KR102226144B1 - Method of preparing fermented soybean paste having anti-inflammatory and immunosuppressive activity - Google Patents
Method of preparing fermented soybean paste having anti-inflammatory and immunosuppressive activity Download PDFInfo
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- KR102226144B1 KR102226144B1 KR1020180151320A KR20180151320A KR102226144B1 KR 102226144 B1 KR102226144 B1 KR 102226144B1 KR 1020180151320 A KR1020180151320 A KR 1020180151320A KR 20180151320 A KR20180151320 A KR 20180151320A KR 102226144 B1 KR102226144 B1 KR 102226144B1
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- South Korea
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- namunjae
- doenjang
- salinity
- powder
- inflammatory
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Classifications
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- 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
- A23L11/00—Pulses, i.e. fruits of leguminous plants, for production of food; Products from legumes; Preparation or treatment thereof
- A23L11/50—Fermented pulses or legumes; Fermentation of pulses or legumes based on the addition of microorganisms
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- 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
- A23L11/00—Pulses, i.e. fruits of leguminous plants, for production of food; Products from legumes; Preparation or treatment thereof
- A23L11/70—Germinated pulse products, e.g. from soy bean sprouts
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- 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
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- 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
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- 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
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- 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
- A23V2250/00—Food ingredients
- A23V2250/20—Natural extracts
- A23V2250/21—Plant extracts
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- 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
- A23V2300/00—Processes
- A23V2300/14—Extraction
Abstract
본 발명은 1) 메주, 나문재 분말, 소금 및 물을 혼합하는 단계; 및 2) 상기 단계 1)의 혼합물을 발효하는 단계;를 포함하는 항염증 또는 면역 억제 활성을 갖는 나문재 된장의 제조방법에 관한 것으로, 본 발명에 따라 제조된 나문재 된장은 염증이 유도된 대식세포에서 산화질소 및 사이토카인 IL-6의 생성량을 감소시키고, COX-2 및 iNOS의 발현을 단백질 및 mRNA 수준에서 억제하므로, 항염증용 및 면역 억제 용도로 유용하게 사용될 수 있다. The present invention comprises the steps of 1) mixing meju, namunjae powder, salt and water; And 2) fermenting the mixture of step 1); It relates to a method for producing Namunjae Doenjang having anti-inflammatory or immunosuppressive activity, comprising: Namunjae Doenjang prepared according to the present invention in macrophages in which inflammation is induced. It reduces the production of nitric oxide and the cytokine IL-6, and suppresses the expression of COX-2 and iNOS at the protein and mRNA levels, so it can be usefully used for anti-inflammatory and immune suppression purposes.
Description
본 발명은 항염증 또는 면역 억제 활성을 갖는 나문재 된장의 제조방법에 관한 것이다. The present invention relates to a method for producing Namunjae Doenjang having anti-inflammatory or immunosuppressive activity.
된장(doenjang, fermented soybean paste)은 대두, 쌀, 보리, 밀 또는 탈지대두 등을 주원료로 하여 누룩균 등을 배양한 후 식염을 혼합하여 발효 숙성시킨 것 또는 메주를 식염수에 담가 발효하고 여액을 분리하여 가공한 것을 말한다(식품공전, 2014년 10월 21일 고시). 된장은 단백질 등 영양소의 함량이 높고 특유의 맛과 향을 지니고 있어 우리 식생활에서 중요한 단백질 공급원 중의 하나일 뿐 아니라 조미료로서도 중요한 역할을 하고 있다. 된장은 메주의 발효과정 중에 미생물이 생산하는 효소에 의해 원료인 대두의 영양소가 분해되어 인체에서 소화 흡수되기 쉬운 형태로 전환되고 건강에 유익한 기능을 가진 물질이 생성된다. 된장에는 대두에 존재하는 이소플라본, 트립신 저해제(inhibitor), 폴리페놀, 글로블린 등과 된장의 발효숙성 과정 중에 생산되는 리놀레산과 펩타이드 등의 다양한 기능성 성분을 함유하고 있으며, 항암효과, 항고혈압 효과, 항산화 효과, 혈전용해 효과, 혈당 강하 효과 등을 나타내는 것으로 알려져있다. 이와 같은 된장의 알려진 기능성 외에 새로운 기능성 물질을 첨가하여 영양 성분이 보다 증진된 된장을 제조하고자 하는 노력이 지속되고 있다. Doenjang (fermented soybean paste) is made from soybean, rice, barley, wheat, or skim soybeans as the main ingredients, and then fermented and aged by mixing salt after fermentation or fermentation by soaking meju in saline and separating the filtrate. It refers to the processed product (Food Code, announced on October 21, 2014). Doenjang is not only one of the important sources of protein in our diet, but also plays an important role as a seasoning because it has a high content of nutrients such as protein and has a unique taste and aroma. During the fermentation process of soybean paste, the nutrients of soybean, the raw material, are decomposed by enzymes produced by microorganisms, and it is converted into a form that is easy to digest and absorb in the human body, and substances with beneficial functions are produced. Doenjang contains various functional ingredients such as isoflavones, trypsin inhibitors, polyphenols, globulins, etc. that are present in soybeans, such as linoleic acid and peptides produced during the fermentation and aging process of soybean paste, and has anticancer, antihypertensive, and antioxidant effects. , Thrombolytic effect, blood sugar lowering effect, etc. are known to exhibit. In addition to the known functionality of doenjang, efforts to manufacture doenjang with improved nutritional content by adding new functional substances are continuing.
한편, 염생식물은 염류가 함유된 토지에서 생육할 수 있는 식물로, 바닷물과 담수가 반복적으로 교차하는 지역에서 생육하게 되면서 육상식물과 해상식물이 가진 다양한 생리 활성 물질들을 동시에 포함하고 있다. 국내의 경우 퉁퉁마디(Saliconia herbacea), 나문재(Suaeda asparagoides , 또는 S. glauca Bunge), 칠면초(Suaeda japonica), 갯질갱(Limonium tetragonum), 해당화(Rosa rugosa), 통보리사초(Carex kobomugi), 수송나물(Salsola kormarovi Iljin), 해홍나물(Suaeda maritima) 등이 대표적인 염생식물로 알려져있다. 이러한 염생 식물은 최근 식품 가공기술의 발달로 인해, 다양한 먹거리로 이용되며, 또한 식물성 소금의 제조원으로 각광받고 있다. 현재까지는 주로 퉁퉁마디(함초)를 이용한 식품 원재료 이용 및 소금 제조가 가장 일반적이다. On the other hand, salt plants are plants that can grow on land containing salts, and as they grow in areas where seawater and fresh water repeatedly cross, they contain various physiologically active substances of land plants and marine plants at the same time. In Korea, Saliconia herbacea , Suaeda asparagoides , or S. glauca Bunge), Suaeda japonica , Limonium tetragonum ), glycosyllium (Rosa rugosa ), gorse sedge ( Carex kobomugi ), transport herbs ( Salsola) kormarovi Iljin ), sea red sprouts ( Suaeda maritima ) are known as representative salt plants. Due to the recent development of food processing technology, these salted plants are used as various foods, and are in the spotlight as a manufacturer of vegetable salt. Until now, the most common use of raw materials for food and salt production is mainly using green onions (green tea).
본 발명의 목적은 항염증 또는 면역 억제 활성을 갖는 나문재 된장의 제조방법을 제공하는 것이다. It is an object of the present invention to provide a method for producing Namunjae Doenjang having anti-inflammatory or immune suppressing activity.
본 발명의 다른 목적은 항염증 또는 면역 억제 활성을 갖는 나문재 된장을 제공하는 것이다. Another object of the present invention is to provide a namunjae doenjang having anti-inflammatory or immunosuppressive activity.
본 발명의 또 다른 목적은 항염증용 및 면역 억제용 조성물을 제공하는 것이다. Another object of the present invention is to provide a composition for anti-inflammatory and immune suppression.
상기 목적을 달성하기 위하여, 본 발명은 1) 메주, 나문재 분말, 소금 및 물을 혼합하는 단계; 및 2) 상기 단계 1)의 혼합물을 발효하는 단계;를 포함하는 항염증 또는 면역 억제 활성을 갖는 나문재 된장의 제조방법을 제공한다. In order to achieve the above object, the present invention 1) mixing meju, namunjae powder, salt and water; And 2) fermenting the mixture of step 1). It provides a method for producing Namunjae miso having anti-inflammatory or immunosuppressive activity.
또한, 본 발명은 상기 제조방법으로 제조된 항염증 또는 면역 억제 활성을 갖는 나문재 된장을 제공한다. In addition, the present invention provides a namunjae doenjang having anti-inflammatory or immunosuppressive activity prepared by the above manufacturing method.
또한, 본 발명은 상기 나문재 된장의 추출물을 유효성분으로 포함하는 항염증용 및 면역 억제용 조성물을 제공한다. In addition, the present invention provides a composition for anti-inflammatory and immune suppression comprising the extract of Namunjae doenjang as an active ingredient.
본 발명에 따라 제조된 나문재 된장은 염증이 유도된 대식세포에서 산화질소 및 사이토카인 IL-6의 생성량을 감소시키고, COX-2 및 iNOS의 발현을 단백질 및 mRNA 수준에서 억제하므로, 항염증용 및 면역 억제 용도로 유용하게 사용될 수 있다. Namoonjae doenjang prepared according to the present invention reduces the production of nitric oxide and cytokine IL-6 in macrophages in which inflammation is induced, and inhibits the expression of COX-2 and iNOS at the protein and mRNA levels, so it is for anti-inflammatory and It can be usefully used for immune suppression purposes.
도 1은 나문재 된장(염도 8% 및 12%) 발효 기간에 따른 수분 함량을 나타낸 도이다
(8G: 염도 8% 및 나문재 무첨가 된장; 85N: 염도 8% 및 나문재 분말 5% 첨가 된장; 810N: 염도 8% 및 나문재 분말 10% 첨가 된장;
12G: 염도 12% 및 나문재 무첨가 된장; 125N: 염도 12% 및 나문재 분말 5% 첨가 된장; 1210N: 염도 12% 및 나문재 분말 10% 첨가 된장).
도 2는 나문재 된장(염도 8% 및 12%) 발효 기간에 따른 산도를 나타낸 도이다
(8G: 염도 8% 및 나문재 무첨가 된장; 85N: 염도 8% 및 나문재 분말 5% 첨가 된장; 810N: 염도 8% 및 나문재 분말 10% 첨가 된장;
12G: 염도 12% 및 나문재 무첨가 된장; 125N: 염도 12% 및 나문재 분말 5% 첨가 된장; 1210N: 염도 12% 및 나문재 분말 10% 첨가 된장).
도 3은 나문재 된장(염도 8% 및 12%) 발효 기간에 따른 아미노태 질소의 함량을 나타낸 도이다
(8G: 염도 8% 및 나문재 무첨가 된장; 85N: 염도 8% 및 나문재 분말 5% 첨가 된장; 810N: 염도 8% 및 나문재 분말 10% 첨가 된장;
12G: 염도 12% 및 나문재 무첨가 된장; 125N: 염도 12% 및 나문재 분말 5% 첨가 된장; 1210N: 염도 12% 및 나문재 분말 10% 첨가 된장).
도 4는 나문재 된장(염도 8% 및 12%) 발효 기간에 따른 환원당의 함량을 나타낸 도이다
(8G: 염도 8% 및 나문재 무첨가 된장; 85N: 염도 8% 및 나문재 분말 5% 첨가 된장; 810N: 염도 8% 및 나문재 분말 10% 첨가 된장;
12G: 염도 12% 및 나문재 무첨가 된장; 125N: 염도 12% 및 나문재 분말 5% 첨가 된장; 1210N: 염도 12% 및 나문재 분말 10% 첨가 된장).
도 5는 나문재 된장(염도 8% 및 12% 발효 기간에 따른 Na의 함량을 나타낸 도이다
(8G: 염도 8% 및 나문재 무첨가 된장; 85N: 염도 8% 및 나문재 분말 5% 첨가 된장; 810N: 염도 8% 및 나문재 분말 10% 첨가 된장;
12G: 염도 12% 및 나문재 무첨가 된장; 125N: 염도 12% 및 나문재 분말 5% 첨가 된장; 1210N: 염도 12% 및 나문재 분말 10% 첨가 된장).
도 6은 나문재 된장(염도 8% 및 12%) 발효 기간에 따른 K의 함량을 나타낸 도이다
(8G: 염도 8% 및 나문재 무첨가 된장; 85N: 염도 8% 및 나문재 분말 5% 첨가 된장; 810N: 염도 8% 및 나문재 분말 10% 첨가 된장;
12G: 염도 12% 및 나문재 무첨가 된장; 125N: 염도 12% 및 나문재 분말 5% 첨가 된장; 1210N: 염도 12% 및 나문재 분말 10% 첨가 된장).
도 7은 나문재 된장(염도 8% 및 12%) 발효 기간에 따른 Mg의 함량을 나타낸 도이다
(8G: 염도 8% 및 나문재 무첨가 된장; 85N: 염도 8% 및 나문재 분말 5% 첨가 된장; 810N: 염도 8% 및 나문재 분말 10% 첨가 된장;
12G: 염도 12% 및 나문재 무첨가 된장; 125N: 염도 12% 및 나문재 분말 5% 첨가 된장; 1210N: 염도 12% 및 나문재 분말 10% 첨가 된장).
도 8은 나문재 된장(염도 8% 및 12%) 발효 기간에 따른 Ca의 함량을 나타낸 도이다
(8G: 염도 8% 및 나문재 무첨가 된장; 85N: 염도 8% 및 나문재 분말 5% 첨가 된장; 810N: 염도 8% 및 나문재 분말 10% 첨가 된장;
12G: 염도 12% 및 나문재 무첨가 된장; 125N: 염도 12% 및 나문재 분말 5% 첨가 된장; 1210N: 염도 12% 및 나문재 분말 10% 첨가 된장).
도 9는 LPS 처리된 대식세포에서 나문재 된장 추출물의 산화질소(NO) 생성 억제 효과를 확인한 도이다
(CON: 무처리군; LPS: LPS 처리군;
8G: 염도 8% 및 나문재 무첨가 된장 처리군; 85N: 염도 8% 및 나문재 분말 5% 첨가 된장 처리군; 810N: 염도 8% 및 나문재 분말 10% 첨가 된장 처리군;
12G: 염도 12% 및 나문재 무첨가 된장 처리군; 125N: 염도 12% 및 나문재 분말 5% 첨가 된장 처리군; 1210N: 염도 12% 및 나문재 분말 10% 첨가 된장 처리군).
도 10은 LPS 처리된 대식세포에서 나문재 된장 추출물의 사이토카인 IL-6 생성 억제 효과를 확인한 도이다
(CON: 무처리군; LPS: LPS 처리군;
8G: 염도 8% 및 나문재 무첨가 된장 처리군; 85N: 염도 8% 및 나문재 분말 5% 첨가 된장 처리군; 810N: 염도 8% 및 나문재 분말 10% 첨가 된장 처리군;
12G: 염도 12% 및 나문재 무첨가 된장 처리군; 125N: 염도 12% 및 나문재 분말 5% 첨가 된장 처리군; 1210N: 염도 12% 및 나문재 분말 10% 첨가 된장 처리군).
도 11은 LPS 처리된 대식세포에서 나문재 된장 추출물의 COX-2 및 iNOS 단백질의 발현 억제 효과를 확인한 도이다
(CON: 무처리군; LPS: LPS 처리군;
8G: 염도 8% 및 나문재 무첨가 된장 처리군; 85N: 염도 8% 및 나문재 분말 5% 첨가 된장 처리군; 810N: 염도 8% 및 나문재 분말 10% 첨가 된장 처리군;
12G: 염도 12% 및 나문재 무첨가 된장 처리군; 125N: 염도 12% 및 나문재 분말 5% 첨가 된장 처리군; 1210N: 염도 12% 및 나문재 분말 10% 첨가 된장 처리군).
도 12는 LPS 처리된 대식세포에서 나문재 된장 추출물의 COX-2 및 iNOS mRNA의 발현 억제 효과를 확인한 도이다
(CON: 무처리군; LPS: LPS 처리군;
8G: 염도 8% 및 나문재 무첨가 된장 처리군; 85N: 염도 8% 및 나문재 분말 5% 첨가 된장 처리군; 810N: 염도 8% 및 나문재 분말 10% 첨가 된장 처리군;
12G: 염도 12% 및 나문재 무첨가 된장 처리군; 125N: 염도 12% 및 나문재 분말 5% 첨가 된장 처리군; 1210N: 염도 12% 및 나문재 분말 10% 첨가 된장 처리군).
도 13은 맛 분석기를 이용하여 염도 8% 나문재 된장의 맛을 분석한 결과이다
(CON: 무처리군; 8G0: 염도 8% 및 나문재 무첨가 된장(발효 0일); 85N0: 염도 8% 및 나문재 분말 5% 첨가 된장(발효 0일); 810N0: 염도 8% 및 나문재 분말 10% 첨가 된장(발효 0일); 8G24: 염도 8% 및 나문재 무첨가 된장(발효 24일); 85N24: 염도 8% 및 나문재 분말 5% 첨가 된장(발효 24일); 810N24: 염도 8% 및 나문재 분말 10% 첨가 된장(발효 24일)).
도 14는 맛 분석기를 이용하여 염도 12% 나문재 된장의 맛을 분석한 결과이다
(CON: 무처리군; 8G0: 염도 8% 및 나문재 무첨가 된장(발효 0일); 85N0: 염도 8% 및 나문재 분말 5% 첨가 된장(발효 0일); 810N0: 염도 8% 및 나문재 분말 10% 첨가 된장(발효 0일); 8G24: 염도 8% 및 나문재 무첨가 된장(발효 24일); 85N24: 염도 8% 및 나문재 분말 5% 첨가 된장(발효 24일); 810N24: 염도 8% 및 나문재 분말 10% 첨가 된장(발효 24일)).1 is a diagram showing the moisture content according to the fermentation period of Namunjae miso (
(8G: Doenjang with 8% salinity and no Namunjae powder; 85N: Doenjang with 8% salinity and 5% Namunjae powder; 810N: Doenjang with 8% salinity and 10% Namunjae powder;
12G: Doenjang with 12% salinity and no Namunjae; 125N: Doenjang with 12% salinity and 5% Namunjae powder added; 1210N: Doenjang with 12% salinity and 10% Namunjae powder).
Figure 2 is a diagram showing the acidity according to the fermentation period of namunjae miso (
(8G: Doenjang with 8% salinity and no Namunjae powder; 85N: Doenjang with 8% salinity and 5% Namunjae powder; 810N: Doenjang with 8% salinity and 10% Namunjae powder;
12G: Doenjang with 12% salinity and no Namunjae; 125N: Doenjang with 12% salinity and 5% Namunjae powder added; 1210N: Doenjang with 12% salinity and 10% Namunjae powder).
Figure 3 is a diagram showing the content of amino nitrogen according to the fermentation period of Namunjae Doenjang (
(8G: Doenjang with 8% salinity and no Namunjae powder; 85N: Doenjang with 8% salinity and 5% Namunjae powder; 810N: Doenjang with 8% salinity and 10% Namunjae powder;
12G: Doenjang with 12% salinity and no Namunjae; 125N: Doenjang with 12% salinity and 5% Namunjae powder added; 1210N: Doenjang with 12% salinity and 10% Namunjae powder).
Figure 4 is a diagram showing the content of reducing sugar according to the fermentation period of Namunjae miso (
(8G: Doenjang with 8% salinity and no Namunjae powder; 85N: Doenjang with 8% salinity and 5% Namunjae powder; 810N: Doenjang with 8% salinity and 10% Namunjae powder;
12G: Doenjang with 12% salinity and no Namunjae; 125N: Doenjang with 12% salinity and 5% Namunjae powder added; 1210N: Doenjang with 12% salinity and 10% Namunjae powder).
Figure 5 is a diagram showing the content of Na according to the fermentation period of Namunjae miso (8% salinity and 12%)
(8G: Doenjang with 8% salinity and no Namunjae powder; 85N: Doenjang with 8% salinity and 5% Namunjae powder; 810N: Doenjang with 8% salinity and 10% Namunjae powder;
12G: Doenjang with 12% salinity and no Namunjae; 125N: Doenjang with 12% salinity and 5% Namunjae powder added; 1210N: Doenjang with 12% salinity and 10% Namunjae powder).
6 is a diagram showing the content of K according to fermentation period of Namunjae miso (
(8G: Doenjang with 8% salinity and no Namunjae powder; 85N: Doenjang with 8% salinity and 5% Namunjae powder; 810N: Doenjang with 8% salinity and 10% Namunjae powder;
12G: Doenjang with 12% salinity and no Namunjae; 125N: Doenjang with 12% salinity and 5% Namunjae powder added; 1210N: Doenjang with 12% salinity and 10% Namunjae powder).
7 is a diagram showing the content of Mg according to the fermentation period of Namunjae miso (
(8G: Doenjang with 8% salinity and no Namunjae powder; 85N: Doenjang with 8% salinity and 5% Namunjae powder; 810N: Doenjang with 8% salinity and 10% Namunjae powder;
12G: Doenjang with 12% salinity and no Namunjae; 125N: Doenjang with 12% salinity and 5% Namunjae powder added; 1210N: Doenjang with 12% salinity and 10% Namunjae powder).
Figure 8 is a diagram showing the content of Ca according to the fermentation period of Namunjae miso (
(8G: Doenjang with 8% salinity and no Namunjae powder; 85N: Doenjang with 8% salinity and 5% Namunjae powder; 810N: Doenjang with 8% salinity and 10% Namunjae powder;
12G: Doenjang with 12% salinity and no Namunjae; 125N: Doenjang with 12% salinity and 5% Namunjae powder added; 1210N: Doenjang with 12% salinity and 10% Namunjae powder).
9 is a diagram confirming the inhibitory effect of nitric oxide (NO) production of Namunjae doenjang extract in LPS-treated macrophages
(CON: no treatment group; LPS: LPS treatment group;
8G: Doenjang treated group with 8% salinity and no addition of Namunjae; 85N: Doenjang-treated group with 8% salinity and 5% Namunjae powder added; 810N: Doenjang-treated group with 8% salinity and 10% Namunjae powder added;
12G:
Figure 10 is a diagram confirming the cytokine IL-6 production inhibitory effect of Namunjae Doenjang extract in LPS-treated macrophages
(CON: no treatment group; LPS: LPS treatment group;
8G: Doenjang treated group with 8% salinity and no addition of Namunjae; 85N: Doenjang-treated group with 8% salinity and 5% Namunjae powder added; 810N: Doenjang-treated group with 8% salinity and 10% Namunjae powder added;
12G:
11 is a diagram confirming the effect of inhibiting the expression of COX-2 and iNOS proteins of Namunjae Doenjang extract in LPS-treated macrophages
(CON: no treatment group; LPS: LPS treatment group;
8G: Doenjang treated group with 8% salinity and no addition of Namunjae; 85N: Doenjang-treated group with 8% salinity and 5% Namunjae powder added; 810N: Doenjang-treated group with 8% salinity and 10% Namunjae powder added;
12G:
12 is a diagram confirming the effect of inhibiting the expression of COX-2 and iNOS mRNA of Namunjae Doenjang extract in LPS-treated macrophages
(CON: no treatment group; LPS: LPS treatment group;
8G: Doenjang treated group with 8% salinity and no addition of Namunjae; 85N: Doenjang-treated group with 8% salinity and 5% Namunjae powder added; 810N: Doenjang-treated group with 8% salinity and 10% Namunjae powder added;
12G:
13 is a result of analyzing the taste of Namunjae Doenjang with 8% salinity using a taste analyzer
(CON: No treatment group; 8G0: Doenjang with 8% salinity and Namunjae-free (
14 is a result of analyzing the taste of Namunjae Doenjang with a salinity of 12% using a taste analyzer
(CON: No treatment group; 8G0: Doenjang with 8% salinity and Namunjae-free (
이하, 본 발명을 상세히 설명한다. Hereinafter, the present invention will be described in detail.
본 발명은 1) 메주, 나문재 분말, 소금 및 물을 혼합하는 단계; 및 2) 상기 단계 1)의 혼합물을 발효하는 단계;를 포함하는 항염증 또는 면역 억제 활성을 갖는 나문재 된장의 제조방법을 제공한다. The present invention comprises the steps of 1) mixing meju, namunjae powder, salt and water; And 2) fermenting the mixture of step 1). It provides a method for producing Namunjae miso having anti-inflammatory or immunosuppressive activity.
상기 단계 1)은 메주 100 중량부에 대하여 나문재 분말 3 내지 15 중량부, 소금 5 내지 20 중량부, 물 5 내지 25 중량부로 혼합할 수 있다. 구체적으로, 상기 단계 1)의 소금은 메주 100 중량부에 대하여 8 내지 17 중량부, 더 구체적으로는 10 내지 17 중량부, 보다 더 구체적으로는 12 내지 15 중량부로 혼합할 수 있다. 또한, 상기 단계 1)의 나문재 분말은 메주 100 중량부에 대하여 3 내지 15 중량부, 구체적으로 5 내지 15 중량부로 혼합할 수 있다. The step 1) may be mixed with 3 to 15 parts by weight of namunjae powder, 5 to 20 parts by weight of salt, and 5 to 25 parts by weight of water based on 100 parts by weight of meju. Specifically, the salt of step 1) may be mixed in an amount of 8 to 17 parts by weight, more specifically 10 to 17 parts by weight, and even more specifically 12 to 15 parts by weight based on 100 parts by weight of meju. In addition, the namunjae powder of step 1) may be mixed in an amount of 3 to 15 parts by weight, specifically 5 to 15 parts by weight, based on 100 parts by weight of meju.
상기 단계 1)의 메주, 나문재 분말, 소금은 제조한 것 또는 시판되는 것 등 제한없이 사용할 수 있다. Meju, namunjae powder, and salt of step 1) can be used without limitation, such as prepared or commercially available.
상기 단계 1)의 메주는 하기의 단계들을 포함하는 제조방법에 의해 제조될 수 있다: a) 세척한 대두를 물에 침지시키는 단계; b) 상기 단계 a)의 침지시킨 대두를 절수하고 증자하는 단계; 및 c) 상기 b)의 증자한 대두에 종균을 접종하고 발효하는 단계. Meju of step 1) may be prepared by a manufacturing method comprising the following steps: a) immersing the washed soybeans in water; b) saving water and steaming the soybeans soaked in step a); And c) inoculating and fermenting the soybeans grown in b).
본 발명의 일 실시예에서, 상기 단계 a)의 침지는 10 내지 20시간 동안 수행할 수 있고, 구체적으로 12 내지 18시간 동안 수행할 수 있다. 상기 단계 b)의 증자는 100 내지 140℃의 온도, 구체적으로 110 내지 130℃의 온도에서 수행할 수 있고, 10 내지 60분 동안, 구체적으로 20 내지 40분 동안 수행할 수 있다. 상기 단계 c)의 종균은 아스퍼길러스 오리재(Aspergillus oryzae)일 수 있으나, 이에 제한되지 않는다. 구체적으로, 아스퍼길러스 오리재(Aspergillus oryzae) KACC 46471 균주일 수 있다. In an embodiment of the present invention, the immersion in step a) may be performed for 10 to 20 hours, and specifically for 12 to 18 hours. The increase in step b) may be performed at a temperature of 100 to 140°C, specifically 110 to 130°C, and may be performed for 10 to 60 minutes, specifically for 20 to 40 minutes. The seed in step c) may be Aspergillus oryzae , but is not limited thereto. Specifically, Aspergillus duck material (Aspergillus oryzae ) KACC 46471 strain.
상기 단계 2)의 발효는 20 내지 40℃의 온도, 구체적으로 25 내지 35℃의 온도, 더 구체적으로 27 내지 33℃의 온도에서 수행할 수 있다. 또한, 상기 단계 2)의 발효는 70 내지 95%의 습도, 구체적으로 75 내지 90%의 습도, 보다 구체적으로 75 내지 85%의 습도에서 수행할 수 있다. 또한, 상기 단계 2)의 발효는 2 내지 10개월, 구체적으로 4 내지 8개월, 더 구체적으로 5 내지 7개월 동안 수행할 수 있다. The fermentation of step 2) may be performed at a temperature of 20 to 40°C, specifically 25 to 35°C, and more specifically at a temperature of 27 to 33°C. In addition, the fermentation of step 2) may be performed at a humidity of 70 to 95%, specifically 75 to 90%, and more specifically at 75 to 85%. In addition, the fermentation of step 2) may be performed for 2 to 10 months, specifically 4 to 8 months, and more specifically 5 to 7 months.
또한, 본 발명은 상기 제조방법으로 제조된 항염증 또는 면역 억제 활성을 갖는 나문재 된장을 제공한다. In addition, the present invention provides a namunjae doenjang having anti-inflammatory or immunosuppressive activity prepared by the above manufacturing method.
상기 나문재 된장은 산화질소의 생성을 억제하는 활성을 가질 수 있고, COX-2 또는 iNOS의 발현을 억제할 수 있다.The namunjae doenjang may have an activity of inhibiting the production of nitric oxide, and may inhibit the expression of COX-2 or iNOS.
상기 나문재 된장의 염도 5 내지 15%일 수 있고, 구체적으로 7 내지 15%일 수 있고, 더 구체적으로 8 내지 15%, 보다 더 구체적으로는 10 내지 15%일 수 있다. The salinity of the namunjae miso may be 5 to 15%, specifically 7 to 15%, more specifically 8 to 15%, and even more specifically 10 to 15%.
또한, 본 발명은 상기 나문재 된장의 추출물을 유효성분으로 포함하는 항염증용 및 면역 억제용 조성물을 제공한다.In addition, the present invention provides a composition for anti-inflammatory and immune suppression comprising the extract of Namunjae doenjang as an active ingredient.
상기 나문재 된장의 추출물은 하기의 단계들을 포함하는 제조방법에 의해 제조될 수 있다: A) 상기 제조방법으로 제조된 나문재 된장에 추출 용매를 가하여 추출하는 단계; 및 B) 상기 A)의 추출물을 여과하는 단계.The extract of the namunjae doenjang may be prepared by a manufacturing method comprising the following steps: A) extracting by adding an extraction solvent to the namunjae doenjang prepared by the above manufacturing method; And B) filtering the extract of A).
상기 단계 A)의 추출 용매는 물, C1 내지 C2의 저급 알코올 또는 이들의 혼합 용매일 수 있다. 상기 C1 내지 C2의 저급 알코올은 메탄올 또는 에탄올일 수 있고, 구체적으로 에탄올일 수 있다. The extraction solvent of step A) may be water, a lower alcohol of C 1 to C 2 , or a mixed solvent thereof. The lower alcohol of C 1 to C 2 may be methanol or ethanol, and specifically ethanol.
상기 단계 A)의 추출 방법은 Soxhlet 추출 또는 환류추출일 수 있다. 이때, 추출 온도는 15 내지 35℃일 수 있고, 구체적으로 20 내지 30℃일 수 있고, 더 구체적으로, 22 내지 28℃일 수 있으나 이에 한정하지 않는다. 또한, 상기 추출 용매는 나문재 된장의 중량 1 g 당 50 내지 300 ㎖의 양으로 첨가될 수 있고, 구체적으로, 100 내지 250 ㎖의 양으로 첨가될 수 있고, 더 구체적으로 120 내지 180 ㎖의 양으로 첨가될 수 있으나, 이에 한정하지 않는다. 또한, 추출 시간은 1 내지 5시간일 수 있고, 구체적으로 1.5 내지 4시간일 수 있으나, 이에 한정하지 않는다. 아울러, 추출 횟수는 1 내지 4회일 수 있고, 구체적으로 1 내지 3회일 수 있으나 이에 한정하지 않는다. The extraction method of step A) may be Soxhlet extraction or reflux extraction. In this case, the extraction temperature may be 15 to 35°C, specifically 20 to 30°C, and more specifically, 22 to 28°C, but is not limited thereto. In addition, the extraction solvent may be added in an amount of 50 to 300 ㎖ per 1 g of Namunjae Doenjang, specifically, may be added in an amount of 100 to 250 ㎖, more specifically in an amount of 120 to 180 ㎖ It may be added, but is not limited thereto. In addition, the extraction time may be 1 to 5 hours, specifically 1.5 to 4 hours, but is not limited thereto. In addition, the number of extractions may be 1 to 4 times, specifically 1 to 3 times, but is not limited thereto.
상기 추출물은 산화질소의 생성을 억제하는 활성을 가질 수 있고, COX-2 또는 iNOS의 발현을 억제할 수 있다.The extract may have an activity of inhibiting the production of nitric oxide, and may inhibit the expression of COX-2 or iNOS.
본 발명의 구체적인 실시예에서, 본 발명자들은 나문재가 첨가된 된장을 제조하고, 이를 에탄올로 추출하여 LPS 처리된 대식세포에 처리한 결과, 염도 12%의 나문재 된장의 추출물은 산화질소의 생성을 억제하고 COX-2 및 iNOS의 발현을 단백질 및 mRNA 수준에서 억제함을 확인하였다 (도 9 내지 도 12 참조).In a specific embodiment of the present invention, the present inventors prepared soybean paste to which Namunjae was added, extracted with ethanol, and treated with LPS-treated macrophages. As a result, the extract of Namunjae Doenjang with a salinity of 12% inhibited the production of nitric oxide. And it was confirmed that the expression of COX-2 and iNOS was inhibited at the protein and mRNA levels (see FIGS. 9 to 12).
따라서, 본 발명에 따라 제조된 나문재 된장은 항염증용 및 면역 억제 용도로 유용하게 사용될 수 있다. Therefore, namunjae doenjang prepared according to the present invention can be usefully used for anti-inflammatory and immune suppression purposes.
본 발명에 따른 조성물은 조성물 전체 중량에 대하여 유효성분인 나문재 된장을 10 내지 95 중량%로 포함할 수 있다. 또한, 본 발명의 조성물은 상기 유효성분 이외에 추가로 동일 또는 유사한 기능을 나타내는 유효성분을 1종 이상 추가로 포함할 수 있다.The composition according to the present invention may contain 10 to 95% by weight of namunjae miso, which is an active ingredient, based on the total weight of the composition. In addition, the composition of the present invention may further include one or more active ingredients exhibiting the same or similar functions in addition to the active ingredients.
본 발명의 조성물은 생물학적 제제에 통상적으로 사용되는 담체, 희석제, 부형제 또는 이의 혼합물을 포함할 수 있다. 약학적으로 허용가능한 담체는 조성물을 생체 내에 전달하는데 적합한 것이면 모두 사용할 수 있다. 구체적으로, 상기 담체는 Merck Index, 13th ed., Merck & Co. Inc.에 기재된 화합물, 식염수, 멸균수, 링거액, 덱스트로스 용액, 말토덱스트린 용액, 글리세롤, 에탄올 또는 이의 혼합물일 수 있다. 또한, 필요에 따라 항산화제, 완충액, 정균제 등과 같은 통상의 첨가제를 첨가할 수 있다.The composition of the present invention may contain carriers, diluents, excipients or mixtures thereof commonly used in biological preparations. Any pharmaceutically acceptable carrier may be used as long as it is suitable for delivering the composition in vivo. Specifically, the carrier is Merck Index, 13th ed., Merck & Co. Inc., saline, sterile water, Ringer's solution, dextrose solution, maltodextrin solution, glycerol, ethanol, or a mixture thereof. In addition, conventional additives such as antioxidants, buffers, and bacteriostatic agents may be added as needed.
상기 조성물을 제제화하는 경우, 보통 사용하는 충진제, 증량제, 결합제, 습윤제, 붕해제, 계면활성제 등의 희석제 또는 부형제를 첨가할 수 있다.When formulating the composition, diluents or excipients such as commonly used fillers, extenders, binders, wetting agents, disintegrants, and surfactants may be added.
본 발명의 조성물은 경구용 제제 또는 비경구용 제제로 제형화될 수 있다. 경구용 제제로는 고형 제제 및 액상 제제가 포함될 수 있다. 상기 고형 제제는 정제, 환제, 산제, 과립제, 캡슐제 또는 트로키제일 수 있고, 이러한 고형 제제는 상기 조성물에 적어도 하나 이상의 부형제를 첨가하여 조제할 수 있다. 상기 부형제는 전분, 탄산칼슘, 수크로스, 락토오스, 젤라틴 또는 이의 혼합물일 수 있다. 또한, 상기 고형 제제는 윤활제를 포함할 수 있고, 그 예로는 마그네슘 스티레이트, 탈크등이 있다. 한편, 상기 액상 제제는 현탁제, 내용액제, 유제 또는 시럽제일 수 있다. 이때, 상기 액상 제제에는 습윤제, 감미제, 방향제, 보존제 등과 같은 부형제가 포함될 수 있다.The composition of the present invention may be formulated as an oral or parenteral formulation. Oral formulations may include solid formulations and liquid formulations. The solid preparation may be a tablet, a pill, a powder, a granule, a capsule or a troche, and the solid preparation may be prepared by adding at least one excipient to the composition. The excipient may be starch, calcium carbonate, sucrose, lactose, gelatin, or a mixture thereof. In addition, the solid preparation may contain a lubricant, and examples thereof include magnesium stearate and talc. On the other hand, the liquid formulation may be a suspension, an inner solution, an emulsion or a syrup. At this time, the liquid formulation may contain excipients such as wetting agents, sweetening agents, fragrances, and preservatives.
상기 비경구용 제제는 주사제, 좌제, 호흡기 흡입용 분말, 스프레이용 에어로졸제, 파우더 및 크림 등을 포함할 수 있다. 상기 주사제는 멸균된 수용액, 비수성용제, 현탁용제, 유제 등을 포함할 수 있다. 이때, 비수성용제 또는 현탁용제로서는 프로필렌글리콜, 폴리에틸렌글리콜, 올리브 오일과 같은 식물성 기름이나, 에틸올레이트와 같이 주사가능한 에스테르 등이 사용될 수 있다.The parenteral preparation may include injections, suppositories, powders for respiratory inhalation, aerosols for sprays, powders and creams. The injection may include a sterilized aqueous solution, a non-aqueous solvent, a suspension solvent, an emulsion, and the like. At this time, as the non-aqueous solvent or suspension solvent, vegetable oils such as propylene glycol, polyethylene glycol, and olive oil, or injectable esters such as ethyl oleate may be used.
본 발명의 조성물은 목적하는 방법에 따라 경구 또는 비경구로 투여될 수 있다. 비경구 투여는 복강내, 직장내, 피하, 정맥, 근육내 또는 흉부내 주사 방식을 포함할 수 있다.The composition of the present invention may be administered orally or parenterally according to a desired method. Parenteral administration may include intraperitoneal, rectal, subcutaneous, intravenous, intramuscular or intrathoracic injection.
상기 조성물은 약제학적으로 유효한 양으로 투여될 수 있다. 이는 질환의 종류, 중증도, 약물의 활성, 약물에 대한 환자의 민감도, 투여 시간, 투여 경로, 치료기간, 동시에 사용되는 약물 등에 따라 달라질 수 있다. 그러나, 바람직한 효과를 위해서, 본 발명에 따른 조성물에 포함되는 유효성분의 양은 0.0001 내지 100 ㎎/㎏, 구체적으로 0.001 내지 10 ㎎/㎏일 수 있다. 상기 투여는 하루에 1회 수회일 수 있다.The composition may be administered in a pharmaceutically effective amount. This may vary depending on the type of disease, the severity, the activity of the drug, the patient's sensitivity to the drug, the administration time, the administration route, the treatment period, and the drugs used at the same time. However, for a desirable effect, the amount of the active ingredient contained in the composition according to the present invention may be 0.0001 to 100 mg/kg, specifically 0.001 to 10 mg/kg. The administration may be several times a day.
본 발명의 조성물은 단독 또는 다른 치료제와 병용하여 투여될 수 있다. 병용 투여시, 투여는 순차적 또는 동시일 수 있다.The composition of the present invention may be administered alone or in combination with other therapeutic agents. When administered in combination, administration may be sequential or simultaneous.
본 발명의 조성물은 식품에 그대로 첨가하거나, 다른 식품 또는 식품 성분과 함께 사용될 수 있다. 이때, 첨가되는 유효성분의 함량은 목적에 따라 결정될 수 있고, 일반적으로는 전체 식품 중량의 0.01 내지 90 중량부일 수 있다.The composition of the present invention may be added to food as it is, or may be used together with other foods or food ingredients. At this time, the content of the active ingredient to be added may be determined according to the purpose, and generally may be 0.01 to 90 parts by weight of the total food weight.
식품의 형태 및 종류는 특별히 제한되지 않는다. 구체적으로, 상기 식품은 정제, 캅셀, 분말, 과립, 액상 및 환의 형태일 수 있다. 상기 식품은 추가성분으로서 여러 가지 향미제, 감미제 또는 천연 탄수화물을 포함할 수 있다. 상기 감미제는 천연 또는 합성 감미제일 수 있고, 천연 감미제의 예로는 타우마틴, 스테비아 추출물 등이 있다. 한편, 합성 감미제의 예로는 사카린, 아스파르탐 등이 있다. 또한, 상기 천연 탄수화물은 모노사카라이드, 디사카라이드, 폴리사카라이드, 올리고당 및 당알코올 등일 수 있다.The form and type of food are not particularly limited. Specifically, the food may be in the form of tablets, capsules, powders, granules, liquids, and pills. The food may contain various flavoring agents, sweetening agents or natural carbohydrates as additional ingredients. The sweetener may be a natural or synthetic sweetener, and examples of the natural sweetener include taumatin and stevia extract. Meanwhile, examples of synthetic sweeteners include saccharin and aspartame. In addition, the natural carbohydrates may be monosaccharides, disaccharides, polysaccharides, oligosaccharides and sugar alcohols.
본 발명의 조성물은 상기 서술한 추가성분 외에, 영양제, 비타민, 전해질, 풍미제, 착색제, 펙스탄 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알코올 등을 더 포함할 수 있다. 이러한 성분은 독립적으로 또는 조합으로 사용될 수 있다. 상기 첨가제의 비율은 본 발명의 조성물의 100 중량부당 0.01 내지 0.1 중량부의 범위에서 선택될 수 있다.In addition to the above-described additional ingredients, the composition of the present invention includes nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pexane and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin. , Alcohol, etc. may be further included. These ingredients can be used independently or in combination. The ratio of the additive may be selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.
이하, 본 발명을 하기 실시예에 의해 상세히 설명한다. Hereinafter, the present invention will be described in detail by the following examples.
단, 하기 실시예는 본 발명을 예시하는 것일 뿐, 본 발명의 내용이 하기 실시예에 의해 한정되는 것은 아니다. However, the following examples are merely illustrative of the present invention, and the contents of the present invention are not limited by the following examples.
<실시예 1> 나문재가 첨가된 된장의 제조<Example 1> Preparation of Doenjang with Namunjae Added
<1-1> 콩알메주의 제조<1-1> Preparation of soybean almeju
세척한 대두를 수돗물에 15시간 동안 침지시키고 1시간 동안 절수한 후, 온도 121℃, 압력 1,000 atm 조건에서 30분 동안 증자하였다. 증자한 대두에 1×106 CFU/g의 아스퍼길러스 오리재(Aspergillus oryzae) KACC 46471 균주를 2%로 접종하고, 온도 30℃, 습도 80% 조건에서 2~3일 동안 발효하여 콩알메주를 제조하였다. 콩알메주에서 균사체가 발견되면 발효를 중지하였다. The washed soybeans were immersed in tap water for 15 hours and water-saved for 1 hour, and then steamed for 30 minutes at a temperature of 121°C and a pressure of 1,000 atm. 1×10 6 CFU/g of Aspergillus duck ash (Aspergillus oryzae ) KACC 46471 strain was inoculated at 2%, and fermented for 2-3 days at 30°C and 80% humidity to prepare soybean almeju. Fermentation was stopped when mycelium was found in soybean almeju.
<1-2> 나문재 된장의 제조<1-2> Preparation of Namunjae Doenjang
제조한 콩알메주에 나문재 분말(미가식품영농조합법인), 천일염(3년 숙성, CJ) 및 물을 하기 표 1의 배합비율에 따라 첨가하고, 30℃, 습도 80% 조건의 숙성실에서 6개월 동안 발효하여 나문재 된장을 제조하였다. 제조한 나문재 된장은 저온에서 보관하였다. 제조한 된장의 염도는 AgNO3 적정법(식품의약품안전처, Korean Food Standards Codex, 2015) 을 변형하여 측정하였다. 구체적으로, 제조한 된장 2.5 g을 증류수로 20배 희석하고 이를 500 rpm에서 1시간 동안 진탕 배양(SI-IS20, (주)신일, 한국)하여 추출한 후, 여과지(Watman No.2)로 여과하였다. 여과액 10 mL에 지시약(5% K2CrO4)을 1 mL 첨가한 후, 0.1N AgNO3로 적정하여 AgNO3의 첨가량을 기초하여 염도를 산출하였다. 3회 반복 측정한 후 그 평균값을 산출하였다. Namunjae powder (Miga Foods Farming Association), sea salt (3-year aging, CJ), and water were added to the prepared soybean almeju according to the mixing ratio shown in Table 1 below, and for 6 months in a aging room at 30℃ and 80% humidity. Fermented to prepare Namunjae Doenjang. The prepared namunjae miso was stored at low temperature. The salinity of the prepared doenjang was measured by modifying the AgNO 3 titration method (Ministry of Food and Drug Safety, Korean Food Standards Codex, 2015). Specifically, 2.5 g of the prepared soybean paste was diluted 20 times with distilled water and extracted by shaking culture for 1 hour at 500 rpm (SI-IS20, Shinil Co., Ltd., Korea), and then filtered through filter paper (Watman No.2). . After adding 1 mL of an indicator (5% K 2 CrO 4 ) to 10 mL of the filtrate, it was titrated with 0.1N AgNO 3 to calculate the salinity based on the amount of AgNO 3 added. After repeated measurement three times, the average value was calculated.
<< 실험예Experimental example 1> 1> 나문재Moonjae Na 된장의 품질 특성 분석 Analysis of quality characteristics of soybean paste
실시예 1에서 제조한 나문재 된장의 품질 특성을 확인하기 위해, 수분함량, 산도, 아미노태 질소 함량, 환원당 함량 및 무기 성분의 함량을 분석하였다. In order to confirm the quality characteristics of Namunjae Doenjang prepared in Example 1, the moisture content, acidity, amino nitrogen content, reducing sugar content, and content of inorganic components were analyzed.
<1-1> 수분함량<1-1> Moisture content
실시예 1에서 제조한 나문재 된장을 발효 기간에 따라 수분 함량을 측정하였다. 수분 함량은 Association of Official Analytical Chemists법(AOAC, 1990)에 따라 105℃에서 상압가열건조법을 사용하여 측정하였다. 구체적으로, 나문재 된장의 1 g을 취하여 105℃의 드라이 오븐(MOV-112, Sanyo Co., Ltd., 일본))에서 항량이 될 때까지 건조하고, 항량에 달한 평량 접시(weighing dish)를 이용하여 무게를 3회 반복 측정한 후, 평균값을 산출하였다. The moisture content of Namunjae Doenjang prepared in Example 1 was measured according to the fermentation period. Moisture content was measured using an atmospheric pressure drying method at 105°C according to the Association of Official Analytical Chemists method (AOAC, 1990). Specifically, take 1 g of Namunjae miso and dry it in a dry oven at 105°C (MOV-112, Sanyo Co., Ltd., Japan) to a constant weight, and use a weighing dish with a constant weight. Then, the weight was repeatedly measured three times, and the average value was calculated.
그 결과, 실시예 1의 나문재 된장 모두 약 55%의 수분 함량을 유지하는 것으로 나타났으며, 이는 한국 전통식품 규격 기준인 60% 이하를 만족하는 결과이다 (도 1). As a result, it was found that both Namunjae Doenjang of Example 1 maintained a moisture content of about 55%, which is a result that satisfies 60% or less, which is a standard standard for Korean traditional foods (FIG. 1).
<1-2> 산도<1-2> acidity
실시예 1에서 제조한 나문재 된장을 발효 기간에 따라 산도를 측정하였다. 먼저, 용기 내 동일한 위치에서 취한 나문재 된장 약 9 g에 증류수 36 mL를 가하고, 균질화기(Homogenizer, Polytron PT-MR 2100, Kinematica AG, 스위스)로 1분 동안 균질화한 후, 8,000 rpm에서 10분 동안 원심분리하였다. 상층액을 분리하여 이를 여과지(Watman No. 2)로 여과하고, 그 여과액 5 mL에 증류수 20 mL를 첨가한 후, 지시약 1% 페놀프탈레인(phenolphthalein)을 첨가하였다. 0.1N NaOH를 pH 8.3이 될 때까지 첨가하여 소비된 NaOH의 양을 측정하여 산도를 산출하였다. The acidity of Namunjae Doenjang prepared in Example 1 was measured according to the fermentation period. First, 36 mL of distilled water was added to about 9 g of Namunjae Doenjang taken at the same location in the container, homogenized with a homogenizer (Homogenizer, Polytron PT-MR 2100, Kinematica AG, Switzerland) for 1 minute, and then at 8,000 rpm for 10 minutes. Centrifuged. The supernatant was separated, filtered through a filter paper (Watman No. 2), and 20 mL of distilled water was added to 5 mL of the filtrate, followed by addition of 1% phenolphthalein as an indicator. 0.1N NaOH was added until the pH reached 8.3, and the amount of NaOH consumed was measured to calculate the acidity.
그 결과, 실시예 1의 나문재 된장 모두 발효 기간이 길어질수록 산도가 높아지다가 일정 기간이 지나면 산도가 유지 또는 감소하였고, 나문재 무첨가 된장(8G 및 12G)보다 나문재를 첨가한 된장의 산도가 대체로 낮은 것으로 나타났다 (도 2). As a result, the acidity of the Namunjae miso in Example 1 increased as the fermentation period increased, and after a certain period of time, the acidity was maintained or decreased, and the acidity of the Namunjae doenjang was generally lower than that of Namunjae-free doenjang (8G and 12G). Appeared (Fig. 2).
<1-3> 아미노태 질소<1-3> amino nitrogen
실시예 1에서 제조한 나문재 된장을 발효 기간에 따라 아미노태 질소 함량을 측정하였다. 아미노태 질소(NO3-N)는 Formol법을 일부 변형하여 측정하였다. 실험예 1-2와 동일한 방법으로 준비한 여과액 5 mL에 중성 포르말린(formalin, pH 8.3) 10 mL과 증류수 10 mL을 첨가하고, 0.1 N NaOH로 pH 8.3로 중화될 때까지 적정하였다. 이때 소모된 0.1 N NaOH mL를 측정하여 아미노태 질소함량으로 결정하였다. 공시험구(대조군)은 중성 포르말린을 대신하여 증류수를 사용하여 측정하여 아미노태 질소 함량을 측정하였다. The amino nitrogen content of Namunjae Doenjang prepared in Example 1 was measured according to the fermentation period. Amino nitrogen (NO 3 -N) was measured by partially modifying the Formol method. To 5 mL of the filtrate prepared in the same manner as in Experimental Example 1-2, 10 mL of neutral formalin (formalin, pH 8.3) and 10 mL of distilled water were added, and titrated until neutralized to pH 8.3 with 0.1 N NaOH. At this time, 0.1 N NaOH mL consumed was measured and determined as the amino nitrogen content. The blank test group (control group) was measured using distilled water instead of neutral formalin to measure the amino nitrogen content.
발효 전의 아미노태 질소 함량은 나문재 무첨가 된장의 경우, 574.9 내지 707.0 mg%, 나문재 첨가 된장의 경우, 535.0 내지 673.4 mg% 로 나타났으며, 발효 기간 동안 나문재 무첨가 된장보다 나문재 첨가 된장의 아미노태 질소 함량이 낮은 것으로 나타났다 (도 3). The amino nitrogen content before fermentation was 574.9 to 707.0 mg% in the case of bean paste without Namunjae, and 535.0 to 673.4 mg% in the case of bean paste with Namunjae. It was found to be low (Figure 3).
<1-4> 환원당<1-4> reducing sugar
실시예 1에서 제조한 나문재 된장을 발효 기간에 따라 환원당의 함량을 측정하였다. 환원당 함량은 DNS법을 이용하여 측정하였다. 실험예 1-2와 동일한 방법으로 준비한 여과액 1 mL에 DNS 시약 3 mL을 혼합하고, 혼합액을 100℃의 물에서 중탕하였다. 5분 후, 물에서 꺼내어 식히고 분광기(Libra S35, Biochrome Ltd., 영국)를 이용하여 550 nm에서 흡광도를 측정하였다. 포도당을 표준 물질로 하여 작성한 검량곡선을 이용하여 환원당 함량을 산출하였다.The content of reducing sugar was measured according to the fermentation period of Namunjae Doenjang prepared in Example 1. The reducing sugar content was measured using the DNS method. 3 mL of DNS reagent was mixed with 1 mL of the filtrate prepared in the same manner as in Experimental Example 1-2, and the mixture was bathed in water at 100°C. After 5 minutes, it was taken out of water, cooled, and absorbance was measured at 550 nm using a spectrometer (Libra S35, Biochrome Ltd., UK). The reducing sugar content was calculated using a calibration curve prepared with glucose as a standard substance.
그 결과, 나문재 된장은 나문재 무첨가 된장보다 환원당이 더 많은 것으로 나타났으며, 특히 염도 12%의 나문재 된장은 발효 기간 동안 무첨가 된장보다 유의적으로 환원당 함량이 증가한 것으로 나타났다 (도 4). As a result, Namunjae Doenjang was found to have more reducing sugar than Namunjae Doenjang, and Namunjae Doenjang with a salinity of 12% was found to significantly increase the reducing sugar content during fermentation period compared to non-added Doenjang (FIG. 4).
<1-5> 무기성분<1-5> Inorganic components
실시예 1에서 제조한 나문재 된장을 발효 기간에 따라 무기성분 Na, K, Mg 및 Ca의 함량을 측정하였다. 무기물 함량은 식품공전방법에 준하여 검체를 고온에서 탄화시켜 유기물질을 회화시킨 후, 이를 용해하여 유도결합플라즈마법으로 측정하였다. The contents of the inorganic ingredients Na, K, Mg, and Ca were measured according to the fermentation period of the namunjae miso prepared in Example 1. According to the food code method, the sample was carbonized at high temperature to incinerate organic substances, and then dissolved and measured by inductively coupled plasma method.
구체적으로, 나문재 된장 1 g (이용하신 시료는 나문재 된장을 그대로 이용하신 것인지 확인 부탁드립니다.)을 회화용기에 칭량하고 550℃의 온도에서 여러 시간 가열하여 백색의 회분이 얻어질 때까지 회화하였다. 완전히 탄화된 회분을 방냉한 후, 약 10 mL의 염산용액(HCL:H2O=1:1, v/v)을 가해 수욕 상에서 완전 증발 건조시켰다. 건조물에 염산용액 약 10 mL를 가하여 30분 동안 용해시키고 50 mL 메스플라스크에 여과·정용하여 시험용액으로 사용하였다. 시험용액의 무기 성분을 ICP-OES(Optima 8300, Perkin elmer co, 미국)로 측정하였다. 이 때 RF 전력은 1,300W, 플라즈마 아르곤 가스 유입 속도 15L/min, Auxiliary 아르곤 가스 유입 속도 0.2L/min, Nebulizer 아르곤 가스 유입 속도 0.6L/min으로 하였다. Specifically, 1 g of Namunjae Doenjang (Please confirm that Namunjae Doenjang is used as it is) was weighed in an incineration container and heated at 550°C for several hours to incinerate until a white ash was obtained. After the completely carbonized ash was allowed to cool, about 10 mL of a hydrochloric acid solution (HCL:H 2 O=1:1, v/v) was added and completely evaporated to dryness in a water bath. About 10 mL of hydrochloric acid solution was added to the dried product, dissolved for 30 minutes, filtered and fixed in a 50 mL volumetric flask, and used as a test solution. The inorganic components of the test solution were measured by ICP-OES (Optima 8300, Perkin elmer co, USA). In this case, the RF power was 1,300W, plasma argon gas inflow rate 15L/min, auxiliary argon gas inflow rate 0.2L/min, nebulizer argon gas inflow rate 0.6L/min.
그 결과, Na, K 및 Mg의 무기성분은 나문재 무첨가 된장보다 나문재 된장에서 함량이 더 낮았고, Ca 성분은 나문재 된장에서 함량이 더 높았다 (도 5 내지 도 8). As a result, the content of inorganic components of Na, K, and Mg was lower in Namunjae Doenjang than in Namunjae Doenjang, and Ca content was higher in Namunjae Doenjang (FIGS. 5 to 8).
<실시예 2> 나문재 된장의 추출물 제조<Example 2> Preparation of extract of Namunjae Doenjang
실시예 1에서 제조한 나문재 된장의 면역 억제 효과를 확인하기 위해, 나문재 된장을 추출물 형태로 제조하였다. In order to confirm the immunosuppressive effect of Namunjae Doenjang prepared in Example 1, Namunjae Doenjang was prepared in the form of an extract.
구체적으로, 실시예 1의 나문재 된장 5 g에 80% 에탄올 150 ㎖을 첨가하고 진탕배양기(Water bath, Wisebath, DH, 한국)에서 상온 조건에서 2~3시간 동안 추출하였다. 이와 같은 추출 과정을 2회 거쳐 상층액을 수집하고, 상층액을 여과지(Watman No. 2)를 이용하여 여과하였다. 여과액을 회전 증발 농축기(rotary vacuum evaporator, EYELA, 일본)로 감압 농축한 후, 동결건조기(Bondiro, Ilshin, 한국)로 건조하여 나문재 된장의 추출물을 제조하였다. Specifically, 150 ml of 80% ethanol was added to 5 g of Namunjae doenjang of Example 1, and extracted for 2 to 3 hours at room temperature in a shaking incubator (Water bath, Wisebath, DH, Korea). The supernatant was collected through the same extraction process twice, and the supernatant was filtered using a filter paper (Watman No. 2). The filtrate was concentrated under reduced pressure with a rotary vacuum evaporator (EYELA, Japan), and then dried with a freeze dryer (Bondiro, Ilshin, Korea) to prepare an extract of Namunjae Doenjang.
<< 실험예Experimental example 2> 2> 나문재Moonjae Na 된장의 면역억제 효과 확인 Checking the immunosuppressive effect of soybean paste
나문재 된장의 면역 억제 효과를 확인하기 위해, LPS 처리로 염증이 유발된 대식세포에 실시예 2에서 제조한 나문재 된장의 추출물을 처리한 후, 산화질소(nitric oxide, NO)의 생성량, 사이토카인의 생성량 및 염증 관련 인자의 발현을 측정하였다. In order to confirm the immunosuppressive effect of Namoonjae Doenjang, after treating the extract of Namoonjae Doenjang prepared in Example 2 on macrophages induced by inflammation by LPS treatment, the amount of nitric oxide (NO) produced, and cytokine Production and expression of inflammation-related factors were measured.
<2-1> 산화질소의 생성량<2-1> The amount of nitric oxide produced
구체적으로, 대식세포 RAW 264.7 세포를 10% FBS(fetal bovin serum, Gibco BRL, Rockγille, 미국), 1% 페니실린/스트렙토마이신(Gibco, Grand Island, 미국)가 함유된 DMEM(Gibco BRL, 미국) 배지에서 배양하고, 배양한 세포에 LPS(1 ㎍/㎖)를 처리하였다. LPS를 처리한 대식세포에 나문재 된장의 추출물(10, 50 및 100 ㎍/㎖)을 처리하고 CO2 인큐베이터(VS-9160C, Vision Scientific Co., Ltd., 한국)에서 37℃, 5% CO2 조건에서 배양하였다. 계대 배양 횟수는 5~10회 이내이며, 발효 전(0 Week) 또는 발효 후(24 Week)의 나문재 된장을 처리한 대식세포의 배양액을 취하여 산화질소의 생성량을 측정하였다. Specifically, macrophage RAW 264.7 cells were cultured in DMEM (Gibco BRL, USA) containing 10% FBS (fetal bovin serum, Gibco BRL, Rockγille, USA) and 1% penicillin/streptomycin (Gibco, Grand Island, USA). And the cultured cells were treated with LPS (1 µg/ml). LPS-treated macrophages were treated with extracts (10, 50 and 100 μg/ml) of Namunjae Doenjang, and then in a CO 2 incubator (VS-9160C, Vision Scientific Co., Ltd., Korea) at 37° C., 5% CO 2 Incubated under conditions. The number of subcultures was within 5 to 10 times, and the amount of nitric oxide produced was measured by taking the culture solution of macrophages treated with Namunjae Doenjang before (0 Week) or after (24 Week) fermentation.
그 결과, LPS 처리로 증가한 산화질소는 나문재 된장 추출물 처리로 인해 그 양이 유의적으로 감소하는 것으로 나타났다 (도 9). As a result, it was found that the amount of nitric oxide increased by the LPS treatment was significantly decreased due to the treatment with Namunjae doenjang extract (FIG. 9).
<2-2> 사이토카인 IL-6의 생성량<2-2> The amount of cytokine IL-6 produced
실험예 2-1과 동일한 방법으로 LPS 처리한 대식세포에 나문재 된장의 추출물을 처리한 후, 사이토카인 IL-6의 생성량을 측정하였다. After treating the LPS-treated macrophages with the extract of Namunjae Doenjang in the same manner as in Experimental Example 2-1, the amount of cytokine IL-6 produced was measured.
그 결과, LPS 처리로 증가한 IL-6는 나문재 된장 추출물을 처리하였을 때, 염도 12%의 나문재 된장 추출물에 대해서만 IL-6 감소 효과를 나타냄을 확인하였다 (도 10). As a result, it was confirmed that IL-6 increased by LPS treatment showed an IL-6 reduction effect only for Namunjae Doenjang extract having a salinity of 12% when treated with Namunjae Doenjang Extract (FIG. 10).
<2-3> COX-2 및 iNOS의 단백질 발현<2-3> COX-2 and iNOS protein expression
실험예 2-1과 동일한 방법으로 LPS 처리한 대식세포에 나문재 된장의 추출물을 처리한 후, COX-2 및 iNOS 단백질의 발현량을 측정하였다. In the same manner as in Experimental Example 2-1, the LPS-treated macrophages were treated with the extract of Namunjae Doenjang, and the expression levels of COX-2 and iNOS proteins were measured.
구체적으로, 100 mm 디쉬에 대식세포를 3.0×105 cells/mL의 농도로 분주하고, 실험예 2-1과 동일한 배양 조건에서 24시간 동안 배양하였다. 배양한 대식세포에 된장 추출물을 10, 50, 100 ㎍/㎖의 농도로 각각 처리하고 4시간 동안 배양한 다음, LPS(1 ㎍/㎖)를 처리하고 20시간 동안 더 배양하였다. 배양한 세포를 DPBS(Dulbecco’Phosphate Buffered Saline, Gibco, 미국)으로 3회 세척하고 원심분리하여 세포 펠렛을 수득하였다. 펠렛에 RIPA 버퍼(GenDEPOT, Barker, 미국)를 첨가하고 4℃, 13,000×g에서 10분간 원심 분리하여 상등액을 수득하였다. 수득한 상등액의 단백질을 정량하여 동량의 단백질을 10% SDS 폴리아크릴아마이드 겔에서 SDS-PAG를 수행하고 니트로셀롤로오스 멤브레인(NC membrane)으로 단백질을 전사하였다. 멤브레인을 블로킹 버퍼(0.5% BSA in TBS)에서 블로킹한 후 1:200 ~ 1:1,000으로 희석한 1차 항체(COX-2, iNOS, β-actin; Abcam, 영국)를 첨가하였다. 1차 항체와 밤새 배양한 후, 1:1,000으로 희석한 2차 항체(Anti-mouse IgG, Anti-rabbit IgG; Abcam, 영국)를 첨가하고 배양하였다. 1시간 후, ECL 용액(GE Healthcare, Bio-Sciences Co., Piscataway, 미국)을 첨가하여 단백질 발현 정도를 Amersham Imager 600 (GE Healthcare, 독일)를 사용하여 측정하고 정량하였다. Specifically, macrophages were dispensed in a 100 mm dish at a concentration of 3.0×10 5 cells/mL, and cultured for 24 hours under the same culture conditions as in Experimental Example 2-1. The cultured macrophages were treated with doenjang extract at concentrations of 10, 50, and 100 µg/ml, respectively, and incubated for 4 hours, followed by treatment with LPS (1 µg/ml) and further cultured for 20 hours. The cultured cells were washed three times with DPBS (Dulbecco' Phosphate Buffered Saline, Gibco, USA) and centrifuged to obtain a cell pellet. RIPA buffer (GenDEPOT, Barker, USA) was added to the pellet and centrifuged at 4° C. and 13,000×g for 10 minutes to obtain a supernatant. By quantifying the protein of the obtained supernatant, the same amount of the protein was subjected to SDS-PAG on a 10% SDS polyacrylamide gel, and the protein was transferred to a nitrocellulose membrane (NC membrane). After blocking the membrane in a blocking buffer (0.5% BSA in TBS), a primary antibody (COX-2, iNOS, β-actin; Abcam, UK) diluted 1:200 to 1:1,000 was added. After incubation with the primary antibody overnight, secondary antibodies (Anti-mouse IgG, Anti-rabbit IgG; Abcam, UK) diluted 1:1,000 were added and cultured. After 1 hour, ECL solution (GE Healthcare, Bio-Sciences Co., Piscataway, USA) was added to measure and quantify the protein expression level using Amersham Imager 600 (GE Healthcare, Germany).
그 결과, LPS 처리로 발현이 증가한 COX-2 및 iNOS는 나문재 된장 추출물을 처리하였을 때, 염도 12%의 나문재 된장 추출물에 대해서만 COX-2 및 iNOS 발현 감소 효과를 나타냄을 확인하였다 (도 11). As a result, it was confirmed that COX-2 and iNOS, whose expression was increased by LPS treatment, showed the effect of reducing COX-2 and iNOS expression only for Namunjae doenjang extract having a salinity of 12% when treated with Namunjae doenjang extract (FIG. 11).
<2-4> COX-2 및 iNOS의 mRNA 발현<2-4> mRNA expression of COX-2 and iNOS
실험예 2-1과 동일한 방법으로 LPS 처리한 대식세포에 나문재 된장의 추출물을 처리한 후, COX-2 및 iNOS의 mRNA 발현량을 측정하였다. After treating the LPS-treated macrophages with the extract of Namunjae Doenjang in the same manner as in Experimental Example 2-1, the mRNA expression levels of COX-2 and iNOS were measured.
구체적으로, 실시예 2-3과 동일한 방법으로 세포 펠렛을 수득하고, RNeasy mini kit(Qiagen, Valencia, 미국)를 사용하여 전체(total) RNA를 추출한 후, oligo dT primer와 amfiRicert cDNA Synthesis Platinum Master Mix(GenDEPOT, USA)를 사용하여 cDNA를 합성하였다. 합성한 cDNA를 주형으로 하여 iNOS 및 COX-2의 유전자를 RT-PCR로 증폭하였다. 이 때 하우스키핑(housekeeping) 유전자인 β-actin 유전자를 내부 대조군(internal control)으로 사용하였다. 각 PCR 산물들을 1% 아가로스 겔에 로딩하여 100V에서 30분 동안 전기영동하고 safe-pinky DNA 겔 염색 용액을 이용하여 염색한 후 Amersham Imager 600(GE Healthcare, 독일)를 사용하여 UV 하에서 겔을 관찰하였다. COX-2 및 iNOS의 mRNA의 발현량은 β-actin과 비교하여 정량화하였고, Image Lab statistical 소프트웨어(Bio-Rad, 미국)을 사용하여 계산하였다. PCR에 이용한 각 프라이머는 Cosmo Genetech(한국)에서 제작하였으며 하기 표 2에 나타내었다.Specifically, after obtaining a cell pellet in the same manner as in Example 2-3, extracting total RNA using an RNeasy mini kit (Qiagen, Valencia, USA), oligo dT primer and amfiRicert cDNA Synthesis Platinum Master Mix (GenDEPOT, USA) was used to synthesize cDNA. Using the synthesized cDNA as a template, the genes of iNOS and COX-2 were amplified by RT-PCR. At this time, a housekeeping gene, β-actin gene, was used as an internal control. Each PCR product was loaded on a 1% agarose gel, electrophoresed at 100V for 30 minutes, stained using a safe-pinky DNA gel staining solution, and observed the gel under UV using Amersham Imager 600 (GE Healthcare, Germany). I did. The expression levels of COX-2 and iNOS mRNA were quantified by comparing with β-actin, and calculated using Image Lab statistical software (Bio-Rad, USA). Each primer used for PCR was produced by Cosmo Genetech (Korea) and is shown in Table 2 below.
그 결과, LPS 처리로 발현이 증가한 iNOS는 염도 12%의 나문재 된장 추출물 처리로 유의적으로 감소하였다 (도 12). As a result, iNOS, whose expression was increased by LPS treatment, was significantly reduced by treatment with Namunjae Doenjang extract having a salinity of 12% (Fig. 12).
<< 실험예Experimental example 3> 3> 나문재Moonjae Na 된장의 맛 성분 확인 Checking the taste component of miso
국내에서 시판 중인 기업형 된장 제품 중 판매량이 가장 높은 제품을 대조군으로 사용하여, 실시예 1의 나문재 된장과 맛 성분의 변화를 비교하였다. 맛 성분은 나문재 된장의 발효 전후와 비교하였으며, 맛 분석기(Taste sensing system, TS-5000Z, Insent Co., 일본)를 이용하여 측정하였다. 맛 분석기는 인공 지질막의 구조의 센서를 통해 맛을 측정하는 장치로 음식을 입 안에서 씹는 동안 느끼는 5가지 initial taste (신맛(sourness), 짠맛(saltiness), 쓴맛(acidic bitterness), 떫은맛(astringency), 감칠맛(umami))와 음식을 삼킨 후 입안에 남아있는 5가지 after taste (깊은맛(richness), 쓴맛(aftertaste-B), 떫은맛(aftertaste-A))를 측정할 수 있도록 고안된 장비이다. The product with the highest sales volume among the corporate-type doenjang products marketed in Korea was used as a control, and the changes in taste components with Namunjae Doenjang of Example 1 were compared. Taste components were compared with before and after fermentation of Namunjae Doenjang, and measured using a taste analyzer (Taste sensing system, TS-5000Z, Insent Co., Japan). The taste analyzer is a device that measures taste through a sensor of the structure of an artificial lipid membrane.The five initial tastes you feel while chewing food in your mouth (sourness, saltiness, acidic bitterness, astringency, etc.) It is a device designed to measure umami) and 5 kinds of after tastes (richness, bitterness (aftertaste-B), astringency (aftertaste-A)) remaining in the mouth after swallowing.
그 결과, 나문재 된장은 시판 중인 된장과 짠맛 정도는 유사한 값을 보였으며, 된장의 깊은 맛(richness)은 시판 중인 된장과 비교하여 유의하게 높은 값을 나타내었다 (도 13 및 도 14). As a result, Namunjae doenjang showed similar saltiness as commercially available doenjang, and the richness of doenjang was significantly higher than that of commercially available doenjang (FIGS. 13 and 14).
<110> REPUBLIC OF KOREA (MANAGEMENT: RURAL DEVELOPMENT ADMINISTRATION) <120> Method of preparing fermented soybean paste having anti-inflammatory and immunosuppressive activity <130> 2018P-10-027 <160> 6 <170> KoPatentIn 3.0 <210> 1 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> beta-actin_forward primer <400> 1 gtgggccgcc ctaggcacca g 21 <210> 2 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> beta-actin_reverse primer <400> 2 ggaggaagag gatgcggcag t 21 <210> 3 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> COX-2_forward primer <400> 3 cactacatcc tgacccactt 20 <210> 4 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> COX-2_reverse primer <400> 4 atgctcctgc ttgagtatgt 20 <210> 5 <211> 27 <212> DNA <213> Artificial Sequence <220> <223> iNOS_forward primer <400> 5 aatggcaaca tcaggtcggc catcact 27 <210> 6 <211> 27 <212> DNA <213> Artificial Sequence <220> <223> iNOS_reverse primer <400> 6 gctgtgtgtc acagaagtct cgaactc 27 <110> REPUBLIC OF KOREA (MANAGEMENT: RURAL DEVELOPMENT ADMINISTRATION) <120> Method of preparing fermented soybean paste having anti-inflammatory and immunosuppressive activity <130> 2018P-10-027 <160> 6 <170> KoPatentIn 3.0 <210> 1 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> beta-actin_forward primer <400> 1 gtgggccgcc ctaggcacca g 21 <210> 2 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> beta-actin_reverse primer <400> 2 ggaggaagag gatgcggcag t 21 <210> 3 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> COX-2_forward primer <400> 3 cactacatcc tgacccactt 20 <210> 4 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> COX-2_reverse primer <400> 4 atgctcctgc ttgagtatgt 20 <210> 5 <211> 27 <212> DNA <213> Artificial Sequence <220> <223> iNOS_forward primer <400> 5 aatggcaaca tcaggtcggc catcact 27 <210> 6 <211> 27 <212> DNA <213> Artificial Sequence <220> <223> iNOS_reverse primer <400> 6 gctgtgtgtc acagaagtct cgaactc 27
Claims (9)
2) 상기 단계 1)의 혼합물을 20 내지 40℃의 온도 및 70 내지 95%의 습도 조건에서 2 내지 10개월 동안 발효하는 단계;를 포함하는 항염증 또는 면역 억제 활성을 갖는 염도 10 내지 15% 나문재 된장의 제조방법으로써,
상기 단계 1)은 메주 100 중량부에 대하여 나문재 분말 3 내지 15 중량부, 소금 5 내지 20 중량부, 물 5 내지 25 중량부로 혼합하는 것을 특징으로 하는, 나문재 된장의 제조방법.
1) mixing meju, namunjae powder, salt and water; And
2) fermenting the mixture of step 1) at a temperature of 20 to 40° C. and a humidity of 70 to 95% for 2 to 10 months; Namunjae having an anti-inflammatory or immune suppression activity of 10 to 15% As a method of making soybean paste,
The step 1) is characterized in that mixing with 3 to 15 parts by weight of Namunjae powder, 5 to 20 parts by weight of salt, and 5 to 25 parts by weight of water based on 100 parts by weight of meju.
상기 단계 1)은 메주 100 중량부에 대하여 나문재 분말 5 내지 15 중량부, 소금 8 내지 17 중량부, 물 5 내지 25 중량부로 혼합하는, 항염증 또는 면역 억제 활성을 갖는 나문재 된장의 제조방법.
The method of claim 1,
The step 1) is mixed with 5 to 15 parts by weight of Namunjae powder, 8 to 17 parts by weight of salt, and 5 to 25 parts by weight of water based on 100 parts by weight of meju.
Namoonjae doenjang having anti-inflammatory or immunosuppressive activity prepared by the method of claim 1.
상기 나문재 된장은 산화질소의 생성을 억제하는 활성을 갖는, 나문재 된장.
The method of claim 4,
The Namunjae Doenjang has the activity of inhibiting the production of nitric oxide, Namunjae Doenjang.
상기 나문재 된장은 COX-2 또는 iNOS의 발현을 억제하는, 나문재 된장.
The method of claim 4,
The Namunjae Doenjang inhibits the expression of COX-2 or iNOS, Namunjae Doenjang.
A composition for anti-inflammatory or immune suppression comprising the extract of Namunjae miso of claim 4 as an active ingredient.
상기 추출물은 산화질소의 생성을 억제하는 활성을 갖는, 항염증용 또는 면역 억제용 조성물.
The method of claim 7,
The extract has an activity to inhibit the production of nitric oxide, anti-inflammatory or immunosuppressive composition.
상기 추출물은 COX-2 또는 iNOS의 발현을 억제하는, 항염증용 또는 면역 억제용 조성물.The method of claim 7,
The extract is for inhibiting the expression of COX-2 or iNOS, anti-inflammatory or immunosuppressive composition.
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