KR102018536B1 - Saccharomyces cerevisiae ksd-yc which is a novel yeast - Google Patents
Saccharomyces cerevisiae ksd-yc which is a novel yeast Download PDFInfo
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- KR102018536B1 KR102018536B1 KR1020170170264A KR20170170264A KR102018536B1 KR 102018536 B1 KR102018536 B1 KR 102018536B1 KR 1020170170264 A KR1020170170264 A KR 1020170170264A KR 20170170264 A KR20170170264 A KR 20170170264A KR 102018536 B1 KR102018536 B1 KR 102018536B1
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- saccharomyces cerevisiae
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Abstract
본 발명은 술 발효 시 발효비율이 높고, 산도가 낮으며, 관능이 우수하며, 수탁번호 KACC 93276P로 수탁된, 사카로마이세스 세레비지에 KSD-YC에 대한 것이다.The present invention relates to KSD-YC of Saccharomyces cerevisiae, which has a high fermentation rate, low acidity, excellent sensory function, and has been deposited with accession number KACC 93276P.
Description
본 발명은 사카로마이세스 세레비지에 KSD-YC에 대한 것이다. 이는 누룩으로 발효된 술덧에서 분리한 신규 효모이다.The present invention relates to Saccharomyces cerevisiae KSD-YC. This is a new yeast isolated from yeast fermented with yeast.
최근 다양한 소재를 이용한 막걸리가 개발되고 소비되고 있다(한국공개특허 10-2017-0030672, 한국등록특허 10-1723162호 등). 그러나 이렇게 소재 및 맛을 다양하게 하기 위하여, 알코올 농도, 발효 온도 등이 다양하게 되는데, 이 경우 효모의 생존율 및 발효 효율이 원하는 만큼 좋지 못한 경우가 많다. Recently, makgeolli using various materials has been developed and consumed (Korean Patent Publication No. 10-2017-0030672, Korean Patent Registration No. 10-1723162, etc.). However, in order to vary the material and taste, the alcohol concentration, the fermentation temperature is varied, in this case, the survival rate and fermentation efficiency of the yeast is often not as good as desired.
이에 본 발명자들은 막걸리 제조 시 사용 스펙트럼이 높은 효모를 연구하던 중 누룩으로 발효된 술덧에서 분리한 신규 효모가 낮은 산도로 발효가 이루어지고, 발효 비율이 높으면서도 관능 및 풍미가 우수하며, 중온 및 저온에서도 발효가 가능하여 알코올 발효 시 발생되는 발효열을 냉각수로 낮춰 줄 필요가 적기 때문에 발효 시 공정 비용이 적게 들며, 발효의 안정성이 유지되어 알코올 수율과 생산성을 높일 수 있음을 확인하고 본 발명을 완성하였다.The inventors of the present invention, while researching yeast with a high spectrum of use in the production of makgeolli, the new yeast separated from the yeast fermented with yeast is made fermentation with low acidity, high fermentation ratio and excellent sensory and flavor, and medium and low temperature In the fermentation is possible because the fermentation heat generated during the alcohol fermentation is less necessary because the cooling water is less process cost during the fermentation, the stability of the fermentation was confirmed that the alcohol yield and productivity can be improved and completed the present invention .
본 발명의 목적은 막걸리 제조 시 유용한 신규 효모를 제공하는 것이다. It is an object of the present invention to provide novel yeasts useful in the manufacture of makgeolli.
상기 목적을 달성하기 위하여 본 발명은 수탁번호: KACC 93276P로 수탁된, 사카로마이세스 세레비지에 KSD-YC를 제공한다.In order to achieve the above object, the present invention provides KSD-YC to Saccharomyces cerevisiae, deposited with accession number: KACC 93276P.
본 발명의 사카로마이세스 세레비지에 KSD-YC는 양조 발효 시 낮은 산도로 발효가 이루어지고, 발효 비율이 높으면서도 관능 및 풍미가 우수하며, 중온 및 저온에서도 발효가 가능하고, 발효의 안정성이 유지되는 특징이 있다.Saccharomyces cerevisiae KSD-YC of the present invention is fermented with low acidity during brewing fermentation, high fermentation ratio and excellent sensory and flavor, fermentation is possible at medium and low temperatures, stability of fermentation There is a feature to be maintained.
도 1은 S. 세레비지에 KSD-YC와 동일한 종 및 속에 포함되는 표준 ITS1-5.8S rRNA-ITS2 염기서열 부분을 비교한 결과이다.
도 2는 S. 세레비지에 KSD-YC의 ITS 유전자 서열에 따른 분류학적 계통분류도를 나타낸다.
도 3은 15 ℃에서 술 발효 시 술 내 총산 변화를 나타낸다.
도 4는 20 ℃에서 술 발효 시 술 내 총산 변화를 나타낸다.
도 5는 25 ℃에서 술 발효 시 술 내 총산 변화를 나타낸다.1 is a result of comparing the standard ITS1-5.8S rRNA-ITS2 nucleotide sequence included in the same species and genus as S. cerevisiae KSD-YC.
Figure 2 shows the taxonomic phylogeny according to the ITS gene sequence of S. cerevisiae KSD-YC.
Figure 3 shows the total acid change in alcohol upon fermentation at 15 ℃.
Figure 4 shows the total acid change in the alcohol during fermentation at 20 ℃.
Figure 5 shows the total acid change in alcohol upon fermentation at 25 ℃.
본 발명은,The present invention,
복원한 전통누룩을 이용하여 발효시킨 술덧에서 분리된 사카로마이세스 세레비지에 KSD-YC에 대한 것이다. 본 발명의 사카로마이세스 세레비지에 KSD-YC는, 중온 및 저온에서 술 발효 시 발효비율이 높으며, 저온에서 효모 생존율이 높고, 글루코스, 글리세롤, 소르비톨, 메틸-D-글루코사이드, 셀로비오스, 말토스, 사카로스 및 라피노스로 구성되는 군으로부터 선택되는 당을 이용할 수 있고, 수탁번호: KACC 93276P로 수탁되었다.Saccharomyces cerevisiae KSD-YC isolated from fermentation using fermented traditional yeast. Saccharomyces cerevisiae KSD-YC of the present invention has a high fermentation rate at the time of fermentation at high temperature and low temperature, high yeast survival rate at low temperature, glucose, glycerol, sorbitol, methyl-D-glucoside, cellobiose, horse A sugar selected from the group consisting of toss, saccharose and raffinose can be used and accession number: KACC 93276P.
또한 본 발명은 사카로마이세스 세레비지에 사카로마이세스 세레비지에 KSD-YC가 첨가된 식품 조성물에 대한 것이다.In addition, the present invention relates to a food composition in which KSD-YC is added to Saccharomyces cerevisiae.
또한 본 발명은In addition, the present invention
쌀, 물, 누룩 및 본 발명의 사카로마이세스 세레비지에 KSD-YC를 혼합하여 1단 발효하여 1단 발효물을 제조하는 단계; 및Preparing a one-stage fermentation product by mixing KSD-YC with rice, water, yeast and Saccharomyces cerevisiae of the present invention; And
상기 1단 발효물에 쌀, 물 및 누룩을 추가로 첨가하여 2단 발효하여 2단 발효물을 제조하는 단계를 포함하는 막걸리의 제조 방법에 대한 것이다.It relates to a method for producing makgeolli comprising the step of fermenting the two-stage fermentation by adding rice, water and yeast further to the one-stage fermentation.
이하, 본 발명을 자세히 설명한다.Hereinafter, the present invention will be described in detail.
사카로마이세스Saccharomyces 세레비지에Cerevisiae KSDKSD -- YCYC
본 발명의 사카로마이세스 세레비지에 KSD-YC는 복원한 전통누룩으로부터 발효한 술덧에서 분리되었으며, 중온 및 저온에서 술 발효 시 발효비율이 높으며, 저온에서 효모 생존율이 높다. 상기 중온은 18 ℃ 이상 24 ℃ 미만을 의미하며, 상기 저온은 12 ℃ 이상 18 ℃ 미만을 의미한다. 바람직하게는 본 발명의 사카로마이세스 세레비지에 KSD-YC는 15 ℃에서 생존률이 35% 이상 95% 미만이며, 바람직하게는 15 ℃에서 생존률이 40% 이상이다. 또한 본 발명의 사카로마이세스 세레비지에 KSD-YC는, 글루코스, 글리세롤, 소르비톨, 메틸-D-글루코사이드, 셀로비오스, 말토스, 사카로스 및 라피노스로 구성되는 군으로부터 선택되는 당을 이용할 수 있고, 수탁번호: KACC 93276P로 수탁된 것이다. Saccharomyces cerevisiae KSD-YC of the present invention was isolated from the fermentation from fermented traditional yeast, high fermentation rate at the fermentation of alcohol at medium and low temperatures, high yeast survival at low temperatures. The middle temperature means 18 ° C. or more and less than 24 ° C., and the low temperature means 12 ° C. or more and less than 18 ° C. Preferably, the Saccharomyces cerevisiae KSD-YC of the present invention has a survival rate of at least 35% and less than 95% at 15 ° C, preferably at least 40% at 15 ° C. In addition, the saccharomyces cerevisiae KSD-YC can be used a sugar selected from the group consisting of glucose, glycerol, sorbitol, methyl-D-glucoside, cellobiose, maltose, saccharose and raffinose. , Accession number: KACC 93276P.
이 때, 사카로마이세스 세레비지에 KSD-YC는 쌀을 원료로 하여 양조 발효 시 15 ℃에서 40 % 이상의 생존율을 가질 수 있고, 20 ℃에서 5 % 이상의 생존율을 가질 수 있다. 또한 사카로마이세스 세레비지에 KSD-YC는 25 ℃에서 80% 이상, 20℃에서 85% 이상, 15℃에서 95% 이상의 발효 비율을 가지며, 발효 비율은 하기 식 1에 따라 계산한다.At this time, KSD-YC in Saccharomyces cerevisiae may have a survival rate of 40% or more at 15 ° C. and a survival rate of 5% or more at 20 ° C. during fermentation using rice as a raw material. In addition, KSD-YC in Saccharomyces cerevisiae has a fermentation rate of at least 80% at 25 ℃, at least 85% at 20 ℃, 95% or more at 15 ℃, the fermentation rate is calculated according to the following
<식 1><
또한 본 발명의 사카로마이세스 세레비지에 KSD-YC는 2-케토-D-글루코네이트, 아라비노스, 자일로스, 아도니톨, 자일리톨, 갈락토스, 이노시톨, N-아세틸-글루코사민, 락토스, 트레할로스 및 멜레지토스로 구성되는 군으로부터 선택되는 당을 이용하지 않는다.In addition, the Saccharomyces cerevisiae KSD-YC is 2-keto-D-gluconate, arabinose, xylose, adonitol, xylitol, galactose, inositol, N-acetyl-glucosamine, lactose, trehalose and It does not use a sugar selected from the group consisting of melezitose.
상기 사카로마이세스 세레비지에 KSD-YC는 누룩으로부터 발효된 술덧의 유래이다.The Saccharomyces cerevisiae KSD-YC is derived from yeast fermented from yeast.
또한 본 발명의 사카로마이세스 세레비지에 KSD-YC는 저산도 발효가 가능하여 양조용 쌀을 고도 도정하지 않고도 깔끔한 주질을 구현할 수 있어 경제적으로 큰 장점을 지닐 수 있고, 저온에서 술 발효 시 효모 생존율과 발효 비율이 높으며, 발효 온도의 스펙트럼이 넓어 알코올 발효 시 발생되는 발효열을 냉각수로 낮춰 줄 필요가 적기 때문에 발효 시 공정 비용이 적게 들며, 발효의 안정성이 유지되어 알코올 수율과 생산성을 높일 수 있다.In addition, KSD-YC in the Saccharomyces cerevisiae of the present invention is capable of low acidity fermentation, so that it is possible to realize a neat quality without high-quality brewing rice, and it can have economic advantages, and yeast during fermentation at low temperature. The survival rate and fermentation rate are high, and because the spectrum of fermentation temperature is wide, it is not necessary to lower the heat of fermentation produced by alcohol fermentation with cooling water. .
식품 조성물Food composition
본 발명은 사카로마이세스 세레비지에 KSD-YC가 첨가된 식품 조성물에 대한 것이다. 상기 식품의 종류는 제한되지 않으며, 일반적인 식품, 건강보조식품, 기능성 식품 등이 될 수 있다. 바람직하게는 상기 식품 조성물은 효모가 사용되는 식품 조성물이다. 더욱 바람직하게는 상기 식품 조성물은 주류이며, 예컨대, 약주, 탁주, 청주 등 발효주 및 증류주 등이다. The present invention relates to a food composition in which KSD-YC is added to Saccharomyces cerevisiae. The type of the food is not limited, and may be general food, health supplement food, functional food, and the like. Preferably the food composition is a food composition in which yeast is used. More preferably, the food composition is liquor, for example, fermented liquor, distilled liquor, etc., such as medicinal liquor, takju, and sake.
막걸리의 제조 방법Manufacturing method of rice wine
본 발명은 쌀, 물, 누룩 및 본 발명의 사카로마이세스 세레비지에 KSD-YC를 혼합하여 1단 발효하여 1단 발효물을 제조하는 단계; 및 상기 1단 발효물에 쌀, 물 및 누룩을 추가로 첨가하여 2단 발효하여 2단 발효물을 제조하는 단계를 포함하는 막걸리의 제조 방법에 대한 것이다. 이 때, 쌀, 물, 누룩 등을 이용하여 막걸리를 제조하는 일반적인 기술을 이용하여 본 발명의 막걸리의 제조 방법을 수행할 수 있다.The present invention comprises the steps of preparing a single-stage fermentation by mixing KSD-YC in rice, water, yeast and Saccharomyces cerevisiae of the present invention; And adding rice, water, and yeast to the first stage fermentation to produce two stage fermentation by fermenting two stages. At this time, the manufacturing method of the makgeolli of the present invention may be performed using a general technique of preparing makgeolli using rice, water, yeast, and the like.
본 발명의 이점 및 특징, 그리고 그것들을 달성하는 방법은 상세하게 후술되어 있는 실시예들을 참조하면 명확해질 것이다. 그러나, 본 발명은 이하에서 개시되는 실시예들에 한정되는 것이 아니라 서로 다른 다양한 형태로 구현될 것이며, 단지 본 실시예들은 본 발명의 개시가 완전하도록 하며, 본 발명이 속하는 기술분야에서 통상의 지식을 가진 자에게 발명의 범주를 완전하게 알려주기 위해 제공되는 것이며, 본 발명은 청구항의 범주에 의해 정의될 뿐이다.Advantages and features of the present invention and methods for achieving them will be apparent with reference to the embodiments described below in detail. However, the present invention is not limited to the embodiments disclosed below, but may be implemented in various different forms, only the present embodiments to make the disclosure of the present invention complete, and common knowledge in the art to which the present invention pertains. It is provided to fully inform the person having the scope of the invention, which is defined only by the scope of the claims.
<재료 및 방법><Materials and Methods>
비교 균주들로는 국내 시판 중인 사카로마이세스 세레비지에 S1, 사카로마이세스 세레비지에 S2 및 사카로마이세스 세레비지에 S3을 사용하였다(표 1). Comparative strains were S1 in Saccharomyces cerevisiae, S2 in Saccharomyces cerevisiae and S3 in Saccharomyces cerevisiae (Table 1).
<실험예 1> 균주의 분리Experimental Example 1 Isolation of Strains
100 중량부의 통밀을 건식분쇄기로 분쇄한 후 물 40 중량부를 가하여 수분함량이 30~40% 정도로 균일하게 하고 두께 2.5cm, 지름 20cm 크기의 원반상으로 디뎌 볏짚이 깔린 선반 위에 놓고 14일간 실내에서 실온으로 발효하였다. 그 후 7일간 볕에 말려 법제시켜 전통 밀누룩을 제조하였다.100 parts by weight of whole wheat flour are pulverized with a dry mill, and 40 parts by weight of water is added to make the water content 30-30% evenly, and placed on a disc with a thickness of 2.5 cm and a diameter of 20 cm. Fermentation was carried out. After that, it was dried for 7 days to make a traditional wheat yeast.
상기 전통 밀누룩 및 증자된 쌀, 물을 혼합하여 15 ℃, 20 ℃ 및 25 ℃ 조건에서 3일간 발효하여 술덧을 만들었다. 이 후, 술덧에 멸균된 증류수로 희석 도말하여 균주들을 분리하였다. 분리된 균주들로부터 효모를 선별하고, 상기 효모들의 발효 비율 및 알코올 생성능 즉, 양조 적성을 평가하였다. 그 중 총산(total acid)이 낮으면서 발효 비율 및 알코올 생성능과 관능 및 풍미가 우수한 균주를 선택하여 하기 실험예 2를 진행하였다(시험 결과를 기재하지 않음). The traditional wheat malt and steamed rice and water were mixed and fermented at 15 ° C., 20 ° C. and 25 ° C. for 3 days to make drunk. Thereafter, the strains were separated by dilution with distilled water sterilized by distillation. Yeasts were selected from the isolated strains and the fermentation rate and alcohol production capacity of the yeasts were evaluated. Among them, a low total acid (total acid) and the fermentation rate and alcohol production ability, the sensory and flavor was selected to proceed to the following Experimental Example 2 (not described test results).
<실험예 2> 계통발생적 분석을 통한 확인Experimental Example 2 Confirmation through Phylogenetic Analysis
상기 실험예 1에서 양조 적성이 가장 우수한 것으로 확인된, 전통 누룩으로부터 분리된 균주의 종을 확인하기 위하여, ITS 유전자의 염기서열을 분석하였다. 상기 분석은 ㈜솔젠트(대전, 대한민국)에서 ITS1 및 ITS4 프라이머(primer)를 사용하여 시퀀싱(sequencing)하여 수행하였다(대전, 대한민국). 구체적으로는, ITS 유전자 전장서열을 SeqMan 소프트웨어(DNASTAR)를 사용하여 조립하였고, NCBI 데이터베이스에 블라스팅 하였다. 그리고 분류학적으로 연관성이 있는 ITS 유전자 서열들을 GenBank로부터 수득하여 염기서열을 BioEdit 프로그램으로 비교하여 수행하였다.In order to identify the species of the strain isolated from the traditional yeast, which was confirmed to have the best brewing aptitude in Experimental Example 1, the nucleotide sequence of the ITS gene was analyzed. The analysis was performed by sequencing using ITS1 and ITS4 primers from Solgent (Daejeon, Korea) (Daejeon, Korea). Specifically, the ITS gene full sequence was assembled using SeqMan software (DNASTAR) and blasted into the NCBI database. Taxonomy related ITS gene sequences were obtained from GenBank and compared with nucleotide sequences by BioEdit program.
또한 유전자 서열에 따른 균주의 계통학적 위치를 MEGA 3 프로그램을 사용하여, 1000번 레플리케이션에 기초한 bootstrap 값을 갖는 neighbor-joining 방법을 사용함으로써 계통학적 트리를 구축하였다.In addition, the systematic location of the strain according to the gene sequence was constructed using the
그 결과, 상기 균주의 ITS 유전자의 염기서열은 하기 서열번호 1과 같았으며(표 2), 사카로마이세스 세레비지에 종에 속하는 표준 균주들과 ITS1-5.8S rRNA-ITS2 염기서열 부분을 비교한 결과, 본 균주는 사카로마이세스 세레비지에에 속하는 것으로 확인되었다(도 1). ITS 유전자 서열에 따른 분류학적 계통 분류도는 도 2와 같았으며(도 2), 종래의 사카로마이세스 세레비지에와 ITS 유전자 서열이 상당히 차이가 나는바 신규 균주로 확인되었다. 그러므로 이를 사카로마이세스 세레비지에 KSD-YC로 명명하고 2017년 1월 24일 국립농업과학원에 기탁하였다(미생물 수탁번호: KACC 93276P). As a result, the nucleotide sequence of the ITS gene of the strain was as shown in SEQ ID NO: 1 (Table 2), comparing the ITS1-5.8S rRNA-ITS2 nucleotide sequence with the standard strains belonging to Saccharomyces cerevisiae species As a result, the strain was found to belong to Saccharomyces cerevisiae (Fig. 1). The taxonomy of taxonomy according to ITS gene sequence was the same as that of FIG. 2 (FIG. 2), and it was identified as a novel strain because the ITS gene sequence was significantly different from the conventional Saccharomyces cerevisiae. Therefore, it was named as KSD-YC in Saccharomyces cerevisiae and deposited with the National Academy of Agricultural Science on January 24, 2017 (Microbial Accession No .: KACC 93276P).
<실험예 3> 내알코올성 평가Experimental Example 3 Evaluation of Alcohol Resistance
사카로마이세스 세레비지에 KSD-YC의 내알코올성을 평가하였다. 먼저, 배양된 대수기의 효모를 에탄올이 각각 1, 5, 10, 15%(v/v)의 농도로 함유된 YPD 배지 30mL에 초기균수 1.0E+05 cells/mL의 농도로 접종한 후, 30 ℃, 150 rpm의 조건으로 배양하였다. 배양 24시간이 경과한 후 배양액을 채취하여 희석한 후 600nm에서 흡광도를 측정하였다.Saccharomyces cerevisiae was evaluated for the alcohol resistance of KSD-YC. First, the incubated yeast of the log phase was inoculated in 30 mL of YPD medium containing ethanol at a concentration of 1, 5, 10, 15% (v / v), respectively, at a concentration of 1.0E + 05 cells / mL of the initial bacterial count, The cells were cultured at 30 ° C. and 150 rpm. After 24 hours of incubation, the culture solution was collected and diluted, and the absorbance was measured at 600 nm.
일반적으로 일정 농도 이상의 에탄올 농도에서는 효모의 생육이 저해되어 알코올 생성에 부정적인 영향을 미친다. 그러므로 에탄올에 대한 내성을 갖는 것이 양조효모로서 바람직하다. 상기 흡광도 측정 결과, 에탄올을 5%의 농도로 첨가한 배지에서의 효모 생육은 전반적으로 양호하였다. 특히 사카로마이세스 세레비지에 KSD-YC는 다른 균주들에 비하여 18 내지 42% 더 높은 내성을 보였으며, 에탄올을 10%의 농도로 첨가한 배지에서는 다른 균주들에 비해 15 내지 460% 높은 내성을 보였다. 그러므로 사카로마이세스 세레비지에 KSD-YC는 내알코올성이 우수한 것으로 확인되었다(표 3).In general, the ethanol concentration above a certain concentration inhibits the growth of yeast and negatively affects the production of alcohol. Therefore, it is preferable as brewing yeast to have resistance to ethanol. As a result of absorbance measurement, yeast growth was generally good in the medium to which ethanol was added at a concentration of 5%. In particular, KSD-YC showed 18-42% higher resistance to Saccharomyces cerevisiae than other strains, and 15-460% higher resistance than other strains in medium containing 10% ethanol. Showed. Therefore, Saccharomyces cerevisiae KSD-YC was confirmed to have excellent alcohol resistance (Table 3).
<실험예 4> 내당성 평가Experimental Example 4 Evaluation of Tolerability
일반적으로 양조효모는 고농도의 당에서 생육하면서 알코올 발효를 할 수 있어야 한다. 그러므로 사카로마이세스 세레비지에 KSD-YC의 내당성(Glucose Tolerance)을 평가하였다. 먼저, 배양된 대수기의 효모를 포도당이 각각 2, 10, 20, 30, 40%(v/v)의 농도로 함유된 YPD 배지 30mL에 초기균수 1.0E+05 cells/mL의 농도로 접종한 후 30 ℃, 150 rpm 조건에서 배양하였다. 배양 24시간이 경과한 후 배양액을 채취하여 적당히 희석한 후 600nm에서 흡광도를 측정하였다.Generally, brewing yeast should be able to ferment alcohol while growing in high concentration of sugar. Therefore, we evaluated the glucose tolerance of KSD-YC in Saccharomyces cerevisiae. First, the incubated yeast of the log phase was inoculated at 30 mL of YPD medium containing glucose at concentrations of 2, 10, 20, 30 and 40% (v / v), respectively, at a concentration of 1.0E + 05 cells / mL of the initial bacterial count. Then incubated at 30 ℃, 150 rpm conditions. After 24 hours of incubation, the culture solution was collected, diluted appropriately, and the absorbance was measured at 600 nm.
그 결과, 균주들은 대체적으로 포도당이 20% 첨가된 배지에서 양호한 생육을 보였다. 특히, 본 발명의 사카로마이세스 세레비지에 KSD-YC는 다른 균주들에 비하여 19 내지 44%의 높은 내당성을 보였다 (표 4).As a result, the strains generally showed good growth in medium added with 20% glucose. In particular, Saccharomyces cerevisiae KSD-YC of the present invention showed a higher sugar resistance of 19 to 44% compared to other strains (Table 4).
<실험예 5> 양조학적 특성Experimental Example 5 Brewing Characteristics
쌀 900 g, 물 1350 mL, 누룩 8.6g 및 효모 배양액을 혼합하였다. 이 때 밑술 mL 당 6.5E+07 cell/mL의 농도가 되게끔 혼합하여, 1단 담금을 하였다. 그리고 15 ℃, 20 ℃ 및 25 ℃ 조건에서 1일간 발효하여 1단 발효를 수행하였다. 900 g of rice, 1350 mL of water, 8.6 g of yeast and yeast culture were mixed. At this time, the mixture was mixed so as to have a concentration of 6.5E + 07 cells / mL per mL of the lower liquor, and immersion was performed in one step. Then, the fermentation was carried out for 1 day at 15 ° C., 20 ° C. and 25 ° C. to carry out single stage fermentation.
1단 발효 후 쌀 2,100 g, 물 3,150mL 및 누룩 20.1g을 추가로 첨가하여 2단 담금하고 그 후 각 15 ℃, 20 ℃ 및 25 ℃ 조건으로 2단 발효하였다. 이 때, 1단 발효와 2단 발효 시 온도는 동일하게 하였으며, 각 실험은 4종의 효모들을 종류 별로 이용하여 수행하였다(S. 세레비지에 KSD-YC, S. 세레비지에 S1, S. 세레비지에 S2, 및 S. 세레비지에 S3).After one-stage fermentation, 2,100 g of rice, 3,150 mL of water, and 20.1 g of yeast were additionally immersed in two stages, and then fermented in two stages under 15 ° C, 20 ° C, and 25 ° C, respectively. At this time, the temperature of the first and second stage fermentation was the same, each experiment was carried out using four types of yeast by type (S. cerevisiae KSD-YC, S. cerevisiae S1, S. S2 in cerevisiae, and S. S3 in cerevisiae).
그리고 발효 온도에 따른 술 발효시 발효 산물, 즉 술의 총산(total acid) 함량, 알코올 농도 및 발효 비율을 평가하였다. 이 때, 발효 비율은 하기 식 1에 따라 계산하였다. The fermentation products, ie total acid content, alcohol concentration and fermentation ratio of the fermentation products were evaluated. At this time, the fermentation ratio was calculated according to the following formula (1).
한편, 아미노산도는 발효가 끝난 술덧을 여과지로 여과한 시료 10mL에 0.1N NaOH로 중화 시킨 후, 중성 포르말린 용액 5mL를 가하여 혼합하고 여기에 0.1N NaOH 용액으로 적정하여 pH 8.3이 될 때까지 소요된 0.1N NaOH의 양으로 표시하여 평가하였다. On the other hand, the amino acid degree is neutralized with 0.1N NaOH to 10mL of the sample of the fermentation finished with filter paper, and then mixed with 5mL of neutral formalin solution and titrated with 0.1N NaOH solution to pH 8.3 It was evaluated by expressing it in the amount of 0.1N NaOH.
또한 발효 중 효모의 생존율은 하기 식 2에 따라 계산하였다. In addition, the survival rate of yeast during fermentation was calculated according to the following
<식 1><
<식 2><
그 결과, S. 세레비지에 KSD-YC는 다른 효모들에 비하여, 발효물인 술의 총 산도가 낮아, 부드럽고 담백한 주질을 부여할 수 있을 것으로 판단되었다(표 5 내지 8, 도 3 내지 5). 또한 고농도로 알코올이 생성되고, 발효 비율이 높았다. 특히, 저온 조건에서 다른 효모들에 비하여 생성되는 알코올 농도가 높고 효모 생존율도 높은 것으로 확인되었다(표 10, 표 11).As a result, KSD-YC in S. cerevisiae was lower than the other yeasts, it was judged that the total acidity of the liquor, which is a fermentation product, could be given a soft and light liquor (Tables 5 to 8, FIGS. 3 to 5). In addition, alcohol was produced at a high concentration, and the fermentation rate was high. In particular, it was confirmed that the alcohol concentration is higher than the other yeast produced at low temperature conditions, and the yeast survival rate is also high (Table 10, Table 11).
발효 온도에 따른 술 발효시 발효물의 총 산 함량 (%(w/v))Total acid content of fermentation product in liquor fermentation according to fermentation temperature (% (w / v))
15 ℃에서 술 발효 시 발효시간에 따른 총 산 변화Total Acid Variation with Fermentation Time During Alcohol Fermentation at 15 ℃
20 ℃에서 술 발효 시 발효시간에 따른 총 산 변화Total Acid Changes with Fermentation Time at 20 ° C
25 ℃에서 술 발효 시 총 산 변화Total Acid Changes During Alcohol Fermentation at 25 ° C
발효 온도 및 효모 종류에 따른 술 발효 시 아미노산도Amino Acid Degree in Alcohol Fermentation by Fermentation Temperature and Yeast Type
발효 온도 및 효모 종류에 따른 효모의 생존율Survival Rate of Yeast by Fermentation Temperature and Yeast Types
발효 온도 및 효모 종류에 따른 최종 알코올 농도 및 발효 비율Final alcohol concentration and fermentation rate according to fermentation temperature and yeast type
<실험예 6> 관능적 특성Experimental Example 6 Sensory Characteristics
상기 실험예 5에서 20℃에서 발효 및 제조된 양조효모별 최종 발효 술덧에 대하여 관능검사를 실시하였다. 먼저, 훈련된 검사요원 15명을 선발하고 9점 척도법(1-좋지 않다, 3-좋지 않은 편이다, 5-보통이다, 7-좋은 편이다, 9-좋다)을 통해 맛과 향 및 전체적인 선호도를 조사하였다. In Experimental Example 5, the sensory test was carried out for the final fermentation by brewing yeast fermented and manufactured at 20 ° C. First, 15 trained inspectors were selected and the taste, aroma, and overall preferences were determined using a 9-point scale (1- Not Good, 3- Not Good, 5- Normal, 7- Good, 9- Good). Was investigated.
그 결과, S. 세레비지에 KSD-YC가 향, 맛 및 전반적 모든 항목에서 유사 내지는 높은 기호도를 나타냈다(표 12). 그러므로 S. 세레비지에 KSD-YC를 이용하여 양조할 경우 맛과 향 등 관능적으로 우수한 주질을 부여할 수 있을 것으로 판단되었다.As a result, S. cerevisiae KSD-YC showed similar or high preference in all the items of flavor, taste and overall (Table 12). Therefore, it was judged that S. cerevisiae could give sensory excellent quality such as taste and aroma when brewed using KSD-YC.
실시예 6, 양조효모별 20℃ 최종 발효 술덧에 대한 관능검사 결과Example 6, sensory test results for the final fermentation 20 ℃ ℃ by brewing yeast
<실험예 7> 당 이용능 평가Experimental Example 7 Evaluation of Sugar Utilization
사카로마이세스 세레비지에 KSD-YC의 당 이용 특성을 API 20C AUX kit(Biomerieux사, 프랑스)를 이용하여 분석하였다. 이 때 비교 균주들로는 사카로마이세스 세레비지에 S1 및 사카로마이세스 세레비지에 S2를 사용하였다.The sugar utilization characteristics of KSD-YC in Saccharomyces cerevisiae were analyzed using API 20C AUX kit (Biomerieux, France). In this case, S1 was used as Saccharomyces cerevisiae and S2 was used as Saccharomyces cerevisiae.
그 결과, 본 발명의 사카로마이세스 세레비지에 KSD-YC는 글루코스, 글리세롤, 소르비톨, 메틸-D-글루코사이드, 셀로비오스, 말토스, 사카로스 및 라피노스를 이용할 수 있었다. 그러나 본 발명의 사카로마이세스 세레비지에는 2-케토-D-글루코네이트, 아라비노스, 자일로스, 아도니톨, 자일리톨, 갈락토스, 이노시톨, N-아세틸-글루코사민, 락토스, 트레할로스 및 멜레지토스의 이용능은 없었다(표 13).As a result, KSD-YC in the Saccharomyces cerevisiae of the present invention was able to use glucose, glycerol, sorbitol, methyl-D-glucoside, cellobiose, maltose, saccharose and raffinose. However, Saccharomyces cerevisiae of the present invention contains 2-keto-D-gluconate, arabinose, xylose, adonitol, xylitol, galactose, inositol, N-acetyl-glucosamine, lactose, trehalose and melezitose. There was no availability (Table 13).
기탁기관명 : 농업생명공학연구원Depositary Name: Institute of Agricultural Biotechnology
수탁번호 : KACC93276PAccession number: KACC93276P
수탁일자 : 2017124Deposit Date: 2017124
<110> KOOKSOONDANG <120> SACCHAROMYCES CEREVISIAE KSD-YC WHICH IS A NOVEL YEAST <130> KNP170020 <160> 1 <170> KoPatentIn 3.0 <210> 1 <211> 789 <212> DNA <213> Saccharomyces cerevisiae <400> 1 attaaagaaa tttaataatt ttgaaaatgg atttttttgt tttggcaaga gcatgagagc 60 ttttactggg caagaagaca agagatggag agtccagccg ggcctgcgct taagtgcgcg 120 gtcttgctag gcttgtaagt ttctttcttg ctattccaaa cggtgagaga tttctgtgct 180 tttgttatag gacaattaaa accgtttcaa tacaacacac tgtggagttt tcatatcttt 240 gcaacttttt ctttgggcat tcgagcaatc ggggcccaga ggtaacaaac acaaacaatt 300 ttatttattc attaaatttt tgtcaaaaac aagaattttc gtaactggaa attttaaaat 360 attaaaaact ttcaacaacg gatctcttgg ttctcgcatc gatgaagaac gcagcgaaat 420 gcgatacgta atgtgaattg cagaattccg tgaatcatcg aatctttgaa cgcacattgc 480 gccccttggt attccagggg gcatgcctgt ttgagcgtca tttccttctc aaacattctg 540 tttggtagtg agtgatactc tttggagtta acttgaaatt gctggccttt tcattggatg 600 tttttttttc caaagagagg tttctctgcg tgcttgaggt ataatgcaag tacggtcgtt 660 ttaggtttta ccaactgcgg ctaatctttt ttatactgag cgtattggaa cgttatcgat 720 aagaagagag cgtctaggcg aacaatgttc ttaaagtttg acctcaaatc aggtaggagt 780 acccgctga 789 <110> KOOKSOONDANG <120> SACCHAROMYCES CEREVISIAE KSD-YC WHICH IS A NOVEL YEAST <130> KNP170020 <160> 1 <170> KoPatentIn 3.0 <210> 1 <211> 789 <212> DNA <213> Saccharomyces cerevisiae <400> 1 attaaagaaa tttaataatt ttgaaaatgg atttttttgt tttggcaaga gcatgagagc 60 ttttactggg caagaagaca agagatggag agtccagccg ggcctgcgct taagtgcgcg 120 gtcttgctag gcttgtaagt ttctttcttg ctattccaaa cggtgagaga tttctgtgct 180 tttgttatag gacaattaaa accgtttcaa tacaacacac tgtggagttt tcatatcttt 240 gcaacttttt ctttgggcat tcgagcaatc ggggcccaga ggtaacaaac acaaacaatt 300 ttatttattc attaaatttt tgtcaaaaac aagaattttc gtaactggaa attttaaaat 360 attaaaaact ttcaacaacg gatctcttgg ttctcgcatc gatgaagaac gcagcgaaat 420 gcgatacgta atgtgaattg cagaattccg tgaatcatcg aatctttgaa cgcacattgc 480 gccccttggt attccagggg gcatgcctgt ttgagcgtca tttccttctc aaacattctg 540 tttggtagtg agtgatactc tttggagtta acttgaaatt gctggccttt tcattggatg 600 tttttttttc caaagagagg tttctctgcg tgcttgaggt ataatgcaag tacggtcgtt 660 ttaggtttta ccaactgcgg ctaatctttt ttatactgag cgtattggaa cgttatcgat 720 aagaagagag cgtctaggcg aacaatgttc ttaaagtttg acctcaaatc aggtaggagt 780 acccgctga 789
Claims (5)
15~20 ℃에서 쌀을 원료로 하여 양조 발효 시 0.168% 미만의 총산을 갖는,
수탁번호: KACC 93276P로 수탁된, 사카로마이세스 세레비지에 KSD-YC.
Glucose, glycerol, sorbitol, methyl-D-glucoside, cellobiose, maltose, saccharose and raffinose can be used,
It has less than 0.168% total acid in brewing fermentation with rice as raw material at 15 ~ 20 ℃
Accession number: Saccharomyces cerevisiae KSD-YC, deposited under KACC 93276P.
15 ℃에서 쌀을 원료로 하여 양조 발효 시 40 % 이상의 생존율을 가지며, 상기 생존율은 하기 식 2에 따라 계산한 것을 특징으로 하는 사카로마이세스 세레비지에 KSD-YC.
<식 2>
The method of claim 1,
Saccharomyces cerevisiae KSD-YC. Has a survival rate of 40% or more during brewing fermentation using rice as a raw material at 15 ℃, the survival rate was calculated according to the following formula (2).
<Equation 2>
15~25 ℃에서 쌀을 원료로 하여 양조 발효 시 84% 이상의 발효 비율을 가지며, 상기 발효 비율은 하기 식 1에 따라 계산한 것을 특징으로 하는 사카로마이세스 세레비지에 KSD-YC.
<식 1>
The method of claim 1,
Saccharomyces cerevisiae KSD-YC. It has a fermentation rate of 84% or more during fermentation with rice as a raw material at 15 ~ 25 ℃, the fermentation rate was calculated according to the following formula 1.
<Equation 1>
The food composition to which KSD-YC is added to Saccharomyces cerevisiae of Claim 1.
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