately and more efficiently than the conventional'decoction method'.
또한 본 발명은 이러한 유효성분 추출방법에 따라 제조되는 선천면역계를 대표하는 대식세포 활성 증진효과가 강화된 조성물을 제공하는 것을 목적으로 한다.In addition, an object of the present invention is to provide a composition with enhanced macrophage activity enhancing effect representing the innate immune system prepared according to such an active ingredient extraction method.
전술한 목적을 달성하기 위한 본 발명은 The present invention for achieving the above object
(A) 한 종류 또는 복수 종류의 건조된 식물성 생약 분쇄물을 준비하는 단계; (B) 상기 분쇄물을 셀룰라제, 헤미셀룰라제 또는 펙티나제 중 최소한 어느 하나를 포함하는 식물 세포벽 분해효소로 처리하는 단계; (C) 상기 효소를 실활시키는 단계;를 포함하는 식물성 생약의 유효성분 추출방법에 관한 것이다.(A) preparing a pulverized product of one type or a plurality of types of dried herbal medicines; (B) treating the pulverized product with a plant cell wall degrading enzyme containing at least one of cellulase, hemicellulase, or pectinase; It relates to a method for extracting an active ingredient from a plant herbal medicine comprising (C) inactivating the enzyme.
또한 본 발명은 상기 추출방법을 활용하여 제조된 대식세포 활성 증진효과를 가진 식물성 생약 조성물에 관한 것이다.In addition, the present invention relates to a plant-based herbal composition having an effect of enhancing macrophage activity prepared using the above extraction method.
이상과 같이 본 발명에 의하면 종래 달임 방식에 비해 경제적인 방식으로 균일한 품질의 식물성 생약 추출물을 얻을 수 있게 된다.As described above, according to the present invention, it is possible to obtain a plant herbal extract of uniform quality in an economical manner compared to the conventional decoction method.
또한 본 발명에 의하면 달임 방식에 비해 비교적 저온에서 추출이 이루어지므로 휘발성 유효성분도 보존이 가능하게 된다.In addition, according to the present invention, since extraction is performed at a relatively low temperature compared to the decoction method, it is possible to preserve volatile active ingredients.
또한 본 발명에 의하면 독특한 추출방식을 채택함으로써 가격이 저렴한 범용성 있는 식물성 생약재들을 활용하여 선천면역계를 대표하는 대식세포 활성 증진효과가 뛰어난 생약 조성물을 제공할 수 있게 된다.In addition, according to the present invention, by adopting a unique extraction method, it is possible to provide a herbal composition having excellent macrophage activity enhancing effect representing the innate immune system by utilizing universally inexpensive plant herbal medicines.
도 1은 본 발명에 의한 조성물이 대식세포에 대하여 독성이 없음을 보여주는 도표.
도 2 내지 도 5는 각각 본 발명의 실시예 조성물에 의한 대식세포의 활성인자 생성능을 보여주는 도표.1 is a diagram showing that the composition according to the present invention is not toxic to macrophages.
2 to 5 are diagrams showing the ability of macrophages to produce activators according to an example composition of the present invention, respectively.
이하 첨부된 도면과 사전실험 및 실시예를 들어 본 발명을 보다 상세히 설명한다. 그러나 이러한 도면과 실시예는 본 발명의 기술적 사상의 내용과 범위를 쉽게 설명하기 위한 예시일 뿐, 이에 의해 본 발명의 기술적 범위가 한정되거나 변경되는 것은 아니다. 이러한 예시에 기초하여 본 발명의 기술적 사상의 범위 안에서 다양한 변형과 변경이 가능함은 당업자에게는 당연할 것이다. Hereinafter, the present invention will be described in more detail with reference to the accompanying drawings and prior experiments and examples. However, these drawings and embodiments are only examples for easily explaining the content and scope of the technical idea of the present invention, thereby not limiting or changing the technical scope of the present invention. It will be obvious to those skilled in the art that various modifications and changes are possible within the scope of the technical idea of the present invention based on these examples.
(1) 식물성 생약의 유효성분 추출방법(1) Method of extracting active ingredients from herbal medicines
전술하였듯이 본 발명은, 건조되어 딱딱하고 치밀한 식물 세포벽 내부 또는 세포벽과 세포벽 사이에 존재하는 식물성 생약의 유효성분을 고온의 열수로 추출하는 종래의 달임 방식을 개선하고자 도출된 것이다. 즉, 본 발명은 식물 세포벽을 이루는 셀룰로즈, 펙틴, 헤미셀룰로즈 등 거대 다당류를 식물 세포벽 분해효소를 처리하여 온건한 조건에서 딱딱하고 치밀한 세포벽을 부분적으로 분해하여 느슨하게 해체함으로써 세포벽에 존재하는 유효성분을 효과적으로 추출되도록 다음과 같은 단계를 포함하는 추출방법에 관한 것이다.As described above, the present invention was derived to improve the conventional decoction method of extracting the active ingredient of a plant herbal medicine existing inside the dry and hard and dense plant cell wall or between the cell wall and the cell wall with hot water at high temperature. That is, in the present invention, the active ingredient present in the cell wall is effectively dismantled by partially decomposing the hard and dense cell wall under moderate conditions by treating the plant cell wall degrading enzyme with macropolysaccharides such as cellulose, pectin, and hemicellulose constituting the plant cell wall. It relates to an extraction method comprising the following steps to be extracted.
(A) 한 종류 또는 복수 종류의 건조된 식물성 생약 분쇄물을 준비하는 단계(A) preparing a pulverized product of one or more types of dried vegetable herbal medicine
(B) 상기 분쇄물을 셀룰라제, 헤미셀룰라제 또는 펙티나제 중 최소한 어느 하나를 포함하는 식물 세포벽 분해효소로 처리하는 단계(B) treating the pulverized product with a plant cell wall degrading enzyme containing at least one of cellulase, hemicellulase, or pectinase.
(C) 상기 효소를 실활시키는 단계.(C) inactivating the enzyme.
본 발명에 의하면 필요에 따라 하나의 식물성 생약에서 유효성분을 추출하여 그대로 활용하거나, 추출된 소재별 유효성분을 적정한 비율로 혼합하여 활용할 수 있다. 나아가 식물성 생약을 소재별로 미리 적정 비율로 혼합한 상태에서 한꺼번에 유효성분(의 혼합물)을 추출할 수도 있다. According to the present invention, an active ingredient may be extracted from a single plant herbal medicine and used as it is, or may be used by mixing the extracted active ingredients in an appropriate ratio. Furthermore, it is possible to extract the active ingredient (a mixture of) at once in a state in which the vegetable herbal medicine is mixed in an appropriate ratio in advance for each material.
본 발명은 효소반응을 기반으로 하므로 기본적으로 추출대상 물질(식물성 생약)의 분쇄물이 미세할수록 추출효율이 증대할 것이지만, 너무 미세하다면 미세화 비용 및 추후 필요할 때 잔사를 제거하는데 어려움이 있기 때문에 분쇄물 크기는 적절하게 조절하는 것이 좋다. 하기 실시예에서는 분쇄물을 100메쉬로 하였으나 최적 입자 크기는 변경 가능할 것이다. Since the present invention is based on an enzymatic reaction, the extraction efficiency will increase as the pulverized product of the substance to be extracted (vegetable herbal medicine) is finer, but if it is too fine, the pulverization cost and difficulty in removing the residue later when necessary. It is good to adjust the size accordingly. In the following examples, the pulverized material was 100 mesh, but the optimum particle size may be changed.
세포벽 분해효소로 처리하는 단계는, '효소반응' 과정을 거치므로 효소의 종류, 효소와 기질(분쇄물) 및 수분의 비율, 온도 및 pH 등 반응조건 등등을 적절하게 선택하여 결정할 필요가 있다. 하기 실시예에서는 특정 환경에서 효소 처리과정을 진행하였지만, 상황에 따라 각 변수를 적절하게 변경하고 조정할 수 있음은 당연할 것이다. 효소반응 종료를 위해 예를 들면 가열하는 방식으로 효소를 실활시킨다. 이렇듯 본 발명에 의하면 세포벽 성분을 효소적으로 분해하고 해체함으로써 생약의 유효성분이 효과적으로 추출된다. 이와 동시에, 효소분해된 펙틴이나 헤미셀룰로즈 자체 또는 이들이 효소에 의해 분해되는 과정에서 수식된 형태로 된 면역활성 다당류들도 종래 달임 방식에 비해 많이 생성될 것을 추론할 수 있다.Since the step of treating with cell wall degrading enzyme goes through an'enzymatic reaction' process, it is necessary to appropriately select and determine the type of enzyme, the ratio of enzyme and substrate (pulverized product) and moisture, reaction conditions such as temperature and pH, etc. In the following examples, the enzyme treatment was performed in a specific environment, but it will be natural that each variable can be appropriately changed and adjusted according to circumstances. In order to terminate the enzyme reaction, the enzyme is deactivated, for example, by heating. As described above, according to the present invention, the active ingredient of the herbal medicine is effectively extracted by enzymatically decomposing and dismantling the cell wall components. At the same time, it can be inferred that enzymatically degraded pectin or hemicellulose itself, or immunoactive polysaccharides in a modified form in the process of being degraded by the enzymes, will also be produced more than in the conventional decoction method.
이렇게 유효성분 추출방법을 거쳐 얻어진 조성물을 건조하여 산재(散材)나 환약(丸藥)과 같은 고체상으로 활용할 수도 있을 것이고, 잔사를 제거하고 추출액을 분리하여 탕약(湯藥)으로 활용할 수도 있을 것이다.By drying the composition obtained through the method of extracting the active ingredient in this way, it may be used as a solid phase such as scattered ash or pills, and it may be used as a bath medicine by removing the residue and separating the extract.
하기 실시예에서 확인할 수 있듯이, 본 발명에 의한 추출방법에 따라 얻어진 식물성 생약 조성물은 종래의 달임 방식으로 제조되는 조성물(대조구)에 비해 월등히 우수한 대식세포 활성인자 생산능력을 가지고 있는데, 이는 결국 본 발명에 의한 생약 조성물이 종래 달임 방식에 의한 것보다 면역활성이 우수함을 나타낸다. 본 발명에 의한 조성물의 생산 수율이 대조구와 유사하거나 더 적음에도 실시예에 의한 조성물이 월등히 우수한 면역 활성인자 생산능을 가진다는 것은, 식물성 생약의 세포벽에 함유되어 있는 면역활성 유효성분이 종래의 열수추출(달임 방식)에 비해 훨씬 고농도로 추출되기 때문으로 해석된다. 즉, 원료 식물성 생약의 세포벽에 존재하는 면역활성 유효성분이 종래 열수추출 방식에 비해 훨씬 효율적으로 그리고 높은 비율로 추출되기 때문으로 보인다.As can be seen from the following examples, the plant herbal composition obtained according to the extraction method according to the present invention has far superior macrophage activator production capacity compared to the composition (control) prepared by the conventional decoction method, which in turn has the present invention. It shows that the herbal composition by is superior to that of the conventional decoction method. Although the production yield of the composition according to the present invention is similar to or less than that of the control, the fact that the composition according to the example has a remarkably excellent ability to produce an immune activator, is that the immune-active active ingredient contained in the cell wall of a vegetable herbal medicine is extracted with conventional hot water. It is interpreted as because it is extracted at a much higher concentration than (decoction method). In other words, it seems that this is because the immunologically active active ingredient present in the cell wall of the raw material plant herbal medicine is extracted much more efficiently and at a higher ratio than the conventional hot water extraction method.
(2) 식물성 생약 조성물(2) vegetable herbal composition
또한 본 발명은 전술한 식물성 생약의 유효성분 추출방법을 활용하여 면역기능 활성화 기능을 가진 크게 두 종류의 식물성 생약 조성물에 관한 것이다.In addition, the present invention relates to largely two types of plant-based herbal compositions having an immune function activating function by utilizing the above-described method for extracting active ingredients of plant-based herbal medicines.
첫 번째 조성물은, 당귀 4~6중량부, 천궁 2.5~3.5중량부, 산수유, 갈근, 백출 각각 1.5~2.5중량부, 감초, 백작약, 복령, 산약, 계피, 건강 각각 0.8~1.2중량부의 혼합 분쇄물을 원재료로 하여 얻은 고체상 또는 액체상 조성물이다.The first composition is 4-6 parts by weight of Angelicae, 2.5-3.5 parts by weight of Chungoong, 1.5-2.5 parts by weight of Cornus milk, Galgeun and Baekchul, respectively, Licorice, Baek Peony, Bokryeong, Sanyak, Cinnamon, and Health respectively 0.8-1.2 parts by weight It is a solid or liquid composition obtained using water as a raw material.
두 번째 조성물은, 당귀 4~6중량부, 천궁 2.5~3.5중량부, 하수오, 갈근, 사삼, 산약 각각 1.5~2.5중량부, 감초, 복령, 마가목, 건강 각각 0.8~1.2중량부의 혼합 분쇄물을 원재료로 하여 얻은 고체상 또는 액체상 조성물이다.The second composition contains 4-6 parts by weight of Angelicae, 2.5-3.5 parts by weight of Chungoong, 1.5-2.5 parts by weight of Hasu-O, Galgeun, Sasam, Sanyak, respectively, Licorice, Bokryeong, Rowan, and Health 0.8-1.2 parts by weight of mixed pulverized products, respectively. It is a solid or liquid composition obtained as a raw material.
하기 실시예에 두 조성물 중 액상의 것에 대한 구체적 제조과정을 기재하였다. 액상 조성물은 고체상 조성물에서 액상 부분을 따로 분리한 것인데, 따라서 고체상 조성물은 액상 조성물(의 성분과 특성)을 그대로 모두 포함하고 있는 것이다.In the following examples, the specific manufacturing process for the liquid of the two compositions is described. The liquid composition is obtained by separating the liquid part from the solid composition, so the solid composition contains all of the liquid composition (components and properties of) as it is.
이렇게 얻어진 두 액상 조성물은 모두 50~100 μg/mL의 농도에서 대식세포에 독성을 보이지 않아 인체에 독성을 나타내지 않았다. Both of the liquid compositions thus obtained were not toxic to macrophages at a concentration of 50 to 100 μg/mL, and thus were not toxic to the human body.
대식세포는 선천면역계의 대표적인 면역세포로서 외부 항원물질의 탐식작용에 의한 제거에 중요할 뿐만 아니라 후천면역계와 각종 면역계의 중요한 면역반응의 조절에 관여하는 중요한 기능을 수행한다. 본 발명에 의한 두 액상 조성물의 대식세포 활성화 기여 여부를 확인한 바, 대조구(종래 방식에 의한 달임 방식에 의한 추출물)에 비해 유의적으로 높은 대식세포 활성을 나타내었다. 보다 구체적으로 보면, 본 발명에 의한 조성물은 대식세포 활성화 인자로 알려진 nitric oxide(NO)와 interleukin(IL)-6, IL-12와 tumor necrosis factor(TNF)-α 등의 사이토카인 생산능이 대조구에 비해 유의적으로 우수하였다(실시예 참조). Macrophages are representative immune cells of the innate immune system and are not only important for the removal of foreign antigens by phagocytosis, but also perform important functions involved in the regulation of important immune responses of the acquired immune system and various immune systems. As a result of confirming whether the two liquid compositions according to the present invention contribute to macrophage activation, they showed significantly higher macrophage activity compared to the control (extract by the decoction method by the conventional method). More specifically, the composition according to the present invention has the ability to produce cytokines such as nitric oxide (NO) and interleukin (IL)-6, IL-12 and tumor necrosis factor (TNF)-α, which are known as macrophage activating factors. Compared to, it was significantly superior (see Examples).
각 활성인자의 특성은 다음과 같다.The characteristics of each activator are as follows.
NO는 활성화된 대식세포로부터 inducible nitric oxide synthase(iNOS)에 의해 다량으로 생산되어 외부 침입물질이나 암세포를 제거하는데 관여하는 라디칼이다. TNF-α는 활성화된 대식세포에 의해 분비되어 면역세포를 조절하는 중요한 사이토카인으로서 변형된 세포의 자살을 유도하거나 종양 생성 및 바이러스 복제를 억제하는 기능을 한다. IL-6는 활성화된 대식세포에서 분비되어 B 세포 계열의 증식과 항체 분비 등의 획득면역에 영향을 주는 사이토카인이다. IL-12는 활성화된 대식세포로부터 분비되어 자연살해세포(natural killer cell, NK cell) 및 보조 T 세포 등의 성숙 및 분화를 촉진시키는 사이토카인이다.NO is a radical produced from activated macrophages by inducible nitric oxide synthase (iNOS) in large quantities and is involved in removing foreign invading substances or cancer cells. TNF-α is an important cytokine that is secreted by activated macrophages to regulate immune cells, and functions to induce suicide of modified cells or to inhibit tumorigenesis and viral replication. IL-6 is a cytokine that is secreted by activated macrophages and affects acquired immunity such as proliferation of B cell lines and secretion of antibodies. IL-12 is a cytokine that is secreted from activated macrophages and promotes maturation and differentiation of natural killer cells (NK cells) and helper T cells.
따라서 본 발명에 의한 조성물은 인체의 면역기능을 활성화 시킬 수 있음을 확인하였다.Therefore, it was confirmed that the composition according to the present invention can activate the immune function of the human body.
[실시예] [Example]
1. 식물성 생약 액상 조성물의 제조1. Preparation of vegetable herbal liquid composition
하기 표 1에 의한 생약재를 분쇄하여 100메쉬 이하의 입자로 준비하고 20배(w/v)의 증류수를 가하여 원료 혼합물 조성1, 조성2를 준비하였다.The herbal medicines according to Table 1 below were pulverized to prepare particles of 100 mesh or less, and 20 times (w/v) of distilled water was added to prepare a raw material mixture composition 1 and composition 2.
조성1, 2 각각에 대하여 다음 표 2와 같이 효소를 생약재 조성에 대해 0.1%(w/w) 첨가하고 교반하면서 다음 표 3에 기재한 조건에서 12시간 반응시켰다. 이어서 가열방식으로 효소를 실활시킨 후 원심분리(9,000×g, 5℃, 30분)하여 상등액을 회수하였다. 이 상태가 본 발명에 의한 식물성 생약 액상 조성물에 해당하는데, 경제성을 위해서는 원심분리 방법이 아닌 통상의 필터링 방식으로 액상 조성물을 얻을 수도 있을 것이다.For each of the compositions 1 and 2, 0.1% (w/w) of the enzyme was added to the herbal composition, as shown in Table 2 below, and reacted for 12 hours under the conditions shown in Table 3 while stirring. Subsequently, the enzyme was deactivated by heating, and then centrifuged (9,000× g , 5°C, 30 minutes) to recover the supernatant. This state corresponds to the vegetable herbal liquid composition according to the present invention, but for economical efficiency, the liquid composition may be obtained by a conventional filtering method instead of a centrifugation method.
각 효소의 출처 및 처리조건을 표 3에 기재하였다.The sources and treatment conditions of each enzyme are listed in Table 3.
각각 상기 표 1의 조성1, 2를 95℃에서 물과 생약 혼합액 부피가 반으로 줄 때까지 끓인 후 1차 상등액과 잔사를 분리하고 다시 잔사에 20배(w/v) 증류수를 첨가하여 동일 온도에서 끓여 반으로 줄인 다음 2차 상등액을 분리하고 1, 2차 상등액을 합쳐서 대조구1, 2를 준비하였다. Each of the compositions 1 and 2 in Table 1 above was boiled at 95°C until the volume of the mixture of water and herbal medicines was halved, and the first supernatant and the residue were separated, and 20 times (w/v) distilled water was added to the residue at the same temperature. Boil at and cut in half, and then the second supernatant was separated and the first and second supernatant were combined to prepare controls 1 and 2.
한편, 효소처리가 열수추출 공정과 병행되었을 때 시너지 효과가 있는지를 확인하기 위하여 대조구1, 2 각각에 다시 상기 표 2, 3의 효소와 조건으로 처리한 조성물을 준비하여 하기 특성 분석을 수행하였는데, 대조구1, 2와 유사하거나 열등한 결과를 나타내었다. 즉, 열수추출 후 효소처리 하는 것은 별다른 긍정적인 효과가 나타나지 않았다. 따라서 이와 관련된 구체적인 기재를 생략한다. On the other hand, when enzyme treatment is combined with the hot water extraction process, the synergistic effect is In order to confirm whether or not, a composition treated with the enzymes and conditions in Tables 2 and 3 was prepared again in each of Controls 1 and 2, and the following characteristics were analyzed, showing similar or inferior results to Controls 1 and 2. In other words, enzymatic treatment after hot water extraction did not show any positive effects. Therefore, detailed descriptions related thereto are omitted.
2. 식물성 생약 액상 조성물의 특성 분석2. Characterization of the vegetable herbal liquid composition
실시예에서 얻어진 각 액상 조성물을 10 brix 정도로 농축 후 투석(투석막 12 kDa)하고 다시 10 brix 정도로 농축한 후 동결건조하였다.Each liquid composition obtained in Examples was concentrated to about 10 brix, dialyzed (
(1) 수율 비교(1) Yield comparison
열수추출한 대조구 1과 2의 수율이 실시예들에 비해 평균적으로 높게 나타났다. 이는 전술하였듯이 실시예에서는 1회 효소처리 후 잔사를 제거하고 추출물을 얻었지만 대조구에서는 2회에 걸쳐 추출을 수행한 다음 잔사를 제거했기 때문으로 해석된다. The yield of Controls 1 and 2 extracted with hot water was higher on average than in the Examples. This is interpreted as because, as described above, in the Example, the residue was removed after the enzyme treatment once, and the extract was obtained, but in the control group, extraction was performed twice and then the residue was removed.
(2) 조성물의 대식세포에 대한 독성 및 증식활성(2) Toxicity and proliferative activity of the composition against macrophages
본 발명의 실시예에 의한 조성물이 대식세포에 대하여 독성을 나타내는지의 여부를 확인하기 위하여 조성물로 대식세포 배양 후에 대식세포의 생존율을 Ez-Cytox 시약을 이용하여 측정하였다(도 1 참조). 도에서 Saline은 조성물이 함유되지 않은 생리식염수를 나타내며, 회색막대와 검은색막대는 각각 조성물 50 및 100 μg/mL 첨가한 경우이다(이하 도면 모두 동일).In order to confirm whether the composition according to the embodiment of the present invention exhibits toxicity to macrophages, the survival rate of macrophages after culturing macrophages with the composition was measured using the Ez-Cytox reagent (see FIG. 1). In the figure, Saline represents the physiological saline solution that does not contain the composition, and the gray bars and the black bars are when 50 and 100 μg/mL compositions are added, respectively (the same is true for all of the drawings below).
도면에서 볼 수 있듯이, 실시예에 의한 조성물들은 모두 50과 100 μg/mL의 농도에서 독성을 보이지 않았으며, 오히려 두 처리 농도에서 saline 대조군과 비교하여 1.24 ~ 1.36배로 대식세포 증식이 촉진됨을 확인할 수 있었다. 실시예에 의한 조성물들은 대조군1, 2와 마찬가지로 독성이 없을 뿐만 아니라 대조군보다 대식세포의 증식을 촉진하는 효과도 관찰되었다. As can be seen from the figure, the compositions according to the examples did not show toxicity at concentrations of 50 and 100 μg/mL, but rather, it was confirmed that macrophage proliferation was promoted by 1.24 to 1.36 times compared to the saline control group at both treatment concentrations. there was. The compositions according to the examples were not only non-toxic like controls 1 and 2, but also an effect of promoting the proliferation of macrophages than the control group was observed.
(3) 조성물의 대식세포 활성능 측정(3) Measurement of macrophage activity of the composition
본 발명의 실시예에 의한 조성물이 대식세포 활성화를 촉진하는지 여부를 확인하기 위하여 ICR 마우스 복강에서 회수한 대식세포에 조성물을 처리하여 대식세포로부터 분비되는 대표적인 활성화 사이토카인의 생산량을 분석하였다.In order to confirm whether the composition according to an embodiment of the present invention promotes macrophage activation, the production of representative activated cytokines secreted from macrophages was analyzed by treating the composition on macrophages recovered from the peritoneal cavity of ICR mice.
① 실험방법① Experiment method
ICR 마우스에 5% thioglycollate(TG) medium을 복강주사하여 단구의 복강으로의 침출을 통한 대식세포 분화를 유도하고 TG 주입 3~4일 후 fetal bovine serum(FBS) 함유 RPMI-1640 배지를 복강 내에 주사하여 복강에 침출되고 분화된 대식세포를 회수하였다. ICR mice were intraperitoneally injected with 5% thioglycollate (TG) medium to induce differentiation of macrophages through leaching of monocytes into the peritoneal cavity, and 3 to 4 days after TG injection, RPMI-1640 medium containing fetal bovine serum (FBS) was injected intraperitoneally. Thus, the differentiated macrophages leached into the abdominal cavity were recovered.
96 well plate 각 well에 대식세포 현탁액을 200 μL(1×106 cell/mL)씩 분주하고 CO2 배양기에서 37℃로 배양하여 well에 부착된 대식세포의 monolayer를 형성시킨 2시간 후 각 well을 phosphate buffered saline(PBS)으로 세척하여 미흡착된 세포를 제거하였다. 흡착된 대식세포에 일정 농도로 희석한 조성물을 가하고 CO2 배양기에서 37℃로 다시 배양하였다. Dispense 200 μL (1×10 6 cells/mL) of macrophage suspension into each well of a 96 well plate and incubate at 37°C in a CO 2 incubator to form a monolayer of macrophages attached to the wells. Unadsorbed cells were removed by washing with phosphate buffered saline (PBS). A composition diluted to a certain concentration was added to the adsorbed macrophages and cultured again at 37° C. in a CO 2 incubator.
대식세포 배양 24시간 후 배양 상등액을 다른 well plate로 옮기고 Ez-Cytox 용액을 첨가하여 대식세포 증식 및 조성물에 의한 독성을 측정하는 한편, 대식세포 배양액을 이용한 시료의 대식세포 활성화 여부를 확인하기 위해 대식세포에서 생산되는 활성화 인자인 IL-6, IL-12와 TNF-α 등의 사이토카인 농도는 제조사의 지침에 따라 ELISA 방법으로 측정하였고, 라디칼 화합물인 nitric oxide(NO)는 Griess법으로 측정하였다.After 24 hours of macrophage culture, the culture supernatant was transferred to another well plate, and Ez-Cytox solution was added to measure macrophage proliferation and toxicity by the composition, while a macrophage culture solution was used to determine whether the sample was activated with macrophages. The concentrations of cytokines such as IL-6, IL-12, and TNF-α, which are activating factors produced in phagocytes, were measured by ELISA method according to the manufacturer's instructions, and nitric oxide (NO), a radical compound, was measured by the Griess method.
측정된 IL-6, IL-12와 TNF-α 및 NO의 생성 정도를 각각 도 2~5에 도시하였다. saline군과 실시예간 p<0.05 이다.The measured levels of production of IL-6, IL-12, TNF-α, and NO are shown in FIGS. 2 to 5, respectively. It is p<0.05 between the saline group and the examples.
② 분석 결과 : IL-6 (도 2 참조)② Analysis result: IL-6 (see Fig. 2)
IL-6은 모든 실시예 조성물이 대조구1(saline 수준)과 대조구 2(시료 100 μg/mL에서 316.3 pg/mL)에 비해 월등히 우수한 생산을 보였으며, 조성물 농도가 높을수록 생산이 많았다. 50과 100 μg/mL 농도에서 실시예2-1(927.1과 6593.5 pg/mL), 2-3(896.9와 4317.9 pg/mL), 2-4(519.1과 3904.5 pg/mL)는 특히 높은 생산을 나타내었다.IL-6 produced significantly better than the control group 1 (saline level) and control group 2 (316.3 pg/mL in 100 μg/mL sample), and the higher the composition concentration, the higher the production of IL-6. At concentrations of 50 and 100 μg/mL, Examples 2-1 (927.1 and 6593.5 pg/mL), 2-3 (896.9 and 4317.9 pg/mL), and 2-4 (519.1 and 3904.5 pg/mL) produced particularly high production. Indicated.
③ 분석결과 : IL-12 (도 3 참조)③ Analysis result: IL-12 (see Fig. 3)
IL-12는 대조구1, 2(4.3~4.8 pg/mL)에서 생리식염수(4.0 pg/mL)와 비교하여 거의 활성이 나타나지 않았으며, 두 농도(50과 100 μg/mL)에서 실시예1-1(5.5와 6.9 pg/mL), 1-2(5.2와 5.0 pg/mL), 1-3(6.0과 7.9 pg/mL), 1-4(4.3과 5.4 pg/mL), 2-2(6.0과 7.2 pg/mL)도 생리식염수군과 비교하여 다소간의 우수성을 보였다. 그러나 실시예2-1(6.7과 12.8 pg/mL), 2-3(6.8과 10.5 pg/mL), 2-4(.5.3과 9.1 pg/mL)는 대조구1, 2 및 식염수와 비교하여 유의적으로 우수한 활성을 보였다.IL-12 showed little activity compared to physiological saline (4.0 pg/mL) in Controls 1 and 2 (4.3 to 4.8 pg/mL), and Example 1 at both concentrations (50 and 100 μg/mL). 1 (5.5 and 6.9 pg/mL), 1-2 (5.2 and 5.0 pg/mL), 1-3 (6.0 and 7.9 pg/mL), 1-4 (4.3 and 5.4 pg/mL), 2-2 ( 6.0 and 7.2 pg/mL) also showed some superiority compared to the physiological saline group. However, Example 2-1 (6.7 and 12.8 pg/mL), 2-3 (6.8 and 10.5 pg/mL), and 2-4 (.5.3 and 9.1 pg/mL) were significant compared to Controls 1 and 2 and saline. It showed excellent activity.
④ 분석결과 : TNF-α (도 4 참조)④ Analysis result: TNF-α (see Fig. 4)
TNF-α는 대조구1, 2(173.8~2028.7 pg/mL)에서 낮은 함량이었으나 실시예는 실시예1-4를 제외하고는 모두 이보다 매우 우수한 활성을 나타내었다. 특히 두 농도에서 실시예2-3(2453.5와 4208.4 pg/mL)은 TNF-α 생산능이 월등히 우수하였다.TNF-α was a low content in Controls 1 and 2 (173.8 to 2028.7 pg/mL), but Examples showed very superior activity, except for Example 1-4. Particularly, at both concentrations, Example 2-3 (2453.5 and 4208.4 pg/mL) had remarkably excellent TNF-α production capacity.
⑤ 분석결과 : NO (도 5 참조)⑤ Analysis result: NO (see Fig. 5)
NO 생산능을 보면, 대조구1, 2(1.0~2.0 μM)은 식염수구(1.1 μM)와 비교하여 활성이 거의 나타나지 않았지만, 두 농도에서 실시예는 모두 이보다 대략 2~20배의 매우 생산능을 보였다. 실시예2-1(10.6과 19.6 μM), 실시예2-3(13.0과 18.0 μM) 및 실시예2-4(6.3과 19.8 μM)는 두 농도에서 모두 월등히 좋은 활성을 나타내었고, 특히 실시예2-3은 50 μg의 낮은 농도에서 가장 높은 NO 생산을 나타내었다.Looking at the NO production ability, Controls 1 and 2 (1.0 to 2.0 μM) showed little activity compared to the saline solution (1.1 μM), but at both concentrations, the examples showed a very productive ability of about 2 to 20 times more than this. Showed. Example 2-1 (10.6 and 19.6 μM), Example 2-3 (13.0 and 18.0 μM), and Example 2-4 (6.3 and 19.8 μM) exhibited remarkably good activity at both concentrations, especially Example 2-3 showed the highest NO production at a concentration as low as 50 μg.
3. 결론3. Conclusion
결론적으로 본 발명의 실시예에 의한 조성물들이 모두 대식세포에 대한 독성을 나타내지 않아 안전성이 확인되었다.In conclusion, all the compositions according to the examples of the present invention did not show toxicity to macrophages, so safety was confirmed.
또한 실시예에 의한 조성물들은 대식세포의 면역 활성화 인자인 IL-6, IL-12와 TNF-α 등 사이토카인과 라디칼 화합물인 NO의 생산능 측면에서 종래 달임 방식으로 얻어진 대조구1, 2에 비해 대부분에서 월등히 우수한 효과를 나타내었다. In addition, the compositions according to the examples are mostly compared to Controls 1 and 2 obtained by conventional decoction in terms of the ability to produce cytokines such as IL-6, IL-12 and TNF-α, which are immune activating factors of macrophages, and NO, which is a radical compound. Showed a remarkably excellent effect in
따라서 본 발명에 의한 추출방법에 따라 제조된 식물성 생약 추출 조성물들이 종래 열수추출(달임 방식)에 의한 조성물에 비해 우수한 면역활성 증진 기능이 있음이 확인되었다. Therefore, it was confirmed that the plant herbal medicine extraction compositions prepared according to the extraction method according to the present invention have superior immune activity enhancing function compared to the conventional hot water extraction (decoction method) compositions.
이는 본 발명의 효소활용 추출방법에 의해서 단단하고 치밀한 식물성 생약의 세포벽이 달임 방식에 의한 것보다 빠르고 효율적으로 느슨하게 해체되면서 세포 내부 또는 세포벽 사이에 존재하는 생약 유효성분이 빠르게 그리고 다량으로 추출되기 때문으로 해석된다. 또한, 식물 세포나 세포벽에 존재하는 면역활성 성분이 효소에 의해 일부 가수분해되면서 수식되어 일반 열수추출보다 유용한 성분이 추출되는 것도 합리적으로 기대할 수 있다.This is interpreted as because the cell walls of the hard and dense plant-based herbal medicines of the present invention are rapidly and efficiently disassembled loosely than those of the decoction method, and the active ingredients of the herbal medicines present inside the cells or between the cell walls are extracted rapidly and in large quantities. do. In addition, it can be reasonably expected that the immune-active components present in plant cells or cell walls are partially hydrolyzed by enzymes and modified to extract useful components rather than general hot water extraction.
Claims (6)
(B) 상기 분쇄물을 셀룰라제, 헤미셀룰라제 또는 펙티나제 중 최소한 어느 하나를 포함하는 식물 세포벽 분해효소로 처리하는 단계;
(C) 상기 효소를 실활시키는 단계;를
포함하는 것을 특징으로 하는 식물성 생약의 유효성분 추출방법.
(A) preparing a pulverized product of one type or a plurality of types of dried herbal medicines;
(B) treating the pulverized product with a plant cell wall degrading enzyme containing at least one of cellulase, hemicellulase, or pectinase;
(C) inactivating the enzyme;
A method for extracting an active ingredient of a plant herbal medicine comprising:
상기 단계(C) 이후에 식물성 생약 분쇄물 잔사를 제거하여 추출액을 분리하는 단계가 추가되는 것을 특징으로 하는 식물성 생약의 유효성분 추출방법.
The method according to claim 1,
After the step (C), the step of separating the extract by removing the residue of the pulverized plant herbal medicine is added.
(B) 상기 분쇄물을 셀룰라제, 헤미셀룰라제 또는 펙티나제 중 최소한 어느 하나를 포함하는 식물 세포벽 분해효소로 처리하는 단계;
(C) 상기 처리된 분쇄물을 건조하는 단계;를 포함하는 방법에 의해 제조되는 것을 특징으로 하는 대식세포 활성 증진효과를 가진 식물성 생약 고체상 조성물.
(A) 4-6 parts by weight of Angelica Angelica, 2.5-3.5 parts by weight of Chungoong, 1.5-2.5 parts by weight of Cornus milk, Galgeun, Baekchul, respectively, Licorice, Baek Peony, Bokryeong, Sanyak, Cinnamon, respectively 0.8-1.2 parts by weight of mixed crushed product Preparing;
(B) treating the pulverized product with a plant cell wall degrading enzyme containing at least one of cellulase, hemicellulase, or pectinase;
(C) drying the treated pulverized product; a solid plant composition having a macrophage activity enhancing effect, characterized in that prepared by a method comprising.
(B) 상기 셀룰라제, 헤미셀룰라제 또는 펙티나제 중 최소한 어느 하나를 포함하는 식물 세포벽 분해효소로 처리하는 단계;
(C) 상기 효소를 실활시키는 단계;
(D) 상기 처리된 분쇄물의 잔사를 제거하여 추출액을 분리하는 단계;를 포함하는 방법에 의해 제조되는 것을 특징으로 하는 대식세포 활성 증진효과를 가진 식물성 생약 액상 조성물.
(A) 4-6 parts by weight of Angelica Angelica, 2.5-3.5 parts by weight of Chungoong, 1.5-2.5 parts by weight of Cornus milk, Galgeun, Baekchul, respectively, Licorice, Baek Peony, Bokryeong, Sanyak, Cinnamon, respectively 0.8-1.2 parts by weight of mixed crushed product Preparing;
(B) treating with a plant cell wall degrading enzyme containing at least one of the cellulase, hemicellulase, or pectinase;
(C) deactivating the enzyme;
(D) removing the residue of the treated pulverized product to separate the extract; a liquid vegetable herbal composition having an effect of enhancing macrophage activity, characterized in that it is prepared by a method comprising.
(B) 상기 분쇄물을 셀룰라제, 헤미셀룰라제 또는 펙티나제 중 최소한 어느 하나를 포함하는 식물 세포벽 분해효소로 처리하는 단계;
(C) 상기 처리된 분쇄물을 건조하는 단계;를 포함하는 방법에 의해 제조되는 것을 특징으로 하는 대식세포 활성 증진효과를 가진 식물성 생약 고체상 조성물.
(A) 4 to 6 parts by weight of Angelica, 2.5 to 3.5 parts by weight of Chungoong, 1.5 to 2.5 parts by weight of each of Hasuoh, Galgeun, Sasam, Sanyak, Licorice, Bokryeong, Rowan, and Health respectively 0.8 to 1.2 parts by weight step;
(B) treating the pulverized product with a plant cell wall degrading enzyme containing at least one of cellulase, hemicellulase, or pectinase;
(C) drying the treated pulverized product; a solid plant composition having a macrophage activity enhancing effect, characterized in that prepared by a method comprising.
(B) 상기 분쇄물을 셀룰라제, 헤미셀룰라제 또는 펙티나제 중 최소한 어느 하나를 포함하는 식물 세포벽 분해효소로 처리하는 단계;
(C) 상기 효소를 실활시키는 단계;
(D) 상기 처리된 분쇄물의 잔사를 제거하여 추출액을 분리하는 단계;를 포함하는 방법에 의해 제조되는 것을 특징으로 하는 대식세포 활성 증진효과를 가진 식물성 생약 액상 조성물.(A) 4 to 6 parts by weight of Angelica, 2.5 to 3.5 parts by weight of Chungoong, 1.5 to 2.5 parts by weight of each of Hasuoh, Galgeun, Sasam, Sanyak, Licorice, Bokryeong, Rowan, and Health respectively 0.8 to 1.2 parts by weight step;
(B) treating the pulverized product with a plant cell wall degrading enzyme containing at least one of cellulase, hemicellulase, or pectinase;
(C) deactivating the enzyme;
(D) removing the residue of the treated pulverized product to separate the extract; a liquid vegetable herbal composition having an effect of enhancing macrophage activity, characterized in that it is prepared by a method comprising.
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KR101774566B1 (en) | 2015-11-05 | 2017-09-04 | 한국식품연구원 | Polysaccharide fraction isolated from enzyme treated barley grass with immune-enhancing activity and uses thereof |
KR101774564B1 (en) | 2014-07-29 | 2017-09-04 | 한국식품연구원 | Polysaccharide fraction isolated from Panax ginseng treated enzymes with immune-enhancing activity and method for producing the same |
KR101868507B1 (en) | 2017-10-19 | 2018-06-19 | (주)해피바이오 | Cosmetic Composition for Anti-oxidation, Anti-inflammatory and Anti-wrinkling Comprising Enzyme-treated Extracts of Annona Muricata and Method for Manufacturing the Same |
-
2019
- 2019-07-26 KR KR1020190090949A patent/KR20210013482A/en not_active Application Discontinuation
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
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KR101774564B1 (en) | 2014-07-29 | 2017-09-04 | 한국식품연구원 | Polysaccharide fraction isolated from Panax ginseng treated enzymes with immune-enhancing activity and method for producing the same |
KR101774566B1 (en) | 2015-11-05 | 2017-09-04 | 한국식품연구원 | Polysaccharide fraction isolated from enzyme treated barley grass with immune-enhancing activity and uses thereof |
KR101868507B1 (en) | 2017-10-19 | 2018-06-19 | (주)해피바이오 | Cosmetic Composition for Anti-oxidation, Anti-inflammatory and Anti-wrinkling Comprising Enzyme-treated Extracts of Annona Muricata and Method for Manufacturing the Same |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
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CN110846332A (en) * | 2019-11-29 | 2020-02-28 | 怀化学院 | Pectinase artificial sequence and expression method and application thereof |
CN110846294A (en) * | 2019-11-29 | 2020-02-28 | 怀化学院 | Recombinant pectinase, gene thereof, recombinant vector, preparation method and application |
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