KR20170114112A - Bacillus subtilis strain SY07 isolated from traditionally fermented soybean product and uses thereof - Google Patents
Bacillus subtilis strain SY07 isolated from traditionally fermented soybean product and uses thereof Download PDFInfo
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- KR20170114112A KR20170114112A KR1020160041160A KR20160041160A KR20170114112A KR 20170114112 A KR20170114112 A KR 20170114112A KR 1020160041160 A KR1020160041160 A KR 1020160041160A KR 20160041160 A KR20160041160 A KR 20160041160A KR 20170114112 A KR20170114112 A KR 20170114112A
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- bacillus subtilis
- blue
- miso
- soybean
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Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
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- A23L11/09—
-
- A23L11/20—
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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
-
- 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
- A23L33/127—Antibiotics
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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
- A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
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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
- A23V2200/302—Foods, ingredients or supplements having a functional effect on health having a modulating effect on age
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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
- A23V2200/328—Foods, ingredients or supplements having a functional effect on health having effect on glycaemic control and diabetes
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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
- A23V2250/02—Acid
- A23V2250/06—Amino acid
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- C12R1/125—
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/07—Bacillus
- C12R2001/125—Bacillus subtilis ; Hay bacillus; Grass bacillus
Abstract
본 발명은 전통장류로부터 분리한 바실러스 서브틸리스(Bacillus subtilis) SY07 균주(기탁번호: KACC92120P), 상기 균주 또는 이의 배양액을 스타터(starter) 균주로 이용하여 청된장을 제조하는 방법 및 상기 바실러스 서브틸리스 SY07 균주를 유효성분으로 함유하는 청된장 제조용 미생물 제제에 관한 것이다.The present invention relates to a method for producing Bacillus subtilis ( Bacillus subtilis) The present invention relates to a method for producing blue mustard by using the above strain or a culture thereof as a starter strain and a microorganism preparation for producing blue soybean paste containing the Bacillus subtilis SY07 strain as an active ingredient (hereinafter referred to as " subtilis strain SY07 ") (Accession No .: KACC92120P) .
Description
본 발명은 전통장류로부터 분리한 바실러스 서브틸리스(Bacillus subtilis) SY07 균주(기탁번호: KACC92120P) 및 이의 용도에 관한 것이다.The present invention relates to a strain of Bacillus subtilis SY07 (Accession No .: KACC92120P) isolated from a traditional soybean and its use.
청국장은 콩을 삶아 볏짚을 군데군데 꽂고 따뜻한 아랫목에 덮어 두면 하룻밤 사이에 표면이 회백색이 되고 끈적끈적한 실이 나게 띄워진다. 여기에 소금이나 마늘, 고춧가루 등을 섞어 절구통에서 찧어 단지에 다져 넣고 두면서 찌개의 재료로 사용하여 온 것으로 우리의 전래 간장, 된장, 고추장과는 달리 전통장류 중 유일하게 소금을 첨가하지 않고 고온에서 속성으로 발효시킨 식품이기 때문에 전통적으로 콩을 수확한 뒤 늦가을부터 초봄 사이에 각 가정에서 손쉽게 제조하여 왔다. 청국장은 된장보다 콩 단백질과 지방질 함량이 많고 소화 흡수율이 높으며, 칼슘과 비타민 A, B의 중요한 공급원, 청국장균의 정장효과, 섬유질의 변비예방효과, 발암물질과 콜레스테롤의 체외 배출효과, 점질물(mucin)의 알코올 흡수에 의한 해장효과, 사포닌의 혈관강화와 혈액순환 촉진과 젖산분해효과, 레시틴의 뇌 노화와 치매, 고혈압과 동맥경화의 예방 등의 효과가 있다는 것이 많이 알려져 왔다.Chungkukjang boiled soybeans, put the rice straw in place, and wrapped in a warm night, the surface turns grayish white and sticky sticky. It is mixed with salt, garlic, and red pepper powder, and it is used as the material of the stew by putting it in the mortar and putting it in the pot. Unlike our traditional soy sauce, miso, and kochujang, Because it is a fermented food, it has traditionally been harvested from soybeans, and has been easily manufactured at home from late autumn to early spring. Chungkukjang has a higher content of soybean protein and lipid than soybean paste, has higher digestion and absorption rate, and is an important source of calcium and vitamins A and B, a sweetening effect of Chungkukjang, prevention of constipation of fiber, effect of in vitro excretion of carcinogens and cholesterol, ) Has been known to have effects such as alcoholic absorption by alcohol absorption, blood vessel strengthening of saponin, promotion of blood circulation and decomposition of lactic acid, brain aging and dementia of lecithin, prevention of hypertension and arteriosclerosis.
된장은 우리나라의 대표적인 발효식품으로, 주재료로서 대두를 사용하는데 대두는 고단백의 식품소재이면서도 사포닌, 이소플라본, 이피리플라본 등의 생리활성물질을 포함하고 있어 항암작용, 골다공증 예방 등의 효과를 나타내어 대두를 비롯한 된장 등 대두 가공식품들은 건강식품으로 인식되고 있고, 대두 중의 이러한 유효성분들은 메주를 쑤어 발효되는 과정 중에 간장 또는 된장 등으로 상당량 옮겨가는 것으로 알려져 있다. 된장의 제조방법은 전통적인 재래식 방법과 현대의 개량식 방법으로 크게 대별된다. 전통적인 된장의 제조방법은, 지역에 따라 다소 차이가 있으나 기본적으로 원료인 대두를 삶아서 찧고 벽돌형 또는 구형으로 성형하여, 2~3일간 건조한 뒤 균열이 생기면 짚 등으로 매달아 적당한 온도와 습도를 유지하게 한다. 이렇게 하여 주위의 다양한 미생물들이 자연적으로 착생되어 번식하게 한 메주를 깨끗이 씻어 2~3월에 소금물에 담궈 3~6개월간 발효 숙성시킨 후 액상은 달여서 간장으로 하고, 나머지는 고형물로 된장을 만든다.Soybean is a representative fermented food in Korea and uses soybeans as its main material. Soybean is a food material of high protein, but it contains saponin, isoflavone, and ipriflavone as physiologically active substances and has effects such as anticancer action and prevention of osteoporosis. And soybean products including soybean paste are recognized as health foods. It is known that such effective ingredients in soybeans are transferred to soy sauce or soybean paste during fermentation of meju. The manufacturing methods of doenjang are largely classified into the conventional method and the modern method. The traditional method of making miso is somewhat different depending on the region, but basically, the raw soybean is boiled, molded into a brick or spherical shape, dried for 2 to 3 days, and if cracked, it is hung with straw to maintain proper temperature and humidity do. In this way, the various microorganisms around it are naturally mixed and washed, washed meju cleanly, and fermented for 3 ~ 6 months in February ~ March, and the liquid phase is dipped into soy sauce and the rest is made into solid soybean paste.
고추장은 약 200년의 역사를 가진 전통 조미식품으로 된장 및 간장과 더불어 빼놓을 수 없는 중요한 발효식품이다. 주로 가정에서 소규모 단위로 제조되어 왔으나, 최근에는 식생활 양식의 변화에 따라 개량된 방식의 공업적 규모로 생산이 가능해졌고, 이러한 경향은 편리성을 추구하는 소비자 욕구와 더불어 점차 확대되고 있다. 고추장은 통상적으로 쌀, 밀가루, 보리, 수수, 팥 등의 전분질 원료에 메줏가루, 엿기름가루, 고춧가루를 섞은 후 소금으로 간을 하여 담그는 재래식 고추장의 제조방법과, 짧은 시간 안에 대량생산할 수 있는 전분질인 밀가루에 코지 또는 효소제를 사용하여 제조하는 개량형 고추장의 제조방법이 알려져 있다. 고추장은 단백질, 지방, 비타민 A와 C 등의 영양분이 풍부하며, 탄수화물이 가수분해되어 생긴 단맛과 콩 단백에서 나온 아미노산의 감칠맛, 고추의 매운맛, 소금의 짠맛 등이 잘 조화되어, 주로 각종 찌개의 맛을 내고, 생채나 숙채, 조림, 구이 등의 조미료로 이용되고 있다.Kochujang is a traditional seasoned food with a history of about 200 years, and it is an important fermented food indispensable with miso and soy sauce. In recent years, it has become possible to produce on an industrial scale in an improved manner in accordance with changes in dietary patterns, and this trend is gradually expanding along with consumers' desire for convenience. Kochujang usually produces a mixture of raw materials such as rice, wheat flour, barley, sorghum and red bean paste, which is prepared by mixing fermented powder, malt powder, and red pepper powder with salt, A method of producing an improved type of kochujang prepared by using koji or an enzyme agent in flour is known. Kochujang is rich in nutrients such as protein, fat, vitamins A and C. The sweetness of hydrolysis of carbohydrates and the richness of amino acids, hot spices of pepper and saltiness of salt from soybean protein are well harmonized, It is used as a seasoning for raw vegetables, stew, stewed vegetables, and roasted vegetables.
한국등록특허 제1460566호에는 싸카로마이세스 속 AFY-1 균주를 포함하는 장류의 제조방법이 개시되어 있고, 한국등록특허 제0824005호에는 청국장 가미 된장 조성물의 제조방법이 개시되어 있으나, 본 발명의 전통장류로부터 분리한 바실러스 서브틸리스 SY07 균주 및 이의 용도와는 상이하다.Korean Patent No. 1460566 discloses a method for preparing a soybean paste containing a strain of Sakaeromycetes AFY-1, and Korean Patent No. 0824005 discloses a method for producing a soybean paste fermented soybean paste composition. However, It differs from the Bacillus subtilis SY07 strain isolated from the traditional soybean and its uses.
본 발명은 상기와 같은 요구에 의해 안출된 것으로서, 본 발명의 목적은 장류 특유의 불쾌한 냄새는 최소화하고, 감칠맛과 풍미가 우수하면서 품질이 증진된 청된장을 제조하는데 사용하기 위한 균주를 제공하는데 있다. 또한, 본 발명을 통해 분리된 바실러스 서브틸리스 SY07 균주를 이용하여 장류 제조조건 및 배합비 등의 제조방법을 최적화하여 기존의 청된장에 비해 풍미가 향상되어 기호도가 증진될 뿐만 아니라, 아미노태 질소, 총 폴리페놀 및 아미노산 함량이 높고, 항산화 및 항당뇨 활성이 증진된 청된장의 제조방법을 제공하는 데 있다.SUMMARY OF THE INVENTION The present invention has been made in view of the above-mentioned needs, and it is an object of the present invention to provide a strain for minimizing unpleasant odors inherent to the intestinal microflora and for producing a blue soybean paste with improved quality and flavor . In addition, by using the Bacillus subtilis SY07 strain isolated through the present invention, it is possible to optimize the manufacturing method such as the production conditions and the blending ratio of the Bacillus subtilis SYO7 strain to improve flavor and taste, Which is high in total polyphenol and amino acid content, and has enhanced antioxidant and anti-diabetic activity.
상기 과제를 해결하기 위해, 본 발명은 전통장류로부터 분리한 바실러스 서브틸리스(Bacillus subtilis) SY07 균주(기탁번호: KACC92120P)를 제공한다.In order to solve the above-mentioned problems, the present invention provides a method for producing Bacillus subtilis ( Bacillus subtilis) subtilis SY07 strain (Accession No .: KACC92120P).
또한, 본 발명은 상기 균주 또는 이의 배양액을 스타터(starter) 균주로 이용하여 청된장을 제조하는 방법을 제공한다.In addition, the present invention provides a method for producing blueberry miso using the strain or a culture thereof as a starter strain.
또한, 본 발명은 전통장류로부터 분리한 바실러스 서브틸리스(Bacillus subtilis) SY07 균주(기탁번호: KACC92120P)를 유효성분으로 함유하는 청된장 제조용 미생물 제제를 제공한다.In addition, the present invention provides a microorganism preparation for producing blue soybean paste containing, as an active ingredient, Bacillus subtilis SY07 strain (accession number: KACC92120P) isolated from a traditional soybean.
본 발명에서 분리한 바실러스 서브틸리스(Bacillus subtilis) SY07 균주(기탁번호: KACC92120P)를 이용하여 제조된 청된장은 장 특유의 구수한 맛과 감칠맛을 증진시키고, 장류 종류 및 부재료의 맛과 향이 잘 어우러져 기호도가 우수하다. 또한, 아미노태 질소, 총 폴리페놀 및 아미노산 함량이 높고, AGI(α-glucosidase inhibition) 및 항산화 활성이 우수하여 품질이 우수할 뿐만 아니라, 바실러스 세레우스 균주가 검출되지 않아, 소비자들이 더욱 선호하는 청된장을 제공할 수 있어, 관련 식품 산업에 매우 유용하게 이용될 수 있다.Isolated from the invention Bacillus subtilis (Bacillus Blue milks prepared by using subtilis strain SY07 (Accession No .: KACC92120P) improve the unique taste and flavor of the mung bean, and the taste and flavor of the mung bean varieties and the sub ingredient are well mixed. In addition, since the content of amino acid nitrogen, total polyphenol and amino acid is high, and α-glucosidase inhibition and antioxidative activity are excellent, the product is not only excellent in quality but also can not detect Bacillus cereus strain, Miso, and can be very useful for the related food industry.
본 발명의 목적을 달성하기 위하여, 본 발명은 전통장류로부터 분리한 바실러스 서브틸리스(Bacillus subtilis) SY07 균주(기탁번호: KACC92120P)를 제공한다.In order to accomplish the object of the present invention, the present invention provides a method for producing Bacillus subtilis subtilis SY07 strain (Accession No .: KACC92120P).
상기 바실러스 서브틸리스(Bacillus subtilis) SY07 균주(기탁번호: KACC92120P)는 전통 장류에서 분리하였으며, 항산화 및 항당뇨 활성이 우수하고, 아미노태 질소 및 유리 아미노산 함량이 높으며, 식중독균에 대한 항균활성이 우수한 청된장을 제조할 수 있다. 상기 바실러스 서브틸리스(Bacillus subtilis) SY07 균주는 국립농업과학원 농업유전자원센터에 2016년 2월 17일자로 기탁하였다(기탁번호: KACC92120P). 상기 기탁된 특정 균주를 이용하여 청된장을 제조하는 것이 다른 균주를 사용하여 청된장을 제조하는 것에 비해 아미노태 질소, 총 폴리페놀 및 아미노산 함량이 높고, 프로테아제 활성, AGI(α-glucosidase inhibition) 및 항산화 활성이 우수하며, 식중독균이 검출되지 않은 품질이 우수한 청된장을 제조할 수 있었다.The Bacillus subtilis Subtilis strain SY07 (Accession No .: KACC92120P) was isolated from a conventional soybean paste, and was able to produce a blue soybean paste having excellent antioxidant and antidiabetic activity, high amino acid nitrogen and free amino acid content and excellent antimicrobial activity against food poisoning bacteria . The Bacillus subtilis subtilis SY07 strain was deposited on February 17, 2016 (Accession No .: KACC92120P) at the National Institute of Agricultural Science and Technology. The production of blue soybean paste using the above-mentioned deposited strains is characterized in that amino acid nitrogen, total polyphenol and amino acid content are high, protease activity, α-glucosidase inhibition (AGI) and It was possible to produce blue bean paste with excellent antioxidant activity and high quality without detection of food poisoning bacteria.
본 발명의 식중독균으로는 바실러스 세레우스, 살모넬라, 황색포도상구균, 대장균 또는 리스테리아와 같은 병원성 식중독균일 수 있으나, 이에 제한되지 않는다. The food poisoning bacteria of the present invention may be, but are not limited to, pathogenic food poisoning bacteria such as Bacillus cereus, Salmonella, Staphylococcus aureus, Escherichia coli or Listeria.
"항균 (antimicrobial)"의 의미는 어떤 농도에서 미생물의 성장 또는 생존을 감소, 방지, 억제, 또는 제거하는 능력을 의미한다.By "antimicrobial" is meant the ability to reduce, prevent, inhibit, or eliminate microbial growth or survival at any concentration.
또한, 본 발명은 상기 균주 또는 이의 배양액을 스타터(starter) 균주로 이용하여 청된장을 제조하는 방법을 제공한다.In addition, the present invention provides a method for producing blueberry miso using the strain or a culture thereof as a starter strain.
본 발명에 있어서, 청된장 제조용 스타터(starter)란 청된장 제조를 위해 발효에 관여하는 미생물을 포함하는 제제 또는 조성물을 의미한다. 청된장 제조 시에 첨가함으로써 발효된 청된장에서 생장할 수 있는 미생물 또는 우점종으로 생장할 수 있는 미생물을 제공하기 위하여 사용된다. 상기 청된장 발효용 스타터를 사용하여 청된장을 제조하는 경우, 상기 청된장 발효용 스타터에 포함된 미생물에 의하여, 청된장의 품질을 일정하게 조절하거나, 특정한 목적, 일 예로 청된장에서 이취를 발생시키지 않거나, 감소시키는 목적 또는 청된장에서 구수한 맛이나 단맛을 강화시키기 위한 목적을 달성할 수 있다. 본 발명에서는 바실러스 서틸리스 SY07 균주 또는 이의 배양액을 스타터 균주로 이용함으로써, 아미노태 질소, 총 폴리페놀 및 아미노산 함량이 높고, 프로테아제 활성, AGI(α-glucosidase inhibition) 및 항산화 활성이 우수하며, 바실러스 세레우스 균주가 검출되지 않은 품질이 우수한 청된장을 제조할 수 있다.In the present invention, a starter for producing blue mustard refers to a preparation or composition containing microorganisms involved in fermentation for producing blue mustard. It is used to provide a microorganism capable of growing in fermented blueberry miso or a microorganism capable of growing as a dominant species by addition in the production of blue miso. In the case of producing the blue soybean paste using the starter for blue soybean fermentation, the quality of the blue soybean paste is controlled by the microorganisms contained in the blue soybean fermentation starter, or the green soybean paste is used for a certain purpose, For the purpose of enhancing the flavor or sweet taste of the sweetened miso or the purpose of reducing or not reducing the sweetness. In the present invention, by using Bacillus subtilis SY07 strain or its culture as a starter strain, it has high amino acid nitrogen, total polyphenol and amino acid content, excellent protease activity, AGI (alpha-glucosidase inhibition) and antioxidative activity, It is possible to produce a blue miso paste with excellent quality without the detection of the S. cerevisiae strain.
또한, 본 발명은 전통장류로부터 분리한 바실러스 서브틸리스(Bacillus subtilis) SY07 균주(기탁번호: KACC92120P)를 유효성분으로 함유하는 청된장 제조용 미생물 제제를 제공한다. 본 발명의 바실러스 서브틸리스 SY07 균주(기탁번호: KACC92120P)를 이용하여 청된장을 제조하는 것이 다른 균주를 사용하여 청된장을 제조하는 것에 비해 아미노태 질소, 총 폴리페놀 및 아미노산 함량이 높고, 프로테아제 활성, AGI(α-glucosidase inhibition) 및 항산화 활성이 우수하며, 식중독균이 검출되지 않은 품질이 우수한 청된장을 제조할 수 있다.In addition, the present invention provides a microorganism preparation for producing blue soybean paste containing, as an active ingredient, Bacillus subtilis SY07 strain (accession number: KACC92120P) isolated from a traditional soybean. The production of blue soybean paste using the strain Bacillus subtilis SY07 (accession number: KACC92120P) of the present invention is higher than that in producing blue soybean paste using different strains, and the content of amino acid nitrogen, total polyphenol and amino acid is higher than that of protease Can be produced with excellent activity, AGI (α-glucosidase inhibition) and antioxidant activity, and good quality without detection of food poisoning bacteria.
이하, 본 발명의 실시예를 들어 상세히 설명한다. 단, 하기 실시예는 본 발명을 예시하는 것일 뿐, 본 발명의 내용이 하기 실시예에 한정되는 것은 아니다.Hereinafter, embodiments of the present invention will be described in detail. However, the following examples are illustrative of the present invention, and the present invention is not limited to the following examples.
제조예Manufacturing example 1: One: 청된장Blue miso 제조 Produce
(a) 세척한 콩을 90~100℃에서 4~5시간 동안 증자하고 38~40℃에서 36시간 동안 발효한 후 소금을 세척한 콩 중량대비 5% 첨가하고 2~3시간 동안 교반하여 청국장을 제조하였다.(a) The washed soybeans were heated at 90 ~ 100 ℃ for 4 ~ 5 hours and fermented at 38 ~ 40 ℃ for 36 hours. 5% of the salt was added to the washed soybeans and stirred for 2 ~ 3 hours. .
(b) 90~100℃에서 4~5시간 동안 증자한 콩을 메주로 성형한 후 바실러스 서브틸리스(Bacillus subtilis) SY07 균주(기탁번호: KACC92120P)를 접종하여 15℃에서 90일 동안 발효시키고 소금물에 담궈 20~25℃에서 12개월 동안 숙성시킨 후 간장으로부터 된장을 분리하였다.(b) After molding the beans which were grown at 90 to 100 ° C for 4 to 5 hours in Meju, the beans were grown in Bacillus subtilis subtilis strain SY07 (accession number: KACC92120P), fermented at 15 DEG C for 90 days, immersed in brine, aged at 20-25 DEG C for 12 months, and then isolated from soy sauce.
(c) 증자한 찹쌀 25 kg에 고춧가루 25 kg, 천일염 11 kg, 메줏가루 7 kg, 엿기름 5 kg 및 물 27 kg을 첨가한 후 20~25℃에서 2~3개월 동안 숙성시켜 고추장을 제조하였다.(c) 25 kg of glutinous rice added, 25 kg of red pepper powder, 11 kg of sun salt, 7 kg of fermented soybean powder, 5 kg of maltose and 27 kg of water were added and aged at 20 ~ 25 ° C for 2 ~ 3 months to prepare kochujang.
(d) 상기 (a)단계의 제조한 청국장 68 kg, 상기 (b)단계의 분리한 된장 29 kg 및 상기 (c)단계의 제조한 고추장 1.5 kg과 다진 마늘 1.5 kg을 혼합하였다.(d) 68 kg of the prepared chungkukjang prepared in the step (a), 29 kg of the isolated miso obtained in the step (b), 1.5 kg of the prepared kochujang prepared in the step (c) and 1.5 kg of chopped garlic were mixed.
비교예Comparative Example 1: One: 청된장Blue miso 제조 Produce
상기 제조예 1의 방법으로 청된장을 제조하되, 바실러스 서브틸리스 (Bacillus subtilis) SY07 균주를 사용하지 않고 청된장을 제조하였다.Blue bean paste was prepared by the method of Preparation Example 1, except that Bacillus subtilis strain SY07 was not used.
비교예Comparative Example 2: 2: 청된장Blue miso 제조 Produce
(a) 세척한 콩을 90~100℃에서 7~8시간 동안 증자하고 35~36℃에서 3~4일 동안 발효시킨 후, 발효시킨 청국장 95 중량%에 소금 5 중량%를 혼합하고 분쇄하여 청국장을 제조하였다.(a) The washed soybeans were heated at 90 to 100 ° C. for 7 to 8 hours and fermented at 35 to 36 ° C. for 3 to 4 days. Then, 5 wt% of salt was mixed with 95 wt% of fermented chungkukjang, .
(b) 90~100℃에서 7~8시간 동안 증자한 콩을 메주로 성형한 후 20℃에서 60일 동안 발효시키고 소금물에 30일 동안 담그고 30℃에서 6개월 동안 숙성시킨 후 간장으로부터 된장을 분리하였다.(b) The beans were grown at 90 ~ 100 ℃ for 7 ~ 8 hours. The beans were fermented at 20 ℃ for 60 days, immersed in salt water for 30 days, aged at 30 ℃ for 6 months, Respectively.
(c) 증자한 찹쌀 25 kg에 고춧가루 25 kg, 천일염 11 kg, 메줏가루 7 kg, 엿기름 5 kg 및 물 27 kg을 첨가한 후 30℃에서 5~7개월 동안 숙성시켜 고추장을 제조하였다.(c) 25 kg of glutinous rice added, 25 kg of red pepper powder, 11 kg of sun salt, 7 kg of fermented soybean powder, 5 kg of maltose and 27 kg of water were added and then aged at 30 ° C for 5 to 7 months to prepare kochujang.
(d) 상기 (a)단계의 제조한 청국장 68 kg, 상기 (b)단계의 분리한 된장 29 kg 및 상기 (c)단계의 제조한 고추장 1.5 kg과 다진 마늘 1.5 kg을 혼합하였다.(d) 68 kg of the prepared chungkukjang prepared in the step (a), 29 kg of the isolated miso obtained in the step (b), 1.5 kg of the prepared kochujang prepared in the step (c) and 1.5 kg of chopped garlic were mixed.
(e) 상기 (d)단계의 혼합한 혼합물을 냉장고에 넣고 20~28시간 동안 숙성시켰다.(e) The mixed mixture of step (d) was put in a refrigerator and aged for 20 to 28 hours.
실험방법Experimental Method
1. 일반성분 분석1. General composition analysis
(1) pH 및 적정산도(1) pH and titratable acidity
시료 5 g을 취하여 자동적정장치 T50(Mettler Toledo GmbH, Switzerland)로 측정하였다.5 g of the sample was taken and measured with an automatic titrator T50 (Mettler Toledo GmbH, Switzerland).
(2) 아미노태 질소(2) Amino nitrogen
아미노태 질소 함량은 Formol 적정법에 준하여 T50(Mettler Toledo GmbH, Switzerland)를 이용해 분석하였다. 시료 2 g을 취하여 증류수 100 ㎖를 가하고 1시간 동안 교반한 후 0.1N NaOH 용액으로 pH 8.4까지 적정하였다. 여기에 중성 포르말린 용액 20 ㎖를 가하고 다시 0.1N NaOH 용액으로 pH 8.4가 되도록 적정하고, 별도로 증류수에 대한 바탕시험을 실시하여 아미노태 질소 함량을 구하였다.Amino nitrogen content was analyzed by T50 (Mettler Toledo GmbH, Switzerland) according to the Formol titration method. 2 g of the sample was taken and 100 ml of distilled water was added thereto. The mixture was stirred for 1 hour and titrated to pH 8.4 with 0.1 N NaOH solution. To this was added 20 ml of neutral formalin solution, titrated to pH 8.4 with 0.1 N NaOH solution, and basal test was conducted for distilled water separately to determine the amino nitrogen content.
2. 항당뇨 분석2. Anti-diabetic analysis
항당뇨 활성은 '식품의약품안전청 건강기능식품 기능성 평가 가이드 높은 혈당 감소에 도움편'의 기능성 시험 방법에 나와 있는 바이오마커 중 α-글루코시다아제 활성억제 측정법과 Sigma의 α-글루코시다아제(EC 3.2.1.20) 분석법을 이용하여 측정하였다. 67 mM 인산칼륨 완충액(pH 6.8) 500 ㎕를 넣고, 반응구에는 시료액 20 ㎕를 첨가하고, 대조구에는 증류수 20 ㎕를 첨가하였다. 그리고 3 mM GSH(Glutathione Reduced Solution)를 20 ㎕ 첨가하고 α-글루코시다아제 효소 용액(0.15~0.3 unit/ml)을 20 ㎕ 첨가한 후 10 mM p-니트로페닐-α-D-글루코시드 용액(PNP-Gluc) 50 ㎕를 첨가하여 37℃의 항온 수조에서 20분간 반응시켰다. 반응액 200 ㎕에 100 mM 탄산나트륨 용액을 800 ㎕를 첨가하여 400 nm에서 분광광도계를 이용하여 흡광도를 측정하였다.The anti-diabetic activity was measured by the method described in the Functional Testing Methods for High Blood Glucose Reduction Guide of the Korean Food and Drug Administration (KFDA) Functional Food Functional Evaluation Guide and the α-glucosidase activity inhibition assay and Sigma's α-glucosidase (EC 3.2 .1.20) analysis method. 500 μl of 67 mM potassium phosphate buffer solution (pH 6.8) was added, and 20 μl of the sample solution was added to the reaction mixture, and 20 μl of distilled water was added to the control. Then, 20 μl of 3 mM GSH (Glutathione Reduced Solution) was added and 20 μl of α-glucosidase enzyme solution (0.15-0.3 unit / ml) was added thereto. Then, 10 mM p-nitrophenyl-α-D-glucoside solution PNP-Gluc) was added and reacted in a constant temperature water bath at 37 ° C for 20 minutes. 800 [mu] l of 100 mM sodium carbonate solution was added to 200 [mu] l of the reaction solution, and the absorbance was measured at 400 nm using a spectrophotometer.
Unit/ml 효소 = {(실험구 흡광도 값 - 블랭크 흡광도 값)(10)(6.1)(d.f)} / {(18.3)(20)(2)(0.2)}Unit / ml enzyme = {(absorbance value of the experiment - blank absorbance value) (10) (6.1) (d.f)} / {(18.3) (20)
6.1 = 반응 혼합물의 부피(㎖)6.1 = Volume of reaction mixture (ml)
d.f = 희석인자d.f = dilution factor
18.3 = 400 nm에서 p-니트로페놀의 흡광계수(mM)18.3 = extinction coefficient of p-nitrophenol in mM (mM)
20 = 분석 시간(분)20 = Analysis time (minutes)
10 = 비색 측정 부피(㎖)10 = colorimetric measurement volume (ml)
2 = 비색 측정에 사용된 혼합물의 부피(㎖)2 = volume (ml) of mixture used for colorimetric measurement
3. 항산화 분석3. Antioxidant analysis
항산화 활성은 '식품의약품안전청 건강기능식품 기능성 평가 가이드 항산화에 도움편'의 기능성 시험 방법에 나와있는 항산화 기능성 확인을 위한 항산화 시스템의 총 항산화능 바이오 마커 중 DPPH 분석 방법을 이용하여 분석하였다. 각 시료용액 20 ㎕에 0.15 mM로 에탄올 희석한 DPPH(2,2-diphenyl-2-picrylhydrazyl) 용액 180 ㎕를 넣고 교반한 후 30분간 방치한 다음 마이크로플레이트 리더를 이용해 517 nm에서 흡광도를 측정하였다. 전자공여능은 시료용액의 첨가군과 무첨가군의 흡광도 차이를 백분율로 나타내었다. 대조군으로 L-아스코르브산을 사용하였다.The antioxidant activity was analyzed by the DPPH assay method among the total antioxidant biomarkers of the antioxidant system for confirming the antioxidant functionalities described in the functional test method of 'Antioxidant Helps Guide to Functional Food Functional Evaluation Guide of the Food and Drug Administration'. To 20 μl of each sample solution, 180 μl of 0.15 mM ethanol-diluted DPPH (2,2-diphenyl-2-picrylhydrazyl) solution was added, stirred, left for 30 minutes, and absorbance was measured at 517 nm using a microplate reader. The electron donating ability was expressed as a percentage of absorbance difference between the addition group and the no addition group of the sample solution. L-ascorbic acid was used as a control.
항산화 활성(%) = {1 - (시료 흡광도/대조군 흡광도)}×100Antioxidant activity (%) = {1- (sample absorbance / control absorbance)} × 100
4. 총 폴리페놀 함량 분석4. Analysis of total polyphenol content
총 폴리페놀 함량은 Folin-Ciocalteu법, 즉 Folin-Ciocalteu 시약이 시료의 폴리페놀성 화합물에 의해 환원된 결과, 몰리브텐 청색으로 발색하는 것을 원리로 측정하였다. 추출 시료 0.1 ㎖에 2N Folin-Ciocalteu 시약(Sigma, st. Louis, MD, USA) 0.5 ㎖와 증류수 8.4 ㎖를 넣은 뒤, 20% Na2CO3 1 ㎖를 넣고 한 시간 방치한 다음 725 nm에서 흡광도를 측정하였으며, 페놀화합물 함량은 표준물질인 갈산을 이용한 표준곡선으로 양을 환산하였다. 갈산을 이용하여 작성한 표준곡선 (y = 11.3x + 0.0403, R2 = 0.9999)으로부터 함량을 구하였다.The total polyphenol content was determined by the principle that the Folin-Ciocalteu method, that is, the Folin-Ciocalteu reagent, was reduced to a molybdenum blue color as a result of reduction by the polyphenolic compound of the sample. 0.5 ml of 2N Folin-Ciocalteu reagent (Sigma, St. Louis, MD, USA) and 8.4 ml of distilled water were added to 0.1 ml of the extracted sample, and 20% Na 2 CO 3 One And the absorbance was measured at 725 nm. The phenol compound content was converted into a standard curve using a standard curve of gallic acid. The content was determined from a standard curve (y = 11.3x + 0.0403, R2 = 0.9999) prepared using gallic acid.
5. 유리 아미노산5. Free amino acids
유리 아미노산은 시료 2 g을 취하여 3차 증류수 30 mL를 넣고 교반한 후 50 mL로 정용한 후 초음파를 이용하여 20분간 추출하고 원심분리하여 상등액 2 mL에 5% TCA 2 mL를 넣고 원심분리하였다. 이 후 상등액을 취하여 0.02 N-HCl로 희석한 후 0.2 ㎛ 시린지 필터(syringe filter)에 통과시킨 후 아미노산 분석기(Hitachi L-8900, Japan), UV/Vis 검출기(440 nm~570 nm), 컬럼(Hitachi 4.6×60 mm speration, Hitachi 4.6×40 mm Ammonia filtering)을 사용하여 분석하였다. 시료량은 20 ㎕, 이동상 Buffer set(PH-SET KANTO, Japan), 분석 온도는 50℃, buffer 유속은 0.40 mL/min, 닌하이드린(ninhydrin) 유속은 0.35 mL/min으로 분석하였다.For the free amino acid, take 2 g of sample, add 30 mL of tertiary distilled water, stir with 50 mL, extract using ultrasonic for 20 minutes, centrifuge, add 2 mL of 5% TCA to the supernatant and centrifuge. The supernatant was diluted with 0.02 N HCl and passed through a 0.2 μm syringe filter. The supernatant was passed through an amino acid analyzer (Hitachi L-8900, Japan), a UV / Vis detector (440 nm to 570 nm) Hitachi 4.6 x 60 mm speration, Hitachi 4.6 x 40 mm Ammonia filtering). The sample volume was 20 ㎕, mobile phase buffer set (PH-SET KANTO, Japan), analysis temperature was 50 ℃, buffer flow rate was 0.40 mL / min and ninhydrin flow rate was 0.35 mL / min.
6. 식중독균 실험6. Food poisoning bacteria experiment
- 식품위생법 상 식품공전에 준하여 2회 반복하여 실시하였다.- The food hygiene law was repeated twice in accordance with the Food Code.
(1) 바실러스 세레우스(1) Bacillus cereus
검체 25 g 또는 25 ㎖를 취하여 225 ㎖의 희석액을 가하여 균질화한 검액을 MYP한천배지(배지 46)에 접종하여 30℃에서 24시간 배양하였다. 배양 후 혼탁한 환을 갖는 분홍색 집락을 선별하였다. 이때 명확하지 않을 경우 24시간 더 배양하여 관찰하였다.25 g of the sample or 25 ml of the sample was diluted with 225 ml of the homogenized solution and inoculated on a MYP agar medium (medium 46) and cultured at 30 ° C for 24 hours. After culture, pink colonies with turbid rings were selected. If not clear, the cells were cultured for 24 hours.
(2) 대장균(2) Escherichia coli
시험용액 1 ㎖를 대장균 건조필름배지(배지 55) 35~37℃에서 24±2시간 배양하였다. 대장균 건조필름 배지에서는 푸른 집락 중 주위에 기포를 형성한 집락수를 계산하였다.1 ml of the test solution was cultured at 35 to 37 ° C for 24 ± 2 hours in an E. coli dried film medium (medium 55). In E. coli dried film media, the number of colonies forming bubbles around blue colonies was calculated.
(3) 황색포도상구균(3) Staphylococcus aureus
가) 증균배양A) Enrichment culture
검체 25 g 또는 25 ㎖를 취하여 225 ㎖의 10% NaCl을 첨가한 TSB 배지(배지 23)에 가한 후 35~37℃에서 18~24시간 증균배양하였다.25 g of the sample or 25 ml of the sample was added to TSB medium (medium 23) supplemented with 225 ml of 10% NaCl and incubated at 35 to 37 ° C for 18 to 24 hours.
나) 분리배양B) Isolation culture
증균 배양액을 Baird-Parker 한천배지(배지 63)에 접종하여 35~37℃에서 18~24시간 배양하였다. 배양 결과 Baird-Parker 한천배지에서 투명한 띠로 둘러싸인 광택이 있는 검정색 집락은 확인시험을 실시하였다.The enrichment culture was inoculated on a Baird-Parker agar medium (medium 63) and cultured at 35-37 ° C for 18-24 hours. Cultivation results showed that a shiny black colonies surrounded by a transparent band on a Baird-Parker agar medium were subjected to an identification test.
(4) 살모넬라(4) Salmonella
가) 증균배양A) Enrichment culture
검체 25 g 또는 25 ㎖를 취하여 225의 펩톤수(배지 56)에 가한 후 35~37℃에서 24±2시간 증균 배양하였다. 배양액 0.1 ㎖를 취하여 10 ㎖의 Rappaport-Vassiliadis 배지(배지 57)에 접종하여 42±1℃에서 24±2시간 배양하였다.25 g of the sample or 25 ml of the sample was added to 225 peptone (medium 56) and incubated at 35 to 37 ° C for 24 ± 2 hours. 0.1 ml of the culture was inoculated into 10 ml of Rappaport-Vassiliadis medium (medium 57) and cultured at 42 占 폚 for 24 占 2 hours.
나) 분리배양B) Isolation culture
증균 배양액을 XLD 한천배지(배지 58)에 접종하여 35~37℃에서 24±2시간 배양한 후 전형적인 집락은 확인시험을 실시하였다.The cultures were inoculated on XLD agar medium (medium 58) and cultured at 35-37 ° C for 24 ± 2 hours. A typical colonization was confirmed.
(5) 리스테리아(5) Listeria
가) 증균배양A) Enrichment culture
검체 25 g 또는 25 ㎖를 취하여 225 ㎖의 리스테리아 증균배지를 가한 후 30℃에서 48시간 배양하였다.25 g of the sample or 25 ml of the sample was added to 225 ml of the Listeria bloom and cultured at 30 ° C for 48 hours.
나) 분리배양 B) Isolation culture
증균배양액을 멸균된 면봉을 이용하여 Oxford 한천배지(배지 38)에 접종하여 30℃에서 24~48시간 배양하였다. 진한 검은색 집락은 확인시험을 실시하였다.The enrichment culture was inoculated into Oxford agar medium (medium 38) using a sterilized cotton swab and cultured at 30 ° C for 24 to 48 hours. A dark black colony was identified.
7. 관능검사7. Sensory evaluation
발효미생물산업진흥원 직원 10 여명을 선정하여 실험목적과 관능적 품질요소를 인식하도록 설명한 후 9점 척도법을 이용하였다. 시료검사 전 입안을 헹구도록 하였으며, 관능평가는 구수한 맛, 감칠맛, 전반적 기호도로 특성이 강할수록 높은 점수로 평가하였다.We selected about 10 employees of Fermented Microorganism Industry Promotion Institute and explained 9 points scale method to explain experiment purpose and sensory quality factor. The mouth was rinsed before the test. The sensory evaluation was evaluated with a high score with strong taste, richness, overall acceptability.
실시예Example 1: 균주 선정 1: Selection of strain
된장으로부터 분리한 바실러스 속 균주 21종을 각각 이용하여 청된장을 제조한 후, 아미노태 질소 함량, 효소(protease) 활성 및 향미를 비교하였다. 그 결과, 아미노태 질소 함량 및 효소 활성이 높고, 향미가 우수한 청된장 제조에 적합한 7번 균주를 최종 선발하였다. 선발된 7번 균주의 동정을 위하여 16S rRNA 유전자 분석을 실시하였다. 균주의 동정을 위하여 균체를 Nutrient broth (NB,DifcoTM)에 접종하여 30℃에서 24시간 동안 배양한 후 원심 분리하여 균체를 회수한 뒤 ZR Fungal/Bacterial DNA Miniprepkit (Zymo Research Corp.)를 이용하여 DNA를 추출한 후 PCR을 통하여 얻은 16S rRNA 유전자로 BLAST 검색을 수행한 결과, 바실러스 서브틸리스(Bacillus subtilis) 균주로 동정되었고, 바실러스 서브틸리스 SY07 균주로 명명하였으며, 국립농업과학원 농업유전자원센터에 2016년 2월 17일자로 기탁하여(기탁번호: KACC92120P), 본 발명의 청된장 제조에 사용하여 품질 특성을 비교하였다.Amino nitrogen content, protease activity and flavor of soybean paste were prepared by using 21 kinds of Bacillus sp. Isolates isolated from soybean paste. As a result, the 7th strain suitable for producing blue soybean paste with high amino nitrogen content and high enzyme activity and high flavor was finally selected. 16S rRNA gene analysis was performed to identify the selected strain No. 7. For identification of the strains, the cells were inoculated into Nutrient broth (NB, Difco ™ ), cultured at 30 ° C. for 24 hours, centrifuged to recover the cells, and then treated with ZR Fungal / Bacterial DNA Miniprepkit (Zymo Research Corp.) As a result of BLAST search with 16S rRNA gene obtained by PCR after extracting DNA, Bacillus subtilis Subtilis strain SY07 was deposited on Feb. 17, 2016 (Accession No .: KACC92120P) to the Institute of Agricultural Genetic Resources, National Institute of Agricultural Science and Technology, Quality characteristics were compared.
실시예Example 2: 2: 청된장Blue miso 일반성분 분석결과 General composition analysis result
제조예 1의 방법으로 제조된 청된장과 제조예 1의 방법으로 제조하되 바실러스 서브틸리스(Bacillus subtilis) SY07 균주를 사용하지 않고 제조된 청된장(비교예 1)의 pH, 적정 산도 및 아미노태 질소 함량을 비교한 결과는 하기 표 2와 같다. 그 결과, pH와 적정 산도는 큰 차이를 나타내지 않았으나, 아미노태 질소 함량의 경우 제조예 1의 청된장이 비교예 1에 비해 3배 이상 높은 아미노태 질소 함량을 나타내었다.Blue bean paste prepared by the method of Preparation Example 1 and the method of Preparation Example 1 were used, but Bacillus subtilis pH, titratable acidity, and amino nitrogen content of the blue soybean paste (Comparative Example 1) prepared without using the subtilis SY07 strain are shown in Table 2 below. As a result, the pH and the titratable acidity did not show a large difference, but the amino acid nitrogen content of the fermented soybean paste of Preparation Example 1 was three times higher than that of Comparative Example 1.
실시예Example 3: 3: 청된장의Blue-green miso 항산화 활성 Antioxidant activity
제조예 1의 방법으로 제조된 청된장과 제조예 1의 방법으로 제조하되 바실러스 서브틸리스(Bacillus subtilis) SY07 균주를 사용하지 않고 제조된 청된장(비교예 1)의 폴리페놀 함량과 DPPH 라디칼 소거능을 비교한 결과는 하기 표 3과 같다. 그 결과, 비교예 1의 청된장에 비해 제조예 1의 청된장이 총 폴리페놀 함량이 높고 항산화 활성이 증진됨을 확인할 수 있었다.Blue bean paste prepared by the method of Preparation Example 1 and the method of Preparation Example 1 were used, but Bacillus subtilis The results of comparing the polyphenol content and DPPH radical scavenging ability of the blue doenjang (Comparative Example 1) prepared without using the subtilis SY07 strain are shown in Table 3 below. As a result, it was confirmed that the blue soybean paste of Preparation Example 1 had higher total polyphenol content and higher antioxidative activity than the blue soybean paste of Comparative Example 1.
실시예Example 4: 4: 청된장의Blue-green miso 항당뇨Anti-diabetic 활성 activation
제조예 1의 방법으로 제조된 청된장과 제조예 1의 방법으로 제조하되 바실러스 서브틸리스(Bacillus subtilis) SY07 균주를 사용하지 않고 제조된 청된장(비교예 1)의 항당뇨 활성을 비교한 결과는 하기 표 4와 같다. 그 결과, 비교예 1의 청된장은 항당뇨 활성을 나타내지 않았으나, 제조예 1의 청된장은 18.68%의 α-글루코시다아제 저해 활성을 나타내었다.Blue bean paste prepared by the method of Preparation Example 1 and the method of Preparation Example 1 were used, but Bacillus subtilis The results of comparison of anti-diabetic activity of blue-green onion (Comparative Example 1) prepared without using the subtilis SY07 strain are shown in Table 4 below. As a result, the blue soybean paste of Comparative Example 1 did not show antidiabetic activity, but the blue soybean paste of Preparation Example 1 showed an inhibitory activity of? -Glucosidase of 18.68%.
실시예Example 5: 5: 청된장의Blue-green miso 유리 아미노산 함량 Free amino acid content
제조예 1의 방법으로 제조된 청된장과 제조예 1의 방법으로 제조하되 바실러스 서브틸리스(Bacillus subtilis) SY07 균주를 사용하지 않고 제조된 청된장(비교예 1)의 유리 아미노산 함량을 비교한 결과는 하기 표 5와 같다.Blue bean paste prepared by the method of Preparation Example 1 and the method of Preparation Example 1 were used, but Bacillus subtilis Subtilis ) SYO7 strain (Comparative Example 1) was compared with the free amino acid content of the prepared blue soybean paste (Comparative Example 1).
그 결과, 비교예 1의 청된장에 비해 제조예 1의 청된장은 포스포에탄올아민(PEA), 요소, 글루탐산, 라이신, 프롤린, 글리신, 발린, 이소류신, 류신, 티로신, 시트룰린, 시스테인, 페닐알라닌, 오르니틴, 히스티딘 및 아르기닌 등과 같은 유리 아미노산 함량이 증진되었고, 총 유리 아미노산 함량에 있어서도 제조예 1은 2,122.81 ㎎/100 g을 나타낸 반면, 비교예 1은 1,296.09 ㎎/100 g으로 총 유리 아미노산 함량도 비교예 1에 비해 증진됨을 확인할 수 있었다.As a result, the blue hazelnut paste of Preparation Example 1, compared to the blue hazelnut paste of Comparative Example 1, was found to contain a mixture of phosphoethanolamine (PEA), urea, glutamic acid, lysine, proline, glycine, valine, isoleucine, leucine, tyrosine, citrulline, cysteine, The content of free amino acids such as ornithine, histidine and arginine was increased, and the total free amino acid content of Preparation Example 1 was 2,122.81 mg / 100 g, while that of Comparative Example 1 was 1,296.09 mg / 100 g. Compared with Example 1.
실시예Example 6: 6: 청된장의Blue-green miso 병원성 식중독균 검사 Pathogenic food poisoning test
제조예 1의 방법으로 제조된 청된장과 제조예 1의 방법으로 제조하되 바실러스 서브틸리스(Bacillus subtilis) SY07 균주를 사용하지 않고 제조된 청된장(비교예 1)의 병원성 식중독균 검사를 실시한 결과, 제조예 1은 바실러스 세레우스, 살모넬라, 황색포도상구균, 대장균 및 리스테리아와 같은 병원성 식중독균이 검출되지 않았으나, 비교예 1은 바실러스 세레우스 균주가 검출됨을 확인할 수 있었다(표 6).Blue bean paste prepared by the method of Preparation Example 1 and the method of Preparation Example 1 were used, but Bacillus subtilis Subtilis ) SY07 strains (Comparative Example 1), no pathogenic food poisoning bacteria such as Bacillus cereus, Salmonella, Staphylococcus aureus, Escherichia coli and Listeria were detected in Production Example 1 , And in Comparative Example 1, Bacillus cereus strain was detected (Table 6).
ND: 검출되지 않음ND: Not detected
실시예Example 7: 7: 청된장의Blue-green miso 관능검사 Sensory test
제조예 1과 비교예 1 및 2의 방법으로 제조된 청된장을 가지고 구수한 맛, 감칠맛 및 전체적인 기호도를 평가한 결과, 제조예 1의 청된장이 비교예들의 청된장에 비해 구수한 맛 및 감칠맛이 높아 더욱 선호한다는 것을 확인할 수 있었다(표 7).As a result of evaluating the taste, flavor and overall acceptability of the blue miso prepared by the method of Preparation Example 1 and Comparative Examples 1 and 2, it was found that the blue miso of Preparation Example 1 had a higher taste and richness than the blue miso of Comparative Example (Table 7).
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| KR102240109B1 (en) * | 2019-11-28 | 2021-04-14 | 재단법인 전남바이오산업진흥원 | A Novel Bacillus subtilis TSD and methods for preparing Doenjang using the same |
| KR20210076550A (en) * | 2019-12-16 | 2021-06-24 | 재단법인 전남바이오산업진흥원 | Methods for preparing oat doenjang using a novel Bacillus subtilis TSD |
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| KR102240109B1 (en) * | 2019-11-28 | 2021-04-14 | 재단법인 전남바이오산업진흥원 | A Novel Bacillus subtilis TSD and methods for preparing Doenjang using the same |
| KR20210076550A (en) * | 2019-12-16 | 2021-06-24 | 재단법인 전남바이오산업진흥원 | Methods for preparing oat doenjang using a novel Bacillus subtilis TSD |
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