KR20030091103A - Composition for increasing immunity comprising enzyme resistant starch - Google Patents
Composition for increasing immunity comprising enzyme resistant starch Download PDFInfo
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- KR20030091103A KR20030091103A KR1020020028516A KR20020028516A KR20030091103A KR 20030091103 A KR20030091103 A KR 20030091103A KR 1020020028516 A KR1020020028516 A KR 1020020028516A KR 20020028516 A KR20020028516 A KR 20020028516A KR 20030091103 A KR20030091103 A KR 20030091103A
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- South Korea
- Prior art keywords
- starch
- enzyme
- resistant starch
- composition
- cells
- Prior art date
Links
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Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
- A23L2/38—Other non-alcoholic beverages
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
- A23L29/20—Foods or foodstuffs containing additives; Preparation or treatment thereof containing gelling or thickening agents
- A23L29/206—Foods or foodstuffs containing additives; Preparation or treatment thereof containing gelling or thickening agents of vegetable origin
- A23L29/212—Starch; Modified starch; Starch derivatives, e.g. esters or ethers
- A23L29/219—Chemically modified starch; Reaction or complexation products of starch with other chemicals
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
- A61K31/716—Glucans
- A61K31/718—Starch or degraded starch, e.g. amylose, amylopectin
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2200/00—Function of food ingredients
- A23V2200/30—Foods, ingredients or supplements having a functional effect on health
- A23V2200/324—Foods, ingredients or supplements having a functional effect on health having an effect on the immune system
Abstract
Description
본 발명은 효소저항전분(resistant starch)을 유효성분으로 포함하는 면역활성 증강용 조성물에 관한 것으로서, 상기 비교적 저렴하게 생산할 수 있고 면역기능을 증진시키는 효과가 있는 조성물을 제공할 수 있다.The present invention relates to a composition for enhancing immune activity comprising an enzyme resistant starch (resistant starch) as an active ingredient, it can be produced at a relatively low cost and can provide a composition having an effect of enhancing immune function.
최근 들어 사회환경 및 식생활의 변화로 이와 관련된 비만, 당뇨병, 고혈압, 암 등의 성인병이 급속히 증가되면서 성인병을 예방, 치료하는 생리활성물질을 함유한 기능성 식품소재에 대한 연구가 많이 진행되고 있다. 이들 기능성 식품 소재중 식이섬유는 인체에서 소장에서 소화되지 않아 칼로리가 없고 혈당을 상승시키지 않으며 따라서 혈중 콜레스테롤이나 지방질의 축적을 저하시켜 성인병을 예방하는 효과가 있다고 알려져 있다.Recently, due to the changes in the social environment and diet, there are a lot of researches on functional food materials containing bioactive substances that prevent and treat adult diseases as the adult diseases such as obesity, diabetes, hypertension and cancer are rapidly increased. Among these functional foods, dietary fiber is not digested in the small intestine in the human body, so it is known that there is no calorie and does not raise blood sugar, thus reducing the accumulation of cholesterol or fat in the blood and preventing adult diseases.
전분은 인간의 식이에서 가장 중요한 탄수화물 소재로서 기능성 식품소재 개발과 관련해서 지속적인 연구가 진행되고 있으며, 1980년대 초에 식이섬유와 유사한 효소저항전분(Enzyme resistant starch, RS)이 알려지게 되었다. 효소저항전분이란 전분 분해효소 -알파, 베타 아밀라제 등으로 처리하여도 분해되지 않고 남는부분을 말하는데 1990년에 유럽의 EURESTA(European Flair Concerted Action on Resistant Starch)에서는 효소저항전분에 대한 정의를 내렸는데 이에 의하면 사람의 소장에서 흡수되지 않는 전분이나 전분의 분해산물을 모두 포함하는 의미이다.Starch is one of the most important carbohydrates in human diet, and ongoing research is underway on the development of functional food materials. In the early 1980s, enzyme resistant starch (RS) similar to dietary fiber became known. Enzyme-resistant starch refers to a portion that remains undegraded even after treatment with starch degrading enzyme -alpha, beta amylase.In 1990, European Flaster Concerted Action on Resistant Starch (EURESTA) defined the enzyme-resistant starch. This means that it contains both starch and decomposition products of starch that are not absorbed by the human small intestine.
효소저항전분은 식이 섬유소처럼 소장에서 소화 및 흡수되지 못하여 혈중 콜레스테롤이나 지방질의 축적을 저하시켜 성인병을 예방하는 효과가 있다고 알려져 있으며 소화되지 않은 부분은 대장에서 미생물에 의해 발효되어 단쇄지방산을 생성하는데 암세포의 성장을 억제하는 부티릭산(butyric acid)의 생성이 섬유소 섭취시보다 많아 대장암의 억제능력이 다른 식이섬유보다 더 큰 것으로 보고되었다(Alexander, Cereal Food World 40 (6) : p455, 1995 ; Edward 등, J .Sci. Food Agric 71: p209, 1996).Enzyme-resistant starch is not known to be digested and absorbed in the small intestine like dietary fiber, and it is known to reduce the accumulation of cholesterol or fat in the blood, preventing adult diseases. The undigested portion is fermented by microorganisms in the large intestine to produce short-chain fatty acids. The production of butyric acid, which inhibits growth, is higher than that of fibrin intake, and it has been reported that colon cancer has a greater inhibitory ability than other dietary fiber (Alexander, Cereal Food World 40 (6): p455, 1995; Edward Et al., J. Sci. Food Agric 71: p209, 1996).
최근 수용성 식이섬유는 순환기 질환의 예방 및 대장의 기능성 향상뿐만 아니라 면역기능 향상에 긍정적인 영향을 미치는 것으로 보고되고 있다. Lim 등은 수용성 식이섬유의 일종인 펙틴을 SD 흰쥐에 2주간 섭취시킨 결과, 면역활성을 나타내는 성분들이 증가되어 위장관의 면역기능 향상에 기여한다고 하였다 (Lim BO 등 , J. Nutr. 127: p663~667, 1997).이와 같이 수용성 식이섬유의 섭취가 체내에서 위장관 뿐만 아니라 체내의 면역조직까지 영향을 미치는 작용기전에 관해서는 섭취된 식이섬유가 장관내에서 단쇄지방산(SCFA)을 형성하여 단쇄지방산이 위장관내에서 T-세포를 증가시키는 것으로 알려져 있다. 장내점막에 존재하는 림프구(lymphocyte)는 임파조직내의 항원과 반응한 후 림프여포(lymphoid follicle)의 배 중심(germinal center)에서 성숙하게 된다. 성숙된림프구(lymphocyte)는 장관 임파절과 흉관을 통하여 체순환계로 들어오게 되고 체조직 전체의 면역기능에 영향을 미치게 된다.Recently, water-soluble dietary fiber has been reported to have a positive effect on the prevention of circulatory diseases and the improvement of the function of the colon as well as the immune function. Lim et al. Reported that pectin, a water-soluble dietary fiber, was ingested in SD rats for two weeks, resulting in an increase in immune-active components that contribute to the immune system's immune function (Lim BO et al., J. Nutr. 127: p663 ~). 667, 1997) . As described above, the intake of soluble dietary fiber affects not only the gastrointestinal tract but also the immune system in the body, and the ingested fiber forms short chain fatty acid (SCFA) in the intestinal tract and T-cells in the gastrointestinal tract. It is known to increase. Lymphocytes present in the intestinal mucosa react with antigens in lymphoid tissues and mature at the germinal centers of the lymphoid follicles. Mature lymphocytes (lymphocytes) enter the circulatory system through the intestinal lymph nodes and chest tube and affect the immune function of the entire body tissues.
효소저항전분은 수용성 식이섬유는 아니지만 다음과 같이 식이섬유와 유사한 기능성을 나타내는데 분변량의 증가, 대장내의 pH저하, 여러종류의 단쇄지방산 생성등이 있다. (Kirtchevsky , European J. of Cancer Preview , 4 : p345~532, 1995 ; Phillips등 , American Journal of Clinical Nutrition, 62 : p121~130, 1995)Enzyme-resistant starch is not a water-soluble dietary fiber, but has similar functionalities to dietary fiber as follows: an increase in fecal volume, a decrease in pH in the large intestine, and various types of short-chain fatty acids. (Kirtchevsky, European J. of Cancer Preview, 4: p345-532, 1995; Phillips et al., American Journal of Clinical Nutrition, 62: p121-130, 1995)
본 발명의 목적은 제조공정이 간단하고, 비교적 저렴하고, 인체에 무해한 효소저항전분을 유효성분으로 포함하는 면역활성 증강용 조성물을 제공하는 것이다.SUMMARY OF THE INVENTION An object of the present invention is to provide a composition for enhancing immune activity, which comprises a simple, low-cost, harmless human enzyme-resistant starch as an active ingredient.
본 발명의 또다른 목적은 상기 효소저항전분을 포함하는 식품, 식품 첨가제 또는 음료를 제공하는 것이다.Still another object of the present invention is to provide a food, food additive or beverage containing the enzyme resistant starch.
상기와 같은 과제를 달성하고자, 본 발명은 효소저항전분을 유효성분으로 포함하는 면역활성 증강용 조성물을 제공하고자 한다.In order to achieve the above object, the present invention is to provide a composition for enhancing immune activity comprising an enzyme-resistant starch as an active ingredient.
효소저항전분은 다음과 같이 분류할 수 있는데, 첫째 세포형태를 그대로 유지하거나 곡류나 종자를 부분적으로 분쇄한 것으로서 물리적으로 소화효소가 접근하기 어려운 형태(RS1 형태), 둘째 B-타입의 전분입자로 호화되지 않은 감자나 녹색 바나나에 존재하는 전분(RS2 형태), 셋째 호화된 후 노화된 아밀로즈(RS3 형태), 넷째 가교화 등 화학적으로 변성된 전분(RS4 형태)으로 나누어 볼 수 있다.이러한 효소저항전분을 모두 본 발명의 조성물에 적용할 수 있다.Enzyme-resistant starch can be classified as follows. First, the cell type is maintained as it is or the grain or seed is partially crushed. Starch (RS2 form) present in ungelatinized potatoes or green bananas, chemically modified starch (RS4 form), such as third gelatinized and aged amylose (RS3 form), and fourth crosslinking. All of the resistive starch can be applied to the composition of the present invention.
본 발명의 바람직한 일례에서, 전분의 OH기를 인산, 아디핀산 등의 가교화제로 서로 화학적으로 결합시킨 RS4 형태의 가교전분을 효소저항전분으로 사용할 수 있으며, 상기에서 전분은 모든 종류의 전분을 포함하는 의도이며, 예를 들면, 옥수수전분, 찰옥수수 전분, 감자전분, 고구마전분, 타피오카 전분, 사고전분, 쌀전분, 밀전분, 또는 동부전분 등을 사용하여 제조할 수 있다. 상기에서 RS4 형태의 가교전분은 전분내의 인접한 OH기들을 인산 또는 아디핀산 등의 가교화제로 서로 화학적으로 공유결합시킨 전분으로 반응정도에 따라서 가열하여도 호화되지 않으며 따라서 호화된 형태의 전분에 주로 작용하는 소화효소(알파아밀라제, 글루코아밀라제등)의 분해작용에 저항하는 특성이 있다. 효소저항전분의 제조방법은 대한민국 등록특허 제294252호, 제 218232호, 및 대한민국 공개특허 제 2001-88009호 등에 자세히 기재되어 있으며, 이에 한정되지 않는다.In a preferred embodiment of the present invention, crosslinked starch of the RS4 type in which the OH group of the starch is chemically bonded to each other with a crosslinking agent such as phosphoric acid and adipic acid may be used as the enzyme resistance starch, wherein the starch includes all kinds of starch. It is intended and can be prepared using, for example, corn starch, waxy corn starch, potato starch, sweet potato starch, tapioca starch, sago starch, rice starch, wheat starch, or eastern starch. The cross-linked starch of the RS4 type is a starch chemically covalently bonded to adjacent OH groups in the starch with a crosslinking agent such as phosphoric acid or adipic acid. It is resistant to the degradation of digestive enzymes (alpha amylase, glucoamylase, etc.). Method for producing enzyme-resistant starch is described in detail in the Republic of Korea Patent No. 294252, 218232, and Republic of Korea Patent Publication No. 2001-88009, but is not limited thereto.
현재 상업적으로 판매되고 있는 효소저항전분은 고아밀로즈 옥수수전분을 습열처리하거나, 혹은 이것을 호화시킨 후 노화시킨 형태이나 이들 제품은 가격이 비싸고 특유의 물성으로 인해 다양한 식품소재로 사용하기에는 제한이 있다. 그러나 가교화한 RS4 형태의 효소저항전분은 옥수수나 감자, 밀, 타피오카 등 일반적으로 널리 사용되는 전분원료를 이용하여서 쉽게 제조할 수 있을 뿐만 아니라 가격이 비교적 저렴하고, 제조공정이 간단하며 고아밀로즈 옥수수전분을 원료로 한 효소저항 전분에 비해 훨씬 다양한 용도로 식품에 사용될 수 있는 장점이 있어 더욱 바람직하다.Enzyme-resistant starch, which is currently commercially available, is wet-heat treated with high amylose corn starch or aged after it is gelatinized. However, these products are expensive and their physical properties are limited to use them in various food materials. However, cross-linked RS4-type enzyme-resistant starch can be easily prepared using commonly used starch raw materials such as corn, potato, wheat and tapioca, and it is relatively inexpensive, simple to manufacture and high amylose. Compared to enzyme-resistant starch based on corn starch, there is an advantage that it can be used in food for a much wider range of applications.
본 발명의 면역활성 증강용 조성물은 효소저항전분을 1~100 중량%로, 바람직하게는 5~50 중량%로 포함할 수 있다. 최종식품의 물성 예를 들어 빵의 경우 부피나 식감, 국수의 경우 조리시간 및 식감 등에 영향을 주지않는 범위에서는 많이 사용하여도 상관이 없으므로 최종 적용 조성물에 따라 적절히 선택할 수 있다.The composition for enhancing immune activity of the present invention may contain 1 to 100% by weight of enzyme resistance starch, preferably 5 to 50% by weight. The properties of the final food, for example, in the range that does not affect the volume or texture of the bread, the cooking time and texture in the case of noodles can be used appropriately according to the final application composition.
본 발명의 조성물은 식품, 식품 첨가제등일 수 있으며, 구체적인 예로 식육가공품, 어육제품, 두부류, 묵류, 면류(예, 라면류, 국수류 등), 빵류(식빵,케익류)건강보조식품, 소스류, 과자류(예, 스넥류), 기타 가공식품,튀김옷류,천연조미료(예, 라면 스프 등)을 포함하나 이에 한정되지 않는다. 바람직하게는 상기 조성물은 빵이나 국수, 스낵등으로 제조할 수 있다. 상기 식품 또는 식품첨가제는 통상의 제조방법으로 제조될 수 있다.The composition of the present invention may be food, food additives, and the like, and in particular, processed meat products, fish products, tofu, jelly, noodles (eg, ramen, noodles, etc.), breads (bread, cakes), health supplements, sauces, confectionery ( Examples include, but are not limited to, snacks), other processed foods, fried foods, and natural seasonings (eg, ramen soup). Preferably, the composition may be prepared from bread, noodles, snacks and the like. The food or food additives may be prepared by a conventional manufacturing method.
본 발명에 따른 조성물을 흰쥐에 섭취시킨 결과 면역활성이 향상되었음을 알 수 있으며, 구체적으로, CD4+ T-세포/CD8+ T-세포 비율, NK 세포의 증식능 및 Con A로 자극시 IL-4의 농도가 증가하여 흰쥐의 장간막 임파절의 면역기능을 향상시킴을 확인할 수 있다(표 10, 표 11, 및 표 12).As a result of ingesting the composition according to the present invention in rats, it can be seen that the immune activity was improved. Specifically, the concentration of CD4 + T-cells / CD8 + T-cells, the proliferation capacity of NK cells, and the concentration of IL-4 upon stimulation with Con A were increased. Increasingly, the immune function of mesenteric lymph nodes in rats can be improved (Table 10, Table 11, and Table 12).
하기 실시예는 오로지 본 발명을 설명하기 위한 것으로 이들 실시예에 의해 본 발명의 범위가 한정되지 않는다는 것은 본 발명이 속하는 분야에서 통상의 지식을 가진 자들에게 있어서 자명할 것이다.The following examples are only intended to illustrate the present invention, it will be apparent to those skilled in the art that the scope of the present invention is not limited by these examples.
실시예 1: 효소저항 전분 제조Example 1 Preparation of Enzyme Resistance Starch
옥수수전분 100g에 물 150g을 첨가하여 전분유로 만든 다음, 온도를 60 ℃로 가온시켰다. 상기 가온 전분유에 염산을 가하여 전분유의 pH를 2.5로 낮추고 3시간정도 교반하였다. 상기 교반물에 황산나트륨을 5%/st.ds 투입하고 30분간 교반하여 용해시킨 후 NaOH 용액을 첨가하여 pH를 12로 상승시켰다. 여기에 소디움 트리메타포스페이트(Sodium trimeta phosphate):소디움 트리폴리포스페이드(Sodium tripoly phosphate)가 99:1인 혼합분을 전분 ds당 10% 투입하고, 약 5시간 반응시킨 다음 pH 5로 중화하여 반응을 종료시켰다. 반응이 끝난 전분유에 3배의 물을 가하여 정제 및 탈수한 후 50 ℃의 에어 오븐에서 건조시키고 분쇄하여 시료로 사용하였다.100 g of corn starch was added to 150 g of water to make starch milk, and the temperature was warmed to 60 ° C. Hydrochloric acid was added to the warmed starch milk to lower the pH of the starch milk to 2.5 and stirred for about 3 hours. 5% / st.ds of sodium sulfate was added to the stirred solution, and stirred for 30 minutes to dissolve, followed by addition of NaOH solution to raise the pH to 12. Sodium trimeta phosphate: Sodium tripolyphosphate (99: 1) mixed with a mixture of 99: 1 10% starch ds, reacted for about 5 hours and then neutralized to pH 5 to react It was finished. Three times of water was added to the finished starch milk, purified and dehydrated, dried in an air oven at 50 ° C., and ground to use as a sample.
실시예 2: 효소저항전분의 분석Example 2 Analysis of Enzyme Resistance Starch
표 1에 나타난 바와 같이 식품공전 분석법(식품공업협회, 1999)의 272쪽 ~ 273쪽에 기재된 방법에 따라 실시예 1에서 제조된 효소저항전분의 성분을 분석하였고 그 결과를 표 1에 또한 표시하였다.As shown in Table 1, the components of the enzyme-resistant starch prepared in Example 1 were analyzed according to the method described on pages 272 to 273 of the food industry analysis method (Food Industry Association, 1999), and the results are also shown in Table 1.
실시예 3 : 체중변화 및 식이효율 조사Example 3 weight change and dietary efficiency investigation
스프라그-돌리(Sprague-Dawley)계 흰쥐를 이용하여 식이의 종류에 따라 셀룰로즈 투여군(cellulose diet group) 및 본 발명에 따른 효소저항전분 투여군(Enzyme resistant starch group)으로 나누어 4주간 사육하였다. 실험식이는AIN-93G를 기준으로 하였으며 하기의 표 2와 같이 하였다. 식이 기간중에 1일 식이량을 매일 측정하였으며 1 주일에 1회 체중의 변화를 측정하고 평균값을 구하였다. 상기 식이 실험 결과로서 각 시료군의 사료섭취량과 체중 증가율을 표 3 및 표 4에 나타냈다.Sprague-Dawley rats were bred for 4 weeks, divided into cellulose diet group and Enzyme resistant starch group according to the present invention, depending on the type of diet. The experimental diet was based on AIN-93G and was performed as shown in Table 2 below. The daily diet was measured daily during the diet, and the weight change was measured once a week and the mean value was obtained. As a result of the dietary experiment, the feed intake and the weight increase rate of each sample group are shown in Tables 3 and 4.
* 각 식이섬유의 순수함량은 총 식이의 5%가 되도록 조정하였다.* The net content of each dietary fiber was adjusted to 5% of the total diet.
실험결과, 각 시료군간에 사료섭취량에는 유의차가 없었다(표 3). 이는 사료섭취량이나 체중증가율의 차이가 없음에도 불구하고 면역활성이 높았다는 것은 면역활성을 높이는 작용을 하는 인자가 있다는 것이다. 또한 기본적인 식품으로서의 역할을 그대로 할 수 있다는 것을 시사한다.As a result, there was no significant difference in feed intake between the groups (Table 3). This means that despite the fact that there is no difference in feed intake or weight gain rate, high immunity means that there is a factor that increases immunity. It also suggests that it can serve as a basic food.
실시예 4: 혈청 지질 농도, 혈청 중성지방 농도, 및 면역글로불린 농도Example 4: Serum Lipid Concentration, Serum Triglyceride Concentration, and Immunoglobulin Concentration
실시예 3에 나타난 세가지 시료군으로 행한 4주간의 실험식이가 끝나는 시점에서 동물을 18시간 이상 절식시키고, 도살한 후 혈액을 통해 혈청지질농도, 혈청 중성지방농도, 면역글로불린 농도를 측정하였다. 실험결과를 하기 표 5 내지 표 7에 나타냈다.At the end of the four-week experimental diet with three sample groups shown in Example 3, the animals were fasted for 18 hours or more, and then the serum lipid concentration, serum triglyceride concentration, and immunoglobulin concentration were measured through blood. The experimental results are shown in Tables 5 to 7.
혈청 지질 농도, 혈청 중성지방 농도, 혈청내 면역글로불린 농도를 조사한 결과, 혈청지질농도는(표 5) 효소저항전분 투여군이 다른군에 비해서 다소 낮았다.Serum lipid concentration, serum triglyceride concentration, and serum immunoglobulin concentration were examined, and the serum lipid concentration (Table 5) was slightly lower in the enzyme-resistant starch-administered group.
한편, 4주간의 실험식이 사육후 항원으로 자극하지 않은 상태에서 측정한 안정시의 혈청 IgE 농도는 효소저항전분 투여군이 좀 높았다(표 7).On the other hand, the stable serum IgE concentration measured in the state of 4 weeks of experimental diet not stimulated with the antigen after breeding was higher in the enzyme-resistant starch administration group (Table 7).
실시예 5: 장점막 임파절의 T-세포 활성Example 5: T-Cell Activity of Mucosal Lymph Nodes
실시예 3에서 얻은 장관막 임파절을 RPMI1640 배양액(FCS 10%)에 넣은 후 균질화시킨 다음 세포수를 4106/mL로 조정하여 실험에 이용하였다. 각 세포용액 100L에 PBS 완충용액 500L(HEPES 1.191g, 10% FCS 5mL, Na-Az 0.1g, 10 PBS 50mL, 증류수를 이용하여 500mL 조정)를 첨가하여 2000g에서 1분간 원심분리 하였다. 침전물에 200배 희석한 PE(phycoerythrin) 항-래트 CD4(PharMingen, USA)와 바이오틴 항-래트 CD8(PharMingen, USA)를, 또 하나의 튜브에는 FITC 항-래트 CD45RA(PharMingen, USA)와 바이오틴 항-래트 NKRPIA(PharMingen, USA)를 20L씩 첨가하였다. 20분간 얼름위에 방치한 후 PBS 완충용액 700L를 넣고 다시 2000g에서 1분간 원심분리하였다. 침전물에 100배 희석한 스트렙타비딘-FITC(fluorescein isothiocyanate)(Beckton Dickinson, USA) 또는 스트렙타비딘-PE를 10L씩 첨가하고 얼음위에서 다시 20분간 방치한 후 PBS 700L를 첨가하였으며, 2000g에서 1분간 원심분리하였다. 침전물에 PBS 500L를 첨가하여 현탁시킨 후 유동세포분석기를 이용하여 T-세포의 분포를 분석하여, 장간막 임파절의 CD4+ T-세포, CD8+ T-세포의 분포 결과를 하기 표 8, 및 표 9에 나타냈고 이들의 비율을 표 10에 나타냈다(각 표에서 구한 수치의 계산식은 순차적으로 다음과 같다).The intestinal membrane lymph nodes obtained in Example 3 were added to RPMI1640 culture medium (FCS 10%), homogenized, and then used for the experiment by adjusting the cell number to 4106 / mL. To 100 L of each cell solution, 500 L of PBS buffer solution (HEPES 1.191 g, 10% FCS 5 mL, Na-Az 0.1 g, 10 PBS 50 mL, 500 mL adjusted using distilled water) was added and centrifuged at 2000 g for 1 minute. PE (phycoerythrin) anti-rat CD4 (PharMingen, USA) and biotin anti-rat CD8 (PharMingen, USA) diluted 200-fold in the sediment, and another tube was FITC anti-rat CD45RA (PharMingen, USA) and biotin Rat NKRPIA (PharMingen, USA) was added in 20 L portions. After standing on ice for 20 minutes, 700L PBS buffer solution was added and centrifuged at 2000 g for 1 minute. 10 liters of streptavidin-FITC (Beckton Dickinson, USA) or streptavidin-PE diluted 100-fold in the precipitate was added and left for 20 minutes on ice, followed by 700L of PBS. Centrifuged. After suspension by adding PBS 500L to the precipitate and analyzing the distribution of T-cells using a flow cytometer, the distribution results of CD4 + T-cells and CD8 + T-cells of mesenteric lymph nodes are shown in Tables 8 and 9 below. And the ratio thereof is shown in Table 10 (calculations of the values obtained in each table are sequentially as follows).
CD4+T-세포(%) = CD4+T-세포 수 / 전체 면역세포수 x 100CD4 + T-cells (%) = CD4 + T-cells / total immune cells x 100
CD8+T-세포(%) = CD8+T-세포 수 /전체 면역세포수 x 100CD8 + T-cells (%) = CD8 + T-cells / total immune cells x 100
CD4+/CD8+ (비율) = CD4+T-세포 수 / CD8+T-세포 수CD4 + / CD8 + (ratio) = CD4 + T-cells / CD8 + T-cells
CD8+ T-세포 분포는 주로 헬퍼(helper) T 세포의 마커(maker)로서 B 세포의 성장 및 분화를 촉진하며, 사이토카인을 분비하여 대식세포를 활성화시키는 기능을 지니고 따라서 안정시 CD4+ T-세포 분포가 증가하는 경우 면역기능이 증가된 것으로 해석된다.CD8 + T-cell distribution is primarily a marker of helper T cells, which promotes the growth and differentiation of B cells, and has the function of activating macrophages by secreting cytokines and thus resting CD4 + T-cell distribution Increasing is interpreted as an increase in immune function.
실험결과, 장간막 임파절의 CD4+ T-세포, CD8+ T-세포의 분포는 세가지 군에서 유의한 차이가 없었으나, CD4+ T-세포/CD8+ T-세포 비율에서는 효소저항전분 투여군이 셀룰로즈 투여군(P<0.05), 옥수수 수용성 식이섬유 투여군(P<0.001)군 보다 높게 나타냈다.As a result, the distribution of CD4 + T-cells and CD8 + T-cells in mesenteric lymph nodes was not significantly different in the three groups, but in the CD4 + T-cell / CD8 + T-cell ratios, the enzyme-resistant starch-treated group was the cellulose-treated group (P <0.05 ), Was higher than the corn soluble dietary fiber group (P <0.001).
구체적으로, 표 8에서 알 수 있는 바와 같이, CD4+T-세포의 함량은 효소저항전분 투여군의 경우 다소 증가하는 경향이 있다. 표 9에서 나타낸 바와 같이, CD8+ T-세포는 주로 세포독성 T 림프구의 마커로서 바이러스에 감염된 세포 및 암세포을 분해시키는 작용을 갖는 세포로서, 안정시 CD8+ T-세포의 분포가 높을수록 면역기능이 저하된 것으로 해석된다. 셀룰로즈 투여군에 비해서 효소저항전분 투여군이 낮은 경향을 보였다.Specifically, as can be seen in Table 8, the content of CD4 + T-cells tend to increase somewhat in the case of the enzyme-resistant starch-administered group. As shown in Table 9, CD8 + T-cells mainly act as markers of cytotoxic T lymphocytes and have a function of degrading virus-infected cells and cancer cells, and the higher the distribution of CD8 + T-cells at rest, the lower the immune function. It is interpreted as. The enzyme-resistant starch-treated group showed a lower tendency than the cellulose-administered group.
흉선 또는 비장의 CD4+/CD8+ 비율은 면역기능의 중요한 지표로서 이용되고 있다. CD8+ T-세포는 CD4+ T-세포중에서도 T-헬퍼 세포 1의 성장을 촉진하나 T-헬퍼 세포2의 성장은 억제하는 것으로 알려져 있다. 따라서 CD4+/CD8+ 비가 증가하는 것은 CD4+ T-세포의 서브세트(subset)인 T-헬퍼 세포1과 T-헬퍼 세포2의 합이 증가된 결과이므로 전체적인 면역기능의 향상을 의미한다. 장간막 임파절에서의 면역세포비율은 효소저항전분 투여군이 셀룰로즈 군에 비해서 유의하게 높았다.The CD4 + / CD8 + ratio of the thymus or spleen is used as an important indicator of immune function. CD8 + T-cells are known to promote the growth of T-helper cell 1 but inhibit the growth of T-helper cell 2 among CD4 + T-cells. Therefore, increasing the CD4 + / CD8 + ratio means that the sum of T-helper cell 1 and T-helper cell 2, which is a subset of CD4 + T-cells, is an increase in overall immune function. The percentage of immune cells in mesenteric lymph nodes was significantly higher in the enzyme-resistant starch group than in the cellulose group.
상기의 장간막 임파절에서의 면역세포 분포결과를 종합해볼 때, CD4+ T-세포, 및 CD8+ T-세포의 분포비율에 있어서는 각 그룹간에 유의한 차가 나타나지 않았으나 면역조절 기능의 지표로 이용되고 있는 CD4+/CD8+ 비율에서는 효소저항전분이 다른 군에 비해서 유의하게 증가한 것으로 나타났다. 따라서 흰쥐의 면역조절기능이 저항성 전분의 섭취에 의하여 유의하게 향상된 것으로 평가된다.In summary, the distribution ratios of CD4 + T-cells and CD8 + T-cells in the mesenteric lymph nodes were not significantly different between the groups, but CD4 + / CD8 + is used as an indicator of immune regulation. In the ratio, the enzyme resistance starch was significantly increased compared to the other groups. Therefore, rat immunomodulatory function was evaluated to be significantly improved by ingestion of resistant starch.
실시예 6: 효소저항전분이 NK세포의 분포Example 6: Distribution of NK Cells with Enzyme Resistance Starch
실시예 3에서 얻은 장관막 임파절을 RPMI1640 배양액(FCS 10%)에 넣은 후 균질화시킨 다음 세포수를 4106/mL로 조정하여 실험에 이용하였다. 각 세포용액 100L에 PBS 완충용액 500L(HEPES 1.191g, 10% FCS 5mL, Na-Az 0.1g, 10 PBS 50mL, 증류수를 이용하여 500mL 조정)를 첨가하여 2000g에서 1분간 원심분리 하였다. 침전물에 200배 희석한 PE(phycoerythrin) 항-래트 CD4(PharMingen, USA)와 바이오틴 항-래트 CD8(PharMingen, USA)를, 또 하나의 튜브에는 FITC 항-래트 CD45RA(PharMingen, USA)와 바이오틴 항-래트 NKRPIA(PharMingen, USA)를 20L씩 첨가하였다. 20분간 얼름위에 방치한 후 PBS 완충용액 700L를 넣고 다시 2000g에서 1분간 원심분리하였다. 침전물에 100배 희석한 스트렙타비딘-FITC(fluorescein isothiocyanate)(Beckton Dickinson, USA) 또는 스트렙타비딘-PE를 10L씩 첨가하고얼음위에서 다시 20분간 방치한 후 PBS 700L를 첨가하였으며, 2000g에서 1분간 원심분리하였다. 침전물에 PBS 500L를 첨가하여 현탁시킨 후 유동세포분석기를 이용하여 NK세포의 분포를 분석하여 표 11에 나타내었다.The intestinal membrane lymph nodes obtained in Example 3 were added to RPMI1640 culture medium (FCS 10%), homogenized, and then used for the experiment by adjusting the cell number to 4106 / mL. To 100 L of each cell solution, 500 L of PBS buffer solution (HEPES 1.191 g, 10% FCS 5 mL, Na-Az 0.1 g, 10 PBS 50 mL, 500 mL adjusted using distilled water) was added and centrifuged at 2000 g for 1 minute. PE (phycoerythrin) anti-rat CD4 (PharMingen, USA) and biotin anti-rat CD8 (PharMingen, USA) diluted 200-fold in the sediment, and another tube was FITC anti-rat CD45RA (PharMingen, USA) and biotin Rat NKRPIA (PharMingen, USA) was added in 20 L portions. After standing on ice for 20 minutes, 700L PBS buffer solution was added and centrifuged at 2000 g for 1 minute. 100-fold diluted streptavidin-FITC (fluorescein isothiocyanate) (Beckton Dickinson, USA) or streptavidin-PE was added in 10 L portions, and the mixture was left on ice for 20 minutes and PBS 700L was added. Centrifuged. Suspended by adding PBS 500L to the precipitate was analyzed in the distribution of NK cells using a flow cytometer shown in Table 11.
NK 세포(%) = NK 세포수 / 전체면역세포 x 100NK cells (%) = NK cell count / total immune cells x 100
효소저항전분 투여군이 셀룰로즈 투여군(p<0.01)에 비해 높게 나타났다. 자연독성세포라고 불리는 NK(Natural killer)세포는 선천성 면역 시스템(innate immune system)에 속한다. 즉 NK 세포는 표적세포에 직접 작용하여 표적세포를 파괴시키는 비특이적 면역기능을 지닌다. 따라서, 효소저항전분은 타시료에 비해서 흰쥐에서 NK 세포의 수치가 유의하게 높아서 비특이적인 면역조절 기능을 향상시키는 효과가 있었다.The enzyme-resistant starch group was higher than the cellulose group (p <0.01). Natural killer (NK) cells, called natural toxic cells, belong to the innate immune system. In other words, NK cells have a nonspecific immune function that directly acts on target cells and destroys them. Therefore, the enzyme resistance starch was significantly higher in NK cells in rats than other samples, thereby improving the nonspecific immunomodulatory function.
실시예 7: 장관막 임파절의 감마-인터페론과 인터루킨-4의 농도Example 7: Concentrations of Gamma-Interferon and Interleukin-4 in Intestinal Lymph Nodes
실시예 3에서 얻은 흰쥐의 비장과 장관막 임파절을 RPMI1640 배양액(FCS 10%)에 넣어 균질화시킨 후 세포수를 4106/mL로 조정하여 실험에 이용하였다. 12-웰 플레이트에 각 세포용액을 300L씩 넣은 후에 100L의 Con A(25g/mL)와 LPS(50g/mL)로 자극하거나 또는 자극하지 않은 채로 72시간 동안 CO2 5%, 온도 37C의 환경하에 배양하였다. 인터루킨-4 농도는 OptEIATM 래트 인터루킨-4 세트(PharMingen, USA)를 이용하여, 그리고 감마-인터페론의 농도는 래트 감마-인터페론 ELISA(Endogen, USA)상업용 키트를 이용하여 측정하였다. 실험식이로 4주간 사육한 흰쥐에서 채취한 장관막 임파절 세포를 72시간동안 37 ℃에서 배양하고 Con A로 자극한 경우와 자극하지 않은 경우에 분비된 인터루킨-4 및 감마-인터페론 농도를 측정하여 표 12에 나타냈다.The spleen and intestinal lymph nodes of the rat obtained in Example 3 were homogenized in RPMI1640 culture medium (FCS 10%), and then used for the experiment by adjusting the cell number to 4106 / mL. Add 300 L of each cell solution to a 12-well plate and incubate in 100% Con A (25 g / mL) and LPS (50 g / mL) with or without stimulation for 5 hours at 5% CO2 and 37C It was. Interleukin-4 concentrations were measured using the OptEIATM rat interleukin-4 set (PharMingen, USA), and gamma-interferon concentrations were measured using the rat gamma-interferon ELISA (Endogen, USA) commercial kit. Intestinal membrane lymph node cells harvested from rats incubated for 4 weeks in an experimental diet were cultured at 37 ° C. for 72 hours, and the levels of interleukin-4 and gamma-interferon secreted when stimulated with Con A and when not stimulated were measured. Shown at 12.
Con A로 장관막 임파절세포를 자극하지 않은 상태에서의 세포증식능은 세 개 군간에서 별 차이가 없는 반면에, Con A로 자극한 경우에는 면역세포 증식능이 효소저항 전분에서 가장 높은 경향을 보였다.Cell proliferation was not significantly different among the three groups when Con A was not stimulated with mesenteric lymph node cells. However, Con A stimulated immune cell proliferation was highest in enzyme-resistant starch.
이상의 결과를 종합한 결과, 셀룰로즈에 비해 효소저항전분을 급여한 흰쥐의 동물실험 결과, 효소저항전분을 섭취할 경우 셀룰로즈 군에 비해 면역기능 향상효과가 탁월한 것이 판명되었다.Based on the above results, animal experiments of rats fed enzyme-resistant starch compared to cellulose showed that ingestion of enzyme-resistant starch showed superior immune function improvement compared to cellulose group.
실시예 8: 효소저항전분을 포함하는 식빵Example 8: Bread containing enzyme resistance starch
하기 표 13에 나타난 성분비에 따라 밀가루 대비 10%로 효소저항전분을 첨가하고 straight법에 의해 식빵을 제조하였으며 완성된 식빵은 기존 식빵대비 맛이나 외관, 볼륨 등에서 차이점이 없었다.According to the component ratios shown in Table 13, the enzyme-resistant starch was added to 10% of the flour, and the bread was prepared by the straight method.
실시예 9: 효소저항전분을 포함한 국수Example 9 Noodles Containing Enzyme Resistance Starch
하기 표 14에 나타난 성분비에 따라 압연법에 의한 생면제조후 식감 평가 결과 기존 국수에 비해서 조리시간, 국수를 삶은 후 면의 퍼짐성, 및 맛 등에서 차이점이 없었다.According to the ingredient ratios shown in Table 14 below, after the raw noodles were prepared by the rolling method, there was no difference in cooking time, spreadability of noodles, and taste after boiling noodles compared to the existing noodles.
실시예 10: 효소저항전분을 포함하는 튀김옷Example 10 Frying Clothes Containing Enzyme-Resistant Starch
하기 표 15에 나타난 성분비에 따라 튀김옷을 제조하여 탕수육의 튀김옷으로 사용한 결과 기존의 전분에 비해 기름 흡수가 적으며 바삭한 맛이 향상되었다.According to the ingredient ratios shown in Table 15 below, the fried cloth was prepared and used as a fried cloth of sweet and sour pork, resulting in less oil absorption and improved crispy taste than conventional starch.
본 발명은 효소저항전분을 유효성분으로 포함하는 면역활성 증강용 조성물을 제공하여, 상기 효소저항전분은 제조공정이 간단하고, 옥수수,찰옥수수, 감자, 밀, 타피오카 등으로부터 비교적 저렴하게 생산할 수 있고, 면역기능을 증진시키는 효과가 있으므로 유용한 기능성 식품소재로서 널리 이용될 수 있다.The present invention provides a composition for enhancing immune activity comprising enzyme-resistant starch as an active ingredient, the enzyme-resistant starch is a simple manufacturing process, can be produced relatively inexpensively from corn, waxy corn, potatoes, wheat, tapioca, etc. In addition, since it has an effect of enhancing immune function, it can be widely used as a useful functional food material.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| US7812221B2 (en) | 2003-06-30 | 2010-10-12 | Commonwealth Scientific And Industrial Research Organization | Wheat with altered branching enzyme activity and starch and starch containing products derived therefrom |
| KR20180121038A (en) * | 2017-04-28 | 2018-11-07 | (주)산돌식품 | A preparation method of cold noodle using waty corn resistant starch |
| KR20200077191A (en) * | 2018-12-20 | 2020-06-30 | 전남대학교산학협력단 | Composition for preparing high dietary fiber sweet potato Muk |
| US10973826B2 (en) | 2015-10-29 | 2021-04-13 | Novartis Ag | Antibody conjugates comprising toll-like receptor agonist |
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| KR101068986B1 (en) | 2011-02-18 | 2011-09-29 | 대상 주식회사 | Resistant starch with pasting properties and manufacturing method of the same |
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| JP4282763B2 (en) * | 1996-03-20 | 2009-06-24 | ザ、ユニバーシティ、オブ、ニュー、サウス、ウェイルズ | Changes in the microbial population in the digestive tract |
| JPH10279487A (en) * | 1997-04-01 | 1998-10-20 | Nippon Shokuhin Kako Co Ltd | Lipid metabolism improver |
| US5855946A (en) * | 1997-06-06 | 1999-01-05 | Kansas State University Research Foundation | Food grade starch resistant to α-amylase and method of preparing the same |
| KR100415308B1 (en) * | 2001-03-02 | 2004-01-16 | 대한민국 | Cross-linked resistant starch and process for preparation thereof |
| KR20030072094A (en) * | 2002-03-05 | 2003-09-13 | 동아제분 주식회사 | Wheat flour premix composition for noodle and rice cake |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| US7812221B2 (en) | 2003-06-30 | 2010-10-12 | Commonwealth Scientific And Industrial Research Organization | Wheat with altered branching enzyme activity and starch and starch containing products derived therefrom |
| US8115087B2 (en) | 2003-06-30 | 2012-02-14 | Commonwealth Scientific And Industrial Research Organisation | Wheat with altered branching enzyme activity and starch and starch containing products derived therefrom |
| US8829315B2 (en) | 2003-06-30 | 2014-09-09 | Commonwealth Scientific And Industrial Research Organisation | Wheat with altered branching enzyme activity and starch containing products derived therefrom |
| US10973826B2 (en) | 2015-10-29 | 2021-04-13 | Novartis Ag | Antibody conjugates comprising toll-like receptor agonist |
| KR20180121038A (en) * | 2017-04-28 | 2018-11-07 | (주)산돌식품 | A preparation method of cold noodle using waty corn resistant starch |
| KR20200077191A (en) * | 2018-12-20 | 2020-06-30 | 전남대학교산학협력단 | Composition for preparing high dietary fiber sweet potato Muk |
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