KR20170022516A - Seed culture for soybean sauce and preparation method of soybean sauce using the same - Google Patents
Seed culture for soybean sauce and preparation method of soybean sauce using the same Download PDFInfo
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- KR20170022516A KR20170022516A KR1020150117608A KR20150117608A KR20170022516A KR 20170022516 A KR20170022516 A KR 20170022516A KR 1020150117608 A KR1020150117608 A KR 1020150117608A KR 20150117608 A KR20150117608 A KR 20150117608A KR 20170022516 A KR20170022516 A KR 20170022516A
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- soy sauce
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- acid
- fermented
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Abstract
Description
본 발명은 간장발효 종균 및 이를 이용한 간장의 제조방법에 관한 것이다.The present invention relates to a soybean fermentation broth and a method for producing soy sauce using the same.
메주는 전통적인 한국의 콩의 누룩이다. 메주는 식종을 위한 특정한 미생물에 의존하지 않고 자연발효에 의해 제조된다. 콩의 누룩의 발효를 위한 대부분의 미생물들은 건조 볏짚, 제조자의 손, 기구, 발효실, 및 주위 공기로부터 비롯되며, 메주 성분은 주된 미생물 종과 시간, 장소, 제조자, 기구에 따라 우발적으로 변하는 미생물 다양성 때문에 환경조건에 따라 잘 변한다.Meju is the yeast of traditional Korean beans. Meju is produced by natural fermentation without relying on specific microorganisms for breeding. Most microorganisms for the fermentation of soybean koji are derived from dry rice straw, the manufacturer's hand, appliance, fermentation room, and ambient air, and the meju ingredient contains microorganism varieties that vary accidentally depending on the main microorganism species and time, place, manufacturer, So it changes according to the environmental conditions.
메주를 제조하기 위해선 찐 콩을 으깨서 성형하여 놓아두어 공기 중에 있는 여러 미생물이 부착하여 증식되도록 하여, 이들 미생물이 분비하는 여러 효소들에 의하여 메주 원료인 콩이 분해되는 발효숙성 과정을 거쳐야한다. 메주 표면에 증식하는 리조푸스속(Rhizopus속), 아스퍼질러스속(Aspergillus속), 뮤코속(Mucor속) 등 αw 0.75 내외를 선호하는 호기성 곰팡이가 분비하는 전분질 분해효소(amylase), 단백질 분해효소(protease), 지방분해효소(lipase) 등에 의하여 콩이 분해되고, 메주 내부에서는 αw 0.9 내외를 선호하는 바실러스 서브틸리스(Bacillus subtilis), 바실러스 퓨밀러스(B. pumilus) 등이 분비하는 단백질 분해효소, 전분질 분해효소, 지방분해효소 등이 분해에 관여한다. In order to produce meju, steamed beans are crushed and molded so that various microorganisms in the air adhere and proliferate, and fermentation aging process in which soybean as a raw material of meju is decomposed by various enzymes secreted by these microorganisms should pass through. Amylase and proteolytic enzyme secreted by aerobic fungus, which prefer αw 0.75 such as Rhizopus spp., Aspergillus spp. And Mucor spp. protease and lipase. In Meju, the proteolytic enzymes secreted by Bacillus subtilis, B. pumilus and the like, which are preferred to be within about 0.9 w / Starcholytic enzymes, and lipolytic enzymes are involved in degradation.
한편, 간장은 한국에서 6 달 내지 1 년 동안 소금물에 메주 담금에 의해 제조된다. 찐 대두의 단백질, 탄화수소, 및 지질과 같은 일부 유기화합물들은 발효메주에서 자란 미생물의 세포 외 효소에 의해 분해되어 가용성 유기화합물로 전환된 아미노산, 그밖에 저급화 분해산물이 가용성화 되어 소금물에 의해 화학적, 물리적으로 용출되고, 내염성세균, 효모 등에 의해서 풍미물질이 생성되면서 숙성된 것이다. 그러나 고염분 상태 하에 생성된 삼투압에 의해 미생물 활성이 멈출 수 있기 때문에 미생물들에 의해 생물학적으로 영향을 미치지 않는다. 그러므로 간장의 질은 오직 발효 메주 내의 미생물 활성에 의해 생성된 가용성 유기화합물의 종류와 특성에 의해서만 결정된다.On the other hand, soy sauce is produced in Korea for 6 months to 1 year by salt-in-water. Some organic compounds such as proteins, hydrocarbons, and lipids in steamed soybeans are converted to soluble organic compounds by degradation by extracellular enzymes of microorganisms grown in fermented Meju, and other degradation products are solubilized, Physically leached, and matured as a flavor substance was produced by salt-resistant bacteria and yeast. However, osmotic pressure produced under high salinity conditions does not biologically affect microbial activity because it can stop microbial activity. Therefore, the quality of the liver is determined solely by the type and nature of the soluble organic compounds produced by the microbial activity in the fermented meju.
종래 전통간장용 메주를 만드는 방법은 실온에서 12 시간 동안 물에 담근 콩을 삶고, 절구에 찧어 성형을 한 후, 2 내지 3 일 동안 겉말림을 한 후, 4 주 동안 짚을 포개어 씌워 재운 후, 햇볕에 말리고, 다시 2 개월 동안 재우는 과정으로 진행되었다. 이러한 종래메주의 제조공정을 거쳐야하기 때문에, 전통간장의 제조는 복잡하고, 오랜 기간이 걸린다는 단점이 있어, 간장의 단기 제조방법 개발이 필요한 실정이다. 또한, 종래 전통간장은 암모니아 성분과 불필요한 잡균의 과다 발생으로 인해 불쾌한 냄새를 풍긴다는 단점이 있다. 대한민국 등록특허 제 10-1399276호에 단기숙성 간장의 제조방법 및 상기 방법으로 제조된 단기숙성 간장, 및 대한민국 등록특허 제10-2090770호에 불쾌취가 제거된 재래식 간장 및 분말간장 제조방법이 개시되어 있으나, 본 발명의 발효간장의 제조방법과는 상이하다.Conventional traditional soy sauce meju is prepared by boiling soybeans dipped in water for 12 hours at room temperature, molding on a mortar, pestling for 2 to 3 days, covering the straw for 4 weeks, And dried again for 2 months. Since the conventional process for preparing meju is required to be performed, conventional soy sauce production is complicated and takes a long period of time. Therefore, it is necessary to develop a short-term production method of soy sauce. In addition, the conventional soy sauce has a disadvantage in that an unpleasant odor is produced due to excessive production of ammonia component and unnecessary germs. Korean Patent No. 10-1399276 discloses a process for producing a short-term ripe soy sauce, a short-term ripe soy sauce prepared by the above method, and a method for producing a conventional soy sauce and powder sauce in which unpleasant odor is removed in Korean Patent No. 10-2090770 However, this method is different from the method for producing fermented soy sauce according to the present invention.
본 발명의 목적은 간장발효를 위한 신규의 균주로서, 염도가 높은 환경에서도 잘 생육하며, 단백분해활성이 있는 신규의 균주를 제공하는 것이다.It is an object of the present invention to provide a novel strain for soybean fermentation which is well-grown in a high saltiness environment and has a proteolytic activity.
또한, 본 발명의 다른 목적은 메주의 제조과정 없이, 상기 간장발효를 위한 신규의 균주의 배양에 의해 직접 제조된 간장을 제공하는 것이다. Another object of the present invention is to provide a soy sauce which is directly produced by culturing a new strain for soy sauce fermentation without the process of preparing meju.
또한, 본 발명의 또 다른 목적은 상기 신규의 균주로 발효된 간장을 제조하는 방법을 제공하는 것이다.It is still another object of the present invention to provide a method for producing soy sauce fermented with the new strain.
본 발명은 내염성이 있으며, 단백분해활성이 있는 오셔노바실러스 김치(Oceanobacillus kimchii sosab) 균주[기탁번호: KFCC11607P]를 제공한다.The present invention provides a strain of Oceanobacillus kimchii sosab (Accession No: KFCC11607P) which is resistant to salt and has proteolytic activity.
또한 본 발명은 내염성이 있으며, 단백분해활성이 있는 바실러스 퓨밀러스 (Bacillus pumilus sosad) 균주[기탁번호: KFCC11608P]를 제공한다.The present invention also provides a Bacillus pumilus sosad strain having a salt resistance and proteolytic activity [Deposit number: KFCC11608P].
또한 본 발명은 오셔노바실러스 김치(Oceanobacillus kimchii sosab) 균주[기탁번호: KFCC11607P], 및 바실러스 퓨밀러스 (Bacillus pumilus sosad) 균주[기탁번호: KFCC11608P], 또는 두 균주의 혼합균주를 포함하는 간장 발효 종균을 제공한다.The present invention also relates to a soybean fermented soybean comprising a strain of Oceanobacillus kimchii sosab [Accession No .: KFCC11607P], a strain of Bacillus pumilus sosad [Accession No .: KFCC11608P], or a mixed strain of two strains .
또한 본 발명은 오셔노바실러스 김치(Oceanobacillus kimchii sosab) 균주[기탁번호: KFCC11607P], 및 바실러스 퓨밀러스 (Bacillus pumilus sosad) 균주[기탁번호: KFCC11608P], 또는 두 균주의 혼합균주를 포함하는 발효식품을 제공한다.The present invention also relates to a fermented food comprising a strain of Oceanobacillus kimchii sosab [Accession No .: KFCC11607P], a strain of Bacillus pumilus sosad [Accession No .: KFCC11608P], or a mixture of two strains to provide.
상기 발효식품은 간장일 수 있다.The fermented food may be a soy sauce.
상기 발효식품은 아스파르트산, 트레오닌, 세린, 글루탐산, 프롤린, 글리신, 알라닌, 발린, 메티오닌, 이소류신, 류신, 타이로신, 페닐알라딘, 히스티딘, 라이신, 및 아르기닌 중에서 선택되는 1종 이상의 유리아미노산을 함유할 수 있다.The fermented food may contain one or more free amino acids selected from aspartic acid, threonine, serine, glutamic acid, proline, glycine, alanine, valine, methionine, isoleucine, leucine, tyrosine, phenylalanine, histidine, lysine, have.
상기 발효식품은 유기 질소화합물 및 암모늄, 또는 두 가지 모두 함유할 수 있다.The fermented food may contain an organic nitrogen compound and ammonium, or both.
상기 발효식품은 마그네슘, 칼슘, 나트륨, 및 칼륨 중에서 선택되는 1종 이상의 무기물을 함유할 수 있다.The fermented food may contain one or more minerals selected from magnesium, calcium, sodium, and potassium.
상기 발효식품은 시트르산, 말산, 숙신산, 락트산, 아세트산, 및 피롤리돈카복실산 중에서 선택되는 1종 이상의 유기산을 함유할 수 있다.The fermented food may contain one or more organic acids selected from citric acid, malic acid, succinic acid, lactic acid, acetic acid, and pyrrolidonecarboxylic acid.
또한 본 발명은 콩 원료에 오셔노바실러스 김치(Oceanobacillus kimchii) sosab 균주[기탁번호: KFCC11607P], 및 바실러스 퓨밀러스 (Bacillus pumilus) sosad 균주[기탁번호: KFCC11608P], 또는 두 균주의 혼합균주를 포함하는 간장발효 종균을 이용한 간장의 제조방법을 제공한다.The present invention also relates to a method for producing soybean oil comprising the steps of: (a) cultivating a soybean material comprising a strain of Oceanobacillus kimchii sosab [Accession No .: KFCC11607P] and a strain of Bacillus pumilus sosad [accession number: KFCC11608P] A method for producing soy sauce using soybean fermentation seeds is provided.
또한 본 발명은 (a) 염수 및 콩 원료가 함유된 액상배지에 종균을 접종하고, 배양하여 배양물을 수득하는 단계; (b) 상기 수득한 배양물을 여과하여 여과액을 수득하는 단계; (c) 상기 수득한 여과액을 침전하여 간장을 수득하는 단계;를 포함하는 것을 특징으로 하는 간장발효 종균을 이용한 간장의 제조방법을 제공한다.The present invention also relates to a method for producing a culture medium, comprising the steps of: (a) inoculating a seed medium into a liquid medium containing a salt water and a soybean material and culturing to obtain a culture; (b) filtering the obtained culture to obtain a filtrate; and (c) precipitating the obtained filtrate to obtain a soy sauce. The present invention also provides a process for producing soy sauce using the fermented soy sauce.
상기 염수의 염도는 10 내지 18%일 수 있다. The salinity of the saline solution may be 10 to 18%.
상기 액상배지는 콩 원료 대 소금 대 물의 중량비가 0.13-0.20 : 0.10-0.18 : 1일 수 있다.The weight ratio of the soybean raw material to the salt to water may be 0.13-0.20: 0.10-0.18: 1.
상기 종균은 오셔노바실러스 김치(Oceanobacillus kimchii) sosab 균주[기탁번호: KFCC11607P], 및 바실러스 퓨밀러스(Bacillus pumilus) sosad 균주[기탁번호: KFCC11608P], 또는 두 균주의 혼합균주인 것을 특징으로 하는 간장발효 종균일 수 있다. Wherein the seed is a mixed strain of Oceano Bacillus kimchii sosab [Accession No .: KFCC11607P], Bacillus pumilus sosad strain [Accession No .: KFCC11608P], or two strains. It may be a seed.
상기 배양은 2 내지 3 달 동안, 30 내지 40 ℃에서 수행될 수 있다.The cultivation may be carried out at 30 to 40 DEG C for 2 to 3 months.
상기 (b)단계 이전에 상기 (a)단계의 배양물을 55 내지 65 분 동안, 65 내지 75 ℃에서 살균하는 단계를 추가할 수 있다.The step (a) may be performed at a temperature of 65 to 75 ° C for 55 to 65 minutes before the step (b).
본 발명에 따른 간장발효 종균을 이용한 간장의 제조방법을 통해 종래의 메주의 제조과정을 생략하고, 염도가 높은 환경에서 잘 생육하며, 단백분해활성이 있는 신규의 종균으로 간장을 발효하여, 간장의 제조시간을 단축할 수 있다.The process for preparing soy sauce using the soy sauce fermentation broth according to the present invention omits the conventional process for preparing meju, and grows well in a high salinity environment. The soy sauce is fermented by a new seed having proteolytic activity, The manufacturing time can be shortened.
또한, 상기 신규의 종균으로 발효된 간장은 맛, 향미, 및 성분 면에서 한국의 전통 간장과 매우 유사하거나, 우위의 풍미를 나타낼 수 있다. In addition, the soy sauce fermented with the new seeds may exhibit a flavor similar to that of Korean traditional soy sauce in terms of taste, flavor, and ingredient, or a superior flavor.
도 1은 200g/L 탈지대두박을 함유한 18% 천일염 용액에서의 오셔노 바실러스 김치 균주와 바실러스 퓨밀러스 균주의 대두 단백질에 대한 가수분해활성을 나타낸 사진이다.
도 2는 200g/L 탈지대두박을 함유한 18% 천일염 용액에서의 (A) 오셔노바실러스 김치, (B) 바실러스 퓨밀러스, 및 (AB) 오셔노바실러스 김치와 바실러스 퓨밀러스의 혼합물의 액침배양에 의한 탈지대두박의 발효과정을 나타낸 사진이다.
도 3은 (A) 오셔노바실러스 김치, (B) 바실러스 퓨밀러스, 및 (C) 오셔노바실러스 김치와 바실러스 퓨밀러스의 혼합물의 탈지대두박 액침배양으로부터 생성된 간장의 사진이다. (D), (E), 및 (F)는 발효 대두박으로부터 간장을 추출하고 남은 찌꺼기를 나타낸 사진이다.FIG. 1 is a photograph showing hydrolytic activity of soybean protein of Oshino Bacillus Kimchi strain and Bacillus subtilis strain in an 18% salting solution containing 200 g / L defatted soybean meal.
Fig. 2 shows the results of immersion culture of a mixture of (A) Oshino Bacillus Kimchi, (B) Bacillus pumilus, and (AB) Oshino Bacillus Kimchi and Bacillus pumilus in 18% salting solution containing 200 g / L defatted soybean meal Fig. 2 is a photograph showing the fermentation process of defatted soybean meal.
FIG. 3 is a photograph of soy sauce produced from defatted soybean immersion culture of (A) Oshino Bacillus Kimchi, (B) Bacillus pumilus, and (C) a mixture of Oshino Bacillus Kimchi and Bacillus pumilus. (D), (E), and (F) are photographs showing residual debris obtained by extracting soy sauce from fermented soybean meal.
본 발명은 내염성이 있으며, 단백분해활성이 있는 오셔노바실러스 김치(Oceanobacillus kimchii) sosab 균주[기탁번호: KFCC11607P]에 관한 것이고, 또한 내염성이 있으며, 단백분해활성이 있는 바실러스 퓨밀러스 (Bacillus pumilus) sosad 균주[기탁번호: KFCC11608P]에 관한 것이다. The present invention relates to an Oceanobacillus kimchii sosab strain [Accession No .: KFCC11607P] which is resistant to salt and has a proteolytic activity and is also resistant to salt and has a proteolytic activity, such as Bacillus pumilus sosad Strain [Accession No .: KFCC11608P].
본 발명의 간장 발효용 균주 중 첫 번째 균주는 동정 결과 오셔노바실러스 김치(Oceanobacillus kimchii)에 속했으며, 이를 오셔노바실러스 김치(Oceanobacillus kimchii) sosab (서열번호 1)으로 명명하고, 2015년 1월 28일자로 한국미생물보존센터에 기탁하여 기탁번호를 KFCC11607P로 부여받았다. 또한, 두 번째 균주는 동정 결과 바실러스 퓨밀러스(Bacillus pumilus)에 속했으며, 이를 바실러스 퓨밀러스(Bacillus pumilus) sosad (서열번호2)로 명명하고, 2015년 1월 28일자로 한국미생물보존센터에 기탁번호를 KFCC11608P로 부여받았다. The first strain of the soybean fermentation strain of the present invention belongs to Oceano bacillus kimchii as a result of identification and is named as Oceanobacillus kimchii sosab (SEQ ID NO: 1) And deposited it with KFCC 11607P as the deposit number. Also, the second strain belongs to Bacillus pumilus as a result of identification and is designated as Bacillus pumilus sosad (SEQ ID NO: 2), and deposited on January 28, 2015 with the Korean Society for Microbiology As KFCC11608P.
또한 본 발명은 오셔노바실러스 김치(Oceanobacillus kimchii) sosab 균주[기탁번호: KFCC11607P], 및 바실러스 퓨밀러스 (Bacillus pumilus) sosad 균주[기탁번호: KFCC11608P], 또는 두 균주의 혼합균주를 포함하는 간장 발효용 종균에 관한 것이고, 또한 본 발명은 오셔노바실러스 김치(Oceanobacillus kimchii) sosab 균주[기탁번호: KFCC11607P], 및 바실러스 퓨밀러스 (Bacillus pumilus) sosad 균주[기탁번호: KFCC11608P], 또는 두 균주의 혼합균주를 포함하는 발효식품에 관한 것이다.The present invention also relates to a method for soybean fermentation which comprises a strain of Oceanobacillus kimchii sosab [Accession No .: KFCC11607P] and a strain of Bacillus pumilus sosad [Accession No .: KFCC11608P], or a mixture of two strains And the present invention relates to a method for producing a yeast strain according to the present invention, which comprises mixing a strain of Oceanobacillus kimchii sosab [Accession No .: KFCC11607P] and a strain of Bacillus pumilus sosad [Accession No .: KFCC11608P] ≪ / RTI >
상기 본 발명의 간장 발효용 균주는, 제조되는 발효간장의 맛, 향미, 및 성분 면에서 한국의 전통 간장과 매우 유사하거나, 우위의 풍미를 나타낼 수 있고, 종래에 메주의 제조과정을 생략하고, 상기 균주로 간장을 발효하여, 간장의 제조시간을 단축할 수 있다.The soybean fermentation strain of the present invention can exhibit a flavor superior to that of Korean traditional soy sauce in terms of taste, flavor, and ingredients of the fermented soy sauce to be produced, By fermenting soy sauce with the above-mentioned strain, the production time of soy sauce can be shortened.
상기 발효식품은 간장, 고추장, 된장, 청국장일 수 있고, 바람직하게는 상기 발효식품은 간장이다.The fermented food may be soy sauce, hot pepper paste, soybean paste, and chongkukjang. Preferably, the fermented food is soy sauce.
상기 발효식품은 아스파르트산, 트레오닌, 세린, 글루탐산, 프롤린, 글리신, 알라닌, 발린, 메티오닌, 이소류신, 류신, 타이로신, 페닐알라딘, 히스티딘, 라이신, 및 아르기닌 중에서 선택되는 1종 이상의 유리아미노산을 함유할 수 있다. 또한 발효식품은 유기 질소화합물 및 암모늄, 또는 두 가지 모두 함유할 수 있다. 또한 마그네슘, 칼슘, 나트륨, 및 칼륨 중에서 선택되는 1종 이상의 무기물을 함유할 수 있다. 또한 시트르산, 말산, 숙신산, 락트산, 아세트산, 및 피롤리돈카복실산 중에서 선택되는 1종 이상의 유기산을 함유할 수 있다. The fermented food may contain one or more free amino acids selected from aspartic acid, threonine, serine, glutamic acid, proline, glycine, alanine, valine, methionine, isoleucine, leucine, tyrosine, phenylalanine, histidine, lysine, have. The fermented food may also contain organic nitrogen compounds and ammonium, or both. It may also contain at least one inorganic substance selected from magnesium, calcium, sodium, and potassium. It may also contain at least one organic acid selected from citric acid, malic acid, succinic acid, lactic acid, acetic acid, and pyrrolidone carboxylic acid.
이는 종래의 전통간장, 및 양조간장에 함유된 성분들로서, 본 발명에 따른 간장발효 종균으로 발효된 발효식품 또한 15 가지 유리아미노산, 4 가지 무기물, 및 4 가지 유기산이 검출되어, 상기의 성분들을 함유하는 것을 실험예 2 내지 5를 통해서 확인하였다. 또한 두 균주의 혼합 균주의 혼합배양액에 의해 제조된 간장 내의 유리아미노산과 총 가용성 질소 화합물, 구연산, 젖산 성분은 한 가지 균주에 의해 제조된 것보다 약간 더 높게 나타났다. 이러한 결과들은 전통간장이 메주의 제조과정 없이, 탈지대두박으로부터 직접 제조될 수 있는 가능성을 보여준다.This is a component contained in conventional traditional soy sauce and brewed soy sauce, and 15 free amino acids, 4 inorganic acids, and 4 organic acids were detected as the fermented food fermented with the soy sauce fermentation seed according to the present invention, Was confirmed through Experimental Examples 2 to 5. The free amino acids and total soluble nitrogen compounds, citric acid and lactic acid contents in soy sauce prepared by the mixed culture of mixed strains of two strains were slightly higher than those produced by one strain. These results show the possibility that traditional soy sauce can be prepared directly from defatted soybean meal without Meju production process.
또한 본 발명은 콩 원료에 오셔노바실러스 김치(Oceanobacillus kimchii) sosab 균주[기탁번호: KFCC11607P], 및 바실러스 퓨밀러스 (Bacillus pumilus) sosad 균주[기탁번호: KFCC11608P], 또는 두 균주의 혼합균주를 포함하는 간장 발효용 종균을 접종하는 발효간장의 제조방법에 관한 것이고, 또한 (a) 염수 및 콩 원료가 함유된 액상배지에 종균을 접종하고, 배양하여 배양물을 수득하는 단계; (b) 상기 수득한 배양물을 여과하여 여과액을 수득하는 단계; (c) 상기 수득한 여과액을 침전하여 간장을 수득하는 단계;를 포함하는 것을 특징으로 하는 간장발효 종균을 이용한 간장의 제조방법에 관한 것이다.The present invention also relates to a method for producing soybean oil comprising the steps of: (a) cultivating a soybean material comprising a strain of Oceanobacillus kimchii sosab [Accession No .: KFCC11607P] and a strain of Bacillus pumilus sosad [accession number: KFCC11608P] The present invention also relates to a method for producing fermented soy sauce which is inoculated with soy sauce fermentation seeds, which method comprises the steps of: (a) inoculating the seed medium to a liquid medium containing a salt water and a soybean material and culturing to obtain a culture; (b) filtering the obtained culture to obtain a filtrate; (c) precipitating the obtained filtrate to obtain a soy sauce. The present invention also relates to a method for producing soy sauce using the fermented soy sauce.
상기 염수의 염도는 10 내지 18%일 수 있다.The salinity of the saline solution may be 10 to 18%.
보편적으로 일반 종속영양세균은 염도가 3% 이상이면 생장이 크게 저해되고, 5% 이상이면 생장이 정지된다. 이에 따라서, 종래의 전통간장의 경우 고체배지의, 염분이 존재하지 않는 상태에서 종균을 배양한 다음에, 소금물을 혼합함으로써, 제조되는 간장의 염도를 맞춰주었다. Generally, in the case of general heterotrophic bacteria, growth of 3% or more is greatly inhibited, and growth of 5% or more stops growth. Accordingly, in the case of conventional traditional soy sauce, the salt content of the soy sauce is adjusted by culturing the seed culture in the absence of saline in the solid medium, followed by mixing with salt water.
이에 반해 본 발명은 10 내지 18%의 높은 염도에서도 내염성이 있고, 단백분해활성을 갖는 종균을 이용함으로써, 콩 원료, 종균, 및 소금물을 한 번에 혼합하고, 방치하는 방식의 편리한 단기숙성 발효간장의 제조방법을 제공할 수 있다. 18% 이하의 염도에서도 종균을 이용하여 간장을 제조할 수 있으나, 3% 이하의 낮은 염도에서는 잡균이 발생하여, 부패 및 불쾌취를 유발할 수 있으며, 간장의 염도를 맞춰주기 위해 따로 소금을 추가 첨가해야하기 때문에, 바람직하게는 15 내지 18% 소금물을 이용하는 것이 적합하다.On the other hand, the present invention provides a convenient short-term aged fermented soy sauce having a salt resistance even at a high salt content of 10 to 18% and a method of mixing soybean raw material, Can be provided. Although soy sauce can be prepared using seeds at a salinity of 18% or less, it is possible to produce germs at a low salinity of 3% or less, which may lead to decay and unpleasant odor. To further adjust the salinity of the soy sauce, , It is preferable to use salt water of preferably 15 to 18%.
상기 액상배지는 콩 원료 대 소금 대 물의 중량비가 0.13-0.20 : 0.10-0.18 : 1이 되도록 혼합한 액상배지일 수 있다. The liquid medium may be a liquid medium in which the weight ratio of the soybean raw material to the salt to water is 0.13-0.20: 0.10-0.18: 1.
보편적으로 된장에서 생존하는 세균은 내염성이지만 단백질을 분해하여 생장하는 능력이 없기 때문에 고체평판배지에서 생장하지 못하며, 액상배지에는 초기 종균의 빠른 생장을 위해 효모추출물 0.1%가 추가될 수도 있으며, 효모추출물이 필수요소는 아니다. In general, the bacteria that survive in doenjang are salt tolerant but do not grow in solid plate culture medium because they do not have the ability to decompose and grow proteins. In the liquid medium, 0.1% yeast extract may be added for early growth of early seeds, This is not an essential element.
또한 소금의 중량비가 상기 하한치 미만인 경우에는 잡균이 발생하여 부패 및 불쾌취를 유발할 수 있으며, 상기 상한치 초과인 경우에는 발효균의 생장이 제한되어 발효의 효율이 낮아지거나, 발효가 중단되는 문제가 발생할 수 있다. 또한 탈지대두박의 중량비가 상기 하한치 미만인 경우에는 발효식품에서 나타나는 성분의 감소가 나타나거나, 풍미를 약화시킬 수 있으며, 상기 상한지 초과인 경우에는 고농도의 탈지대두박에 대한 효소의 단백질 분해활성이 부분적으로 일어날 수 있어, 간장의 발효가 제대로 이루어지지 않는 문제가 발생할 수 있다. If the weight ratio of the salt is less than the lower limit value, germs may be generated to cause spoilage and unpleasant odor. If the salt is above the upper limit value, the growth of the fermenting bacteria may be limited and the fermentation efficiency may be lowered, have. In addition, when the weight ratio of defatted soybean is below the lower limit, the component appearing in the fermented food may appear to decrease or the flavor may be weakened. If the weight of the defatted soybean is above the upper limit, And the fermentation of the soy sauce may not be properly performed.
상기 종균은 오셔노바실러스 김치(Oceanobacillus kimchii) sosab 균주[기탁번호: KFCC11607P], 및 바실러스 퓨밀러스(Bacillus pumilus) sosad 균주[기탁번호: KFCC11608P], 또는 두 균주의 혼합균주인 것을 특징으로 하는 간장발효 종균을 이용할 수 있다. Wherein the seed is a mixed strain of Oceano Bacillus kimchii sosab [Accession No .: KFCC11607P], Bacillus pumilus sosad strain [Accession No .: KFCC11608P], or two strains. Seeds can be used.
도 1을 통해 오셔노바실러스 김치 균주, 및 바실러스 퓨밀러스 균주가 지니고 있는 단백질 가수분해 활성을 확인할 수 있다. 두 균주는 18% 염분 조건에서 콩 단백질을 가용성 펩티드와 아미노산으로 분해할 수 있음을 확인하였다.1, the protein hydrolysis activity of Oshino Bacillus Kimchi strain and Bacillus subtilis strain can be confirmed. Both strains were found to be capable of digesting soy protein into soluble peptides and amino acids under 18% salt conditions.
상기 배양은 2 내지 3 달 동안, 30 내지 40 ℃에서 수행될 수 있다. 통상적으로 액침배양의 온도는 상기 온도에서 수행되는데, 이는 상기 온도에서 골고루 접종하여 균체가 균일하게 성장할 수 있기 때문이다.The cultivation may be carried out at 30 to 40 DEG C for 2 to 3 months. Typically, the temperature of the immersion culture is carried out at this temperature, since the cells can grow uniformly evenly by inoculation at this temperature.
상기 (b) 단계 이전에 상기 (a) 단계의 배양물을 55 내지 60 분 동안, 65 내지 75 ℃에서, 바람직하게는 60 분 동안 70 ℃에서 살균하는 단계를 추가할 수 있다. 배양물을 살균하지 않는 경우에는 종균이 지속적으로 생장하여 간장이 탁해지거나 영양성분이 변할 수 있다. The step of sterilizing the culture of step (a) at 55 占 폚 for 60 minutes and at 65 占 폚 to 75 占 폚, preferably for 60 minutes before the step (b), at 70 占 폚 may be added. If the culture is not sterilized, the seeds will grow continuously and the liver may become cloudy or the nutrients may change.
상기 여과는 한랭사를 사용할 수 있다. 상기 한랭사(cheese cloth)는 매우 얇게 짜진 사와 비슷한 느낌이 있는 평직의 면직물로서, 원래 얇은 마직물로 되어 있었는데, 그 감촉을 흉내 내어 촉감이 약간 질긴 면직물이 만들어졌으며 이후 목면의 것이 늘어나게 된 것임을 나타낸다. 여과에 사용되는 여과제는 특별히 한정할 필요는 없다. The filtration may use cold yarn. The cheese cloth was a plain weave cotton fabric with a similar feel to a very thinly woven fabric. It was originally made of a thin woven fabric, which mimics the texture of the cotton fabric, creating a somewhat tacky cotton fabric, which in turn indicates that the cotton fabric is stretched. The filtration agent used for filtration is not particularly limited.
본 명세서에서, 용어 '액침배양(submerged culture)'이란 호기성의 미생물을 액 중에 생육시키기 위해 배양액에 통기 또는 통기 교반하면서 하는 배양 방법을 의미하는 것으로서, 발효조 바닥으로부터 배양액 중에 높은 압력으로 주입된 공기를 교반기에 의해서 미립화하여 산소용해를 촉진하며, 배양액 중에서 미생물을 균일하게 분산시키는 특징이 있다. 또한 액내배양 또는 심부배양 또는 통기교반 발효법이라고도 알려져 있다.The term " submerged culture " as used herein refers to a culture method in which a culture solution is aerated or aerated while agitating aerobic microorganisms for growth in a liquid, It is atomized by a stirrer to accelerate dissolution of oxygen, and microorganisms are uniformly dispersed in a culture solution. It is also known as an in-liquid culture or deep culture or aeration and stirring fermentation method.
이하에서 실시예 등을 통해 본 발명을 더욱 상세히 설명하고자 하며, 다만 이하에 실시예 등에 의해 본 발명의 범위와 내용이 축소되거나 제한되어 해석될 수 없다. 또한, 이하의 실시예를 포함한 본 발명의 개시 내용에 기초한다면, 구체적으로 실험 결과가 제시되지 않은 본 발명을 통상의 기술자가 용이하게 실시할 수 있음은 명백한 것이며, 이러한 변형 및 수정이 첨부된 특허청구범위에 속하는 것도 당연하다.Hereinafter, the present invention will be described in more detail with reference to Examples and the like, but the scope and content of the present invention can not be construed to be limited or limited by the following Examples. It will be apparent to those skilled in the art that various modifications and variations can be made in the present invention without departing from the spirit and scope of the present invention as set forth in the following claims. It is natural that it belongs to the claims.
또한 이하에서 제시되는 실험 결과는 상기 실시예 및 비교예의 대표적인 실험 결과만을 기재한 것이며, 아래에서 명시적으로 제시하지 않은 본 발명의 여러 구현예의 각각의 효과는 해당 부분에서 구체적으로 기재하도록 한다.
In addition, the experimental results presented below only show representative experimental results of the embodiments and the comparative examples, and the respective effects of various embodiments of the present invention which are not explicitly described below will be specifically described in the corresponding part.
실시예Example
제조예 1-1: 내염성 및 단백질 분해활성이 우수한 균주의 분리Production Example 1-1: Isolation of strains having excellent salt tolerance and proteolytic activity
간장의 직접발효에 적합한 균주를 스크리닝(screening)하기 위한 조건은 다음과 같았다.Conditions for screening strains suitable for direct fermentation of soy sauce were as follows.
1. 식품에서 분리되어야 한다.1. Must be separated from food.
2. 고농도의 염도에 대한 내염성을 지니고 있어야 한다.2. It should have salt resistance against high salt concentration.
3. 고농도의 염도에서 대두단백질을 분해하는 세포외 단백질 분해효소를 분비해야 한다.3. Extracellular proteolytic enzymes that break down soy protein at high salt intensities should be secreted.
4. 섭씨 30 ℃ 내외의 온도에서 최적 생장하는 생리특성이 있어야 한다.4. Optimal growth physiological characteristics should be available at temperatures around 30 ℃.
상기의 조건 1 내지 4를 충족하는 식품유래 균주를 분리하기 위해서, 약 1 년 동안 68 종류의 재래식 된장을 수집하였으며, 180 g/L의 천일염을 포함하는 대두박 분말 고체평판배지를 제조하여 상기 재래식 된장에서 균을 스크리닝 하였다. In order to isolate the food-derived strains satisfying the above-mentioned conditions 1 to 4, 68 types of conventional soybean paste were collected for about one year, and a soybean meal powder solid plate culture medium containing 180 g / L of austenitic salt was prepared, To screen for bacteria.
스크리닝 시행 결과, 4 종류의 세균 집락(colony)이 생성되었고, 이 중 2 종류가 상기 조건 1 내지 4를 충족하는 것으로 확인하였다. As a result of the screening, four kinds of colonies of bacteria were produced, and it was confirmed that two of them met the conditions 1 to 4 above.
제조예 1-2: 선별 균주의 동정Production Example 1-2: Identification of Screening Strain
분리된 2 종류의 균주는 16S rDNA 염기서열 분석을 통해 오셔노바실러스 김치(Oceanobacillus kimchii) sosab 균주[기탁번호 KFCC11607P](서열번호 1) 및 바실러스 퓨밀러스 (Bacillus pumilus) sosad 균주[기탁번호 KFCC11608P](서열번호 2)로 최종 동정하였다.The isolated two strains were analyzed by 16S rDNA sequencing to identify the strains of Oceanobacillus kimchii sosab (Accession No. KFCC11607P) (SEQ ID NO: 1) and Bacillus pumilus sosad strain (Accession No. KFCC11608P) SEQ ID NO: 2).
제조예 2Production Example 2
된장으로부터 오셔노바실러스 김치(Oceanobacillus kimchii) sosab 균주[기탁번호 KFCC11607P] 및 바실러스 퓨밀러스 (Bacillus pumilus) sosad 균주[기탁번호 KFCC11608P]를 분리하여 두 가지 종균을 준비하였다. Two seeds were prepared from soybean by separating Oceano Bacillus kimchii sosab strain [Accession No. KFCC11607P] and Bacillus pumilus sosad strain [Accession No. KFCC11608P].
실시예 1: 오셔노바실러스 김치 종균으로 발효된 간장 제조Example 1: Preparation of soy sauce fermented with Oshino Bacillus Kimchi seeds
제조예 2의 오셔노바실러스 김치(Oceanobacillus kimchii) sosab 균주[기탁번호 KFCC11607P]을 탈지대두박 200 g 및 천일염 180 g을 함유한 1L 용액에 첨가하여 접종하였다. 초기 약 15 일 동안은 RPM 60으로 교반배양하였고, 이후 90일 동안 30 ℃에서 액침정치배양한 후, 배양물은 60 분 동안, 70 ℃에서 살균하였고, 한랭사로 여과하여 여과액을 수득하였다. 여과액은 마이크로 입자가 침전 될 때까지 자연침전하여 수득한 상청액인 간장을 제조하였다.An Oceanobacillus kimchii sosab strain (Accession No. KFCC11607P) of Preparation Example 2 was added to a 1 L solution containing 200 g of defatted soybean meal and 180 g of sunflower, and inoculated. After initial incubation for about 15 days with RPM 60, the cells were incubated at 30 ° C for 90 days. The cultures were then sterilized at 70 ° C for 60 minutes and filtered to obtain a filtrate. The filtrate was spontaneously precipitated until the microparticles were precipitated, to prepare liver supernatant.
실시예 2: 바실러스 퓨밀러스 종균으로 발효된 간장 제조Example 2: Preparation of soy sauce fermented with Bacillus subtilis seeds
오셔노바실러스 김치(Oceanobacillus kimchii) sosab 균주[기탁번호 KFCC11607P]를 사용하는 대신 제조예 2의 바실러스 퓨밀러스 (Bacillus pumilus) sosad 균주[기탁번호 KFCC11608P]를 사용하는 것을 제외하고는 실시예 1과 동일하게 간장을 제조하였다.Same as Example 1 except that Bacillus pumilus sosad strain of Production Example 2 (Accession No. KFCC11608P) was used instead of Oceanobacillus kimchii sosab strain [Accession No. KFCC11607P] Soy sauce was prepared.
실시예 3: 오셔노바실러스 김치 종균 및 바실러스 퓨밀러스 종균의 혼합물로 발효된 간장 제조Example 3: Preparation of soy sauce fermented with a mixture of Oishino Bacillus sp. Kimchi and Bacillus fumillus seedlings
오셔노바실러스 김치(Oceanobacillus kimchii) sosab 균주[기탁번호 KFCC11607P]를 사용하는 대신 제조예 2의 바실러스 퓨밀러스 (Bacillus pumilus) sosad 균주[기탁번호 KFCC11608P] 및 오셔노바실러스 김치(Oceanobacillus kimchii) sosab 균주[기탁번호 KFCC11607P]의 혼합물을 사용하는 것을 제외하고는 실시예 1과 동일하게 간장을 제조하였다. Bacillus pumilus sosad strain [Accession No. KFCC11608P] and Oceanobacillus kimchii sosab strain of Preparation Example 2 (Deposit No. KFCC11608P) were used instead of Oceano Bacillus kimchii sosab strain [Accession No. KFCC11607P] No. KFCC11607P] was used as the soy sauce.
비교예 1: 전통간장Comparative Example 1: Traditional soy sauce
종래의 자연발효된 메주를 이용하여 제조된 전통간장을 준비하였다.Traditional soy sauce prepared using conventional fermented meju was prepared.
비교예 2: 양조간장Comparative Example 2: Brewed soy sauce
시중에 판매되고 있는 양조간장을 준비하였다.
Brewing soy sauce sold on the market was prepared.
실험예Experimental Example
실험예 1: 단백질 분해활성 확인Experimental Example 1: Identification of proteolytic activity
상기 제조예 1-1의 조건 3을 만족하는 균주의 단백질 분해활성을 확인하기 위해, 탈지대두 분말 박 플레이크(굵은 입자) 200 g/L, 및 천일염 180 g/L로 구성된 액상배지를 이용하여 5일 간 오셔노바실러스 김치 균주와 바실러스 퓨밀러스 균주를 배양하고, 그 배양액을 백금루프를 이용하여 탈지대두박 분말(고운 입자) 3 g/L, 천일염 160 g/L, 및 한천 20g/L로 구성된 한천평판배지(두께 5 mm, 직경 90 mm)에 스트리킹하여 35 ℃에서, 5 일간 배양하였다. 이는 도1을 통해 오셔노바실러스 김치 균주, 및 바실러스 퓨밀러스 균주가 지니고 있는 단백질 가수분해 활성을 확인할 수 있다. 두 균주는 18% 염분 조건에서 콩 단백질을 가용성 펩티드와 아미노산으로 분해할 수 있음을 확인할 수 있다.In order to confirm the proteolytic activity of the strain satisfying Condition 3 of Production Example 1-1, a liquid medium consisting of 200 g / L of defatted soybean powder thin flakes (coarse particles) and 180 g / L of sodium chloride was used to prepare 5 The broth of Oshino Bacillus Kimchi and Bacillus fumillus was cultured and the culture broth was cultivated in a platinum loop using an agar composed of 3 g / L of defatted soybean meal powder (fine particles), 160 g / L of sun salt and 20 g / L of agar Streaked on a plate medium (thickness 5 mm, diameter 90 mm) and incubated at 35 ° C for 5 days. The results are shown in FIG. 1, which shows the protein hydrolysis activity of Oshino Bacillus Kimchi strain and Bacillus subtilis strain. Both strains are able to decompose soy protein into soluble peptides and amino acids under 18% salt conditions.
또한 도 2를 통해 종균배양 동안의 탈지대두박의 발효과정을 확인할 수 있으며, 도 3에는 본 발명의 간장 제조방법에 따라서, 오셔노바실러스 김치 균주, 바실러스 퓨밀러스 균주, 및 두 균주의 혼합균주의 탈지대두박의 배양으로부터 제조된 간장 및 간장을 추출하고 남은 찌꺼기를 확인할 수 있다.FIG. 3 shows the fermentation process of the defatted soybean during the seed culture through the method of FIG. 2, and FIG. 3 shows the results of the defatting of the mixed strains of Oshino Bacillus Kimchi strain, Bacillus subtilis strain, Soy sauce and soy sauce produced from soybean meal culture can be extracted to identify residual debris.
고체배지 상에서 생장하는 세균은 생장을 위해 세포외 단백분해효소를 분비하고, 불용성의 탈지대두단백질을 분해하여 수용성의 아미노산이나 펩티드로 전환하기 때문에 고체배지는 반투명하게 된다. 또한 탈지대두박 분말을 사용한 이유는 한천배지에 고르게 분산하기 위한 것으로서, 효소에 의한 분해효율도 증가한다. 3 g/L의 낮은 농도를 사용한 이유는 고농도를 적용하면 효소에 의한 분해활성에 의해 부분적으로 단백질이 분해되면서 투명해지는 현상을 관찰할 수 없기 때문이고, 투명해지지 않고 반투명해지는 이유는 탈지대두박 분말에 단백질 외에도 불용성의 탄수화물이 존재하기 때문이다. Bacteria growing on the solid medium secrete extracellular proteolytic enzymes for growth and convert the insoluble defatted soy protein into soluble amino acids or peptides, so that the solid medium becomes translucent. The reason for using the defatted soybean meal powder is to disperse evenly on the agar medium, and the efficiency of the degradation by the enzyme is also increased. The reason why low concentration of 3 g / L is used is because, when applied at a high concentration, the protein is partially decomposed due to the decomposition activity by the enzymatic activity, and the phenomenon of transparency can not be observed. The reason why the translucent is not transparent is that the defatted soybean meal powder In addition to proteins, insoluble carbohydrates exist.
실험예 2: 유리아미노산 함량 측정Experimental Example 2: Measurement of free amino acid content
식품분석에 사용되는 일반적인 기술에 기초하여 간장의 유리아미노산(Free amino acids, AAs)의 함량을 분석하였다. 간장은 구연산나트륨 완충용액으로 20 배 희석하였고, 0.45μm 필터(Sartorius, GmbH, Gottingen, German)를 통해 여과하여 유리아미노산 분석에 사용하였다. 유리아미노산은 S433 암모니아 자동분석기(Sykam)가 내장된 암모니아 여과 칼럼(LCA, k04/Na, 4.6 × 100mm, Sykam, GmbH, Eresing, Germany)을 사용하여 분석하였다. 모든 완충용액은 Sykam으로부터 구매한 가수분해물 프로그램에 의한 유리아미노산 분석에 사용하였다. The contents of free amino acids (AAs) in liver were analyzed based on the general techniques used in food analysis. The liver was diluted 20-fold with sodium citrate buffer and used for free amino acid analysis by filtration through a 0.45 μm filter (Sartorius, GmbH, Göttingen, Germany). Free amino acids were analyzed using an ammonia filtration column (LCA, k04 / Na, 4.6 x 100 mm, Sykam, GmbH, Eresing, Germany) equipped with S433 ammonia automatic analyzer (Sykam). All buffer solutions were used for free amino acid analysis by the hydrolyzate program purchased from Sykam.
상기 실시예 1 내지 3 및 비교예 1의 유리아미노산 함량을 하기 표 1에 나타내었다.The free amino acid contents of Examples 1 to 3 and Comparative Example 1 are shown in Table 1 below.
상기 표 1을 통해 볼 수 있듯이, 전통간장인 비교예 1에서 검출된 15 가지 유리아미노산이, 본 발명의 발효간장인 실시예 1 내지 3에서도 검출됨을 확인하였으며, 두 균주의 혼합배양액에 의해 제조된 간장 내의 유리아미노산의 함량이 한 가지 균주에 의해 제조된 간장보다 약간 더 높은 것을 확인하였다. 또한 전통간장인 비교예 1보다도 본 발명의 발효간장인 실시예 1 내지 3의 유리아미노산 함량이 현저히 높은 것을 확인할 수 있다. 특히, 구수한 맛을 내는 아스파르트산과 단맛을 내는 세린과 알라닌이 전통간장보다 본 발명의 발효간장에 더 많이 함유되어 있어, 전통간장에 비해 구수하고, 단맛을 내어 풍미가 좋은 발효간장을 제공할 수 있다는 것을 확인하였다. As can be seen from the above Table 1, it was confirmed that the 15 free amino acids detected in Comparative Example 1, which is a conventional soy sauce, were also detected in the fermented soy sauce examples 1 to 3 of the present invention. The content of free amino acids in soy sauce was slightly higher than that of soy sauce prepared by one strain. In addition, it can be confirmed that the content of free amino acids in Examples 1 to 3, which are fermented soy sauce of the present invention, is significantly higher than that of Comparative Example 1 which is a conventional soy sauce. In particular, the fermented soy sauce of the present invention contains more abundant asparagus acid and sweet sour serine and alanine than the traditional soy sauce, so that it is possible to provide a fermented soy sauce that is more salty and sweet than the traditional soy sauce, Respectively.
실험예 3: 유기질소화합물 및 암모늉 함량 측정Experimental Example 3: Measurement of organic nitrogen compounds and ammonia content
간장에 함유된 유기 질소 화합물은 기본적으로 가용성이며, 마이크로케달법(micro-Kjeldahl method)을 통해 분석하였다. 유기질소(유리아미노산과 펩타이드)는 질소-단백질 환산계수(6.25)에 의해 측정되었다. 암모늄은 양이온 칼럼(IonPac, Dionex CS12A)이 내장된 이온 크로마토그래피(Dionex DX-500)를 사용하여 분석하였다.Organic nitrogen compounds in the liver were basically soluble and were analyzed by the micro-Kjeldahl method. Organic nitrogen (free amino acids and peptides) was measured by a nitrogen-to-protein conversion factor (6.25). Ammonium was analyzed using ion chromatography (Dionex DX-500) with a cation column (IonPac, Dionex CS12A) embedded.
메탄술폰산(20 mM 100%)은 이동상으로 사용하였다. 칼럼의 온도 및 유속은 35 ℃및 0.8 ml/분으로 조정하였다.Methanesulfonic acid (20
상기 실시예 1 내지 3 및 비교예 1의 유기질소화합물 및 암모늄의 함량을 하기 표 2에 나타내었다.The contents of the organic nitrogen compounds and ammonium in Examples 1 to 3 and Comparative Example 1 are shown in Table 2 below.
상기 표 2를 통해 볼 수 있듯이, 본 발명의 발효간장인 실시예 1 내지 3이 전통간장인 비교예 1보다 유기질소화합물의 함량이 높은 것을 확인하였으며, 암모늄 함량은 전통간장이 본 발명의 발효간장에 비해 월등히 높은 수치를 보였다.As can be seen from the above Table 2, the fermented soy sauce according to Examples 1 to 3 of the present invention had higher contents of organic nitrogen compounds than those of Comparative Example 1 with a conventional soy sauce. The ammonium content was higher than that of the conventional soy sauce fermented soy sauce Compared with the previous year.
암모니아성 질소는 단백질 분해과정에서 탈아미노화(deamination)에 의해 생성되며, 암모니아성 질소가 식품에 과량 축적되면 부패취로 작용하므로 일반적으로 장류 제품의 변채 또는 이상발효의 지표로 사용된다. 이에 따라, 암모늄 함량이 전통간장에 비해 현저히 적은 본 발명의 발효간장은 불쾌취가 비교적 적어, 소비자들에게 향미가 좋은 발효간장을 제공할 수 있다.Ammonia nitrogen is produced by deamination in the proteolytic process. When ammonia nitrogen accumulates excessively in food, it acts as a decay, and is generally used as an index of irreversible or abnormal fermentation of the product. Accordingly, the fermented soy sauce according to the present invention, in which the ammonium content is significantly lower than that of the conventional soy sauce, has a relatively low discomfort and can provide fermented soy sauce having good flavor to consumers.
실험예 4: 무기물 및 중금속 함량 측정Experimental Example 4: Measurement of inorganic and heavy metal content
간장에 함유된 무기물의 함량은 ICPOE(inductively-coupled plasma optic emission) 분광기(SPECTRO Analytical Instrument, Kleve, Germany)를 사용하여 분석하였다. 간장은 0.45 μm 멤브레인 필터(Satorius)를 통해 여과하였고, 이중 증류수로 20 배 희석하였다. 희석된 여과용액은 마그네슘(279.553 nm), 나트륨(589.592 nm), 칼륨(766.491 nm), 칼슘(396.847 nm), 망간(257.61 nm), 아연(213.856 nm), 및 구리(324.745 nm)의 특정한 파장 하에 ICPOE 주입기에 주입하였다. The content of inorganic substances in the liver was analyzed using an inductively coupled plasma optical emission (ICPOE) spectrometer (SPECTRO Analytical Instrument, Kleve, Germany). The liver was filtered through a 0.45 μm membrane filter (Satorius) and diluted 20 times with double distilled water. The diluted filtrate solution contained specific wavelengths of magnesium (279.553 nm), sodium (589.592 nm), potassium (766.491 nm), calcium (396.847 nm), manganese (257.61 nm), zinc (213.856 nm) 0.0 > ICPOE < / RTI >
무기물의 농도는 표준물질(AccuTraceTM Reference Standard, AccuStandard, NewHaven, CT, USA)을 사용하여 얻은 흡광도와 희석률을 기초하여 계산하였다.Concentrations of minerals were calculated based on the absorbance and dilution rates obtained using the standard (AccuTrace TM Reference Standard, AccuStandard, New Haven, CT, USA).
상기 실시예 1 내지 3 및 비교예 1의 무기물 및 중금속의 함량을 하기 표 3에 나타내었다.The contents of the minerals and heavy metals in Examples 1 to 3 and Comparative Example 1 are shown in Table 3 below.
상기 표 3을 통해 볼 수 있듯이, 전통간장이 본 발명의 발효간장에 비해 Mg의 함량이 높으나, Ca, Na, K의 함량은 본 발명의 발효간장에서 더 높게 나타나, 무기물 함량이 향상된 발효간장을 제공 할 수 있음을 확인하였다. 또한 실시예 1 내지 3, 및 비교예 1 모두에서 중금속인 Pb, As, Cu, Zn, 및 Mn 함량은 모두 기준치 이하로 미량 검출되었다.As can be seen from Table 3, the content of Mg, Na and K is higher in the fermented soy sauce of the present invention than in the fermented soy sauce of the present invention, whereas the content of Mg, Na and K is higher in the fermented soy sauce of the present invention. And that it is possible to provide it. In both of Examples 1 to 3 and Comparative Example 1, the contents of Pb, As, Cu, Zn, and Mn, which are heavy metals, were all detected in a trace amount below the reference value.
실험예 5: 유기산 함량 측정Experimental Example 5: Measurement of organic acid content
간장에 함유된 유기산의 함량은 이온배제 칼럼(Shodex, Rspak KC-811, Showa Denko, Tokyo, Japan) 및 시차굴절 검출기(Shodex, RI-101)가 내장된 고속액체크로마토그래피(HPLC) (Beckman, Coulter System Gold, Brea, CA, USA)를 사용하여 분석하였다.The content of organic acids contained in the liver was determined by high-performance liquid chromatography (HPLC) (Beckman, R) using an ion exclusion column (Shodex, Rspak KC-811, Showa Denko, Tokyo, Japan) and a differential refraction detector (Shodex, Coulter System Gold, Brea, CA, USA).
칼럼과 검출기는 40 ℃로 조정하였다. 이동상은 과염소산(6mM)을 사용하였고, 유속은 0.8 ml/분으로 조정하였다. 샘플은 12,000 × g의 힘으로 30 분 동안 원심분리 하여 제조하였고, 0.45 μm 멤브레인 필터를 통해 여과하였으며, 그 다음 이온 교환 칼럼(Amberlite IR120, Rohm and Haas, Philadelphia, PA, USA)에서 탈염하였다. 그 다음, 여과 및 탈염된 샘플 30 μl를 고속액체크로마토그래피에 주입하였다. 유기산 농도는 표준물질을 사용하여 생성된 크로마토그램의 피크 넓이를 기초하여 계산하였다. The column and detector were adjusted to 40 < 0 > C. The mobile phase used was perchloric acid (6 mM) and the flow rate was adjusted to 0.8 ml / min. Samples were prepared by centrifugation at 12,000 x g for 30 min, filtered through a 0.45 μm membrane filter, and then desalted on an ion exchange column (Amberlite IR120, Rohm and Haas, Philadelphia, PA, USA). Then, 30 μl of the filtered and demineralized sample was injected into high performance liquid chromatography. The organic acid concentration was calculated based on the peak area of the chromatogram generated using the standard material.
상기 실시예 1 내지 3 및 비교예 1의 유기산의 함량을 하기 표 4에 나타내었다.The contents of the organic acids in Examples 1 to 3 and Comparative Example 1 are shown in Table 4 below.
상기 표 4를 통해 볼 수 있듯이, 전통간장에 비해 본 발명의 발효간장에서 검출된 유기산 성분의 함량이 모두 현저히 높게 나타났다. 한편, 발효간장에서는 말산과 숙신산이 검출되지 않았는데 이는 발효과정에서 작용한 세균의 종류가 단순하고 종균으로 사용한 세균이 말산과 숙신산을 발효하는 기능이 없기 때문이다. As shown in Table 4, the content of organic acid components detected in the fermented soy sauce of the present invention was significantly higher than that of the conventional soy sauce. On the other hand, malic acid and succinic acid were not detected in the fermented soy sauce, because the kinds of bacteria acting in the fermentation process are simple and the bacterium used as the seed bacteria has no function of fermenting malic acid and succinic acid.
실험예 6: 관능 검사Experimental Example 6: Sensory Evaluation
상기의 실시예 1 내지 3 및 비교예 1 내지 비교예 2의 간장들의 관능검사를 실시하였다. 남녀 구분없이 대학교 3, 4 학년의 학생(21~25 세) 15 명에게 다섯 가지 간장의 특성을 알려주지 않고 냄새와 맛을 비교하게 하였다.The sensory evaluation of the soy sauce of Examples 1 to 3 and Comparative Examples 1 to 2 was carried out. Fifteen students (21 ~ 25 years old) in university 3 and 4 years without any discrimination of men and women were asked to compare the odor and taste without informing the characteristics of five kinds of soy sauce.
향은 하루 전에 하루 전에 50 mL 코니칼 튜브에 10 mL씩 담아 실온에 놓아둔 간장 시료의 뚜껑을 개방한 후 10 초 내에, 약 20 cm의 거리에서 감지하게 하였으며 시료 간의 검사 시간은 5 분 간격으로 조정하였다. 또한 향은 불쾌한 향과 달콤한 향으로 구분하였다 (불쾌한 향= 지린 내, 달콤한 향= 카라멜 또는 유사한 냄새).The fragrance was sensed at a distance of about 20 cm within 10 seconds after opening the lid of the soy sauce sample which was placed at room temperature in a 50 mL conical tube one day before the day before, and the inspection time between samples was 5 minutes Respectively. In addition, fragrance was divided into unpleasant fragrance and sweet fragrance (unpleasant fragrance = fragrant fragrance, sweet fragrance = caramel or similar smell).
맛은 10 배 희석한 간장 0.1 mL을 대상 학생의 혀의 상부 중앙에 떨어뜨린 후 감지하게 하였으며, 시료 간의 검사 시간은 식수로 입을 헹구는 시간을 포함하여 10 분 간격으로 조정하였다. 또한 맛은 쓴맛, 고소한 맛, 담백한 맛으로 구분하였다 (쓴맛=떫고 불편한 맛, 고소한 맛=콩 고유의 풍미가 느껴지는 맛, 달콤한 맛). The taste was determined by dropping 0.1 mL of 10-fold diluted soy sauce in the middle of the upper part of the tongue of the target student, and the test time between the samples was adjusted at intervals of 10 minutes including the time for rinsing the mouth with drinking water. In addition, the taste was divided into bitter taste, sweet taste, and light taste (bitter taste = uncomfortable taste, sweet flavor = flavor of bean flavor, sweet taste).
또한, 1점 매우 나쁨, 2점 나쁨, 3점 보통, 4점 좋음, 5점 매우 좋음으로 나타나는 5점 기호척도법으로 실시하였다. 간장의 투명도와 색 등은 제조방법에 따른 차이가 있기 때문에 관능검사에 포함하지 않았다.In addition, the 5-point symbol scale method was used, which indicated that 1 point was very bad, 2 points were bad, 3 points were good, 4 points were good, and 5 points were very good. The transparency and color of soy sauce were not included in the sensory evaluation because of differences in production methods.
상기 실시예 1 내지 3 및 비교예 1 내지 2의 관능검사 결과를 하기 표 5에 나타내었다.The results of the sensory evaluation of Examples 1 to 3 and Comparative Examples 1 to 2 are shown in Table 5 below.
맛
flavor
표 5에서 제시한 바와 같이, 본 발명의 발효간장인 실시예 1 내지 3은 전통적인 메주를 이용하여 제조한 간장인 비교예 1에 비하면 향에 의한 불쾌감은 상대적으로 낮은 것으로 나타났으며, 양조간장인 비교예 2와는 유의적 차이는 나타나지 않았고, 달콤한 향은 불쾌한 향과 대조적인 것으로 나타났다. 또한 본 발명의 발효간장 실시예 1 내지 3을 비교해보면, 실시예 3에서 불쾌한향은 가장 적고, 달콤한 향이 가장 높게 나타나는 것을 확인하였다.As shown in Table 5, Examples 1 to 3 of the fermented soy sauce according to the present invention showed relatively low discomfort due to incense compared with Comparative Example 1 which was prepared using conventional meju, No significant difference was observed with Comparative Example 2, and the sweet aroma was in contrast to the unpleasant aroma. Further, when the fermented soy sauce examples 1 to 3 of the present invention were compared, it was confirmed that the unpleasant aroma was the least and the sweet aroma was the highest in Example 3.
한편 쓴맛은 양조간장인 비교예 2의 점수가 가장 높게 나타났고, 고소한 맛 및 달콤한 맛은 본 발명의 발효간장인 실시예 3의 점수가 가장 높게 나타났다.On the other hand, the bitter taste of the fermented soy sauce according to the present invention was the highest in Comparative Example 2, which was brewed soy sauce, and the fermented soy sauce having the highest taste and sweet taste showed the highest score.
또한, 본 발명의 발효간장인 실시예 1 내지 3의 종합적인 기호도는 전통간장인 비교예 1보다 월등히 높은 점수를 받았으며. 본 발명의 발효간장인 실시예 3이 소비자들의 기호에 가장 맞는다는 결과를 보였으며, 실시예 2가 그 다음으로 높은 기호도를 나타냈다. In addition, the overall preference of the fermented soy sauce according to Examples 1 to 3 of the present invention was significantly higher than that of Comparative Soy sauce 1. Example 3 of the fermented soy sauce according to the present invention showed the best fit with consumers' preferences, and Example 2 showed the next highest preference.
이러한 결과는 메주를 제조하는 과정에서 다양한 미생물에 의해 무작위로 생산되는 대사산물에 의한 영향으로 확인하였다. 반면 발효간장은 두 종류의 내염성 바실러스가 대두박의 단백질을 분해하여 생산한 아미노산과 호흡대사산물(주로 이산화탄소, 질산염)을 포함하기 때문에 향과 맛에 미치는 부정적인 요인을 제한적으로 함유할 것이다. These results were confirmed by the effects of metabolic products randomly produced by various microorganisms in the process of making meju. On the other hand, fermented soy sauce will contain a limited number of negative factors on flavor and taste, since the two types of salt-tolerant bacillus contain amino acids and respiratory metabolites (mainly carbon dioxide, nitrate) produced by decomposition of soybean protein.
따라서 메주의 제조과정 없이 직접 발효과정을 적용하여 생산한 간장은 양조간장에 맛과 향이 단순하지만 불필요한 미생물의 대사산물을 포함하고 있지 않아 재래식 전통간장을 대체하기에 적합한 것으로 확인하였다.Therefore, the soy sauce produced by direct fermentation process without meju production process was simple and delicious in brewing soy sauce, but it did not contain unnecessary microbial metabolites and was confirmed to be suitable for replacing traditional traditional soy sauce.
<110> PARK, Doo Hyun KIM, kwang Nam SEOL, Bo Min <120> Seed culture for soybean sauce and preparation method of soybean sauce using the same <130> HPC6154 <160> 2 <170> KopatentIn 2.0 <210> 1 <211> 1262 <212> DNA <213> Oceanobacillus kimchii <400> 1 ctacaagggg gctccagcct aatacatgca agtcgagcgc aggaagttat ctgatcctct 60 tttagaggtg acgataatgg aatgagcggc ggacgggtga gtaacacgta ggcaacctgc 120 ctgtaagact gggataactc gtggaaacgc gagctaatac cggataacac ttttcatctc 180 ctgatgagaa gttgaaaggc ggcttttagc tgtcacttac agatgggcct gcggcgcatt 240 agctagttgg taaggtaatg gcttaccaag gcgacgatgc gtagccgacc tgagagggtg 300 atcggccaca ctgggactga gacacggccc agactcctac gggaggcagc agtagggaat 360 cttccgcaat ggacgaaagt ctgacggagc aacgccgcgt gagtgatgaa ggttttcgga 420 tcgtaaaact ctgttgttag ggaagaacaa gtgccatagt aagctgatgg caccttgacg 480 gtacctaacc agaaagccac ggctaactac gtgccagcag ccgcggtaat acgtaggtgg 540 caagcgttgt ccggaattat tgggcgtaaa gcgctcgcag gcggttcttt aagtctgatg 600 tgaaatctta cggctcaacc gtaaacgtgc attggaaact ggggaacttg agtgcagaag 660 aggagagtgg aattccacgt gtagcggtga aatgcgtaga gatgtggagg aacaccagtg 720 gcgaaggcga ctctctggtc tgtaactgac gctgaggagc gaaagcgtgg ggagcgaaca 780 ggattagata ccctggtagt ccacgccgta aacgatgagt gctaggtgtt agggggtttc 840 cgccccttag tgctgaagtt aacgcattaa gcactccgcc tggggagtac ggccgcaagg 900 ctgaaactca aaagaattga cgggggcccg cacaagcggt ggagcatgtg gtttaattcg 960 aagcaacgcg aagaacctta ccaggtcttg acatcctctg aacactcgta gagatagagt 1020 tttcccttcg gggacagagt gacaggtggt gcatggttgt cgtcagctcg tgtcgtgaga 1080 tgttgggtta agtcccgcaa cgagcgcaac ccttgatctt agttgccagc attcagttgg 1140 gcactctaag gtgactgccg gtgacaaacc cggaggaagg ttggggacga cgtcaaatca 1200 tcatgcccct tatgacctgg gctacacacg tagctacaat aggatggaac aaagggcacc 1260 ca 1262 <210> 2 <211> 1203 <212> DNA <213> Bacillus pumilus <400> 2 aacaccagcc ggtgccctat tcatgcagtt cgagcgcgag atattctatg atctcttcgg 60 agtgattagg atggatcgag cggcggacgg gtgagtaaca cgtgggtaac ctgcctgtaa 120 gactaggata actccgggaa accggagcta ataccggata gttccttgaa ccgcatggtt 180 caaggatgaa agacggtttc ggctgtcact tacagatgga cccgcggcgc attagctagt 240 tggtggggta atggctcacc aaggcgacga tgcgtagccg acctgagagg gtgatcggcc 300 acactgggac tgagacacgg cccagactcc tacgggaggc agcagtaggg aatcttccgc 360 aatggacgaa agtctgacgg agcaacgccg cgtgagtgat gaaggttttc ggatcgtaaa 420 gctctgttgt tagggaagaa caagtgcgag agttaactgc tcgcaccttg acggtaccta 480 accagaaagc cacggctaac tacgtgccag cagccgcggt aatacgtagg tggcaagcgt 540 tgtccggaat tattgggcgt aaagggctcg caggcggttt cttaagtctg atgtgaaagc 600 ccccggctca accggggagg gtcattggaa actgggaaac ttgagtgcag aagaggagag 660 tggaattcca cgtgtagcgg tgaaatgcgt agagatgtgg aggaacacca gtggcgaagg 720 cgactctctg gtctgtaact gacgctgagg agcgaaagcg tggggagcga acaggattag 780 ataccctggt agtccacgcc gtaaacgatg agtgctaagt gttagggggt ttccgcccct 840 tagtgctgca gctaacgcat taagcactcc gcctggggag tacggtcgca agactgaaac 900 tcaaaggaat tgacgggggc ccgcacaagc ggtggagcat gtggtttaat tcgaagcaac 960 gcgaagaacc ttaccaggtc ttgacatcct ctgacaaccc tagagatagg gctttcccat 1020 tccggggaca gagtgacagg tggtgcatgg ttgtcgtcag ctcgtgtcgt gagatgttgg 1080 gttaagtccc gcaacgagcg caacccttga tcttagttgc cagcatttag ttgggcactc 1140 taaggtgact gccggtgaca aaccggagga aggtggggat gacgtcaaat catcatggcc 1200 cct 1203 <110> PARK, Doo Hyun KIM, kwang Nam SEOL, Bo Min <120> Seed culture for soybean sauce and preparation method of soybean sauce using the same <130> HPC6154 <160> 2 <170> Kopatentin 2.0 <210> 1 <211> 1262 <212> DNA <213> Oceanobacillus kimchii <400> 1 ctacaagggg gctccagcct aatacatgca agtcgagcgc aggaagttat ctgatcctct 60 tttagaggtg acgataatgg aatgagcggc ggacgggtga gtaacacgta ggcaacctgc 120 ctgtaagact gggataactc gtggaaacgc gagctaatac cggataacac ttttcatctc 180 ctgatgagaa gttgaaaggc ggcttttagc tgtcacttac agatgggcct gcggcgcatt 240 agctagttgg taaggtaatg gcttaccaag gcgacgatgc gtagccgacc tgagagggtg 300 atcggccaca ctgggactga gacacggccc agactcctac gggaggcagc agtagggaat 360 cttccgcaat ggacgaaagt ctgacggagc aacgccgcgt gagtgatgaa ggttttcgga 420 tcgtaaaact ctgttgttag ggaagaacaa gtgccatagt aagctgatgg caccttgacg 480 gtacctaacc agaaagccac ggctaactac gtgccagcag ccgcggtaat acgtaggtgg 540 caagcgttgt ccggaattat tgggcgtaaa gcgctcgcag gcggttcttt aagtctgatg 600 tgaaatctta cggctcaacc gtaaacgtgc attggaaact ggggaacttg agtgcagaag 660 aggagagtgg aattccacgt gtagcggtga aatgcgtaga gatgtggagg aacaccagtg 720 gcgaaggcga ctctctggtc tgtaactgac gctgaggagc gaaagcgtgg ggagcgaaca 780 ggattagata ccctggtagt ccacgccgta aacgatgagt gctaggtgtt agggggtttc 840 cgccccttag tgctgaagtt aacgcattaa gcactccgcc tggggagtac ggccgcaagg 900 ctgaaactca aaagaattga cgggggcccg cacaagcggt ggagcatgtg gtttaattcg 960 aagacacgcg aagaacctta ccaggtcttg acatcctctg aacactcgta gagatagagt 1020 tttcccttcg gggacagagt gacaggtggt gcatggttgt cgtcagctcg tgtcgtgaga 1080 tgttgggtta agtcccgcaa cgagcgcaac ccttgatctt agttgccagc attcagttgg 1140 gcactctaag gtgactgccg gtgacaaacc cggaggaagg ttggggacga cgtcaaatca 1200 tcatgcccct tatgacctgg gctacacacg tagctacaat aggatggaac aaagggcacc 1260 ca 1262 <210> 2 <211> 1203 <212> DNA <213> Bacillus pumilus <400> 2 aacaccagcc ggtgccctat tcatgcagtt cgagcgcgag atattctatg atctcttcgg 60 agtgattagg atggatcgag cggcggacgg gtgagtaaca cgtgggtaac ctgcctgtaa 120 gactaggata actccgggaa accggagcta ataccggata gttccttgaa ccgcatggtt 180 caaggatgaa agacggtttc ggctgtcact tacagatgga cccgcggcgc attagctagt 240 tggtggggta atggctcacc aaggcgacga tgcgtagccg acctgagagg gtgatcggcc 300 acactgggac tgagacacgg cccagactcc tacgggaggc agcagtaggg aatcttccgc 360 aatggacgaa agtctgacgg agcaacgccg cgtgagtgat gaaggttttc ggatcgtaaa 420 gctctgttgt tagggaagaa caagtgcgag agttaactgc tcgcaccttg acggtaccta 480 accagaaagc cacggctaac tacgtgccag cagccgcggt aatacgtagg tggcaagcgt 540 tgtccggaat tattgggcgt aaagggctcg caggcggttt cttaagtctg atgtgaaagc 600 ccccggctca accggggagg gtcattggaa actgggaaac ttgagtgcag aagaggagag 660 tggaattcca cgtgtagcgg tgaaatgcgt agagatgtgg aggaacacca gtggcgaagg 720 cgactctctg gtctgtaact gacgctgagg agcgaaagcg tggggagcga acaggattag 780 ataccctggt agtccacgcc gtaaacgatg agtgctaagt gttagggggt ttccgcccct 840 tagtgctgca gctaacgcat taagcactcc gcctggggag tacggtcgca agactgaaac 900 tcaaaggaat tgacgggggc ccgcacaagc ggtggagcat gtggtttaat tcgaagcaac 960 gcgaagaacc ttaccaggtc ttgacatcct ctgacaaccc tagagatagg gctttcccat 1020 tccggggaca gagtgacagg tggtgcatgg ttgtcgtcag ctcgtgtcgt gagatgttgg 1080 gttaagtccc gcaacgagcg caacccttga tcttagttgc cagcatttag ttgggcactc 1140 taaggtgact gccggtgaca aaccggagga aggtggggat gacgtcaaat catcatggcc 1200 cct 1203
Claims (16)
상기 발효식품은 간장인 것을 특징으로 하는 발효식품.5. The method of claim 4,
Wherein the fermented food is a liver.
상기 발효식품은 아스파르트산, 트레오닌, 세린, 글루탐산, 프롤린, 글리신, 알라닌, 발린, 메티오닌, 이소류신, 류신, 타이로신, 페닐알라딘, 히스티딘, 라이신, 및 아르기닌 중에서 선택되는 1종 이상의 유리아미노산을 함유하는 것을 특징으로 하는 발효식품.6. The method of claim 5,
Wherein the fermented food contains at least one free amino acid selected from aspartic acid, threonine, serine, glutamic acid, proline, glycine, alanine, valine, methionine, isoleucine, leucine, tyrosine, phenylalanine, histidine, lysine and arginine Features fermented food.
상기 발효식품은 유기 질소화합물 및 암모늄, 또는 두 가지 모두 함유하는 것을 특징으로 하는 발효식품.6. The method of claim 5,
Wherein the fermented food comprises an organic nitrogen compound and ammonium, or both.
상기 발효식품은 마그네슘, 칼슘, 나트륨, 및 칼륨 중에서 선택되는 1종 이상의 무기물을 함유하는 것을 특징으로 하는 발효식품.6. The method of claim 5,
Wherein the fermented food contains one or more minerals selected from magnesium, calcium, sodium, and potassium.
상기 발효식품은 시트르산, 말산, 숙신산, 락트산, 아세트산, 및 피롤리돈카복실산 중에서 선택되는 1종 이상의 유기산을 함유하는 것을 특징으로 하는 발효식품.6. The method of claim 5,
Wherein the fermented food comprises at least one organic acid selected from the group consisting of citric acid, malic acid, succinic acid, lactic acid, acetic acid, and pyrrolidonecarboxylic acid.
(b) 상기 수득한 배양물을 여과하여 여과액을 수득하는 단계;
(c) 상기 수득한 여과액을 침전하여 간장을 수득하는 단계;를 포함하는 것을 특징으로 하는 간장발효 종균을 이용한 간장의 제조방법.(a) inoculating the seed medium into a liquid medium containing the salt water and the soybean material and culturing to obtain a culture;
(b) filtering the obtained culture to obtain a filtrate;
(c) precipitating the obtained filtrate to obtain a soy sauce. The method of producing soy sauce using the fermented soy sauce according to claim 1,
상기 염수의 염도는 10 내지 18%인 것을 특징으로 하는 간장발효 종균을 이용한 간장의 제조방법.12. The method of claim 11,
Wherein the salinity of the saline solution is 10 to 18%.
상기 액상배지는 콩 원료 대 소금 대 물의 중량비가 0.13-0.20 : 0.10-0.18 : 1인 것을 특징으로 하는 간장발효 종균을 이용한 간장의 제조방법.12. The method of claim 11,
Wherein the weight ratio of the soybean raw material to the salt to water is 0.13-0.20: 0.10-0.18: 1.
상기 종균은 오셔노바실러스 김치(Oceanobacillus kimchii) sosab 균주[기탁번호: KFCC11607P], 및 바실러스 퓨밀러스(Bacillus pumilus) sosad 균주[기탁번호: KFCC11608P], 또는 두 균주의 혼합균주인 것을 특징으로 하는 간장발효 종균을 이용한 간장의 제조방법.12. The method of claim 11,
Wherein the seed is a mixed strain of Oceano Bacillus kimchii sosab [Accession No .: KFCC11607P], Bacillus pumilus sosad strain [Accession No .: KFCC11608P], or two strains. A method for producing soy sauce using seeds.
상기 배양은 2 내지 3 달 동안, 30 내지 40 ℃에서 수행되는 것을 특징으로 하는 발효간장의 제조방법.12. The method of claim 11,
Wherein the culturing is performed at 30 to 40 占 폚 for 2 to 3 months.
상기 (b) 단계 이전에 상기 (a) 단계의 배양물을 55 내지 65 분 동안, 65 내지 75 ℃에서 살균하는 단계를 추가하는 것을 특징으로 하는 발효간장의 제조방법.12. The method of claim 11,
Wherein the step (b) further comprises the step of sterilizing the culture of step (a) at 55 to 65 minutes and at 65 to 75 占 폚 before the step (b).
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109825456A (en) * | 2019-03-15 | 2019-05-31 | 青海省农林科学院 | A kind of Manas' bacillus marinus E40208a1 and its application |
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|---|---|---|---|---|
| KR100290770B1 (en) | 1994-12-30 | 2001-06-01 | 박종섭 | Method for forming contact hole |
| KR101399276B1 (en) | 2012-10-25 | 2014-05-30 | 재단법인 전라북도생물산업진흥원 | Method for producing short-ripened soy sauce and short-ripened soy sauce produced by same method |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR100290770B1 (en) | 1994-12-30 | 2001-06-01 | 박종섭 | Method for forming contact hole |
| KR101399276B1 (en) | 2012-10-25 | 2014-05-30 | 재단법인 전라북도생물산업진흥원 | Method for producing short-ripened soy sauce and short-ripened soy sauce produced by same method |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109825456A (en) * | 2019-03-15 | 2019-05-31 | 青海省农林科学院 | A kind of Manas' bacillus marinus E40208a1 and its application |
| CN109825456B (en) * | 2019-03-15 | 2023-04-18 | 青海省农林科学院 | Marina marine bacillus E40208a1 and application thereof |
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