KR102092528B1 - Method for producing soy sauce using Bacillus subtilis SRCM103716 and Aspergillus oryzae strain - Google Patents
Method for producing soy sauce using Bacillus subtilis SRCM103716 and Aspergillus oryzae strain Download PDFInfo
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- KR102092528B1 KR102092528B1 KR1020190116335A KR20190116335A KR102092528B1 KR 102092528 B1 KR102092528 B1 KR 102092528B1 KR 1020190116335 A KR1020190116335 A KR 1020190116335A KR 20190116335 A KR20190116335 A KR 20190116335A KR 102092528 B1 KR102092528 B1 KR 102092528B1
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- sesame
- bacillus subtilis
- fermented
- sesame seeds
- soy sauce
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Images
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
- A23L27/00—Spices; Flavouring agents or condiments; Artificial sweetening agents; Table salts; Dietetic salt substitutes; Preparation or treatment thereof
- A23L27/50—Soya sauce
-
- 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
- A23L25/00—Food consisting mainly of nutmeat or seeds; Preparation or treatment thereof
- A23L25/30—Mashed or comminuted products, e.g. pulp, pastes, meal, powders; Products made therefrom, e.g. blocks, flakes, snacks; Liquid or semi-liquid products
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
- A23L29/065—Microorganisms
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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
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
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- A23Y2220/00—
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- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Nutrition Science (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Microbiology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
Description
본 발명은 (1) 분쇄한 참깨박에 물을 가수하여 증자한 후 냉각한 참깨박에 바실러스 서브틸리스(Bacillus subtilis) SRCM103716 균주를 접종한 후 발효하여 총 폴리페놀 및 총 플라보노이드 함량과 항산화 활성 및 β-글루코시다아제(β-glucosidase) 활성이 증진된 참깨박 고초균 발효물을 제조하는 단계; (2) 분쇄한 참깨박에 물을 가수하여 증자한 후 냉각한 참깨박에 아스퍼질러스 오리재(Aspergillus oryzae) 균주를 접종한 후 발효하여 참깨박 발효물을 제조하는 단계; (3) 상기 (1)단계의 제조한 참깨박 고초균 발효물과 상기 (2)단계의 제조한 참깨박 발효물을 혼합하여 리그난 함량 및 항산화 활성이 증진된 참깨박 혼합 발효물을 제조하는 단계; 및 (4) 상기 (3)단계의 제조한 참깨박 혼합 발효물에 염수를 첨가하여 숙성시킨 후 여과하는 단계를 포함하여 제조하는 것을 특징을 하는 참깨박 발효 간장의 제조방법 및 상기 방법으로 제조된 참깨박 발효 간장에 관한 것이다.The present invention (1) by inoculating the strain of Bacillus subtilis SRCM103716 after fermentation by increasing water by adding water to the crushed sesame seeds and fermenting them to inoculate the total polyphenol and total flavonoid content and antioxidant activity and preparing β-glucosidase (β-glucosidase) activity-enhanced sesame seed archaea fermentation; (2) After adding water to the crushed sesame seeds, increase the amount, and then cool the cooled sesame seeds with Aspergillus oryzae ) producing a sesame leaf fermentation product by inoculating the strain and then fermenting it; (3) preparing a mixed fermented sesame seeds with enhanced lignan content and antioxidant activity by mixing the fermented sesame seeds produced in step (1) and the fermented sesame seeds prepared in step (2); And (4) adding a brine to the sesame-bak mixed fermentation product prepared in step (3), followed by aging and filtering, thereby producing a method of producing sesame-bak fermented soy sauce and prepared by the above method. It is about fermented soy sauce in sesame seeds.
간장은 우리나라 사람의 식생활에서 없어서는 안 되는 중요한 식료품이며, 콩을 주원료로 하는 장류 중 하나로 발효, 숙성 과정을 거쳐 만들어진 전통 발효식품으로서, 식물성 단백질이 높은 소금농도에서 미생물의 작용으로 분해하여 구수한 향미를 나타내게 하였기 때문에 조미료가 되는 동시에 저장성이 우수한 가공식품이다.Soy sauce is an important food that is indispensable in the diet of Korean people, and is a traditional fermented food made through fermentation and aging process as one of the main ingredients of soybeans. It is a processed food that is excellent in storage at the same time as a seasoning.
간장은 매우 특성이 있는 식품으로 여기에 관여되는 발효미생물의 종류 및 제조기술의 차이에 따라 달라지므로 맛과 향기에 특성이 생긴다. 이러한 장류는 각 가정에서 제조하여 이용해 왔으나 경제성장 및 식생활 양식의 변화로 가정에서 제조하는 것보다 시장에서 구매하는 경향으로 바뀜에 따라 장류의 소비량은 물론 제조방법도 크게 발전하고 있다.Soy sauce is a very characteristic food, and it depends on the type of fermentation microorganism involved in it and the difference in manufacturing technology. Although these sauces have been manufactured and used in each household, the consumption and consumption of the sauces have been greatly improved as the trend of purchasing them in the market rather than those manufactured at home has changed due to economic growth and changes in dietary habits.
세사몰 및 세사미놀은 저분자 식물성 화합물인 리그난의 일종으로서 항산화 활성 및 콜레스테롤혈증, 죽상동맥경화증 및 고혈당을 저해한다고 알려져 있다. 또한 여성 호르몬인 에스트로겐과 비슷한 구조를 가진 유효성분으로 체내에서 에스트로겐 수용체와 결합하여 에스트로겐의 활성으로 여성 갱년기 증상을 완화시키고 심혈관계질환 예방, 암 등의 만성 퇴행성 질환을 예방한다고 알려져 있다. 특히 참깨에 리그난 배당체인 세사미놀 배당체는 생체막 등의 산화적 장해를 방어하며 α-토코페롤의 상승작용을 하여 체내의 항노화 활성을 나타낸다고 알려져 있다. 한편 세사미놀 배당체는 항산화 활성을 나타내는 페놀성 수산기가 당과 결합되어 있는 참깨 종실 자체에서 나타나는 항산화 활성보다 식품으로 섭취하여 장내세균의 β-글루코시다아제(β-glucosidase)의 작용으로 아글리콘(aglycon) 형태로 가수분해되어 장내 흡수를 도와 체내의 산화를 방어한다는 연구보고가 있다. 세사몰은 볶은 참기름의 대표적인 성분으로 알려져 있으며, 참기름의 리그난 성분의 항산화 활성이 참기름의 산화 안정성에 크게 기여하는 것으로 알려져 있으며, 착유하고 남은 탈지참깨박에는 수용성 리그난 성분이 잔존하며 대부분 종실 외피에 세사미놀배당체(sesaminol glucoside) 형태로 존재하고 있다. 이러한 이점에도 불구하고 아직까지 참깨박을 이용한 연구는 미비한 실정이며 식품소재의 활용 또한 낮은 실정이다.Sesamol and sesaminol are low-molecular vegetable compounds, and are known to inhibit antioxidant activity and cholesterol, atherosclerosis, and hyperglycemia. It is also known as an active ingredient with a structure similar to that of the female hormone estrogen, which binds to the estrogen receptor in the body to relieve female menopausal symptoms and prevent chronic degenerative diseases such as cancer and cancer by activating estrogen. In particular, sesaminol glycosides, which are lignan glycosides in sesame, are known to defend against oxidative disorders such as biomembranes and show synergistic action of α-tocopherol, thereby exhibiting anti-aging activity in the body. On the other hand, sesaminol glycosides are consumed as food rather than antioxidant activity in the sesame seed seed itself, which has a phenolic hydroxyl group that exhibits antioxidant activity and is associated with sugar, and thus aglycone (aglycon) under the action of β-glucosidase of intestinal bacteria ) There is a research report that it is hydrolyzed in the form to help absorb in the intestine and prevent oxidation in the body. Sesamol is known as a representative component of roasted sesame oil, and it is known that the antioxidant activity of sesame oil's lignan component greatly contributes to the oxidative stability of sesame oil. It exists in the form of sesaminol glucoside. Despite these advantages, studies using sesame seeds are still insufficient, and the utilization of food materials is also low.
한국등록특허 제1714417호에는 토마토 발효액을 이용한 간장의 제조방법이 개시되어 있고, 한국등록특허 제1399276호에는 단기숙성 간장의 제조방법이 개시되어 있으나, 본 발명의 바실러스 서브틸리스 SRCM103716 및 아스퍼질러스 오리재 균주를 이용한 간장의 제조방법과는 상이하다.Korean Registered Patent No. 1714417 discloses a method for producing soy sauce using a tomato fermentation broth, and Korean Registered Patent No. 1399276 discloses a method for producing short-term mature soy sauce, but Bacillus subtilis SRCM103716 and Aspergillus of the present invention. It is different from the method for producing soy sauce using the duck strain.
본 발명은 상기와 같은 요구에 의해 안출된 것으로서, 본 발명에서는 총 폴리페놀 및 총 플라보노이드 함량과 항산화 활성이 우수하고, 효소 분비능을 지니는 고초균을 분리하고, 상기 분리한 균주를 이용하여 고품질의 참깨박 발효 간장의 제조방법을 제공하는 데 있다.The present invention was devised by the above-mentioned demands, and in the present invention, it is excellent in total polyphenol and total flavonoid content and antioxidant activity, isolates archaea bacteria having enzyme secretion ability, and uses the separated strain to produce high-quality sesame seeds. It is to provide a method of manufacturing fermented soy sauce.
상기 과제를 해결하기 위해, 본 발명은 (1) 분쇄한 참깨박에 물을 가수하여 증자한 후 냉각한 참깨박에 바실러스 서브틸리스(Bacillus subtilis) SRCM103716 균주를 접종한 후 발효하여 총 폴리페놀 및 총 플라보노이드 함량과 항산화 활성 및 β-글루코시다아제(β-glucosidase) 활성이 증진된 참깨박 고초균 발효물을 제조하는 단계; (2) 분쇄한 참깨박에 물을 가수하여 증자한 후 냉각한 참깨박에 아스퍼질러스 오리재(Aspergillus oryzae) 균주를 접종한 후 발효하여 참깨박 발효물을 제조하는 단계; (3) 상기 (1)단계의 제조한 참깨박 고초균 발효물과 상기 (2)단계의 제조한 참깨박 발효물을 혼합하여 리그난 함량 및 항산화 활성이 증진된 참깨박 혼합 발효물을 제조하는 단계; 및 (4) 상기 (3)단계의 제조한 참깨박 혼합 발효물에 염수를 첨가하여 숙성시킨 후 여과하는 단계를 포함하여 제조하는 것을 특징으로 하는 참깨박 발효 간장의 제조방법을 제공한다. In order to solve the above problems, the present invention is (1) by increasing water by adding water to the crushed sesame seeds, and then inoculating Bacillus subtilis SRCM103716 strain on the cooled sesame seeds to ferment and inoculate the total polyphenols. Preparing a sesame seed archaea fermentation product having enhanced total flavonoid content, antioxidant activity, and β-glucosidase activity; (2) After adding water to the crushed sesame seeds, increase the amount, and then cool the cooled sesame seeds with Aspergillus oryzae ) producing a sesame leaf fermentation product by inoculating the strain and then fermenting it; (3) preparing a mixed fermented sesame seeds with enhanced lignan content and antioxidant activity by mixing the fermented sesame seeds produced in step (1) and the fermented sesame seeds prepared in step (2); And (4) provides a method for producing sesame seeds fermented soy sauce, characterized in that it comprises a step of aging after aging by adding brine to the mixed fermented sesame seeds prepared in step (3).
또한, 본 발명은 상기 방법으로 제조된 참깨박 발효 간장을 제공한다.In addition, the present invention provides a fermented soy sauce sesame seeds prepared by the above method.
본 발명의 간장은 리그난 함량 및 항산화 활성이 증진된 참깨박 혼합 발효물을 주성분으로 이용하기 때문에 품질이 우수하고, 유리 아미노산 함량과 향기 성분이 증진되고, 기호도가 우수한 고기능성 간장을 제공할 수 있다.The soy sauce of the present invention can provide a high-functional soy sauce having excellent quality, enhanced free amino acid content and aroma components, and excellent palatability because it uses the mixed fermented sesame seeds with enhanced lignan content and antioxidant activity as the main ingredients. .
도 1은 본 발명의 바실러스 서브틸리스 SRCM103716의 계통수(phyrogenetic tree)이다.
도 2는 참깨박 고초균 발효물의 제조공정을 도식화한 것이다.
도 3은 고초균으로 발효 전과 후, 참깨박 발효물의 생균수를 비교한 그래프이다.
도 4는 고초균으로 발효 전과 후, 참깨박 발효물의 DPPH 라디칼 소거능을 비교한 그래프이다.
도 5는 고초균으로 발효 전과 후, 참깨박 발효물의 총 폴리페놀 함량을 비교한 그래프이다.
도 6은 고초균으로 발효 전과 후, 참깨박 발효물의 총 플라보노이드 함량을 비교한 그래프이다.
도 7은 혼합비율에 따른 참깨박 발효 혼합물의 DPPH 라디칼 소거능을 비교한 그래프이다.
도 8은 참깨박 발효 혼합물을 이용한 간장의 제조공정을 도식화한 것이다.1 is a phylogenetic tree of the Bacillus subtilis SRCM103716 of the present invention.
Figure 2 is a schematic diagram showing the manufacturing process of sesame seed archaea fermentation.
Figure 3 is a graph comparing the number of live bacteria before and after fermentation with sesame leaf fermentation as an archaebacteria.
Figure 4 is a graph comparing the DPPH radical scavenging ability of sesame seed fermentation before and after fermentation with archaea.
Figure 5 is a graph comparing the total polyphenol content of sesame leaf fermentation before and after fermentation with archaea.
Figure 6 is a graph comparing the total flavonoid content of sesame leaf fermentation before and after fermentation with archaea.
7 is a graph comparing the DPPH radical scavenging ability of the sesame seed fermentation mixture according to the mixing ratio.
8 is a schematic diagram of a process for producing soy sauce using a fermented mixture of sesame seeds.
본 발명의 목적을 달성하기 위하여, 본 발명은In order to achieve the object of the present invention, the present invention
(1) 분쇄한 참깨박에 물을 가수하여 증자한 후 냉각한 참깨박에 바실러스 서브틸리스(Bacillus subtilis) SRCM103716 균주를 접종한 후 발효하여 총 폴리페놀 및 총 플라보노이드 함량과 항산화 활성 및 β-글루코시다아제(β-glucosidase) 활성이 증진된 참깨박 고초균 발효물을 제조하는 단계;(1) After adding water to the crushed sesame seeds and increasing the amount, the cooled sesame seeds were inoculated with Bacillus subtilis SRCM103716 strain and fermented to give total polyphenol and total flavonoid content, antioxidant activity, and β-gluco Preparing a fermented sesame-pseudomonas aeruginosa fermentation with enhanced cydase (β-glucosidase) activity;
(2) 분쇄한 참깨박에 물을 가수하여 증자한 후 냉각한 참깨박에 아스퍼질러스 오리재(Aspergillus oryzae) 균주를 접종한 후 발효하여 참깨박 발효물을 제조하는 단계;(2) After adding water to the crushed sesame seeds, increase the amount, and then cool the cooled sesame seeds with Aspergillus oryzae ) producing a sesame leaf fermentation product by inoculating the strain and then fermenting it;
(3) 상기 (1)단계의 제조한 참깨박 고초균 발효물과 상기 (2)단계의 제조한 참깨박 발효물을 혼합하여 리그난 함량 및 항산화 활성이 증진된 참깨박 혼합 발효물을 제조하는 단계; 및(3) preparing a mixed fermented sesame seeds with enhanced lignan content and antioxidant activity by mixing the fermented sesame seeds produced in step (1) and the fermented sesame seeds prepared in step (2); And
(4) 상기 (3)단계의 제조한 참깨박 혼합 발효물에 염수를 첨가하여 숙성시킨 후 여과하는 단계를 포함하여 제조하는 것을 특징을 하는 참깨박 발효 간장의 제조방법을 제공한다.(4) provides a method for producing a sesame seed fermented soy sauce, characterized in that it is prepared by adding brine to the fermented sesame meal prepared in the step (3), followed by filtration.
본 발명의 참깨박 발효 간장의 제조방법에서, 상기 바실러스 서브틸리스(Bacillus subtilis) SRCM103716 균주는 β-글루코시다아제(β-glucosidase) 활성이 우수하고, 바실러스 세레우스(Bacillus cereus), 스타필로코커스 아우레우스(Staphylococcus aureus) 및 리스테리아 모노시토게네스(Listeria monocytogenes)의 유해미생물에 대한 항균활성 및 β-용혈 활성이 있고, 페닐알라닌(phenylalanine)의 유해물질, 우레아제(urease) 및 β-글루쿠로니다아제(β-glucuronidase)의 유해효소 및 티라민 및 히스타민의 바이오제닉 아민을 생성하지 않으면서, 프로테아제(protease), 아밀라아제(amylase) 및 셀룰라제(cellulase) 분비능을 지니면서, 참깨박 발효시 총 폴리페놀 및 총 플라보노이드 함량과 항산화 활성을 증가율이 높은 균주를 선발한 것으로, 2019년 07월 10일 한국미생물보존센터(KCCM)에 기탁하여 기탁번호 KFCC11832P를 부여받았다.In the method of preparing the fermented soy sauce of the present invention, the Bacillus subtilis SRCM103716 strain has excellent β-glucosidase activity, and Bacillus cereus , Staphylococcus Aureus ( Staphylococcus aureus ) and Listeria monocytogenes ( Listeria monocytogenes ) have antimicrobial and β-hemolytic activity against harmful microorganisms, harmful substances of phenylalanine, urease and β-glucuro Total poly during fermentation of sesame seeds, having the ability to secrete proteases, amylases and cellulases without producing harmful enzymes of β-glucuronidase and biogenic amines of tyramine and histamine A strain with a high increase rate of phenol and total flavonoid content and antioxidant activity was selected, and deposited with the Korea Microbial Conservation Center (KCCM) on July 10, 2019. Endowed with panoramic number KFCC11832P.
또한, 본 발명의 참깨박 발효 간장의 제조방법에서, 상기 (1)단계의 참깨박 고초균 발효물은 바람직하게는 분쇄한 참깨박에 물을 1.2~1.8배 가수하여 110~130℃에서 10~20분 동안 증자한 후 냉각한 참깨박에 바실러스 서브틸리스(Bacillus subtilis) SRCM103716 균주를 접종한 후 34~40℃에서 20~28시간 동안 발효하여 제조할 수 있으며, 더욱 바람직하게는 분쇄한 참깨박에 물을 1.5배 가수하여 121℃에서 15분 동안 증자한 후 냉각한 참깨박에 바실러스 서브틸리스(Bacillus subtilis) SRCM103716 균주를 접종한 후 37℃에서 24시간 동안 발효하여 제조할 수 있다. 상기와 같은 조건으로 제조한 참깨박 고초균 발효물은 총 폴리페놀 및 총 플라보노이드 함량과 항산화 활성 및 β-글루코시다아제(β-glucosidase) 활성이 우수한 이점이 있다.In addition, in the method of manufacturing the fermented soy sauce of sesame seeds of the present invention, the sesame leaves Bacillus fermentation product of step (1) is preferably 1.2 to 1.8 times water is added to the crushed sesame leaves to 10 to 20 at 110 to 130 ° After inoculating the cooled sesame seeds for a minute and then inoculating Bacillus subtilis SRCM103716 strain, fermentation can be performed at 34-40 ° C. for 20-28 hours, more preferably crushed sesame seeds. It can be prepared by inoculating Bacillus subtilis SRCM103716 strain with cooled sesame seeds after increasing the water for 1.5 minutes by adding water 1.5 times at 121 ° C. and fermenting at 37 ° C. for 24 hours. The sesame seed archaea fermentation product prepared under the above conditions has an advantage of total polyphenol and total flavonoid content and excellent antioxidant activity and β-glucosidase activity.
또한, 본 발명의 참깨박 발효 간장의 제조방법에서, 상기 (2)단계의 참깨박 발효물은 바람직하게는 분쇄한 참깨박에 물을 0.4~0.6배 가수하여 110~130℃에서 10~20분 동안 증자한 후 냉각한 참깨박에 아스퍼질러스 오리재(Aspergillus oryzae) 균주를 접종한 후 28~32℃에서 68~76시간 동안 발효하여 제조할 수 있으며, 더욱 바람직하게는 분쇄한 참깨박에 물을 0.5배 가수하여 121℃에서 15분 동안 증자한 후 냉각한 참깨박에 아스퍼질러스 오리재(Aspergillus oryzae) 균주를 접종한 후 30℃에서 72시간 동안 발효하여 제조할 수 있다.In addition, in the method for producing fermented soy sauce of sesame seeds of the present invention, the fermented sesame seeds of step (2) is preferably 0.4 to 0.6 times water is added to the crushed sesame seeds for 10 to 20 minutes at 110 to 130 ° C. It can be prepared by inoculating strains of Aspergillus oryzae on cooled sesame seeds after increasing the amount, and fermenting at 28-32 ° C for 68-76 hours, more preferably water in crushed sesame seeds. Was added 0.5 times and increased for 15 minutes at 121 ℃, and then cooled to Aspergillus or Aspergillus oryzae ) can be prepared by inoculating the strain and fermenting at 30 ° C for 72 hours.
또한, 본 발명의 참깨박 발효 간장의 제조방법에서, 상기 (3)단계의 혼합은 바람직하게는 참깨박 고초균 발효물과 참깨박 발효물을 4~6:94~96 중량비율로 혼합할 수 있으며, 더욱 바람직하게는 참깨박 고초균 발효물과 참깨박 발효물을 5:95 중량비율로 혼합할 수 있다. 상기와 같은 조건으로 혼합한 발효물을 이용하여 간장을 제조하는 것이 향기 성분이 증가하고, 유리 아미노산 함량이 높으면서 기호도가 우수한 간장 제조에 적합한 참깨박 혼합 발효물로 제조할 수 있었다.In addition, in the method of preparing the fermented sesame soy sauce of the present invention, the mixing of step (3) is preferably a sesame gourd bacillus fermentation product and a sesame leaf fermentation product can be mixed in a weight ratio of 4-6: 94-96, , More preferably, the fermentation of sesame seeds and Bacillus spp. May be mixed at a weight ratio of 5:95. The production of soy sauce using the fermented product mixed under the above conditions increased the fragrance component, and the high amino acid content was high, and it could be prepared as a mixed fermented sesame seeds suitable for soy sauce production.
또한, 본 발명의 참깨박 발효 간장의 제조방법에서, 상기 (4)단계는 바람직하게는 참깨박 혼합 발효물에 23~25%(w/v) 염수를 2~3배(v/w) 첨가하여 30~35℃에서 3~4주 동안 숙성시킨 후 여과할 수 있으며, 더욱 바람직하게는 참깨박 혼합 발효물에 24%(w/v) 염수를 2.5배(v/w) 첨가하여 30~35℃에서 3~4주 동안 숙성시킨 후 여과할 수 있다. 상기와 같은 조건으로 숙성시켜 간장을 제조하는 것이 총 질소 함량이 높고 감칠맛 및 구수한 맛이 증진되어 품질이 우수한 간장으로 제조할 수 있었다.In addition, in the method of preparing the fermented soy sauce of sesame seeds of the present invention, step (4) preferably adds 23 to 25% (w / v)
본 발명의 참깨박 발효 간장의 제조방법은, 보다 구체적으로는The method for producing the fermented soy sauce of sesame seeds of the present invention, more specifically
(1) 분쇄한 참깨박에 물을 1.2~1.8배 가수하여 110~130℃에서 10~20분 동안 증자한 후 냉각한 참깨박에 바실러스 서브틸리스(Bacillus subtilis) SRCM103716 균주를 접종한 후 34~40℃에서 20~28시간 동안 발효하여 총 폴리페놀 및 총 플라보노이드 함량과 항산화 활성 및 β-글루코시다아제(β-glucosidase) 활성이 증진된 참깨박 고초균 발효물을 제조하는 단계;(1) Water is added to the crushed sesame seeds 1.2 ~ 1.8 times, and the water is increased at 110 ~ 130 ℃ for 10 ~ 20 minutes. After cooling, the cooled sesame seeds are inoculated with Bacillus subtilis SRCM103716 strain, and then 34 ~ Fermentation at 40 ° C. for 20-28 hours to produce a sesame-pseudomonas aeruginosa fermentation product with enhanced total polyphenol and total flavonoid content, antioxidant activity, and β-glucosidase activity;
(2) 분쇄한 참깨박에 물을 0.4~0.6배 가수하여 110~130℃에서 10~20분 동안 증자한 후 냉각한 참깨박에 아스퍼질러스 오리재(Aspergillus oryzae) 균주를 접종한 후 28~32℃에서 68~76시간 동안 발효하여 참깨박 발효물을 제조하는 단계;(2) Water is added to the crushed sesame seeds 0.4 ~ 0.6 times, and the water is increased at 110 ~ 130 ℃ for 10 ~ 20 minutes, and then cooled to the cooled sesame seeds. Aspergillus oryzae ) producing a sesame leaf fermentation product by inoculating the strain and fermenting at 28-32 ° C for 68-76 hours;
(3) 상기 (1)단계의 제조한 참깨박 고초균 발효물과 상기 (2)단계의 제조한 참깨박 발효물을 4~6:94~96 중량비율로 혼합하여 리그난 함량 및 항산화 활성이 증진된 참깨박 혼합 발효물의 제조하는 단계; 및(3) The sesame leaf fermentation product produced in the step (1) and the sesame leaf fermentation product prepared in the step (2) are mixed at a weight ratio of 4 to 6:94 to 96 to improve the lignan content and antioxidant activity. Preparing a sesame-bak mixed fermentation product; And
(4) 상기 (3)단계의 제조한 참깨박 혼합 발효물에 23~25%(w/v) 염수를 2~3배(v/w) 첨가하여 30~35℃에서 3~4주 동안 숙성시킨 후 여과하는 단계를 포함할 수 있으며,(4) 23 ~ 25% (w / v) brine is added 2-3 times (v / w) to the mixed fermented sesame leaf prepared in the step (3), aged at 30 ~ 35 ℃ for 3-4 weeks After filtering it may include a step,
더욱 구체적으로는More specifically
(1) 분쇄한 참깨박에 물을 1.5배 가수하여 121℃에서 15분 동안 증자한 후 냉각한 참깨박에 바실러스 서브틸리스(Bacillus subtilis) SRCM103716 균주를 접종한 후 37℃에서 24시간 동안 발효하여 총 폴리페놀 및 총 플라보노이드 함량과 항산화 활성 및 β-글루코시다아제(β-glucosidase) 활성이 증진된 참깨박 고초균 발효물을 제조하는 단계;(1) Water was added 1.5 times to the crushed sesame seeds and increased at 121 ° C for 15 minutes, followed by inoculation of Bacillus subtilis SRCM103716 strain on the cooled sesame leaves and fermentation at 37 ° C for 24 hours. Preparing total sesame seed archaea fermentation with enhanced polyphenol and total flavonoid content, antioxidant activity, and β-glucosidase activity;
(2) 분쇄한 참깨박에 물을 0.5배 가수하여 121℃에서 15분 동안 증자한 후 냉각한 참깨박에 아스퍼질러스 오리재(Aspergillus oryzae) 균주를 접종한 후 30℃에서 72시간 동안 발효하여 참깨박 발효물을 제조하는 단계;(2) increase the water to ground Sesame at 121 ℃ to 0.5 times the singer for 15 minutes and then cooled to Aspergillus Sesame's duck material (Aspergillus oryzae ) preparing a sesame leaf fermentation product by inoculating the strain and fermenting at 30 ° C. for 72 hours;
(3) 상기 (1)단계의 제조한 참깨박 고초균 발효물과 상기 (2)단계의 제조한 참깨박 발효물을 5:95 중량비율로 혼합하여 리그난 함량 및 항산화 활성이 증진된 참깨박 혼합 발효물의 제조하는 단계; 및(3) The sesame leaf fermentation product produced in step (1) and the sesame leaf fermentation product prepared in step (2) are mixed at a weight ratio of 5:95 to improve the lignan content and antioxidant activity. Preparing water; And
(4) 상기 (3)단계의 제조한 참깨박 혼합 발효물에 24%(w/v) 염수를 2.5배(v/w) 첨가하여 30~35℃에서 3~4주 동안 숙성시킨 후 여과하는 단계를 포함할 수 있다.(4) 24% (w / v) brine is added 2.5 times (v / w) to the mixed fermentation product prepared in step (3), aged at 30-35 ° C. for 3-4 weeks, and filtered. It may include steps.
본 발명은 또한, 상기 방법으로 제조된 참깨박 발효 간장을 제공한다.The present invention also provides a fermented soy sauce sesame seeds prepared by the above method.
이하, 본 발명을 실시예에 의해 상세히 설명한다. 단, 하기 실시예는 본 발명을 예시하는 것일 뿐, 본 발명의 내용이 하기 실시예에 한정되는 것은 아니다.Hereinafter, the present invention will be described in detail by examples. However, the following examples are only to illustrate the present invention, the content of the present invention is not limited to the following examples.
제조예Manufacturing example 1. One. 참깨박Sesame seeds 발효 혼합물 Fermentation mixture
(1) 분쇄한 참깨박에 물을 1.5배(v/w) 가수하여 121℃에서 15분 동안 증자한 후 60℃로 냉각한 참깨박에 바실러스 서브틸리스(Bacillus subtilis) SRCM103716 균주를 초기균수가 3~4 log CFU/mL가 되도록 접종한 후 37℃에서 24시간 동안 발효하여 참깨박 고초균 발효물을 제조하였다.(1) Water was added 1.5 times (v / w) to the crushed sesame seeds and increased for 15 minutes at 121 ° C, and then Bacillus subtilis SRCM103716 strain was first added to the sesame seeds cooled to 60 ° C. After inoculation to be 3 to 4 log CFU / mL, fermentation was performed at 37 ° C. for 24 hours to prepare a sesame seed archaea fermentation product.
(2) 분쇄한 참깨박에 물을 0.5배(v/w) 가수하여 121℃에서 15분 동안 증자한 후 60℃로 냉각한 참깨박에 아스퍼질러스 오리재(Aspergillus oryzae) 균주를 0.3%(v/w) 접종한 후 30℃에서 72시간 동안 발효하여 참깨박 발효물을 제조하였다.(2) 0.5 times (v / w) water was added to the crushed sesame seeds, and the mixture was increased at 121 ° C for 15 minutes, and then cooled to 60 ° C in the sesame leaves. Aspergillus oryzae ) was inoculated with 0.3% (v / w) strain, and fermented at 30 ° C for 72 hours to prepare a sesame leaf fermentation product.
(3) 상기 (1)단계의 제조한 참깨박 고초균 발효물과 상기 (2)단계의 제조한 참깨박 발효물을 5:95 중량비율로 혼합하였다.(3) The fermented sesame seeds produced in step (1) and the fermented sesame seeds prepared in step (2) were mixed at a weight ratio of 5:95.
제조예Manufacturing example 2. 2. 참깨박Sesame seeds 발효 간장 Fermented soy sauce
상기 제조예 1의 제조한 참깨박 발효 혼합물에 24%(w/v) 염수를 2.5배(v/w) 첨가하여 30~35℃에서 3~4주 동안 숙성시킨 후 여과하여 발효액을 분리하였다. 상기 분리한 발효액을 100℃에 3분 동안 살균한 후 필터페이퍼(Advantec No.2,Toyo Roshi Kaisha Ltd., japan)로 여과하였다.To the fermentation mixture prepared in Preparation Example 1, 24% (w / v) brine was added 2.5 times (v / w), aged at 30-35 ° C. for 3-4 weeks, filtered and separated from the fermentation broth. The separated fermentation broth was sterilized at 100 ° C. for 3 minutes, and then filtered with filter paper (Advantec No. 2, Toyo Roshi Kaisha Ltd., Japan).
실시예Example 1. 고초균별 특성 1. Characteristics by archaea
간장, 고추장, 된장, 젓갈 등에서 분리한 진흥원 기보유중인 바실러스 200여 종 중 β-글루코시다아제 활성을 보유한 고초균 15종을 선발하여 실험을 진행하였다. 실험 균주 리스트는 표 1과 같다. Among the 200 Bacillus species that were previously held by the Promotion Agency isolated from soy sauce, red pepper paste, miso, and salted fish, 15 Bacillus subtilis possessing β-glucosidase activity were selected and tested. Table 1 shows the list of experimental strains.
1) 세포외분비 효소활성(Amylase, Protease, Cellulase) 분석1) Analysis of extracellular secretion enzyme activity (Amylase, Protease, Cellulase)
한천 확산법 방법에 준하여 바실러스 균주를 5 ml의 LB broth에 접종하여 24시간 동안 30℃, 150 rpm으로 진탕 배양기에서 배양하였으며, 배양액을 13,000 rpm에서 원심분리한 후 0.45 ㎛ 필터를 이용하여 제균한 시료를 시험액으로 사용하였다. Bacillus strains were inoculated in 5 ml of LB broth according to the agar diffusion method, and cultured in a shaking incubator at 30 ° C and 150 rpm for 24 hours. After centrifuging the culture solution at 13,000 rpm, the sample sterilized using a 0.45 μm filter It was used as a test solution.
바실러스 균주의 균체 외로 방출하는 세포의 효소 중 아밀라아제(amylase), 프로테아제(protease), 셀룰라아제(cellulase) 활성을 검증하기 위해 한천 확산법(agar diffusion method)를 사용하였으며, 각 효소와 특이적으로 반응하는 기질의 성분이 포함된 선발배지를 제조하여 사용하였다. 아밀라아제 활성을 측정하기 위해 가용성 전분(starch soluble, Junsei)를 기질로 선택하여 가용성 전분 1%가 포함된 2% 한천을 첨가한 배지를 사용하였으며, 프로테아제 활성을 측정하기 위해 2% 탈지유(BD, Difco)가 첨가된 1.5% 한천 배지를 제조하여 사용하였다. 셀룰라아제 활성을 검증하기 위해 CMC(carboxylmethyl cellulose, Junsei)를 기질로 선택하여 1% CMC가 첨가된 1.5% 한천 배지를 제조하였다. 각 기질이 포함된 배지에 8 mm의 홀(hole)을 형성한 후 각 웰(well)에 균주 시험액 100 ㎕씩을 분주하여 37℃에서 24시간 배양하여 형성된 클리어 존(clear zone)의 크기를 측정하여 균주의 효소활성을 측정하였으며, 아밀라아제 활성을 측정하기 위해서 Lugol staining 방법을, 셀룰라아제 활성을 측정하기 위해서는 congo red staining을 이용하였다.Among the enzymes of cells released into the cells of the Bacillus strain, amylase, protease, and cellulase activities were used to verify the agar diffusion method, and substrates that specifically react with each enzyme The starting medium containing the components of was prepared and used. In order to measure amylase activity, a medium containing 2% agar containing 1% of soluble starch was used by selecting soluble starch (starch soluble, Junsei) as a substrate, and 2% skim milk (BD, Difco) to measure protease activity ) Was added and used to prepare 1.5% agar medium. To verify the cellulase activity, CMC (carboxylmethyl cellulose, Junsei) was selected as a substrate to prepare 1.5% agar medium with 1% CMC added. After forming 8 mm holes in the medium containing each substrate, 100 μl of the strain test solution was dispensed into each well, and the size of the clear zone formed by incubating at 37 ° C. for 24 hours was measured. The enzyme activity of the strain was measured, Lugol staining method was used to measure amylase activity, and congo red staining was used to measure cellulase activity.
*+; positive, -; negative * +; positive,-; negative
2) 식품유해미생물에 대한 항균활성 측정2) Measurement of antibacterial activity against food harmful microorganisms
한천 확산법 방법에 준하여 바실러스 균주를 5 ml의 LB broth에 접종하여 24시간 동안 30℃, 150 rpm 진탕 배양기에서 배양하였으며, 배양액을 13,000 rpm에서 원심분리한 후 0.45 ㎛ 필터를 이용하여 제균한 시료를 시험액으로 사용하였다.According to the agar diffusion method, the Bacillus strain was inoculated into 5 ml of LB broth and cultured in a shaking culture incubator at 30 ° C and 150 rpm for 24 hours. It was used as.
식품유해활성을 측정하기 위해 지시균주를 KCCM(Korean Culture Center of Microorganisms)와 KCTC(Korean Collection for Type Cultures)에서 분양받아 사용하였다. 각 지시균주를 적절한 배지에 배양하여 O.D(optical density)가 600 nm에서 0.4~0.6이 되도록 배양한 후 배지에 0.8% 한천 배지에 1% 혼합하여 배지를 제조하였으며, 한천 확산법(agar diffusion method)에 준하여 각 지시균주가 포함된 배지에 8 mm의 홀(hole)을 형성한 후 각 웰(well)에 균주 시험액 100 ㎕씩을 분주하여 30~37℃에서 24시간 배양한 후 홀(hole) 주변에 형성된 억제환의 크기를 측정하여 선발된 균주의 항균활성을 측정하였다.In order to measure food harmful activity, indicator strains were pre-distributed from KCCM (Korean Culture Center of Microorganisms) and KCTC (Korean Collection for Type Cultures). Each indicator strain was cultivated in an appropriate medium, and OD (optical density) was incubated to be 0.4 to 0.6 at 600 nm, followed by mixing with 1% in 0.8% agar medium in the medium to prepare a medium. After forming a hole of 8 mm in the medium containing each indicator strain, 100 μl of the strain test solution was dispensed into each well and cultured at 30 to 37 ° C. for 24 hours, and then formed around the hole. The size of the inhibitory ring was measured to measure the antibacterial activity of the selected strain.
그 결과, SRCM103716 균주는 바실러스 세레우스(Bacillus cereus), 스타필로코커스 아우레우스(Staphylococcus aureus) 및 리스테리아 모노시토게네스(Listeria monocytogenes)에 대한 항균 활성을 확인하였다.As a result, the SRCM103716 strain confirmed antibacterial activity against Bacillus cereus , Staphylococcus aureus , and Listeria monocytogenes .
3) 선별균주의 용혈활성(Hemolysis type) 분석3) Analysis of hemolysis type of selected strains
바실러스 균주의 용혈활성을 분석하기 위해 Blood agar(Kisanbio, Korea)를 구입하여 각 blood agar 배지에 선별 균주를 획선도말한 후 24시간 동안 30℃에서 배양하였으며, 콜로니 주변 한천에 변화가 없는 경우 γ-hemolysis, 적혈구를 완전히 파괴하여 집락 주변으로 투명한 환(clear zone)이 형성된 경우 β-hemolysis, 일부 적혈구만 파괴하여 콜로니 주변에 녹색으로 관찰되는 경우 α-hemolysis로 판정하였다.In order to analyze the hemolytic activity of Bacillus strains, blood agar (Kisanbio, Korea) was purchased, and the selected strains were streaked on each blood agar medium and cultured at 30 ° C. for 24 hours. Hemolysis, when red blood cells were completely destroyed and a clear zone was formed around the colony, β-hemolysis was observed, and when only some red blood cells were destroyed and observed in green around the colonies, it was determined as α-hemolysis.
그 결과, SRCM103716 균주는 β-용혈 활성이 있으면서, 페닐알라닌(phenylalanine), 우레아제(urease) 및 β-글루쿠로니다아제(β-glucuronidase)의 유해대사산물을 생성하지 않음을 확인하였다.As a result, it was confirmed that the SRCM103716 strain has β-hemolytic activity, and does not produce harmful metabolites of phenylalanine, urease, and β-glucuronidase.
4) 바이오제닉 아민 생성 유무 확인4) Check if biogenic amine is generated
바이오제닉 아민 생성 유무를 확인한 결과, BCH3520 균주를 제외한 모든 균주에서 티라민 및 히스타민을 생성하지 않음을 확인하였다.As a result of confirming the presence or absence of biogenic amine production, it was confirmed that tyramine and histamine were not produced in all strains except the BCH3520 strain.
ND: 검출되지 않음ND: not detected
5) β-글루코시다아제(β-glucosidase) 활성5) β-glucosidase activity
실험 균주 15종의 β-글루코시다아제(β-glucosidase) 활성을 측정하기 위하여 β-glucosidase activity assay kit(Sigma)를 이용하였다. 실험 균주 15종을 전배양한 다음, 본배양액은 LB broth(Difco)에 1%(v/v) 접종하여 37℃에서 24시간 진탕 배양하였다. 본배양액의 상등액을 수거한 다음, 균체를 50 mM phosphate buffer(pH 7.0)에 풀어준 후, 이를 시료로 이용하였다. 1 웰에 220 ㎕ 증류수, 1 웰에는 20 ㎕ 증류수, 200 ㎕ calibrator를 첨가하였고, 샘플의 각각의 웰에 assay buffer 200 ㎕, β-NPG substrate 8 ㎕를 넣어준 다음, 시료를 20 ㎕ 첨가하였다. 이후 초기 흡광도를 405 nm에서 측정한 다음 37℃에서 20분 동안 방치한 다음 최종 흡광도를 405 nm에서 측정하였다. 이 후, 아래의 계산식을 이용하여 β-글루코시다아제 활성 값을 확인하였다. 실험 균주 15종에 대한 β-글루코시다아제 활성값은 표 2에 나타내었다.The β-glucosidase activity assay kit (Sigma) was used to measure the β-glucosidase activity of 15 types of experimental strains. After pre-culturing 15 kinds of experimental strains, the main culture solution was inoculated with 1% (v / v) in LB broth (Difco) and shake cultured at 37 ° C for 24 hours. After collecting the supernatant of the main culture solution, the cells were released in 50 mM phosphate buffer (pH 7.0) and used as a sample. 220 µl distilled water to 1 well, 20 µl distilled water, 200 µl calibrator was added to 1 well, and 200 µl of assay buffer and 8 µl of β-NPG substrate were added to each well of the sample, and then 20 µl of the sample was added. Thereafter, the initial absorbance was measured at 405 nm, then left at 37 ° C for 20 minutes, and then the final absorbance was measured at 405 nm. Thereafter, the value of β-glucosidase activity was confirmed using the following formula. The β-glucosidase activity values for 15 test strains are shown in Table 2.
β-글루코시다아제 활성(Unit/L) = (최종 흡광도 - 최초 흡광도 × 250 units/L)/(calibrator 흡광도 - 물 흡광도)β-glucosidase activity (Unit / L) = (final absorbance-initial absorbance × 250 units / L) / (calibrator absorbance-water absorbance)
β-글루코시다아제 활성은 BCH3711, SRCM103716에서 각각 9.63 unit/L, 12.6 unit/L로 가장 높게 나타났다. β-glucosidase activity was highest in BCH3711 and SRCM103716, 9.63 unit / L and 12.6 unit / L, respectively.
실시예Example 2. 고초균 종류에 따른 2. According to the type of archaea 참깨박Sesame seeds 발효물의Fermentation 품질특성 Quality characteristics
참깨박의 고초균 발효를 위하여 도 2와 같은 방법으로 발효를 진행하였다.The fermentation was performed in the same manner as in FIG. 2 for the fermentation of Sesame seeds.
1) 참깨박 소재 고초균 15종 발효물 생균수 1) The number of live bacteria of 15 types of archaea bacteria from sesame seeds
발효하기 전과 후의 생균수의 결과는 도 3에 나타내었다. 그 결과, 전체 발효한 시료에서 8 log CFU/mL 이상 측정되었다. 15종 모두 참깨박 소재에서 잘 자라는 것을 확인하였다.The results of the viable cell count before and after fermentation are shown in FIG. 3. As a result, 8 log CFU / mL or more was measured in the whole fermented sample. All 15 species were confirmed to grow well in sesame seeds.
2) β-글루코시다아제 활성 2) β-glucosidase activity
균주에 따른 참깨박 발효물에 대하여 β-글루코시다아제 활성을 측정하였다. 시료는 상등액을 수거하여 50 mM phosphate buffer (pH 7.0)에 풀어준 후, 이를 시료로 이용하였다. β-glucosidase activity assay kit (Sigma)을 이용하여 전과 같은 방법으로 측정한 결과는 표 6과 같다. 그 결과, 바실러스 서브틸리스 SRCM103716로 발효한 참깨박에서 β-글루코시다아제 활성이 16.88 unit/L로 가장 높게 측정되었다.Β-glucosidase activity was measured for the fermented sesame seeds according to the strain. The supernatant was collected and released in 50 mM phosphate buffer (pH 7.0), and used as a sample. Table 6 shows the results measured in the same manner as before using the β-glucosidase activity assay kit (Sigma). As a result, Bacillus subtilis In sesame seeds fermented with SRCM103716, β-glucosidase activity was the highest at 16.88 unit / L.
3) DPPH 라디칼 소거능 측정3) DPPH radical scavenging ability measurement
DPPH (1,1-diphenyl-2-picrylhydrazyl) 라디칼 소거능은 Blois (1958)의 방법을 변형하여 전자공여작용(electron donationg abilities, EDA)에 대한 효과로 각 시료의 환원력을 측정하였다. 즉, 시료 50 ㎕에 0.15 mM DPPH 용액(99% MeOH에 용해) 150 ㎕을 가한 후 30분간 상온에서 방치한 후 분광광도계를 사용하여 흡광도 517 nm에서 흡광도의 변화를 측정하였다. 각 시료의 라디칼 소거능은 아래의 식에 의해 시료 첨가구 및 무첨가구간의 흡광도 차이를 백분율로 나타내었다.DPPH (1,1-diphenyl-2-picrylhydrazyl) radical scavenging ability was modified by the method of Blois (1958) to measure the reducing power of each sample as an effect on electron donation ability (EDA). That is, after adding 150 µl of 0.15 mM DPPH solution (dissolved in 99% MeOH) to 50 µl of the sample and standing at room temperature for 30 minutes, the change in absorbance at 517 nm was measured using a spectrophotometer. The radical scavenging ability of each sample was expressed as a percentage of the difference in absorbance between the sample addition and non-addition zones by the following equation.
전자공여능(%) = (1 - 시료첨가구의 흡광도/무처리구의 흡광도) × 100Electron donating ability (%) = (1-absorbance of sample addition / absorption of no treatment) × 100
참깨박 소재의 발효 전과 후에 따른 DPPH 라디칼 분석 결과는 도 4와 같다. 발효전에는 43.24~45.34%의 활성값을 가졌으나, 발효 후 모두 활성이 증가하였다. 이 중 BCH3711, SRCM103716를 이용한 참깨박 발효물이 각각 63.82%, 66.45%로 가장 높게 증가하였다.The results of DPPH radical analysis according to before and after fermentation of sesame seeds are shown in FIG. 4. Before fermentation, it had an activity value of 43.24 to 45.34%, but the activity increased after fermentation. Among them, fermented sesame seeds using BCH3711 and SRCM103716 were the highest at 63.82% and 66.45%, respectively.
4) 총 폴리페놀(polyphenol) 함량4) Total polyphenol content
총 페놀 함량은 Folin-Denis 변법에 따라 추출된 시료를 농도별로 적절히 희석한 후 측정하였다. 각 농도별 시료 200 ㎕에 Folin-Ciocalteu 시약 12.5 ㎕를 혼합하여 교반한 뒤, 120분 동안 상온에서 방치하여 반응시켰다. 반응액의 흡광도 값은 UV-Vis 분광광도계(DU 800, Beckman coulter, Fullerton, CA, USA)를 사용하여 750 nm에서 분석하였다. 총 페놀 함량은 갈산을 분석시료와 동일한 방법으로 분석하여 얻은 표준 검량선으로부터 시료 추출물의 총 페놀 함량을 산출하였고, 총 폴리페놀 함량은 1 L 중의 mg gallic acid equivalent로 나타내었다.The total phenol content was measured after appropriately diluting samples extracted according to the Folin-Denis method by concentration. After mixing and stirring 12.5 µl of Folin-Ciocalteu reagent in 200 µl of each concentration sample, the mixture was allowed to stand at room temperature for 120 minutes to react. The absorbance value of the reaction solution was analyzed at 750 nm using a UV-Vis spectrophotometer (
참깨박 소재의 발효 전과 후에 따른 폴리페놀 함량을 측정한 결과는 도 5와 같다. 발효 후 대조구를 제외한 모든 균주에서 폴리페놀 함량이 증가하는 것을 확인하였으며, 이 중 특히 BCH3637, BCH3678, SRCM103716의 3종은 각각 2490.2, 2559.4, 2803.3 mg/L로 가장 높게 증가하였다.The results of measuring the polyphenol content before and after fermentation of the sesame seeds are shown in FIG. 5. After fermentation, it was confirmed that the polyphenol content increased in all strains except the control group. Among them, 3 types of BCH3637, BCH3678, and SRCM103716 were the highest, respectively, 2490.2, 2559.4, and 2803.3 mg / L.
5) 총 플라보노이드(flavonoid) 함량5) Total flavonoid content
총 플라보노이드 함량은 Davis 변법에 따라 추출된 시료를 농도별로 적절히 희석한 후 측정하였다. 각 농도별 시료 0.5 mL에 10% 질산알루미늄(aluminum nitrate) 0.1 mL, 1M 초산 칼륨(potassium acetate) 0.1 mL 및 에탄올 4.3 mL를 차례로 가하여 혼합한 후 상온에서 40분간 방치하여 반응시켜 450 nm에서 흡광도를 측정하였다. 표준물질로는 갈산을 시료와 동일한 방법으로 분석하여 얻은 표준 검량선으로부터 시료 추출물의 총 플라보노이드 함량을 산출하였고, 총 플라보노이드 함량은 1 L 중의 mg gallic acid equivalent로 나타내었다.The total flavonoid content was measured after appropriately diluting the sample extracted according to the Davis method. To 0.5 mL of samples of each concentration, 0.1 mL of 10% aluminum nitrate, 0.1 mL of 1M potassium acetate, and 4.3 mL of ethanol were added in sequence, mixed, and allowed to stand at room temperature for 40 minutes to react, resulting in absorbance at 450 nm. It was measured. As a standard substance, the total flavonoid content of the sample extract was calculated from a standard calibration curve obtained by analyzing gallic acid in the same way as the sample, and the total flavonoid content was expressed as mg gallic acid equivalent in 1 L.
참깨박 소재의 발효 전과 후에 따른 플라보노이드 함량을 측정한 결과는 도 6과 같다. 발효 후 대조구를 제외한 모든 균주에서 플라보노이드 함량이 증가하는 것을 확인하였으며, 이 중 특히 BCH3637, BCH3678, SRCM103716의 3종은 각각 1423.27, 1688.67, 1553.08 mg/L로 가장 높게 증가하는 것으로 확인되었다.The results of measuring the content of flavonoids before and after fermentation of sesame seeds are shown in FIG. 6. After fermentation, it was confirmed that the flavonoid content was increased in all strains except the control group. Among these, 3 types of BCH3637, BCH3678, and SRCM103716 were found to be the highest at 1423.27, 1688.67, and 1553.08 mg / L, respectively.
5. 참깨박 고초균 발효 균주 선발5. Sesame leaf archaea fermentation strain selection
이에 참깨박 소재의 발효에 대한 균주로 β-글루코시다아제 활성 및 DPPH 라디칼 소거능 활성이 우수하고 발효 이후 총 폴리페놀 함량과 플라보노이드 함량의 증가율이 높은 바실러스 서틸리스(Bacillus subtilis) SRCM103716로 선발하였으며, 선발한 균주의 계통도는 도 1과 같으며, 2019년 07월 10일 한국미생물보존센터(KCCM)에 기탁하여 기탁번호 KFCC11832P를 부여받았다.Thus, as a strain for fermentation of sesame seeds, it was selected as Bacillus subtilis SRCM103716, which has excellent β-glucosidase activity and DPPH radical scavenging activity, and has a high increase rate of total polyphenol content and flavonoid content after fermentation. The schematic diagram of the selected strain is as shown in FIG. 1, and was deposited with the Korea Microbial Conservation Center (KCCM) on July 10, 2019, and was assigned a deposit number KFCC11832P.
실시예Example 3. 3. 참깨박Sesame seeds 발효물의Fermentation 혼합비율 설정 Mixing ratio setting
참깨박 바실러스 서브틸리스(Bacillus subtilis) SRCM103716 발효물과 참깨박 아스퍼질러스 오리재(Aspergillus oryzae) 발효물의 혼합 비율을 설정하기 위하여 표 7과 같이 샘플을 제조하였다. DPPH 라디칼 소거능을 확인하여 가장 높은 항산화활성을 갖는 비율로 참깨박 발효 간장 제조 구간을 설정하기로 하였다.Sesame leaf Bacillus subtilis SRCM103716 Fermentation and Sesame leaf Aspergillus oryzae ) Samples were prepared as shown in Table 7 to set the mixing ratio of the fermentation product. By confirming the DPPH radical scavenging ability, it was decided to set the fermented soy sauce production section at the rate with the highest antioxidant activity.
1) One) 참깨박Sesame seeds 발효물Fermentation 혼합비율에 따른 According to the mixing ratio DPPHDPPH 라디칼Radical 소거능Erasing ability 결과 result
참깨박 발효물 혼합비율에 따른 DPPH 라디칼 소거능 결과는 도 7과 같다. 아스퍼질러스 오리재(A. oryzae) 95%, 바실러스 서브틸리스(B. subtilis) SRCM103716 5% 혼합 비율에서 DPPH 라디칼 활성 결과 84.12%로 가장 높게 측정되었으며, 다음으로 아스퍼질러스 오리재(A. oryzae) 100%에서 77.28%로 측정되었다. 이에 참깨박 발효 간장 제조를 위하여 아스퍼질러스 오리재(A. oryzae) 95%, 바실러스 서브틸리스(B. subtilis) SRCM103716 5%가 가장 적합함을 확인하였다.Results of DPPH radical scavenging ability according to the mixing ratio of sesame seeds are shown in FIG. 7. DPPH radical activity at 84.12% was highest in Aspergillus oridae ( A. oryzae ) 95%, B. subtilis SRCM103716 5%, followed by Aspergillus ori . oryzae ) from 100% to 77.28%. In order to Sesame fermented soy sauce production Aspergillus duck material (A. oryzae) 95%, Bacillus subtilis (B. subtilis) it was confirmed that the best SRCM103716 5%.
2) 2) 참깨박Sesame seeds 비발효물과Non-fermented 참깨박Sesame seeds 발효 혼합물의 Of the fermentation mixture DPPHDPPH 라디칼Radical 소거활성 및 Scavenging activity and 리그League 난 성분을 측정I measure the ingredients
최종적으로 참깨박 아스퍼질러스 오리재 발효물 95%와 바실러스 서브틸리스 SRCM103716 발효물 5%를 혼합하여 간장 제조에 사용하기로 결정하였다. 이에 참깨박 비발효물과 참깨박 발효 혼합물의 DPPH 라디칼 소거활성 및 리그난 성분을 측정하였다.Finally, it was decided to mix 95% of the fermented sesame seed Aspergillus orijae with 5% of the Bacillus subtilis SRCM103716 fermentation for use in the production of soy sauce. Accordingly, DPPH radical scavenging activity and lignan components of the non-fermented sesame seeds and the fermented mixture of sesame seeds were measured.
(2-1) (2-1) 참깨박Sesame seeds 발효 혼합물의 Of the fermentation mixture DPPHDPPH 라디칼Radical 소거활성 결과 Scavenging activity result
바실러스 서틸리스(Bacillus subtilis) SRCM103716 균주와 아스퍼질러스 오리재(Aspergillus oryzae)를 이용한 참깨박 발효 혼합물과 비발효물의 DPPH 라디칼 소거활성 결과는 표 8에 나타내었다. 측정 결과 100배 희석에서 참깨박 비발효물과 참깨박 발효 혼합물이 각각 28.4%, 73.3%로 발효 후 약 160% 항산화 활성이 증가하는 것으로 확인되었다.The results of DPPH radical scavenging activity of the fermentation mixture of sesame seeds and non-fermentation using Bacillus subtilis SRCM103716 strain and Aspergillus oryzae are shown in Table 8. As a result of the measurement, it was confirmed that at 100-fold dilution, the fermented mixture of non-fermented sesame and fermented sesame seeds was 28.4% and 73.3%, respectively, and the antioxidant activity increased after fermentation.
(2-2) (2-2) 참깨박Sesame seeds 발효 혼합물의 Of the fermentation mixture 리그난Lignan 성분 분석 결과 Ingredient analysis results
발효물의 리그난 성분을 추출 및 분석하였으며 방법은 다음과 같다. 발효물 시료 2.5 g에 50 mL 메탄올을 첨가하여 소니케이터(JAC4020)를 이용하여 1시간 동안 추출하였다. 추출 후 원심분리기(Hanil MF80)를 이용하여 3,000 rpm에서 10분간 원심분리하여 고형분을 제거하였다. 0.45 ㎛ 실린지 필터로 여과 후 분석하였으며 분석 조건은 표 9와 동일하다.The lignan component of the fermentation was extracted and analyzed, and the method is as follows. 50 mL methanol was added to 2.5 g of the fermentation product sample, followed by extraction using a sonicator (JAC4020) for 1 hour. After extraction, solids were removed by centrifugation at 3,000 rpm for 10 minutes using a centrifuge (Hanil MF80). After filtering with a 0.45 μm syringe filter, the analysis conditions are the same as in Table 9.
(4.6 × 150mm ID, 3)Capcell PAK MGII C18 Column
(4.6 × 150mm ID, 3)
Sesamin (3), Sesamolin (4)Sesamol (1), Sesaminol (2),
Sesamin (3), Sesamolin (4)
바실러스 서틸리스(Bacillus subtilis) SRCM103716 균주와 아스퍼질러스 오리재(Aspergillus oryzae)를 이용한 참깨박 발효 혼합물과 비발효물의 리그난 함량 분석 결과를 표 10에 나타내었다. 분석 결과 전체 리그난 함량이 57.94% 증가하였으며, 각각의 리그난 조성의 변화로는 세사미놀 3 배당체(Sesaminol triglucoside)의 함량이 발효 후 18.93% 감소하였고, 세사미놀 2 배당체(Sesaminol diglucoside) 및 세사미놀(Sesaminol)의 함량은 각각 1340.44%, 2071.43% 증가하는 것으로 확인되었다. 이는 전체적인 리그난의 함량 증가뿐만 아니라 발효대사에 의해 세사미놀 배당체 결합이 분해되어 고분자 배당체 결합은 감소하는 반면 저분자 세사미놀 배당체의 함량은 증가하는 것으로 확인되었다.Table 10 shows the results of analysis of the lignan content of the fermented mixture of sesame seeds and non-fermentation using Bacillus subtilis SRCM103716 strain and Aspergillus oryzae . As a result of the analysis, the total lignan content increased by 57.94%, and the content of sesaminol triglucoside decreased by 18.93% after fermentation, and sesaminol diglucoside and sesaminol as the change in each lignan composition. ) Was found to increase by 1340.44% and 2071.43%, respectively. This was confirmed that the sesaminol glycoside bond is decomposed by fermentation metabolism as well as the overall lignan content increase, and the polymer glycoside bond decreases while the content of the low molecular sesaminol glycoside increases.
triglucosideSesaminol
triglucoside
diglucosideSesaminol
diglucoside
실시예Example 4. 4. 참깨박Sesame seeds 발효 혼합물을 이용한 간장의 품질특성 Quality Characteristics of Soy Sauce Using Fermented Mixture
참깨박 발효 혼합물을 이용한 간장의 제조공정은 도 8과 같다.The manufacturing process of soy sauce using the fermented mixture of sesame seeds is shown in FIG. 8.
1) 참깨박 간장의 일반성분 분석1) Analysis of general ingredients of sesame seeds soy sauce
pH는 pH 미터(pH/mV/TEMP Meter P25, (주)이스텍, 한국)를 이용하여 측정하였다. 염도는 전처리한 시료를 10 ml를 취해 0.1N-AgNO3(F=1.000)로 적정(End point: 적갈색)하고, 하기식으로 계산하였다.The pH was measured using a pH meter (pH / mV / TEMP Meter P25, ISTEK, Korea). Salinity was calculated by the following formula (End point: reddish brown) titrated with 0.1N-AgNO 3 (F = 1.000) by taking 10 ml of the pre-treated sample.
NaCl(%) = (0.1N - AgNO3의 소비량(ml) × 0.00585 × D × F × 100)/SNaCl (%) = (0.1N-AgNO3 consumption (ml) × 0.00585 × D × F × 100) / S
D: 희석배수(250/10)D: dilution factor (250/10)
S: 시료채취량(g)S: sampling amount (g)
0.00585: 0.1N-AgNO3의 ml수에 해당하는 NaCl의 양(g)0.00585: amount of NaCl (g) corresponding to the number of ml of 0.1N-AgNO 3
F: 0.1N-AgNO3의 FactorF: Factor of 0.1N-AgNO 3
또한, 총 질소 함량(T.N)은 마이크로 켈달법(Micro Kjeldahl Method)에 의해 총 질소 함량을 측정하였다. 간장 1 ml을 분해 플라스크에 넣고 분해촉매제 7 g과 황산 12 ml를 가하여 50분간 가열, 분해시켰다. 냉각 후 켈달(Kjeldahl) 증류장치를 이용하여 증류하고 이때 분리되어 나오는 암모니아를 4% 붕산(Boric acid) 용액에 포집하고 0.1N HCl 표준용액으로 적정하였다.In addition, the total nitrogen content (T.N) was measured by the total nitrogen content by the Micro Kjeldahl Method. 1 ml of soy sauce was placed in a decomposition flask, 7 g of a decomposition catalyst and 12 ml of sulfuric acid were added, followed by heating and decomposition for 50 minutes. After cooling, distillation was performed using a Kjeldahl distillation apparatus, and the ammonia separated at this time was collected in a 4% boric acid solution and titrated with a 0.1N HCl standard solution.
총 질소량(%) = (T-B)Total nitrogen content (%) = (T-B)
0.1N - AgNO3의 소비량(ml) × 0.00585 × D × F × 100)/S0.1N-AgNO3 consumption (ml) × 0.00585 × D × F × 100) / S
T: 시료 0.1N HCl 소모량T: Sample 0.1N HCl consumption
B: Blank 0.1N HCl 소모량B: Blank 0.1N HCl consumption
S: 시료 채취량S: sampling amount
순고형분은 간장 5 ml를 취해 해사와 잘 섞고 105℃ 건조기에서 4시간 건조 후 방냉, 칭량하여 전고형물의 함량을 구하여 순고형물 함량을 구하였다.The net solid content was taken 5 ml of soy sauce, mixed well with seaweed, dried for 4 hours in a 105 ° C dryer, then cooled and weighed to obtain the total solid content to obtain the net solid content.
순고형물 함량(%) = (건조 후 무게 - Blank 용기무게 × 20) - 염도(%)Net solids content (%) = (Weight after drying-Blank container weight × 20)-Salinity (%)
색도(ABS값)는 시료 10 ml를 50배 희석하여 UV-VIS 분광광도계(Shimadzu, UV mini-1240)를 이용하여 500 nm에서 흡광도를 측정하였다.As for the chromaticity (ABS value), 10 ml of the sample was diluted 50-fold, and absorbance was measured at 500 nm using a UV-VIS spectrophotometer (Shimadzu, UV mini-1240).
ABS값 = 500 nm ABS값 × 50(희석배수)ABS value = 500 nm ABS value × 50 (dilution multiple)
바실러스 서틸리스(Bacillus subtilis) SRCM103716 균주와 아스퍼질러스 오리재(Aspergillus oryzae)를 이용한 참깨박 간장의 일반분석 결과를 표 11에 나타내었다. 숙성시간이 경과함에 따라 pH는 소폭 낮아지는 경향을 보였고 이는 발효과정에서 생성되는 유기산에 의한 것으로 숙성기간이 길어짐에 따라 유기산의 함량이 증가되어 pH가 소폭 낮아지는 것으로 볼 수 있다. 또한 총 질소 함량, 순추출물, 갈색도는 숙성기간에 따라 증가하는 것으로 나타났으며, 밝기를 나타내는 L값의 경우 숙성이 진행됨에 따라 감소하는 모습을 보였다.Table 11 shows the general analysis results of sesame soy sauce using Bacillus subtilis SRCM103716 strain and Aspergillus oryzae . As the aging time elapsed, the pH tended to decrease slightly. This is due to the organic acid produced in the fermentation process. As the aging period increases, it can be seen that the pH of the organic acid increases and the pH decreases slightly. In addition, the total nitrogen content, net extract, and brownness were found to increase with the aging period, and the L value indicating brightness decreased as aging progressed.
2) 2) 참깨박Sesame seeds 간장의 향기성분 분석 Analysis of aroma components in soy sauce
참깨박 간장으로부터 휘발성 향기 성분을 분리, 동정하였으며, 분석 조건은 표 12와 같다.Volatile fragrance components were separated and identified from sesame seed soy sauce, and the analysis conditions are shown in Table 12.
(60m length × 0.25mm I.D × 0.50㎛)DB-WAX (Agilent)
(60m length × 0.25mm ID × 0.50㎛)
4℃/min ~ 150℃ - 5min
5℃/min ~ 230℃ - 10min40 ℃-3min
4 ℃ / min ~ 150 ℃-5min
5 ℃ / min ~ 230 ℃-10min
바실러스 서틸리스(Bacillus subtilis) SRCM103716 균주와 아스퍼질러스 오리재(Aspergillus oryzae)를 이용한 참깨박 발효 혼합물 비율에 따른 참깨박 간장의 향기성분 분석 결과를 표 13 내지 15에 나타내었다. 참깨박 발효 간장의 휘발성 성분을 분석한 결과 총 61종 화합물이 동정되었다. 이 중에서 알데히드류 5종, 방향족화합물 10종, 알코올류 10종, 산류 3종, 케톤류 10종, 에스테르류 4종, 기타 화합물 19종이 동정되었다. 특징적으로는 단일균주를 이용한 간장에 비해 복합균주를 이용한 간장에서 과일향, 엿기름향을 가진 3-메틸 부타날(3-Methyl butanal)의 함량이 5.28%, 훈연, 나무향, 베이컨향을 갖는 2-메톡시 페놀(2-methoxy phenol)이 17.01%, 과일향, 크림향, 버터향을 갖는 2,3-부탄디올(2,3-Butanediol)은 1.19% 각각 증가함을 확인하였다. 또한, 방향성을 보이지 않는 기타 휘발성 화합물의 함량이 34.3%에서 20.83%로 13.47% 감소하는 경향을 보여 결과적으로 SRCM103716 균주와 아스퍼질러스 오리재(Aspergillus oryzae)를 활용한 복합균주의 간장에서 아스퍼질러스 오리재(Aspergillus oryzae)만을 접종한 단일균주의 간장보다 향기성분의 화합물이 증가하는 것으로 확인되었다.Table 13 to 15 show the results of the analysis of the fragrance components of sesame seeds soy sauce according to the ratio of Bacillus subtilis SRCM103716 strain and Aspergillus oryzae using Aspergillus oryzae . As a result of analyzing the volatile components of the fermented soybean sesame seeds, a total of 61 compounds were identified. Among them, 5 aldehydes, 10 aromatic compounds, 10 alcohols, 3 acids, 10 ketones, 4 esters, and 19 other compounds were identified. Characteristically, the content of 3-methyl butanal with fruity and malt scent in the soy sauce using the complex strain is 5.28%, smoked, wood flavor, and bacon flavor compared to the soy sauce using the single strain. -It was confirmed that 2,3-butanediol (2,3-butanediol) having 2-methoxy phenol (17.01%), fruit flavor, cream flavor, and butter flavor increased by 1.19%. In addition, the content of other non-aromatic volatile compounds showed a tendency to decrease by 13.47% from 34.3% to 20.83%. As a result, aspergillus in the soy sauce of the complex strain using SRCM103716 strain and Aspergillus oryzae It was confirmed that the compound of the aroma component was increased more than the soy sauce of a single strain inoculated with Orijae ( Aspergillus oryzae ) alone.
3) 3) 참깨박Sesame seeds 간장의 유리 아미노산 측정 Measurement of free amino acids in the liver
유리 아미노산은 시료 2 g을 취하여 3차 증류수 30 mL를 넣고 교반하고 50 mL로 정용한 후 초음파를 이용하여 20분간 추출한 후 원심분리(3000 rpm, 10분)한 다음 상등액 2 mL에 5% TCA 2 mL를 넣고 원심분리(10,000 rpm, 10분)한 후 상등액을 취하여 0.02N-HCl로 희석한 후 0.2 ㎛ 실린지 필터에 통과시킨 후 하기 표 16의 분석조건으로 아미노산 분석기로 분석하였다.For free amino acids, take 2 g of sample, add 30 mL of tertiary distilled water, stir, 50 mL, extract for 20 minutes using ultrasonic waves, centrifuge (3000 rpm, 10 min), and 2 mL of 5% TCA in 2 mL of supernatant. After inserting and centrifuging (10,000 rpm, 10 minutes), the supernatant was taken, diluted with 0.02N-HCl, passed through a 0.2 μm syringe filter, and analyzed with an amino acid analyzer under the analysis conditions in Table 16 below.
Hitachi 4.6×40mm (Ammonia filtering)Hitachi 4.6 × 60mm (speration)
Hitachi 4.6 × 40mm (Ammonia filtering)
발효과정 중에 효소작용으로 생성되는 유리 아미노산의 함량이 높을수록 맛과 영양이 뛰어난 우수식품으로 평가된다. 본 실험의 바실러스 서브틸리스(Bacillus subtilis) SRCM103716 균주와 아스퍼질러스 오리재(Aspergillus oryzae)를 이용한 참깨박 발효 혼합물 비율에 따른 발효 간장의 유리 아미노산 결과는 표 17과 같다. 아스퍼질러스 오리재 100% 발효 간장과 아스퍼질러스 오리재 95%, 바실러스 서브틸리스 SRCM103716 5% 발효 간장의 전체 유리 아미노산 함량은 각각 319.3 mg/L, 333.6 mg/L로 측정되었고, 장류의 글루탐산(glutamic acid)은 아스퍼질러스 오리재 100% 발효 간장과 아스퍼질러스 오리재 95%+바실러스 서브틸리스 SRCM103716 5% 발효 간장에서 각각 47.82 mg/L, 70.44 mg/L로 나타났다. 또한, 아스파르트산(aspartic acid)의 함량은 20.99 mg/L, 25.29 mg/L로 측정되어 구수한 맛, 간장 고유의 맛에 밀접한 영향을 미치는 성분들이 아스퍼질러스 오리재 95%+바실러스 서브틸리스 SRCM103716 5% 발효 간장에서 더 높게 나타났다. 쓴맛을 나타내는 유리 아미노산인 이소류신(isoleucine), 류신(leucine), 페닐알라닌(phenylalanine) 3종 모두 결과적으로 SRCM103716 균주+아스퍼질러스 오리재(Aspergillus oryzae)를 활용한 복합 균주의 간장에서 아스퍼질러스 오리재(Aspergillus oryzae)만을 접종한 단일 균주의 간장보다 낮게 나타나, 구수한 맛, 간장 고유의 맛이 증가하고, 쓴맛이 감소함을 확인하였다.The higher the content of free amino acids produced by enzymatic action during the fermentation process, the better the taste and nutrition. The free amino acid results of fermented soy sauce according to the ratio of the Bacillus subtilis SRCM103716 strain and the sesame leaf fermentation mixture using Aspergillus oryzae are shown in Table 17. 100% fermented soy sauce and 95% aspergillus raw material, Bacillus subtilis SRCM103716 The total free amino acid content of 5% fermented soy sauce was measured to be 319.3 mg / L and 333.6 mg / L, respectively, and the glutamic acid of the enteric acid was 100% aspergillus ory and 95% of aspergillus ory + Bacillus subtilis SRCM103716 was found to be 47.82 mg / L and 70.44 mg / L in 5% fermented soy sauce, respectively. In addition, the content of aspartic acid (aspartic acid) was measured as 20.99 mg / L, 25.29 mg / L, and the ingredients that closely affect the savory taste and the unique taste of soy sauce were Aspergillus ducks 95% + Bacillus subtilis SRCM103716 was higher in 5% fermented soy sauce. As a result, all three of the free amino acids, isoleucine, leucine, and phenylalanine, which are bitter tastes, result in SRCM103716 strain + Aspergillus orijae ( Aspergillus oryzae ) in the soy sauce of a complex strain using Aspergillus ( Aspergillus) oryzae ) showed lower than the soy sauce of the single strain inoculated only, and it was confirmed that the savory taste, soy sauce-specific taste increased, and the bitter taste decreased.
<110> CHARM GO EUL CO., LTD. <120> Method for producing soy sauce using Bacillus subtilis SRCM103716 and Aspergillus oryzae strain <130> PN19312 <160> 1 <170> KoPatentIn 3.0 <210> 1 <211> 478 <212> DNA <213> Bacillus subtilis <400> 1 cttctcagga cgaacgctgg cggcgtgcct aatacatgca agtcgagcgg acagatggga 60 gcttgctccc tgatgttagc ggcggacggg tgagtaacac gtgggtaacc tgcctgtaag 120 actgggataa ctccgggaaa ccggggctaa taccggatgg ttgtttgaac cgcatggttc 180 aaacataaaa ggtggcttcg gctaccactt acagatggac ccgcggcgca ttagctagtt 240 ggtgaggtaa cggctcacca aggcaacgat gcgtagccga cctgagaggg tgatcggcca 300 cactgggact gagacacggc ccagactcct acgggaggca gcagtaggga atcttccgca 360 atggacgaaa gtctgacgga gcaacgccgc gtgagtgatg aaggttttcg gatcgtaaag 420 ctctgttgtt agggaagaac aagtaccgtt cgaatagggc ggtaccttga cggtacct 478 <110> CHARM GO EUL CO., LTD. <120> Method for producing soy sauce using Bacillus subtilis SRCM103716 and Aspergillus oryzae strain <130> PN19312 <160> 1 <170> KoPatentIn 3.0 <210> 1 <211> 478 <212> DNA <213> Bacillus subtilis <400> 1 cttctcagga cgaacgctgg cggcgtgcct aatacatgca agtcgagcgg acagatggga 60 gcttgctccc tgatgttagc ggcggacggg tgagtaacac gtgggtaacc tgcctgtaag 120 actgggataa ctccgggaaa ccggggctaa taccggatgg ttgtttgaac cgcatggttc 180 aaacataaaa ggtggcttcg gctaccactt acagatggac ccgcggcgca ttagctagtt 240 ggtgaggtaa cggctcacca aggcaacgat gcgtagccga cctgagaggg tgatcggcca 300 cactgggact gagacacggc ccagactcct acgggaggca gcagtaggga atcttccgca 360 atggacgaaa gtctgacgga gcaacgccgc gtgagtgatg aaggttttcg gatcgtaaag 420 ctctgttgtt agggaagaac aagtaccgtt cgaatagggc ggtaccttga cggtacct 478
Claims (5)
(2) 분쇄한 참깨박에 물을 0.4~0.6배 가수하여 110~130℃에서 10~20분 동안 증자한 후 냉각한 참깨박에 아스퍼질러스 오리재(Aspergillus oryzae) 균주를 접종한 후 28~32℃에서 68~76시간 동안 발효하여 참깨박 발효물을 제조하는 단계;
(3) 상기 (1)단계의 제조한 참깨박 고초균 발효물과 상기 (2)단계의 제조한 참깨박 발효물을 4~6:94~96 중량비율로 혼합하여 리그난 함량 및 항산화 활성이 증진된 참깨박 혼합 발효물의 제조하는 단계; 및
(4) 상기 (3)단계의 제조한 참깨박 혼합 발효물에 23~25%(w/v) 염수를 2~3배(v/w) 첨가하여 30~35℃에서 3~4주 동안 숙성시킨 후 여과하는 단계를 포함하여 제조하는 것을 특징으로 하는 참깨박 발효 간장의 제조방법.(1) Water is added to the crushed sesame seeds 1.2 to 1.8 times, and the water is increased at 110 to 130 ° C for 10 to 20 minutes, and the cooled sesame seeds have excellent β-glucosidase activity, and Bacillus Bacillus cereus , Staphylococcus aureus and Listeria monocytogenes have antimicrobial and β-hemolytic activity against harmful microorganisms, harmful substances of phenylalanine, The ability to secrete protease, amylase and cellulase without producing harmful enzymes of urease and β-glucuronidase and biogenic amines of tyramine and histamine Bacillus subtilis (Bacillus subtilis) SRCM103716 strain (Accession No .: KFCC11832P) was inoculated and fermented at 34-40 ° C. for 20-28 hours to total polyphenol and total flavonoid content and β-glucosidase The (β-glucosidase) comprising: an activity of promoting the production Sesame Bacillus subtilis fermentation;
(2) Water is added to the crushed sesame seeds 0.4 ~ 0.6 times, and the water is increased at 110 ~ 130 ℃ for 10 ~ 20 minutes. After cooling, the cooled sesame seeds are inoculated with Aspergillus oryzae strain, and then 28 ~ Preparing fermented sesame seeds by fermenting at 32 ° C. for 68 to 76 hours;
(3) The sesame leaf fermentation product produced in the step (1) and the sesame leaf fermentation product prepared in the step (2) are mixed at a weight ratio of 4 to 6:94 to 96 to improve the lignan content and antioxidant activity. Preparing a sesame-bak mixed fermentation product; And
(4) 23 ~ 25% (w / v) brine is added 2-3 times (v / w) to the mixed fermented sesame leaf prepared in the step (3), aged at 30 ~ 35 ℃ for 3 ~ 4 weeks Method of producing a fermented soy sauce with sesame seeds, characterized in that it comprises a step of filtering after the let.
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Citations (4)
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US3914436A (en) * | 1972-12-11 | 1975-10-21 | Noda Inst For Scientific Res | Process for manufacturing soy sauce using enzymatic preparation(s) |
JP2007319157A (en) * | 2006-06-02 | 2007-12-13 | Food Industry Research & Development Institute | Sesame-fermented composition and method for producing the same |
KR20120001875A (en) * | 2010-06-30 | 2012-01-05 | 대상 주식회사 | Process for preparing sesame sauce |
KR20190020359A (en) * | 2017-08-21 | 2019-03-04 | 김길수 | Method for Preparing Sesami Sauce Using Sesami Cake |
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Publication number | Priority date | Publication date | Assignee | Title |
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US3914436A (en) * | 1972-12-11 | 1975-10-21 | Noda Inst For Scientific Res | Process for manufacturing soy sauce using enzymatic preparation(s) |
JP2007319157A (en) * | 2006-06-02 | 2007-12-13 | Food Industry Research & Development Institute | Sesame-fermented composition and method for producing the same |
KR20120001875A (en) * | 2010-06-30 | 2012-01-05 | 대상 주식회사 | Process for preparing sesame sauce |
KR20190020359A (en) * | 2017-08-21 | 2019-03-04 | 김길수 | Method for Preparing Sesami Sauce Using Sesami Cake |
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