KR20230149157A - Saccharomyces cerevisiae strain nibrfgc000501771 for dry-typt rice wine and process for preparing method of dry-typt rice wine using same - Google Patents
Saccharomyces cerevisiae strain nibrfgc000501771 for dry-typt rice wine and process for preparing method of dry-typt rice wine using same Download PDFInfo
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- KR20230149157A KR20230149157A KR1020220048519A KR20220048519A KR20230149157A KR 20230149157 A KR20230149157 A KR 20230149157A KR 1020220048519 A KR1020220048519 A KR 1020220048519A KR 20220048519 A KR20220048519 A KR 20220048519A KR 20230149157 A KR20230149157 A KR 20230149157A
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- strain
- liquor
- ethyl
- yeast
- dry type
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Abstract
Description
본 발명은 드라이 타입의 약주 제조용 사카로마이세스 세레비지애 NIBRFGC000501771 균주 및 이를 이용한 드라이 타입의 약주 제조 방법에 관한 것이다.The present invention relates to the Saccharomyces cerevisiae NIBRFGC000501771 strain for producing dry type herbal liquor and a method for producing dry type herbal liquor using the same.
우리나라는 예로부터 가가호호 술을 빚어 마시는 풍습이 뿌리를 내려왔다. 지방에 따라, 가문에 따라 또 빚는 사람의 솜씨에 따라 갖가지 방법과 기술을 발휘한 가양주들이 등장하여 맛과 향기를 자랑했는 바, 이러한 배경에서 향토성을 띤 토속주가 명주로 등장하였고 현재에도 많은 양조장에서 전통적인 가양주를 살리고자 하는 노력이 진행되고 있으며 품질을 위해 현대적인 기술을 접목한 술들이 출시되고 있다. In Korea, the custom of making and drinking alcohol has been rooted since ancient times. Depending on the region, family, and craftsmanship of the brewer, gayangjus using various methods and techniques appeared, boasting taste and aroma. Against this background, local liquors with local characteristics emerged as famous liquors, and are still used in many breweries today. Efforts are being made to revive traditional gayangju, and alcohol that incorporates modern technology to improve quality is being released.
다만, 양조 과정에서 누룩만을 사용하고 효모는 누룩에 존재하는 야생효모를 그대로 이용하여 만든 술은 발효와 공정 관리에 어렵고 품질이 안정적이지 않다. 일부 전통주 양조장에서는 안정적인 품질관리를 위해서 효모를 구매하여 사용하는데, 우리 전통주의 주조에 활용할 수 있는 효모는 매우 제한적이다. 실제로 효모 제품의 국내시장 규모는 연 230억이나, 대부분 수입에 의존하는 실정이며, 이로 인해 국산 주류의 정통성이 담보되지 못할 뿐 아니라, 외화 낭비도 심각한 실정이다. However, alcohol made by using only yeast during the brewing process and wild yeast present in the yeast is difficult to manage and control the fermentation process, and the quality is not stable. Some traditional liquor breweries purchase and use yeast for stable quality control, but the yeast that can be used for brewing our traditional liquor is very limited. In fact, the domestic market for yeast products is KRW 23 billion per year, but most of them depend on imports. As a result, not only is the authenticity of domestically produced alcoholic beverages not guaranteed, but there is also a serious waste of foreign currency.
특히, 양조에 사용되는 효모는 많은 양조장에서는 수입 제빵효모를 사용하는 경우가 많은데 이는 수입 효모가 배양된 환경이 외국이기에 우리나라 국산 원재료를 사용하는 양조와는 그 환경이 맞지 않는다. 따라서, 정통성을 확보하고 우수한 풍미를 발현하는 전통주의 양조를 위한 우수한 효모 균주의 개발이 필요한 실정이다.In particular, many breweries use imported baker's yeast for the yeast used in brewing, but this is because the environment in which the imported yeast is cultivated is a foreign country, and the environment does not match that of brewing using domestic raw materials. Therefore, there is a need to develop excellent yeast strains for brewing traditional liquor that secures authenticity and exhibits excellent flavor.
이러한 국내 현실을 반영하여 다양한 풍미를 발현하는 자생 효모 균주를 개발하고자, 본 발명자들은 전국의 꽃, 열매, 누룩, 토양 등에서 야생효모 1,700여 균주를 순수 분리·배양하여 보존 중이며, 이중 배양이 쉽고 알코올 발효 능력이 있어 양조에 활용이 가능한 88개 균주를 대상으로 양조 품질과 적합성을 평가하였다. 그 결과, 알코올 생성력과 응집성이 좋은 드라이 타입의 약주 제조에 적합한 우수한 양조효모의 특성을 규명하여 본 발명을 완성하게 되었다.In order to develop native yeast strains that express various flavors by reflecting this domestic reality, the present inventors have isolated, cultured, and preserved over 1,700 strains of wild yeast from flowers, fruits, yeast, soil, etc. across the country. Double cultivation is easy and alcohol Brewing quality and suitability were evaluated for 88 strains that have fermentation ability and can be used for brewing. As a result, the present invention was completed by identifying the characteristics of excellent brewing yeast suitable for the production of dry type herbal liquor with good alcohol production ability and cohesiveness.
따라서, 본 발명의 주된 목적은 드라이 타입의 약주 제조용 사카로마이세스 세레비지애 신균주 NIBRFGC000501771(기탁번호 KACC 83061BP)을 제공하는 데 있다.Therefore, the main purpose of the present invention is to provide a new strain of Saccharomyces cerevisiae NIBRFGC000501771 (accession number KACC 83061BP) for producing dry type medicinal liquor.
본 발명의 다른 목적은 상기 사카로마이세스 세레비지애 신균주 NIBRFGC000501771(기탁번호 KACC 83061BP)을 이용하여 드라이 타입 약주의 제조방법과 이에 따라 제조된 드라이 타입의 약주를 제공하는데 있다.Another object of the present invention is to provide a method for manufacturing dry type herbal liquor using the new Saccharomyces cerevisiae strain NIBRFGC000501771 (accession number KACC 83061BP) and the dry type herbal liquor prepared thereby.
본 발명의 다른 목적 및 이점은 하기의 발명의 상세한 설명, 청구범위 및 도면에 의해 보다 명확하게 된다.Other objects and advantages of the present invention will become clearer from the following detailed description, claims, and drawings.
본 발명의 한 양태에 따르면, 본 발명은 드라이 타입의 약주 제조용 사카로마이세스 세레비지애 신균주 NIBRFGC000501771(기탁번호 KACC 83061BP)을 제공한다.According to one aspect of the present invention, the present invention provides a new strain of Saccharomyces cerevisiae NIBRFGC000501771 (accession number KACC 83061BP) for producing dry type medicinal liquor.
본 발명자들은 약주 제조에 사용되는 수입 효모를 대체하는 국산 효모 균주를 개발하기 위하여 예의 연구 노력한 결과, 사카로마이세스 세레비지애 신균주 NIBRFGC000501771(기탁번호 KACC 83061BP)이 시중에서 판매되는 상용 효모보다 관능성에서 뛰어난 특성을 나타내며, 향기 성분을 고농도로 생산하고, 드라이 타입의 약주 생산에 유용함을 확인하고, 본 발명을 완성하게 되었다.As a result of intensive research efforts by the present inventors to develop a domestic yeast strain to replace the imported yeast used in the manufacture of medicinal liquor, the new Saccharomyces cerevisiae strain NIBRFGC000501771 (accession number KACC 83061BP) has a higher sensory function than commercially available yeast. It was confirmed that it exhibits excellent properties, produces aroma components at high concentrations, and is useful in the production of dry type herbal liquor, leading to the completion of this invention.
본 발명의 드라이 타입의 약주 제조용 사카로마이세스 세레비지애 신균주 NIBRFGC000501771은 국립생물자원관에서 분리·배양한 야생효모 1,700여 균주 중 배양이 쉽고, 알코올 발효 능력이 있어 양조에 활용이 가능한 88개 균주를 대상으로, 크기, 발효력, 응집성(Flocculation), Cu 배지 성장성, 포자 생성(Sporulation), Melibiose 이용 능력 등의 연구 및 약주 시험양조 연구를 통해 양조용으로 사업화 가능성이 높은 33종을 1차 선발한 후, 시험양조를 통해 알코올 생성력과 효모의 응집성이 좋은 드라이 타입의 효모 4종을 2차 선발하고, 최종적으로 알코올 생성력, 당함량 축적 정도, 발효의 안정성, 유기산 함량, 불휘발분 잔류량, 효모 응집성(Flocculation), 향기 성분 분석 및 관능 평가를 통해 도출된 드라이 타입의 약주 양조용 효모이다. 상기 균주는 2022년 02월 17일자로 국립농업과학원 농업유전자원센터(KACC)에 사카로마이세스 세레비지애 NIBRFGC000501771 균주로 기탁하여 KACC 83061BP를 부여받았다. 본 발명에서 "사카로마이세스 세레비지애 NIBRFGC000501771 균주", "NIBRFGC000501771 균주" 및 "501771 균주"는 동일한 의미로 서로 혼용하여 사용할 수 있다.The new strain of Saccharomyces cerevisiae for producing dry type herbal liquor NIBRFGC000501771 of the present invention is one of 88 strains that are easy to culture and have alcohol fermentation ability, which can be used in brewing among over 1,700 wild yeast strains isolated and cultured at the National Institute of Biological Resources. Through research on size, fermentation power, flocculation, growth on Cu medium, spore production, and Melibiose usage ability, as well as medicinal liquor test brewing research, 33 types with high potential for commercialization for brewing were initially selected. Afterwards, through test brewing, four types of dry type yeast with good alcohol production ability and yeast cohesion were selected for the second time, and finally alcohol production ability, degree of sugar content accumulation, fermentation stability, organic acid content, non-volatile matter residual amount, and yeast cohesion ( Flocculation), a dry type medicinal liquor brewing yeast derived through aroma component analysis and sensory evaluation. The strain was deposited as Saccharomyces cerevisiae NIBRFGC000501771 strain at the Agricultural Genetic Resources Center (KACC) of the National Academy of Agricultural Sciences on February 17, 2022, and was assigned KACC 83061BP. In the present invention, “Saccharomyces cerevisiae NIBRFGC000501771 strain”, “NIBRFGC000501771 strain”, and “501771 strain” have the same meaning and can be used interchangeably.
본 발명의 용어 "드라이 타입의 약주"는 알코올 함량이 높은 약주를 의미하며, 본 발명에서는 발효가 완료된 원주의 알코올 도수가 14.0%(v/v) 내지 20.0%(v/v)의 높은 알코올 함량을 갖는 약주를 의미한다. The term "dry type herbal liquor" in the present invention refers to herbal liquor with a high alcohol content, and in the present invention, the alcohol content of the fermented raw liquor is 14.0% (v/v) to 20.0% (v/v). It means medicinal liquor with .
따라서, 본 발명의 상기 NIBRFGC000501771 균주로 발효가 완료된 술덧(원주라고도 함)의 최종 알코올 도수는 14.0%(v/v) 내지 20.0%(v/v)인 것을 특징으로 한다. 도 1을 참조하면, 본담금 1일(발효 4일) 경과 시점에서 NIBRFGC000501771 균주를 사용한 제조예 1 술덧의 알코올 도수는 3.5%(v/v)이었고, 본담금 2일(발효 5일) 경과 시점에서 8.2%(v/v)이었으며, 본담금 3일(발효 6일) 경과 시점에서 12.3%(v/v)로 각각 분석되었다. 본담금 초기 에틸 알코올 생성력이 좋은 드라이 약주의 경우, 본담금 직후부터 높은 알코올 함량이 특징이다. 이러한 특징은 불필요한 오염균 생장의 저해 요인으로 작용한다. 따라서, 본담금 초기 발효 진행 속도가 빠른 특징은 높은 알코올 함량으로 다른 미생물 오염을 줄여서 안전한 발효에 유용하고 양조공정 관리에 효율적이다. 또한, 드라이 타입의 약주 제조 과정에서 술덧의 에틸알코올 함량이 높아지면 제품화 과정에서 술의 용량을 더 늘릴 수 있어 제조 수율이 높아지며, 경제적이다. 발효 진행 속도와 술덧의 최종 알코올 함량에 대한 실험 결과를 통해 검토해 본 바, 본 발명의 NIBRFGC000501771 균주는 발효가 완료된 술덧(원주)의 최종 알코올 도수가 14.0%(v/v) 내지 20.0%(v/v)인 드라이 타입의 약주를 제조할 수 있는 것으로 확인되었으며, 상기에서 설명한 바와 같은 맥락에서 드라이 타입의 약주 제조에 특화된 균주이다. Therefore, the final alcohol content of the fermented liquor (also known as Wonju), which has been completely fermented with the NIBRFGC000501771 strain of the present invention, is characterized in that it is 14.0% (v/v) to 20.0% (v/v). Referring to Figure 1, the alcohol content of Preparation Example 1 liquor using the NIBRFGC000501771 strain was 3.5% (v/v) after 1 day of main soak (4 days of fermentation), and 2 days after main soak (5 days of fermentation). It was 8.2% (v/v), and was analyzed as 12.3% (v/v) after 3 days of main deposit (6 days after fermentation). In the case of dry herbal liquor, which has a good ability to produce ethyl alcohol at the beginning of main soak, it is characterized by high alcohol content immediately after main soak. These characteristics act as a factor that inhibits the growth of unnecessary contaminants. Therefore, the characteristic of the rapid initial fermentation speed of main soak is that it reduces other microbial contamination due to the high alcohol content, which is useful for safe fermentation and efficient in brewing process management. In addition, as the ethyl alcohol content of the liquor increases during the manufacturing process of dry type herbal liquor, the capacity of the liquor can be further increased during the commercialization process, increasing the manufacturing yield and being economical. As reviewed through experimental results on the speed of fermentation progress and the final alcohol content of the liquor, the NIBRFGC000501771 strain of the present invention has a final alcohol content of 14.0% (v/v) to 20.0% (v/) of the fermented liquor (Wonju). v) It was confirmed that it can produce dry type herbal liquor, and in the same context as described above, it is a strain specialized for the production of dry type herbal liquor.
또한, 본 발명의 NIBRFGC000501771 균주는 발효 과정에서 시트르산(citric acid) 및 숙신산(Succinic acid)을 다량 생성시키며, 본 발명의 효모 균주를 이용한 약주는 특정 유기산 함량이 높은 특징이 있다(표 4 참조). 술의 관능적 품질에 가장 크게 영향을 미치는 성분은 당류, 산류 및 에틸알코올 성분이고, 당분 함량이 낮은 술에서는 어느 정도 신맛 성분이 높을수록 관능적으로 좋은 평가를 받는 것을 감안하면, 시트르산(citric acid) 및 숙신산(Succinic acid)을 다량 생성시키되, 젖산(Lactic acid) 및 아세트산(Acetic acid)을 소량 생성시키는 본 발명의 NIBRFGC000501771 균주는 드라이 타입의 약주 제조에 적합한 특성을 갖는다. In addition, the NIBRFGC000501771 strain of the present invention produces large amounts of citric acid and succinic acid during the fermentation process, and herbal liquor using the yeast strain of the present invention is characterized by a high content of specific organic acids (see Table 4). The components that have the greatest impact on the sensory quality of alcohol are sugars, acids, and ethyl alcohol. Considering that in alcohol with a low sugar content, the higher the sourness component, the better the sensory evaluation, citric acid and The NIBRFGC000501771 strain of the present invention, which produces a large amount of succinic acid but small amounts of lactic acid and acetic acid, has characteristics suitable for the production of dry type herbal liquor.
본 발명의 NIBRFGC000501771(기탁번호 KACC 83061BP) 균주는 효모의 응집성이 높은 것을 특징으로 한다(표 5 참조). 효모는 발효 후반부에 효모들 사이에서 엉김 현상이 일어나 발효 탱크 하단부에 가라앉는 특성이 있다. 특히, 약주는 혼탁한 형태로 제품화하는 탁주와 달리 발효가 끝난 술덧을 맑고 투명하게 하여 제품화하는 것이 특징인 술이고, 약주 양조공정에서 맑고 투명하게 만드는 여과 공정이 가장 어려운 공정에 해당한다. 드라이 타입의 약주용 효모는 응집성이 높아야 발효가 끝난 후 효모가 빠르게 침강해 이후 공정인 여과 공정을 손쉽게 진행할 수 있다. 본 발명의 바람직한 실시예에서는 수입 라빠리장 효모의 응집도 8.8%와 대비해 본 발명의 사카로마이세스 세레비지애 NIBRFGC000501771 균주가 2배 이상 증가한 18.36의 응집도를 갖는 것을 확인하였으며, 이러한 높은 응집성은 드라이 타입의 약주 제조에 적합한 특성이다.The NIBRFGC000501771 (accession number KACC 83061BP) strain of the present invention is characterized by high yeast aggregation (see Table 5). Yeast has the characteristic of causing agglomeration between yeasts in the latter half of fermentation and settling at the bottom of the fermentation tank. In particular, Yakju is a liquor that, unlike Takju, which is commercialized in a cloudy form, is characterized by commercializing the fermented liquor in a clear and transparent manner. In the Yakju brewing process, the filtration process to make it clear and transparent is the most difficult process. Dry type yeast for medicinal liquor must have high cohesiveness so that the yeast settles quickly after fermentation is completed and the subsequent filtration process can be easily carried out. In a preferred embodiment of the present invention, it was confirmed that the Saccharomyces cerevisiae NIBRFGC000501771 strain of the present invention had an aggregation degree of 18.36, which was more than two-fold increased compared to the aggregation degree of 8.8% for imported Laparijang yeast, and this high aggregation was observed in the dry type. This characteristic is suitable for manufacturing herbal liquor.
또한, 본 발명의 NIBRFGC000501771 균주는 향기 성분을 고함량으로 생성시키는 것을 특징으로 한다(표 6 참조). In addition, the NIBRFGC000501771 strain of the present invention is characterized by producing a high content of aroma components (see Table 6).
본 발명에서, 상기 향기 성분은 메틸 아세테이트(methyl acetate), 에틸 아세테이트(ethyl acetate), 이소-부틸 아세테이트(iso-buthyl acetete), 에틸 부틸레이트(ethyl butylate), 1-프로판올(1-propanol), 이소부탄올(isobutanol), 이소펜틸 아세테이트(isopentyl acetate), 1-부탄올(1-butanol), 이소아밀 알코올(iso-amyl alcohol), 에틸 헥사노에이트(Ethyl hexanoate), 1-헥사놀(1-hexanol), 에틸 옥타노에이트(ethyl octanoate), 푸르푸랄(furfural), 페닐 아세테이트(phenyl acetate), 에틸페닐 아세테이트(ethylphenyl acetate), 및 에틸 도데카노에이트(ethyl dodecanoate) 일 수 있다. In the present invention, the fragrance component includes methyl acetate, ethyl acetate, iso-butyl acetate, ethyl butylate, 1-propanol, Isobutanol, isopentyl acetate, 1-butanol, iso-amyl alcohol, Ethyl hexanoate, 1-hexanol ), ethyl octanoate, furfural, phenyl acetate, ethylphenyl acetate, and ethyl dodecanoate.
본 발명의 바람직한 실시예에서는 NIBRFGC000501771 균주로 제조된 약주의 향기 성분을 면밀하게 분석하였고, 본 발명의 NIBRFGC000501771 균주가 향기 성분의 총합을 가장 많이 생성시키는 효모 균주 중 하나임을 확인한 바 있다. 구체적으로 동 균주로 발효된 제조예 1의 시료는 장미향, 꿀향, 사과향으로 감지되는 ethyl phenyl acetate 성분, 바나나향으로 감지되는 iso butyl acetate 성분, 배향으로 감지되는 iso pentyl acetate 성분, 사과향으로 감지되는 ethyl hexanoate 성분, 그리고 멜론향, 풋사과향, 파인애플향으로 인지되는 ethyl octanoate 성분 함량이 동시에 높게 검출되는 것을 확인하였다. 따라서, 감칠맛과 특유의 신맛을 제공하는 숙신산 및 시트르산 함량이 높고, 향기성분을 다량 생성해 내는 본 발명의 NIBRFGC000501771 효모 균주는 드라이 타입의 약주 주조용 양조효모로 적합한 균주이다. 또한, 관능검사 결과에서도 NIBRFGC000501771 균주가 외관, 향, 맛, 감촉 등 모든 항목에서 가장 좋은 평가를 받은 바 있다. In a preferred embodiment of the present invention, the aroma components of herbal medicine prepared with the NIBRFGC000501771 strain were closely analyzed, and it was confirmed that the NIBRFGC000501771 strain of the present invention is one of the yeast strains that produces the largest amount of aroma components. Specifically, the sample of Preparation Example 1 fermented with the same strain contained ethyl phenyl acetate component detected as rose, honey, and apple aroma, iso butyl acetate component detected as banana aroma, iso pentyl acetate component detected as pear aroma, and ethyl detected as apple aroma. It was confirmed that the contents of hexanoate and ethyl octanoate, which are recognized as melon, green apple, and pineapple aromas, were detected simultaneously. Therefore, the NIBRFGC000501771 yeast strain of the present invention, which has a high content of succinic acid and citric acid, which provides umami and a unique sour taste, and produces a large amount of aromatic components, is a strain suitable as a dry type brewing yeast for brewing herbal liquor. In addition, in the sensory test results, strain NIBRFGC000501771 received the best evaluation in all categories including appearance, aroma, taste, and texture.
본 발명의 다른 양태에 따르면, 본 발명은 정제수에 입국 및 사카로마이세스 세레비지애 신균주 NIBRFGC000501771(기탁번호 KACC 83061BP)을 첨가하고 2일 내지 5일 동안 발효하여 밑술을 제조하는 단계; 및 상기 밑술에 팽화미, 정제효소제 및 정제수를 첨가하고 6일 내지 15일 동안 발효하여 담금하는 단계;를 포함하는 드라이 타입의 약주 제조 방법을 제공한다. 상기 약주 제조방법에서, 전분질 원료인 팽화미 원료 중량의 15% 내지 30%에 해당하는 입국을 밑술 제조단계에 사용할 수 있으며, 급수되는 정제수는 전분 원료 대비 110% 내지 180% 중량으로 투입할 수 있다. 또한, 정제효소제는 전분 원료 대비 0.02% 내지 0.06% 중량부로 사용할 수 있으며, 효모는 1㎖ 당 8×107 내지 8×108 cfu의 균수로 배양하여 입국 100 중량부 대비 1.0 내지 9.3㎖의 부피로 사용할 수 있고, 바람직하게는 1㎖ 당 2×108 cfu의 균수로 배양하여 입국 100 중량부 대비 3.1㎖의 부피로 사용할 수 있다. 밑술 제조 단계의 발효는 2일 내지 5일, 바람직하게는 4일 내지 5일 진행할 수 있고, 담금 단계의 발효는 6일 내지 15일, 바람직하게는 10일 내지 14일 진행할 수 있는데, 담금 단계에서 알코올 도수의 변화가 없는 경우 발효 과정을 종료할 수 있다. 발효 과정이 종료된 술덧은 주대를 이용하여 거른 후, 밀폐 용기에 담아 0℃ 내지 5℃에서 3일 내지 7일간 보관하면서 침전물 제거한 후 1~2㎛ 종이필터 여과기로 정밀하게 여과하여 약주로 제조될 수 있다. According to another aspect of the present invention, the present invention includes the steps of adding a new strain of Saccharomyces cerevisiae NIBRFGC000501771 (accession number KACC 83061BP) to purified water and fermenting for 2 to 5 days to produce a base liquor; and adding puffed rice, purified enzyme, and purified water to the base liquor and fermenting and soaking for 6 to 15 days. It provides a dry type herbal liquor manufacturing method including. In the herbal liquor manufacturing method, 15% to 30% of the weight of the puffed rice raw material, which is a starchy raw material, can be used in the fermentation stage, and the purified water supplied can be added at a weight of 110% to 180% of the starch raw material. . In addition, purified enzymes can be used in an amount of 0.02% to 0.06% by weight compared to the starch raw material, and yeast is cultured at a bacterial count of 8 × 10 7 to 8 × 10 8 cfu per 1 ㎖ to produce a volume of 1.0 to 9.3 ㎖ based on 100 parts by weight of entry. It can be used, and preferably can be cultured at a bacterial count of 2×10 8 cfu per 1 ml and used in a volume of 3.1 ml based on 100 parts by weight of entry. The fermentation of the base liquor manufacturing step can be carried out for 2 to 5 days, preferably 4 to 5 days, and the fermentation of the immersion stage can be carried out for 6 to 15 days, preferably 10 to 14 days. In the immersion stage, If there is no change in alcohol content, the fermentation process can be terminated. After the fermentation process has been completed, the fermentation process is filtered using a strainer, stored in an airtight container at 0℃ to 5℃ for 3 to 7 days, sediment is removed, and then precisely filtered using a 1-2㎛ paper filter to prepare medicinal liquor. You can.
본 발명의 다른 양태에 따르면, 본 발명은 드라이 타입의 약주 제조 방법으로 제조된 향기 성분이 강화된 드라이 타입의 약주를 제공한다. According to another aspect of the present invention, the present invention provides a dry type herbal liquor with enhanced aroma components prepared by a dry type herbal liquor manufacturing method.
본 발명의 드라이 타입의 약주는 향기 성분이 강화된 것을 특징으로 하는데, 상기 향기 성분의 예로는 메틸 아세테이트(methyl acetate), 에틸 아세테이트(ethyl acetate), 이소-부틸 아세테이트(iso-buthyl acetete), 에틸 부틸레이트(ethyl butylate), 1-프로판올(1-propanol), 이소부탄올(isobutanol), 이소펜틸 아세테이트(isopentyl acetate), 1-부탄올(1-butanol), 이소아밀 알코올(iso-amyl alcohol), 에틸 헥사노에이트(Ethyl hexanoate), 1-헥사놀(1-hexanol), 에틸 옥타노에이트(ethyl octanoate), 푸르푸랄(furfural), 페닐 아세테이트(phenyl acetate), 에틸페닐 아세테이트(ethylphenyl acetate), 및 에틸 도데카노에이트(ethyl dodecanoate)를 들 수 있다. The dry type herbal liquor of the present invention is characterized by enhanced aroma components. Examples of the aroma components include methyl acetate, ethyl acetate, iso-butyl acetate, and ethyl. Butylate, 1-propanol, isobutanol, isopentyl acetate, 1-butanol, iso-amyl alcohol, ethyl Ethyl hexanoate, 1-hexanol, ethyl octanoate, furfural, phenyl acetate, ethylphenyl acetate, and ethyl Dodecanoate (ethyl dodecanoate) may be mentioned.
본 발명의 드라이 타입의 약주 제조 방법과 드라이 타입의 약주는 상술한 본 발명의 사카로마이세스 세레비지애 신균주 NIBRFGC000501771(기탁번호 KACC 83061BP)에 의해 제조되는 것이기 때문에, 이들 사이에 공통된 내용은 반복 기재에 따른 명세서의 과도한 복잡성을 피하기 위하여, 그 기재를 생략한다.Since the dry type medicinal liquor manufacturing method and the dry type medicinal liquor of the present invention are manufactured by the new strain of Saccharomyces cerevisiae NIBRFGC000501771 (accession number KACC 83061BP) of the present invention described above, the common content between them is repeated. In order to avoid excessive complexity of the specification due to the description, the description is omitted.
이상 설명한 바와 같이, 본 발명은 드라이 타입의 약주 제조용 사카로마이세스 세레비지애 신균주 NIBRFGC000501771(기탁번호 KACC 83061BP)을 제공한다.As described above, the present invention provides a new strain of Saccharomyces cerevisiae NIBRFGC000501771 (accession number KACC 83061BP) for the production of dry type medicinal liquor.
본 발명의 사카로마이세스 세레비지애 신균주 NIBRFGC000501771(기탁번호 KACC 83061BP)은 발효가 완료된 술덧의 최종 알코올 도수가 14.0%(v/v) 내지 20.0%(v/v)인 드라이 타입의 약주를 제조할 수 있고, 효모의 응집성이 높으며, 향기 성분을 고함량으로 생성시키는 장점이 있으므로, 관능성이 개선된 고품질의 약주 제조에 유용하게 이용될 수 있다.The new Saccharomyces cerevisiae strain NIBRFGC000501771 (accession number KACC 83061BP) of the present invention is a dry type herbal liquor with a final alcohol content of 14.0% (v/v) to 20.0% (v/v) of the fermented liquor. It can be manufactured, has high yeast cohesion, and has the advantage of producing a high content of aroma components, so it can be usefully used in the production of high-quality herbal liquor with improved sensory properties.
도 1은 선발 효모 3종 및 수입 상용 효모 1종을 이용하여 제조되는 약주의 발효 경과에 따른 알코올 함량 분석 결과를 나타낸 그래프이다.
도 2는 사카로마이세스 세레비지애 NIBRFGC000501771 균주를 YPD 배지에서 배양하고, 배양 형태를 촬영한 사진이다.
도 3은 사카로마이세스 세레비지애 NIBRFGC000501771 균주를 YPD 배지에서 배양하고, 광학현미경(왼쪽, 1,000배)과 주사전자현미경(오른쪽)으로 촬영한 사진이다.
도 4는 YT 마이크로플레이트(MicroPlate)를 이용하여 NIBRFGC000501771 균주의 표현형 마이크로어레이(phenotype microarray) 분석을 수행한 결과이다[위의 표는 산화(oxidation), 아래 표는 동화(assimilation) 분석 결과임].
도 5는 NIBRFGC000501771 균주의 유연관계를 분석한 계통도이다. Figure 1 is a graph showing the results of alcohol content analysis according to the fermentation process of Yakju produced using three types of starter yeast and one type of imported commercial yeast.
Figure 2 is a photograph of the Saccharomyces cerevisiae NIBRFGC000501771 strain cultured in YPD medium and the culture form.
Figure 3 is a photograph of Saccharomyces cerevisiae NIBRFGC000501771 strain cultured in YPD medium and taken with an optical microscope (left, 1,000 times) and a scanning electron microscope (right).
Figure 4 shows the results of phenotypic microarray analysis of strain NIBRFGC000501771 using YT MicroPlate [the table above shows the results of oxidation analysis, and the table below shows the assimilation analysis results].
Figure 5 is a schematic diagram analyzing the kinship relationship of the NIBRFGC000501771 strain.
이하, 실시예를 통하여 본 발명을 더욱 상세히 설명하기로 한다. 이들 실시예는 단지 본 발명을 예시하기 위한 것이므로, 본 발명의 범위가 이들 실시예에 의해 제한되는 것으로 해석되지는 않는다.Hereinafter, the present invention will be described in more detail through examples. Since these examples are merely for illustrating the present invention, the scope of the present invention is not to be construed as limited by these examples.
실시예 1. 실험 재료 및 방법Example 1. Experimental materials and methods
1-1. 효모1-1. leaven
국립생물자원관에서 분리·배양하여 보존 중인 효모 균주 88종을 대상으로 효모의 크기, 발효력, 응집성(Flocculation), Cu 배지 성장성, 포자 생성(Sporulation), 멜리바이오스(Melibiose) 이용 능력 등의 평가와, 약주 시험양조를 통해 양조용으로 사용 가능한 33종의 효모 균주를 1차 선발하였다. 이후, 2차 시험양조를 통해 발효가 완료된 술덧의 최종 알코올 도수가 14.0%(v/v) 내지 20.0%(v/v)이고 향기가 뛰어난 드라이 타입의 효모 3종을 선발하여 실험에 사용하였다. Evaluation of yeast size, fermentation ability, flocculation, Cu medium growth, spore production, melibiose utilization ability, etc. of 88 types of yeast strains isolated, cultured and preserved at the National Institute of Biological Resources. Through test brewing of Yakju, 33 types of yeast strains that can be used for brewing were initially selected. Afterwards, through the second test brewing, three types of dry type yeast with a final alcohol content of 14.0% (v/v) to 20.0% (v/v) and excellent aroma were selected and used in the experiment.
2차 선발된 효모 3종은 Saccharomyces cerevisiae NIBRFGC000501771(경기도 용인시 채집, 야생화), Saccharomyces cerevisiae NIBRFGC000503472(경기도 남양주시 채집), Saccharomyces cerevisiae NIBRFGC000503478(강원도 홍천군)의 3종 균주이다. 대조군으로는 시판 중인 라빠리장 효모를 사용하였다.The three types of yeast selected for the second round were Saccharomyces cerevisiae NIBRFGC000501771 (collected from Yongin-si, Gyeonggi-do, wild flowers), Saccharomyces cerevisiae NIBRFGC000503472 (collected from Namyangju-si, Gyeonggi-do), and Saccharomyces cerevisiae NIBRFGC000503478 (Hongcheon-gun, Gangwon-do). As a control, commercially available Laparijang yeast was used.
효모는 glycerol 20% stock으로 -45℃에 보관하였고, YMA 평판배지에 도말하여 콜로니(colony)를 확인한 후, YMB 액체배지에 배양하여 사용하였다. 배지는 Yeast Malt Broth(MB Cell), Yeast Malt Agar(MB Cell) 제품을 사용하였다.Yeast was stored at -45°C with 20% glycerol stock, smeared on YMA plate medium to confirm colonies, and then cultured in YMB liquid medium for use. Yeast Malt Broth (MB Cell) and Yeast Malt Agar (MB Cell) products were used as media.
1-2. 약주 원료1-2. medicinal liquor raw materials
약주 제조에 사용한 전분 원료는 쌀을 고온, 진공, 팽창시켜 만든 팽화미(국내산, ㈜조은곡식)를 사용하였고, 발효제로는 입국(역가 60sp, ㈜조은곡식)과 정제효소제(역가 15,000sp, ㈜바이오랜드)를 사용하였다.The starch raw material used to manufacture Yakju was puffed rice (domestic production, JoEun Grain Co., Ltd.) made by expanding rice at high temperature, vacuum, and the leavening agent was Iryuk (potency 60 sp, Jo Eun Grain Co., Ltd.) and purified enzyme (potency 15,000 sp, Co., Ltd.). Bioland) was used.
1-3. 약주 제조1-3. Herbal liquor manufacturing
발효주 담금 방법은 전분 원료인 팽화미 원료의 20%에 해당하는 입국을 사용하였으며, 급수량은 전분 원료 대비 160%로 하였다.The fermented liquor immersion method used water equivalent to 20% of the starch raw material, and the water supply amount was 160% of the starch raw material.
정제효소제는 전분 원료 대비 0.04% 사용하였으며, 효모는 1㎖ 당 2×108 cfu의 배양액 3.1㎖를 넣어주었다. 알코올 도수의 변화가 없으면 발효를 중지시키고, 알코올 등 품질분석을 바로 진행하였으며, 관능 평가용 시료는 3℃에서 보관하였다.Purified enzyme was used at 0.04% of the starch raw material, and 3.1 ml of yeast culture medium of 2 × 10 8 cfu per 1 ml was added. If there was no change in alcohol content, fermentation was stopped, quality analysis such as alcohol was immediately performed, and samples for sensory evaluation were stored at 3°C.
1-4. 성분 분석1-4. Ingredient analysis
[알코올 도수][Alcohol content]
알코올은 발효주 시료 상등액 100㎖에 증류수 30㎖를 첨가하여 자동증류장치(C.Gerhardt, DE/VAPODEST200)를 통해 증류액 90㎖를 회수하고, 증류수로 총부피 100㎖를 적정한 후 디지털 알코올 측정장치(Multiple Auto Sampler, KEM, JP/DA-645)를 사용하여 15℃에서의 알코올 함량(%(v/v))을 측정하였다.For alcohol, add 30 ml of distilled water to 100 ml of supernatant of the fermented liquor sample, recover 90 ml of distillate through an automatic distillation device (C.Gerhardt, DE/VAPODEST200), titrate the total volume of 100 ml with distilled water, and use a digital alcohol measuring device ( Alcohol content (% (v/v)) at 15°C was measured using Multiple Auto Sampler, KEM, JP/DA-645.
[당도][Sugar content]
당도 측정은 알코올 도수 측정 후 남은 잔액을 이용하여 디지털 당도 측정기(Atago, Japan)를 통해 °Brix 단위로 측정하였다.Sugar content was measured in °Brix using a digital sweetness meter (Atago, Japan) using the remaining balance after measuring the alcohol content.
[총산 및 아미노산도][Total acids and amino acids]
총산 및 아미노산도는 AOAC법을 참고하여 국세청 주류분석규정(국세청 기술 연구소, 2008)에 따라 CO2를 제거한 시료 상층액 10㎖를 전위차 측정기(Metrohm, 855 Robotic Titrosampler)를 사용하여 0.1N NaOH pH 8.2가 될 때까지 적정한 ㎖ 수를 산도로 하고, 여기에 중성 포르말린 용액 5㎖를 가하여 유리된 산을 0.1N NaOH pH 8.2이 될 때까지 적정한 ㎖수를 아미노산도로 하였다. For the total acid and amino acid degrees, refer to the AOAC method and measure 10 ml of sample supernatant from which CO 2 was removed in accordance with the National Tax Service's Alcohol Analysis Regulations (National Tax Service Technology Research Institute, 2008) using a potentiometer (Metrohm, 855 Robotic Titrosampler) with 0.1N NaOH pH 8.2. The acidity was determined by titrating ml of water until pH reached 8.2. 5 ml of neutral formalin solution was added thereto, and the ml of water titrated until pH reached 8.2 was calculated by adding 0.1N NaOH.
초산으로 환산한 값을 총산으로 하여 g/100㎖ 단위로 나타내었다.The value converted to acetic acid was expressed as the total acid in g/100 ml.
(총산)(total acid)
Total acid(as acetic acid) g/100㎖ = 적정 ㎖ 수 × 0.006 × 10Total acid (as acetic acid) g/100 ㎖ = titration ㎖ number × 0.006 × 10
글리신으로 환산한 값을 아미노산도로 하여 g/100㎖ 단위로 나타내었다.The value converted to glycine was expressed as an amino acid diagram in g/100 ml.
(아미노산도)(Amino acids)
Amino acid(as grycine) g/100㎖ = 적정 ㎖ 수 × 0.0075 × 10Amino acid (as grycine) g/100 ㎖ = titration ㎖ number × 0.0075 × 10
[불휘발분][Non-volatile matter]
시료 25㎖(15℃)를 증발접시에 넣고, 물중탕기에서 가열하여 충분히 증발시킨 후, 건조기를 이용하여 105~110℃에서 2시간 건조하였다. 건조 시료를 데시케이터에서 식힌 후(30분 방치) 무게를 달아 증발접시 무게를 제외하여 측정한 후, 측정값에 4배로 곱하여 불휘발분의 그램 수를 산출하였다.25 ml of sample (15°C) was placed in an evaporating dish, heated in a water bath, sufficiently evaporated, and then dried at 105-110°C for 2 hours using a dryer. After cooling the dried sample in a desiccator (left for 30 minutes), it was weighed, excluding the weight of the evaporation dish, and the measured value was multiplied by 4 to calculate the number of grams of non-volatile matter.
[유기산][Organic acid]
시료를 원심분리(5,000×g, 10min)한 다음 0.5㎛의 membrane filter로 여과하여 Ion Chromatograph(Metrohm 850 Professional IC) 장치로 분석하였다. Column은 Metrosep Organic acid 250/7.8 모델을 사용하였고, Column 온도는 35℃, 솔벤트 시스템(Solvent system)은 Organic acid-H2SO4 0.002㎖, 유량(Flow rate)은 0.5mL/min의 조건으로 측정하였다.The sample was centrifuged (5,000 × g, 10 min), filtered through a 0.5㎛ membrane filter, and analyzed using an Ion Chromatograph (Metrohm 850 Professional IC). The column used was the Metrosep Organic acid 250/7.8 model, and the column temperature was measured at 35°C, the solvent system was 0.002 mL of organic acid-H2SO4, and the flow rate was 0.5 mL/min.
[유리당][Yuridang]
시료를 50% Acetonitrile 용매로 10배 희석하고 0.2㎛ membrane filter로 여과한 후, UPLC(ACQUITY UPLC H-Class/RI Detector) 장치를 이용하여 과당(Fructose), 포도당(Glucose), 자당(Sucrose), 맥아당(Maltose), 유당(Lactose)을 분석하였다. Column은 ACQUITY BEH Amide(100mm x 2.1mm, 1.7㎛) 모델을 사용하였고, Column 온도는 35℃, 솔벤트 시스템(Solvent system)은 0.2% TEA in Acetonitrile: 0.2% TEA in DW = 75% : 25%, 유량(Flow rate)은 0.13㎖/min의 조건으로 측정하였다.The sample was diluted 10 times with 50% Acetonitrile solvent and filtered through a 0.2㎛ membrane filter. Then, using UPLC (ACQUITY UPLC H-Class/RI Detector) device, fructose, glucose, sucrose, Maltose and lactose were analyzed. The column used the ACQUITY BEH Amide (100mm Flow rate was measured at 0.13 mL/min.
[주요 향기성분] [Main fragrance ingredients]
Gas Chromatography/Flame Ionization Detector(GC/FID)(Agilent 7890, Agilent Technologies INC., Santa Clara, USA)를 이용하여 향기성분을 분석하였다. Fragrance components were analyzed using Gas Chromatography/Flame Ionization Detector (GC/FID) (Agilent 7890, Agilent Technologies INC., Santa Clara, USA).
분석시료를 20mL 바이알(vial)에 캡(cap)을 닫아 밀봉한 후, 헤드스페이스 샘플러(headspace sampler)(7697Am Agilent Technologies INC.. Santaclara, USA)에 넣고, 오븐(oven) 70℃, 루프(loop)(3㎖) 80℃, 라인(line) 90℃로 유지하여 GC/FID 장치에 주입하여 분석하였다. Column은 Agilent HP-FFAP(30m x 320㎛ x 0.25㎛)를 사용하였고, 캐리어 가스(Carrier gas)는 N2, 오븐(Oven) 온도는 40℃에서 4분간 유지 후 5℃/min의 속도로 100℃까지 상승시켰으며, 이후 6℃/min으로 200℃까지 상승 후 4분 유지하고, 10℃/min으로 220℃까지 상승시켰다. 스플릿 비율(Split ratio)은 10:1, 유량(Flow rate)은 0.8㎖/min의 조건으로 분석하였다.The analysis sample was sealed in a 20mL vial by closing the cap, placed in a headspace sampler (7697Am Agilent Technologies INC.. Santaclara, USA), and placed in an oven at 70°C with a loop. ) (3 mL) was maintained at 80°C and line at 90°C and injected into the GC/FID device for analysis. The column used was Agilent HP-FFAP ( 30m It was raised to ℃, then raised to 200℃ at 6℃/min, maintained for 4 minutes, and raised to 220℃ at 10℃/min. The split ratio was analyzed at 10:1 and the flow rate at 0.8 mL/min.
[효모의 응집 테스트(Flocculation test)][Yeast Flocculation test]
효모를 Yeast Malt Broth 배지에 1.5 × 106 Cells/㎖ 농도로 배양한 다음 배양액을 Tube "A"에 10㎖, Tube "B"에 10㎖로 각각 채취하였다. Tube "A"는 원심분리(900×g, 2.5min)한 다음 상등액을 버리고, 증류수 9.9㎖와 용액 A(PH7 0.5M EDTA용액) 0.1㎖를 넣고 15초 동안 혼합하여 생성된 혼합 용액을 1㎖ 채취한 후, 9㎖ 증류수를 넣어 600㎚에서 흡광도를 측정하였다. Tube "B"는 원심분리(900×g, 2.5min)한 상등액을 버리고 세척액(증류수 1ℓ, CaSo4 0.51g) 10㎖를 넣어 15초 혼합 후 원심분리(900×g, 2.5min) 한 다음, 상등액을 버리고 용액 B(CaSo4 0.51g, CH3COONa 6.8g, CH3COOH 4.05g을 증류수 1ℓ에 녹인 후, pH 4.5로 만듦)를 넣어 15초 혼합하였다. 6분간 정치하여 상등액 1㎖를 채취한 후, 9㎖ 증류수를 넣어 600㎚에서 흡광도를 측정하였다. 효모의 응집력은 하기 [식 1]을 이용하여 측정하였다.Yeast was cultured in Yeast Malt Broth medium at a concentration of 1.5 Tube "A" was centrifuged (900 After collection, 9 mL of distilled water was added and the absorbance was measured at 600 nm. In Tube " B ", discard the supernatant from centrifugation (900 The supernatant was discarded, and solution B (0.51 g of CaSo 4 , 6.8 g of CH 3 COONa, and 4.05 g of CH 3 COOH dissolved in 1 liter of distilled water, adjusted to pH 4.5) was added and mixed for 15 seconds. After standing for 6 minutes and collecting 1 ml of supernatant, 9 ml of distilled water was added and the absorbance was measured at 600 nm. The cohesion of yeast was measured using the following [Equation 1].
[식 1][Equation 1]
실시예 2. 약주 제조Example 2. Preparation of medicinal liquor
상기 실시예 1-1의 2차 선발 과정을 통해 선정된 드라이 타입의 효모 균주 3종[Saccharomyces cerevisiae NIBRFGC000501771(경기도 용인시 채집, 야생화), Saccharomyces cerevisiae NIBRFGC000503472(경기도 남양주시 채집), Saccharomyces cerevisiae NIBRFGC000503478(강원도 홍천군 채집)]의 발효능을 분석하기 위해 약주를 제조하였다. 대조군으로는 시판 중인 라빠리장 효모를 사용하였다.Three dry type yeast strains selected through the secondary selection process of Example 1-1 [ Saccharomyces cerevisiae NIBRFGC000501771 (collected from Yongin-si, Gyeonggi-do, wild flowers), Saccharomyces cerevisiae NIBRFGC000503472 (collected from Namyangju-si, Gyeonggi-do), Saccharomyces cerevisiae NIBRFGC00050347 8 (Collected in Hongcheon-gun, Gangwon-do )], herbal liquor was prepared to analyze its fermentation ability. As a control, commercially available Laparijang yeast was used.
밑술은 입국을 사용하였으며, 전체원료 500g 대비 20%인 100g을 사용하였다. 급수는 입국 중량 대비 160%(w/v)인 160㎖를 사용하였으며, 밑술 발효는 4일간 진행되었다. 밑술의 발효가 완료된 후, 팽화미 400g과 물 792㎖, 정제효소제 0.2g을 첨가하였고, 담금 발효는 13일 정도 진행하여 최종 발효가 멈추는 단계까지 발효를 진행하였다. 발효 기간이 경과한 후, 200 mesh 주대를 이용하여 술을 거른 후, 밀폐 용기에 담아 0℃에서 5일간 보관하여 침전물을 제거하였고, 1~2㎛ 종이필터 여과기로 정밀하게 여과하여 약주를 제조하였다. 제조된 약주의 기본 성분 분석과 기기 분석을 진행하였다. 약주 담금법은 하기 [표 1]에 정리하였다. The root liquor was used in Korea, and 100g, which is 20% of the total raw material of 500g, was used. 160 ml of water, which is 160% (w/v) of the entry weight, was used, and the fermentation of the base liquor was carried out for 4 days. After the fermentation of the base liquor was completed, 400g of puffed rice, 792ml of water, and 0.2g of purified enzyme were added, and the soak fermentation continued for about 13 days until the final fermentation stopped. After the fermentation period had elapsed, the liquor was filtered using a 200 mesh strainer, placed in an airtight container and stored at 0°C for 5 days to remove sediment, and then precisely filtered using a 1-2㎛ paper filter to prepare herbal liquor. . Basic component analysis and instrumental analysis of the manufactured herbal medicine were conducted. The herbal liquor soaking method is summarized in [Table 1] below.
[표 1][Table 1]
실시예 3. 약주의 발효 진행 속도 분석 및 일반성분 분석Example 3. Fermentation progress rate analysis and general component analysis of Yakju
3-1. 발효 진행 속도 분석3-1. Fermentation progress rate analysis
각 실험군별 발효 경과에 따른 에틸알코올 함량 분석을 수행하고, 분석 결과를 도 1에 정리하였다. Ethyl alcohol content analysis was performed according to the fermentation process for each experimental group, and the analysis results are summarized in Figure 1.
도 1을 참조하면, 각 실험군별 17일간 발효시킨 후 알코올 도수는 제조예 2(NIBRFGC000503472 균주) 19.4%(v/v), 제조예 3(NIBRFGC000503478 균주) 19.3%(v/v), 제조예 1(NIBRFGC000501771균주) 18.5%(v/v) 및 비교예 1(라빠리장) 16.9%(v/v)의 순으로 측정되었다. Referring to Figure 1, after fermentation for 17 days for each experimental group, the alcohol content was 19.4% (v/v) in Preparation Example 2 (NIBRFGC000503472 strain), 19.3% (v/v) in Preparation Example 3 (NIBRFGC000503478 strain), and Preparation Example 1. (NIBRFGC000501771 strain) was measured at 18.5% (v/v) and Comparative Example 1 (La Parisjang) at 16.9% (v/v).
효모의 종류를 달리한 3개 제조예와 비교예를 대상으로 밑술 발효 4일 및 담금 발효 13일의 기간별로 에틸알코올 생성 경과를 살펴보았다. 밑술 발효 구간에서는 모든 실험군에서 비교예인 라빠리장 사용군과 발효 속도가 유사한 것으로 분석되었다. 발효 1일째에는 NIBRFGC000503478 균주(7.4%(v/v)), 라빠리장(6.9%(v/v)), NIBRFGC000503472 균주(6.8%(v/v)), NIBRFGC000501771 균주(5.8%(v/v))의 순으로 발효 속도가 빠른 것으로 나타났다.The production of ethyl alcohol was examined over a period of 4 days for fermentation of base liquor and 13 days for soaking fermentation in three production examples and comparative examples using different types of yeast. In the base liquor fermentation section, it was analyzed that the fermentation speed was similar in all experimental groups to the group using Laparijang, which was a comparative example. On the first day of fermentation, NIBRFGC000503478 strain (7.4% (v/v)), Laparijang (6.9% (v/v)), NIBRFGC000503472 strain (6.8% (v/v)), and NIBRFGC000501771 strain (5.8% (v/v) The fermentation speed was found to be faster in the order of )).
2일이 경과한 후에는 NIBRFGC000503478 균주, NIBRFGC000503472 균주 및 NIBRFGC000501771 균주의 순으로 각각 12.8%(v/v), 12.0%(v/v) 및 11.0%(v/v)의 알코올을 생성시키는 것으로 측정되었다. After 2 days, it was measured that the NIBRFGC000503478 strain, NIBRFGC000503472 strain, and NIBRFGC000501771 strain produced 12.8% (v/v), 12.0% (v/v), and 11.0% (v/v) of alcohol, respectively. .
밑술 발효 3일째 술덧의 알코올 도수는 제조예 3(NIBRFGC000503478 균주) 16.2%(v/v), 제조예 1(NIBRFGC000501771 균주) 14.6%(v/v), 비교예 1(라빠리장) 14.3%(v/v), 및 제조예 2(NIBRFGC000503472 균주) 13.9%(v/v)의 순으로 높게 측정되었다. The alcohol content of the wort on the 3rd day of fermentation was 16.2% (v/v) in Preparation Example 3 (NIBRFGC000503478 strain), 14.6% (v/v) in Preparation Example 1 (NIBRFGC000501771 strain), and 14.3% (v/v) in Comparative Example 1 (La Paris Jang). v/v), and Preparation Example 2 (NIBRFGC000503472 strain) was measured to be 13.9% (v/v).
발효 4일째 1단을 담금 하였고, 술덧이 희석되어 알코올 도수는 3.0 ~ 3.7%(v/v) 수준으로 낮아졌다. 발효 6일째(담금 1일째) 알코올 도수를 비교해본 결과, 제조예 1(NIBRFGC000501771 균주)의 알코올 도수는 담금 초기 에틸알코올 발효 안정성이 높아 비교예 1(라빠리장)과 비교하여 2배 이상 높은 8.2%(v/v)로 측정되었고, 발효 7일째(담금 2일째)에는 12.3%(v/v)로 안정적인 에틸알코올 발효능을 보였다.On the 4th day of fermentation, one batch was soaked, and the alcohol content was diluted and the alcohol content was lowered to 3.0 ~ 3.7% (v/v). As a result of comparing the alcohol content on the 6th day of fermentation (1st day of soaking), the alcohol content of Preparation Example 1 (NIBRFGC000501771 strain) was 8.2, which is more than twice as high as that of Comparative Example 1 (Laparijang) due to the high ethyl alcohol fermentation stability at the beginning of soaking. It was measured in % (v/v), and showed a stable ethyl alcohol fermentation ability of 12.3% (v/v) on the 7th day of fermentation (2nd day of soaking).
그러나, 제조예 2(NIBRFGC000503472 균주) 및 제조예 3(NIBRFGC000503478 균주)의 알코올 도수는 각각 4.3%(v/v) 및 4.0%(v/v)로 측정되어, 비교예 1(라빠리장)의 알코올 도수 5.3%(v/v)와 비교하여 낮은 수치로 측정되었다. 제조예 2(NIBRFGC000503472 균주) 및 제조예 3(NIBRFGC000503478 균주)은 담금 2일 차부터 급격한 거품발생과 함께 끓어 넘침 현상이 발생하였는데, 이는 초기 알코올 생성이 더딘 것과도 연결된다. 병행복발효 방식으로 담금 횟수를 증가시키는 방식을 적용하는 전통주의 발효공법에서 덧술 첨가에 따라 초기 발효가 저해되는 NIBRFGC000503472 및 NIBRFGC000503478 효모 균주는 최종 알코올 도수가 19.4%(v/v), 19.5%(v/v)로 높게 나왔음에도 불구하고 약주 발효에 적합하지 않은 효모로 분석되었다.However, the alcohol content of Preparation Example 2 (NIBRFGC000503472 strain) and Preparation Example 3 (NIBRFGC000503478 strain) was measured to be 4.3% (v/v) and 4.0% (v/v), respectively, compared to that of Comparative Example 1 (La Paris Jang). It was measured as a low value compared to the alcohol content of 5.3% (v/v). Preparation Example 2 (NIBRFGC000503472 strain) and Preparation Example 3 (NIBRFGC000503478 strain) showed rapid foaming and boiling over from the second day of soaking, which is also connected to the slow initial alcohol production. In the traditional fermentation method that uses a parallel fermentation method to increase the number of soaks, the NIBRFGC000503472 and NIBRFGC000503478 yeast strains, whose initial fermentation is inhibited by the addition of alcohol, have a final alcohol content of 19.4% (v/v) and 19.5% (v). /v), it was analyzed as a yeast unsuitable for fermenting medicinal liquor.
양조초기에는 발효가 빠르게 진행됨으로써 공정관리가 용이하여야 하는데, NIBRFGC000503472 및 NIBRFGC000503478 효모 균주와 같이 급격한 거품발생과 함께 끓어 넘침 현상이 발생하는 균주는 담금 초기 저조하게 발효가 진행되는 균주를 의미하고, 발효관리, 여과, 제품화 등의 공정 과정에서 적합하지 않은 균주에 해당한다. In the early stages of brewing, fermentation progresses quickly to facilitate process management. Strains that produce rapid foaming and boiling over, such as NIBRFGC000503472 and NIBRFGC000503478 yeast strains, refer to strains that ferment slowly at the beginning of immersion, and require fermentation management. , corresponds to a strain that is not suitable for processing processes such as filtration and commercialization.
한편, 본 발명의 NIBRFGC000501771 효모 균주로 발효한 약주는 최종 알코올 도수가 18.5%(v/v)로 높게 분석되었다. 드라이 타입의 약주 양조효모의 제조에서는 높은 알코올 생성능을 갖는 효모가 적합하며, 이를 이용하여 제조된 약주는 높은 수율로 생산될 수 있다. 즉, 드라이 타입의 약주 효모는 발효가 완료된 술덧의 최종 알코올 도수가 14.0%(v/v) 보다는 높아야 하며, 점진적으로 발효가 진행되면서 최대 20%(v/v)까지 발효되는 것이 드라이용 약주 제조에 적합한 효모이다.Meanwhile, the final alcohol content of Yakju fermented with the NIBRFGC000501771 yeast strain of the present invention was analyzed as high as 18.5% (v/v). For the production of dry type herbal liquor brewing yeast, yeast with a high alcohol production ability is suitable, and herbal liquor manufactured using it can be produced with high yield. In other words, for dry type herbal liquor yeast, the final alcohol content of the fermented liquor must be higher than 14.0% (v/v), and fermentation progresses gradually to a maximum of 20% (v/v). This yeast is suitable for
발효 진행 속도와 술덧의 최종 알코올 함량으로 검토해 본 바, 드라이 타입의 약주 제조에 적합한 양조용 효모로는 NIBRFGC000501771 효모 균주가 적합한 것으로 분석되었다.After reviewing the speed of fermentation and the final alcohol content of the liquor, the NIBRFGC000501771 yeast strain was found to be suitable as a brewing yeast for the production of dry type herbal liquor.
3-2. 일반 성분 분석3-2. General ingredient analysis
상기 발효가 끝난 제조예 및 비교예의 술덧에 대하여 일반성분을 분석하였고, 하기 [표 2]에 정리하였다. The general components of the fermented liquor of the production examples and comparative examples were analyzed and summarized in [Table 2] below.
[표 2][Table 2]
산도(Acidity)와 아미노산도(Amino Acidity), 총산(Total acid) 함량은 모든 실험군에서 유사한 값이 검출되었다. 다만, 라빠리장으로 발효한 비교예 1의 시료는 총산 함량이 0.31g/100mL로 측정되어 다른 제조예에 비해 높게 나타났다. 또한, 불휘발분은 라빠리장으로 발효한 비교예 1의 시료가 6.2g/100㎖로 매우 높게 측정되었고, NIBRFGC000503472 균주로 발효한 제조예 2의 시료가 3.3g/100㎖로 가장 낮게 측정되었다. 술덧에 있는 포도당 2%(v/v)는 효모에 의해 알코올 발효되어 약 1%(v/v)의 에틸알코올로 생성된다. 라빠리장 효모로 발효한 비교예 1의 시료는 알코올 도수가 가장 낮았던 실험군으로, 알코올 도수가 낮으면 발효되지 않은 당류가 술덧에 축적되어 불휘발분이 높아지는 상기 결과와 일치하였다. 반대로 알코올 도수가 가장 높았던 NIBRFGC000503472 균주로 발효한 제조예 2의 시료에서 불휘발분이 가장 낮게 측정된 것도 상기 결과와 일치하였다.Similar values for acidity, amino acidity, and total acid content were detected in all experimental groups. However, the sample of Comparative Example 1 fermented with Laparijang had a total acid content of 0.31g/100mL, which was higher than that of other production examples. In addition, the non-volatile matter of the sample of Comparative Example 1 fermented with Laparijang was measured very high at 6.2 g/100 ml, and the sample of Preparation Example 2 fermented with strain NIBRFGC000503472 was measured the lowest at 3.3 g/100 ml. 2% (v/v) of glucose in the liquor is fermented into alcohol by yeast to produce about 1% (v/v) ethyl alcohol. The sample of Comparative Example 1 fermented with Laparijang yeast was the experimental group with the lowest alcohol content, which was consistent with the above results in that when the alcohol content was low, unfermented sugars accumulated in the liquor and the non-volatile matter increased. Conversely, the lowest non-volatile content measured in the sample of Preparation Example 2 fermented with the NIBRFGC000503472 strain, which had the highest alcohol content, was consistent with the above results.
3-3. 유리당 함량 분석3-3. Free sugar content analysis
발효가 끝난 제조예 및 비교예의 술덧에 대하여 유리당 중 단당류와 이당류 함량을 분석하였고, 하기 [표 3]에 정리하였다. The contents of monosaccharides and disaccharides among free sugars were analyzed for the fermented liquor of the production examples and comparative examples, and are summarized in [Table 3] below.
[표 3][Table 3]
유리당 함량은 모든 제조예와 비교예에서 과당, 자당, 맥아당, 유당은 검출되지 않았고, 포도당만 라빠리장으로 발효한 비교예 1의 시료에서 1.8%(w/v)로 높게 검출되었다. NIBRFGC000501771 균주로 발효한 제조예 1의 시료에서 0.3%(w/v), NIBRFGC000503472 균주로 발효한 제조예 2의 시료에서 0.1%(w/v), NIBRFGC000503478 균주로 발효한 제조예 3의 시료에서 0.1%(w/v) 순으로 낮게 검출되었다. As for the free sugar content, fructose, sucrose, maltose, and lactose were not detected in all preparation examples and comparative examples, and only glucose was detected as high as 1.8% (w/v) in the sample of Comparative Example 1 fermented with Laparijang. 0.3% (w/v) in the sample of Preparation Example 1 fermented with the NIBRFGC000501771 strain, 0.1% (w/v) in the sample of Preparation Example 2 fermented with the NIBRFGC000503472 strain, and 0.1% in the sample of Preparation Example 3 fermented with the NIBRFGC000503478 strain. It was detected in the order of %(w/v).
라빠리장으로 발효한 비교예 1의 시료는 알코올 도수가 가장 낮았던 실험군으로 알코올 도수가 낮으면 발효되지 않은 당류가 술덧에 축적되어 포도당 함량이 높아지는 상기 결과와 일치하였고, NIBRFGC000503472 균주로 발효한 제조예 2와 NIBRFGC000503478 균주로 발효한 제조예 3 시료는 당류가 적고 알코올 도수가 높아 드라이 약주 제조용으로 적합한 듯 보이지만, 담금 초기에 급격한 거품발생과 함께 끓어 넘침 현상이 발생하는 것은 제조관리가 어려워 실용성이 떨어지기 때문에 드라이 약주 제조용으로 부적합하다.The sample of Comparative Example 1 fermented with Laparijang was the experimental group with the lowest alcohol content, and was consistent with the above results in that when the alcohol content was low, unfermented sugars accumulated in the liquor and the glucose content increased. Production example fermented with strain NIBRFGC000503472 Preparation Example 3 samples fermented with strains 2 and NIBRFGC000503478 appear to be suitable for manufacturing dry herbal liquor due to their low sugar content and high alcohol content, but the rapid foaming and boiling over in the early stages of immersion make it difficult to manage and impractical. Therefore, it is not suitable for manufacturing dry herbal liquor.
이에 반해 NIBRFGC000501771 균주로 발효한 제조예 1의 시료는 당류가 적고 알코올 도수가 높아 약주 제조용으로 적합한 것으로 분석된다.On the other hand, the sample of Preparation Example 1 fermented with the NIBRFGC000501771 strain was analyzed to be suitable for manufacturing herbal liquor because it had low sugar content and high alcohol content.
실시예 4. 유기산 함량 분석Example 4. Organic acid content analysis
발효가 완료된 드라이 타입의 발효 술덧의 유기산 함량 분석 결과를 하기 [표 4]에 정리하였다. The organic acid content analysis results of the dry type fermented liquor after completion of fermentation are summarized in [Table 4] below.
[표 4][Table 4]
효모의 종류별로 발효시킨 드라이 타입의 약주에 함유된 유기산은 사용한 효모의 종류에 따라 개별 유기산 함량에 차이가 있는 것으로 측정되었다.Organic acids contained in dry type herbal liquor fermented by type of yeast were measured to have differences in individual organic acid content depending on the type of yeast used.
구체적으로 아세트산(Acetic acid)은 NIBRFGC000503472 균주로 발효한 제조예 2의 경우 469mg/L, NIBRFGC000503478 균주로 발효한 제조예 3의 경우 383mg/L, NIBRFGC000501771 균주로 발효한 제조예 1의 경우 358mg/L, 그리고, 라빠리장으로 발효한 비교예 1의 경우 165mg/L로 측정되었고, 라빠리장으로 발효한 비교예 1이 가장 적은 양을 함유하는 것으로 측정되었다. 다만, 젖산(Lactic acid)은 라빠리장으로 발효한 비교예 1이 3,404mg/L로 측정되었고, NIBRFGC000503472 균주로 발효한 제조예 2는 1,714mg/L, NIBRFGC000503478 균주로 발효한 제조예 3은 1,694mg/L, NIBRFGC000501771 균주로 발효한 제조예 1은 929mg/L로 측정되었다. Specifically, acetic acid was 469 mg/L in Preparation Example 2 fermented with the NIBRFGC000503472 strain, 383 mg/L in Preparation Example 3 fermented with the NIBRFGC000503478 strain, and 358 mg/L in Preparation Example 1 fermented with the NIBRFGC000501771 strain. In addition, in the case of Comparative Example 1 fermented with Laparijang, it was measured to be 165 mg/L, and Comparative Example 1 fermented with Laparijang was measured to contain the lowest amount. However, lactic acid was measured at 3,404 mg/L in Comparative Example 1 fermented with Lapari paste, 1,714 mg/L in Preparation Example 2 fermented with the NIBRFGC000503472 strain, and 1,694 mg/L in Preparation Example 3 fermented with the NIBRFGC000503478 strain. mg/L, Preparation Example 1 fermented with strain NIBRFGC000501771 was measured at 929 mg/L.
NIBRFGC000501771 균주로 발효한 제조예 1의 경우, 젖산(Lactic acid), 아세트산(Acetic acid)의 낮은 수치를 통해 정상적인 알코올 발효가 진행되었음을 확인할 수 있었고, NIBRFGC000503472 균주로 발효한 제조예 2와 NIBRFGC000503478 균주로 발효한 제조예 3의 경우, 담금 초기에 급격한 끓어오름 현상이 높은 수치의 젖산(Lactic acid) 및 아세트산(Acetic acid)의 생성에 의한 것임을 확인할 수 있었다. 최 등의 연구(2005)에 따르면, 발효 도중에 생성된 알코올이 초산균(acetic acid bacteria)에 의해 아세트산(acetic acid)으로 전환되기도 한다. 즉, 젖산(Lactic acid) 및 아세트산(Acetic acid) 함량은 유산균, 초산균 및 다른 효모균의 오염 척도가 된다고 볼 수 있다. 유산균, 초산균 및 다른 효모균 등으로 술덧이 오염되면 이들이 생성한 부산물에 의해 최종 제품에 나쁜 향취가 이행되어 술의 품질이 저하되기도 한다. In the case of Preparation Example 1 fermented with the NIBRFGC000501771 strain, it was confirmed that normal alcohol fermentation occurred through low levels of lactic acid and acetic acid, and Preparation Example 2 fermented with the NIBRFGC000503472 strain and NIBRFGC000503478 strain. In the case of Preparation Example 3, it was confirmed that the rapid boiling phenomenon at the beginning of immersion was caused by the production of high levels of lactic acid and acetic acid. According to research by Choi et al. (2005), alcohol produced during fermentation is converted to acetic acid by acetic acid bacteria. In other words, the content of lactic acid and acetic acid can be considered a measure of contamination by lactic acid bacteria, acetic acid bacteria, and other yeast bacteria. If alcoholic beverages are contaminated with lactic acid bacteria, acetic acid bacteria, and other yeast bacteria, the by-products they produce may impart a bad flavor to the final product, deteriorating the quality of the alcohol.
한편, 라빠리장으로 발효된 비교예 1은 젖산 함량이 높아 이상발효 현상을 의심해 볼 수 있겠으나, 이상발효 시 함께 증가하는 아세트산 함량은 낮은 것으로 보아 이상발효 현상보다는 효모의 개별 특성으로 사료된다. On the other hand, Comparative Example 1 fermented with Laparijang had a high lactic acid content, so abnormal fermentation could be suspected. However, the acetic acid content that increases during abnormal fermentation was low, which is believed to be an individual characteristic of the yeast rather than an abnormal fermentation phenomenon.
특히, 발효가 끝난 발효 원주의 에틸알코올(ethyl alcohol) 생성량이 높은 NIBRFGC000503472 균주로 발효된 제조예 2, 및 NIBRFGC000503478 균주로 발효한 제조예 3의 술덧에서 담금 초기에 급격한 끓어오름이 발생하는 현상이 관찰되었고, 발효 초기 알코올 생성이 낮은 현상이 관찰되었으며, 젖산(Lactic acid) 및 아세트산(Acetic acid)이 유의적으로 많이 검출된 결과를 통해, 상기 NIBRFGC000503472 및 NIBRFGC000503478 균주가 약주 제조에 바람직하지 못한 효모인 것으로 분석되었다.In particular, a phenomenon of rapid boiling occurring at the beginning of immersion was observed in Preparation Example 2, which was fermented with the NIBRFGC000503472 strain, and Preparation Example 3, which was fermented with the NIBRFGC000503478 strain. The phenomenon of low alcohol production in the early stage of fermentation was observed, and significantly more lactic acid and acetic acid were detected, showing that the NIBRFGC000503472 and NIBRFGC000503478 strains are undesirable yeast for manufacturing herbal liquor. analyzed.
또한, 시트르산(Citric acid), 숙신산(Succinic acid), 말산(Malic acid) 함량은 제조예 1(NIBRFGC000501771 균주)의 시료에서 가장 높은 수치로 검출되었다. 숙신산은 당류가 효모에 의해 발효될 때 부산물로 생성되는 산으로, 특유의 신맛, 감칠맛, 풍미를 제공하고, 시트르산의 경우 감귤류에 많이 함유되어 있으며, 말산의 경우 사과산이라고도 불리며 청량한 과일에 많이 포함되어 있어, 그 함량이 높을수록 관능적인 측면에서 좋은 평가를 받을 수 있다.In addition, the highest levels of citric acid, succinic acid, and malic acid were detected in the sample of Preparation Example 1 (NIBRFGC000501771 strain). Succinic acid is an acid produced as a by-product when sugars are fermented by yeast, and provides a unique sourness, umami, and flavor. Citric acid is contained in large quantities in citrus fruits, and malic acid is also called malic acid and is abundant in refreshing fruits. Therefore, the higher the content, the better it is evaluated in terms of sensuality.
총산(Total acid) 함량은 비교예 1(라빠리장)의 실험군에서 6,606mg/L로 가장 높게 검출되었고, 다음으로 제조예 2(NIBRFGC000503472 균주) 및 제조예 3(NIBRFGC000503478 균주), 제조예 1(NIBRFGC000501771 균주)의 순으로 높게 검출되었다. 제조예 2 및 제조예 3의 총산 함량이 높은 것은 젖산(Lactic acid) 및 아세트산(Acetic acid) 함량이 높은 것에서 기인한 것으로 바람직한 효모라 할 수 없다.Total acid content was detected highest at 6,606 mg/L in the experimental group of Comparative Example 1 (La Paris Jang), followed by Preparation Example 2 (NIBRFGC000503472 strain) and Preparation Example 3 (NIBRFGC000503478 strain), Preparation Example 1 ( NIBRFGC000501771 strain) was detected in the highest order. The high total acid content of Preparation Example 2 and Preparation Example 3 is due to the high content of lactic acid and acetic acid and cannot be considered a desirable yeast.
실시예 5. 효모의 응집 테스트Example 5. Yeast agglutination test
효모는 발효 후반부에 효모들 사이에서 엉김 현상이 일어나 발효 탱크 하단부에 가라앉는 특성이 있다. 응집 테스트(Flocculation test)는 효모의 가라앉는 정도를 분석하는 것이다. 특히, 약주는 혼탁한 형태로 제품화하는 탁주와 달리 발효가 끝난 술덧을 맑고 투명하게 하여 제품화하는 것이 특징인 술이고, 약주 양조공정에서 맑고 투명하게 만드는 여과 공정이 가장 어려운 공정에 해당한다. 드라이 타입의 약주용 효모는 응집성이 높아야 발효가 끝난 후 효모가 빠르게 침강해 이후 공정인 여과 공정을 손쉽게 진행할 수 있다. Yeast has the characteristic of causing agglomeration between yeasts in the latter half of fermentation and settling at the bottom of the fermentation tank. The flocculation test analyzes the degree to which yeast settles. In particular, Yakju is a liquor that, unlike Takju, which is commercialized in a cloudy form, is characterized by commercializing the fermented liquor in a clear and transparent manner. In the Yakju brewing process, the filtration process to make it clear and transparent is the most difficult process. Dry type yeast for medicinal liquor must have high cohesiveness so that the yeast settles quickly after fermentation is completed and the subsequent filtration process can be easily carried out.
실시예 1-4의 응집 테스트 방법에 따라 실험하여 측정한 결과를 하기 [표 5]에 정리하였다. The results of the experiment and measurement according to the agglutination test method of Examples 1-4 are summarized in [Table 5] below.
[표 5][Table 5]
[표 5]를 참조하면, NIBRFGC000503472 효모로 발효한 제조예 2 및 NIBRFGC000503478 효모로 발효한 제조예 3의 시료는 각각 41.75% 및 40.62%로 응집도가 높은 것으로 측정되어, 여과 공정만을 고려할 때 약주 제조용 효모로 유용하게 사용될 수 있는 것으로 볼 수 있는 여지가 있다. 다만, 전술한 발효 경과에 따른 에틸알코올 및 유기산 생성 프로파일을 참고하면 전반적으로 발효 과정에 문제가 있었고, 과도하게 응집성이 강한 특성은 발효 중 급격한 끓어 넘침 현상이 발생하는 원인 중 하나로 작용하는 것으로 볼 수도 있으므로, 드라이용 약주 효모로 부적합한 것으로 판정되었다. 따라서, NIBRFGC000503472 및 NIBRFGC000503478 효모를 제외하고, 응집성 및 침강성이 우수한 NIBRFGC000501771 효모가 드라이 타입의 약주 제조용 효모 후보로 선택되었다. Referring to [Table 5], the samples of Preparation Example 2 fermented with NIBRFGC000503472 yeast and Preparation Example 3 fermented with NIBRFGC000503478 yeast were measured to have high aggregation rates of 41.75% and 40.62%, respectively, and considering only the filtration process, the yeast for medicinal liquor production There is room for it to be seen as something that can be usefully used. However, referring to the ethyl alcohol and organic acid production profile during the fermentation process described above, there was a problem in the overall fermentation process, and the excessively cohesive nature may be seen as one of the causes of rapid boiling over during fermentation. Therefore, it was judged to be unsuitable as dry herbal liquor yeast. Therefore, except for NIBRFGC000503472 and NIBRFGC000503478 yeast, NIBRFGC000501771 yeast, which has excellent coagulation and sedimentation properties, was selected as a dry type yeast candidate for manufacturing herbal liquor.
실시예 6. 향기성분 함량 분석Example 6. Analysis of fragrance ingredient content
실시예 1-5의 향기성분 분석 방법에 따라 발효가 완료된 드라이 타입 약주의 발효 술덧에 함유된 주요 향기성분 분석 결과를 하기 [표 6]에 정리하였다. The analysis results of the main aroma components contained in the fermented liquor of dry type herbal liquor, which was completely fermented according to the aroma component analysis method of Examples 1-5, are summarized in [Table 6] below.
[표 6][Table 6]
효모의 종류별로 발효시킨 드라이 타입 약주의 주요 향기 성분의 총 함량은 제조예 1(NIBRFGC000501771 균주), 비교예 1(라빠리장)에서 각각 2,833.6mg/L, 2,414.0mg/L의 순으로 높게 측정되었다. The total content of major aroma components of dry type herbal liquor fermented by type of yeast was measured to be as high as 2,833.6 mg/L and 2,414.0 mg/L in Preparation Example 1 (NIBRFGC000501771 strain) and Comparative Example 1 (Laparijang), respectively. .
세부 성분별로 살펴보면, 과일향, 꽃향, 바나나향, 배향 등의 향기로 감지되는 methyl acetate, ethyl acetate 및 iso-buthyl acetete 성분의 함량 합계는 제조예 1(NIBRFGC000501771) 383.6mg/L, 비교예 1(라빠리장) 253.5mg/L, 제조예 3(NIBRFGC000503478) 207.7mg/L 및 제조예 2(NIBRFGC000503472) 201mg/L의 순으로 검출되었다.Looking at the detailed components, the total content of methyl acetate, ethyl acetate, and iso-butyl acetate components detected as fruity, floral, banana, and pear scents is 383.6 mg/L in Preparation Example 1 (NIBRFGC000501771), and Comparative Example 1 ( Laparijang) 253.5 mg/L, Preparation Example 3 (NIBRFGC000503478) 207.7 mg/L, and Preparation Example 2 (NIBRFGC000503472) 201 mg/L.
멜론향, 풋사과향, 파인애플향 등으로 감지되는 ethyl octanoate 성분 함량은 제조예 1(NIBRFGC000501771) 3.7mg/L, 제조예 3(NIBRFGC000503478) 0.5mg/L, 및 제조예 2(NIBRFGC000503472) 0.4mg/L 순으로 높게 검출되었고, 비교예 1(라빠리장)은 1.3mg/L로 검출되었다.The content of ethyl octanoate, which is detected as melon, green apple, and pineapple flavors, is 3.7 mg/L in Preparation Example 1 (NIBRFGC000501771), 0.5 mg/L in Preparation Example 3 (NIBRFGC000503478), and 0.4 mg/L in Preparation Example 2 (NIBRFGC000503472). It was detected at higher levels in that order, and Comparative Example 1 (La Paris Jang) was detected at 1.3 mg/L.
또한, 장미향, 사과향, 꿀향 등으로 감지되는 ethyl phenyl acetate 성분 함량은 제조예 1(NIBRFGC000501771) 2.4mg/L, 제조예 3(NIBRFGC000503478) 2.0mg/L 순으로 높게 검출되었고, 비교예 1(라빠리장)은 2.2mg/L로 검출되었다. In addition, the content of ethyl phenyl acetate, which is detected as rose scent, apple scent, honey scent, etc., was detected as high as 2.4 mg/L in Preparation Example 1 (NIBRFGC000501771), 2.0 mg/L in Preparation Example 3 (NIBRFGC000503478), and Comparative Example 1 (Lappari intestine) was detected at 2.2mg/L.
그리고, 위스키, 브랜디, 사과향 등으로 감지되는 ethyl hexanoate 성분의 함량은 제조예 1(NIBRFGC000501771) 4.4mg/L, 제조예 3(NIBRFGC000503478) 1.8mg/L, 제조예 2(NIBRFGC000503472) 1.6 mg/L 순으로 높은 수치가 검출되었고, 비교예 1(라빠리장)은 1.7mg/L로 검출되었다. In addition, the content of ethyl hexanoate detected as whiskey, brandy, apple flavor, etc. was 4.4 mg/L in Preparation Example 1 (NIBRFGC000501771), 1.8 mg/L in Preparation Example 3 (NIBRFGC000503478), and 1.6 mg/L in Preparation Example 2 (NIBRFGC000503472). A high level was detected, and Comparative Example 1 (La Paris Jean) was detected at 1.7 mg/L.
발효 도중 효모로부터 아미노산 분해로 생성되는 퓨젤유(Fusel oil)에 해당하는 1-propanol, isobutanol, 1-butanol, isoamyl alcohol 성분의 검출량 합계는 제조예 1(2,340.4mg/L)과 비교예 1(2,067.0mg/L)에서 가장 높게 검출되었고, 제조예 2 및 제조예 3에서는 각각 1,554.9mg/L 및 1,540.3mg/L로 검출되었다. The total detected amounts of 1-propanol, isobutanol, 1-butanol, and isoamyl alcohol, which correspond to fusel oil produced by amino acid decomposition from yeast during fermentation, were Preparation Example 1 (2,340.4 mg/L) and Comparative Example 1 (2,067.0 mg). /L), and in Preparation Example 2 and Preparation Example 3, it was detected at 1,554.9 mg/L and 1,540.3 mg/L, respectively.
지방산 성분이 효모 균체 내에서 에틸알코올 성분과 결합하여 주로 생성되는 지방산에스터는 사과향, 파인애플향, 꽃향, 왁스향 등으로 대별되며, 주요 성분인 ethyl octanoate 성분과 ethyl dodecanoate 성분의 함량 합계는 제조예 1(4.6mg/L) 및 비교예 1(2.3 mg/L)에서 높게 검출되었고, 제조예 2 및 제조예 3에서는 각각 1.7mg/L 및 1.1mg/L로 검출되었다. Fatty acid esters, which are mainly produced by combining fatty acid components with ethyl alcohol components within yeast cells, are broadly classified into apple flavor, pineapple flavor, flower flavor, and wax flavor, and the total content of the main components, ethyl octanoate component and ethyl dodecanoate component, is Preparation Example 1 (4.6 mg/L) and Comparative Example 1 (2.3 mg/L), and in Preparation Example 2 and Preparation Example 3, it was detected at 1.7 mg/L and 1.1 mg/L, respectively.
상기 향기 성분 측정 결과를 종합하면, 제조예 2(NIBRFGC000503472)와 제조예 3(NIBRFGC000503478)은 급격한 거품발생 및 끓어 넘침 현상과 초기 발효 속도가 낮은 경향성을 보였는데, 이는 본 실험에서 향기 성분 함량이 낮은 수치로 검출된 것과 일맥상통하였고, 드라이 타입의 약주용으로 사용하기는 부적합한 것으로 평가되었다. Summarizing the results of the aroma component measurement, Preparation Example 2 (NIBRFGC000503472) and Preparation Example 3 (NIBRFGC000503478) showed a tendency for rapid foaming and boiling over and a low initial fermentation rate, which was due to the low aroma component content in this experiment. It was consistent with what was detected numerically, and was evaluated as unsuitable for use as a dry type herbal medicine.
종합하면, 향기 성분 함량의 총합이 높게 검출된 시험군은 제조예 1(NIBRFGC000501771) 및 비교예 1(라빠리장)이었고, 세부적으로 에스터류와 퓨젤 오일, 지방산에스터 함량이 높은 실험군은 제조예 1(NIBRFGC000501771) 및 비교예 1(라빠리장)의 실험군이었다. 따라서, NIBRFGC000501771 균주로 제조된 드라이 타입의 약주는 관능 평가에서도 우수한 성적을 획득할 수 있을 것으로 예상되었다. In summary, the test group in which the total content of fragrance components was detected to be high was Preparation Example 1 (NIBRFGC000501771) and Comparative Example 1 (La Paris Jean), and in detail, the test group with high contents of esters, fusel oils, and fatty acid esters was Preparation Example 1. (NIBRFGC000501771) and Comparative Example 1 (La Paris Jean). Therefore, it was expected that the dry type herbal liquor manufactured with strain NIBRFGC000501771 would achieve excellent results in sensory evaluation as well.
특히, 향기성분을 다량 함유한 NIBRFGC000501771 균주로 제조되는 약주는 장미향, 꿀향, 사과향으로 감지되는 ethyl phenyl acetate 성분, 바나나향으로 감지되는 iso butyl acetate 성분, 배향으로 감지되는 iso pentyl acetate 성분, 사과향으로 감지되는 ethyl hexanoate 성분, 그리고 멜론향, 풋사과향, 파인애플향으로 인지되는 ethyl octanoate 성분 함량이 동시에 높게 검출되는 것으로 확인되었다. In particular, the herbal liquor manufactured with the NIBRFGC000501771 strain, which contains a large amount of aromatic components, includes ethyl phenyl acetate component detected as rose, honey, and apple aroma, iso butyl acetate component detected as banana aroma, iso pentyl acetate component detected as pear aroma, and apple aroma. It was confirmed that the content of ethyl hexanoate, which is recognized as melon, green apple, and pineapple, was detected simultaneously at high levels.
실시예 7. 관능 평가Example 7. Sensory evaluation
효모별로 제조한 드라이 타입의 약주에 대한 관능 평가는 국세청주류면허지원센터의 관능평가 전문 패널(n=10)이 참여하여 외관, 맛, 향기, 감촉 항목에 대하여 묘사분석으로 평가하였다. 외관과 감촉 항목은 최저점 1에서 최고점 3점으로, 맛과, 향기 항목은 최저점 1에서 최고점 6점으로 하여 평가점수의 평균값을 하기 [표 7]에 정리하였다.Sensory evaluation of dry type herbal liquor manufactured by yeast was conducted by a sensory evaluation expert panel (n=10) from the National Tax Service Liquor License Support Center and evaluated appearance, taste, aroma, and texture items through descriptive analysis. Appearance and texture items ranged from a minimum score of 1 to a maximum score of 3 points, and taste and scent items ranged from a minimum score of 1 to a maximum score of 6 points. The average values of the evaluation scores are summarized in [Table 7] below.
[표 7][Table 7]
색상과 여과 정도 등 외관을 평가하는 항목에서, NIBRFGC000501771 균주로 제조한 약주가 높은 점수를 얻었다. 맑고 투명한 외관을 갖는 약주의 특성을 고려하면, 응집성이 뛰어나 빨리 침강되므로 여과를 쉽게 진행할 수 있는 특성을 갖는 효모가 좋은 효모이므로, 전술한 바와 같이 여과 정도가 뛰어난 NIBRFGC000501771 균주는 약주 드라이용 효모로 유리한 조건이라 할 수 있다. 향기와 맛이 모두 좋아 높은 관능평가 점수를 획득한 시료도 NIBRFGC000501771 균주로 제조한 실험군이다. 제조예 2(NIBRFGC000503472) 및 제조예 3(NIBRFGC000503478)의 시료는 모든 항목에서 나쁜 점수를 받았다. 이는 급격한 거품발생 및 끓어 넘침 현상에 따라 초기 발효 진행이 늦어져 충분하게 향기성분을 생산하지 못했기 때문으로 사료되며, 이들 균주는 드라이 약주 제조를 위한 양조용 효모로 사용되기 어려운 것으로 평가되었다. In terms of evaluating appearance, such as color and degree of filtration, herbal liquor made with strain NIBRFGC000501771 received high scores. Considering the characteristics of Yakju, which has a clear and transparent appearance, a good yeast is a yeast that has excellent cohesiveness and quickly settles, allowing easy filtration. As mentioned above, the strain NIBRFGC000501771, which has excellent filtration, is advantageous as a yeast for drying Yakju. It can be said to be a condition. The sample that obtained a high sensory evaluation score due to both good scent and taste was also an experimental group prepared with the NIBRFGC000501771 strain. The samples of Preparation Example 2 (NIBRFGC000503472) and Preparation Example 3 (NIBRFGC000503478) received bad scores in all items. This is believed to be because the initial fermentation progress was delayed due to rapid foaming and boiling over, which did not produce sufficient aroma components, and these strains were evaluated as difficult to use as brewing yeast for manufacturing dry herbal liquor.
라빠리장 효모를 사용한 비교예 1의 실험군은 2순위의 관능 평가점수를 획득하였으며, NIBRFGC000501771 균주로 제조한 제조예 1의 약주가 비교예에 비해 월등하게 높은 관능 평가점수를 획득함에 따라, NIBRFGC000501771 균주가 드라이 타입의 약주 제조를 위한 양조용 효모로 시판용 효모 균주보다 더 우수한 성능을 발휘할 수 있음을 확인하였다.The experimental group of Comparative Example 1 using Laparijang yeast obtained the second highest sensory evaluation score, and as the herbal liquor of Preparation Example 1 prepared with the NIBRFGC000501771 strain obtained a significantly higher sensory evaluation score than the comparative example, the NIBRFGC000501771 strain It was confirmed that brewing yeast for the production of dry type medicinal liquor can perform better than commercial yeast strains.
상기 실험 결과를 종합하면, 알코올 생산력, 당 함량 축적 정도, 발효의 안정성, 유기산 함량, 불휘발분 잔류량 등의 측면에서 NIBRFGC000501771 효모 균주가 드라이 타입의 약주 제조에 우수한 것으로 확인되었다. 또한, 효모의 응집성(Flocculation)이 뛰어나 약주의 여과에 유리한 효모는 NIBRFGC000501771 효모 균주인 것으로 나타났다.Summarizing the above experimental results, it was confirmed that the NIBRFGC000501771 yeast strain was excellent for the production of dry type herbal liquor in terms of alcohol production capacity, degree of sugar content accumulation, fermentation stability, organic acid content, and non-volatile matter residual amount. In addition, the yeast strain NIBRFGC000501771 was found to be advantageous for filtration of medicinal liquor due to its excellent flocculation.
또한, 향기성분 분석 결과, NIBRFGC000501771 균주는 2833.6mg/L의 총 향기성분을 생산하는 것으로 측정되었다. 특히, NIBRFGC000501771 균주로 제조된 약주는 바나나향, 과일향, 꽃향으로 감지되는 iso butyl acetate 성분, 장미향, 사과향, 꿀향으로 감지되는 ethyl phenyl acetate 성분, 그리고 위스키, 브랜디, 사과향 등으로 감지되는 ethyl hexanoate 성분, 및 멜론향, 풋사과향, 파인애플향 등으로 감지되는 ethyl octanoate 성분 함량이 동시에 높게 검출되었다. 이외에도 본 발명의 NIBRFGC000501771 균주로 제조된 약주는 감칠맛과 특유의 신맛을 제공하는 숙신산 및 시트르산 함량이 높았고, 말산 함량이 높은 것으로 분석되었으며, 이를 통해 NIBRFGC000501771 효모 균주가 드라이 타입의 약주 주조용 효모로 바람직하게 이용될 수 있음을 확인할 수 있었다. 관능검사 결과에서도 NIBRFGC000501771 균주는 외관, 향, 맛, 감촉 등 모든 항목에서 가장 좋은 평가를 받았다. In addition, as a result of aroma component analysis, strain NIBRFGC000501771 was measured to produce a total aroma component of 2833.6 mg/L. In particular, the herbal liquor manufactured with strain NIBRFGC000501771 contains iso butyl acetate component detected as banana, fruit, and flower aroma, ethyl phenyl acetate component detected as rose, apple, and honey aroma, and ethyl hexanoate component detected as whiskey, brandy, and apple aroma. , and the content of ethyl octanoate, which is detected as melon, green apple, and pineapple aromas, was simultaneously detected at high levels. In addition, the herbal liquor prepared with the NIBRFGC000501771 strain of the present invention was analyzed to have a high succinic acid and citric acid content, which provides umami and a unique sour taste, and a high malic acid content, which makes the NIBRFGC000501771 yeast strain desirable as a dry type yeast for brewing herbal liquor. It was confirmed that it could be used. In the sensory test results, the NIBRFGC000501771 strain received the best evaluation in all categories, including appearance, aroma, taste, and texture.
상기 결과를 종합하면, NIBRFGC000501771 효모 균주가 드라이 타입의 약주 주조용 효모로 가장 적합한 균주로 평가된다.Considering the above results, the NIBRFGC000501771 yeast strain is evaluated as the most suitable dry type yeast for brewing herbal liquor.
실시예 8. NIBRFGC000501771 균주의 균학적 특성 분석Example 8. Analysis of mycological characteristics of strain NIBRFGC000501771
8-1. 배양 및 형태학적 특징8-1. Culture and morphological characteristics
NIBRFGC000501771 균주가 평판배지에서 생장하는 양상을 YPD(Difco) 한천배지에 접종 후 25℃에서 7일간 정치배양 한 후 관찰하였다.The growth pattern of NIBRFGC000501771 strain on plate medium was observed after inoculation on YPD (Difco) agar medium and culture at 25°C for 7 days.
도 2를 참조하면, YPD 배지에서 배양된 사카로마이세스 세레비지애(S. cerevisiae) NIBRFGC000501771 균주의 콜로니 색깔은 흰색이며 가장자리는 부드러운 양상을 보였다.Referring to Figure 2, the colony color of S. cerevisiae NIBRFGC000501771 strain cultured in YPD medium was white and the edges were smooth.
8-2. 세포 형태8-2. cell shape
평판배지에서 자란 사카로마이세스 세레비지애(S. cerevisiae) NIBRFGC000501771 균주의 세포 형태는 광학현미경 1,000배와 주사전자현미경(Scanning Electron Microscopy)으로 관찰하였다. The cell morphology of Saccharomyces cerevisiae NIBRFGC000501771 strain grown on plate medium was observed using a light microscope at 1,000x magnification and scanning electron microscopy.
광학현미경으로 관찰한 결과, 구형 또는 난형의 모세포에서 딸세포가 출아하는 전형적인 효모 세포의 모습을 보였다(도 3 왼쪽 그림 참조). As a result of observation with an optical microscope, it showed a typical yeast cell appearance in which daughter cells germinate from a spherical or ovoid mother cell (see left figure in Figure 3).
주사전자현미경 하에서도 구형 또는 난형의 세포에서 딸세포가 출아하고, 딸세포가 떨어져 나간 스카가 관찰되었다(도 3 오른쪽 그림 참조). 또한, NIBRFGC000501771 균주에서는 분아해 나온 딸세포가 아닌 세포끼리도 연결되는 양상이 관찰되었다. 이는 세포외 기질 단백질(extracellular matrix protein) 중 어드헤신(adhesin) 등의 단백질들이 세포들의 응집을 촉진하는 것으로 생각된다. 이는 본 발명의 NIBRFGC000501771 균주가 응집력이 좋아 약주용 효모로 적합한 것과 일맥상통한다.Even under a scanning electron microscope, daughter cells budding from spherical or egg-shaped cells and scars where the daughter cells fell off were observed (see the right figure in Figure 3). In addition, in the NIBRFGC000501771 strain, cells other than the daughter cells that had sprouted were observed to be connected to each other. It is thought that proteins such as adhesin among extracellular matrix proteins promote the aggregation of cells. This is consistent with the fact that the NIBRFGC000501771 strain of the present invention has good cohesiveness and is suitable as a medicinal yeast.
8-3. 생리적 특성8-3. physiological characteristics
사카로마이세스 세레비지애(S. cerevisiae) NIBRFGC000501771 균주의 탄소원 이용 능력은 Biolog社의 YT MicroPlate를 이용한 표현형 마이크로어레이 테크놀로지(phenotype microarray technology) 기법으로 확인하였다. 96 웰 마이크로플레이트(96 well microplate)에 담긴 개별 탄소원을 이용하여 자랄 때, 세포 내 호흡의 결과로 무색의 테트라졸륨 바이올렛(tetrazolium violet) 시약이 보라색으로 바뀌는 양상 및 단일 탄소원에서 생장하는 양상을 측정하여 탄소원 이용 유무를 파악하였다. The ability of the S. cerevisiae strain NIBRFGC000501771 to utilize carbon sources was confirmed using phenotype microarray technology using YT MicroPlate from Biolog. When grown using individual carbon sources contained in a 96 well microplate, the colorless tetrazolium violet reagent turns purple as a result of intracellular respiration and the growth pattern from a single carbon source is measured. The use of carbon sources was determined.
실험 결과를 정리한 도 4를 참조하면, NIBRFGC000501771 균주는 산화시키는 기질로 Inulin, Maltose, D-Raffinose, Sucrose, Turanose, α-D-Glucose, D-Galactose를 이용할 수 있었다. D-Trehalose에 대한 산화력은 이용 가능한 다른 기질에 비해 낮은 것으로 조사되었다. 동화(Assimilation)에 이용할 수 있는 탄소원은 Inulin, Maltose, Maltotriose, D-Raffinose, Stachyose, Sucrose, Turanose, α-D-Glucose, D-Galactose, 및 D-Trehalose 성분이었다. D-Melezitose 성분은 다른 탄소원에 비해 이용도가 낮았으며, D-Trehalose 성분은 안정적으로 이용하지 못하는 것으로 나타났다. Maltitol과 D-Ribose, D-Xylose 성분은 이용도가 낮고 이용 안정성 또한 낮은 것으로 나타났다. D-xylose 성분과 혼합 이용도를 알아보는 조사 결과, [Succinic Acid Mono-Methyl Ester + D-Xylose]와 [N-Acetyl-L-Glutamic Acid + D-Xylose] 성분은 D-Xylose 성분 단독 이용 결과와 유사하게 이용도가 낮았고, 안정성이 낮은 양상을 보였으나, [D-Galactose + D-Xylose] 성분은 활발하게 이용하는 것으로 나타났다.Referring to Figure 4, which summarizes the experimental results, strain NIBRFGC000501771 was able to use Inulin, Maltose, D-Raffinose, Sucrose, Turanose, α-D-Glucose, and D-Galactose as oxidizing substrates. The oxidizing power of D-Trehalose was found to be low compared to other available substrates. Carbon sources available for assimilation included Inulin, Maltose, Maltotriose, D-Raffinose, Stachyose, Sucrose, Turanose, α-D-Glucose, D-Galactose, and D-Trehalose. The D-Melezitose component had lower availability compared to other carbon sources, and the D-Trehalose component appeared not to be used stably. Maltitol, D-Ribose, and D-Xylose ingredients were found to have low availability and low stability. As a result of a survey to determine the D-xylose ingredient and its mixing use, [Succinic Acid Mono-Methyl Ester + D-Xylose] and [N-Acetyl-L-Glutamic Acid + D-Xylose] were the results of using D-Xylose alone. Similar to , utilization was low and stability was low, but the [D-Galactose + D-Xylose] ingredient was found to be actively used.
8-4. 유전적 특징8-4. genetic characteristics
사카로마이세스 세레비지애(S. cerevisiae) NIBRFGC000501771 균주의 유전적 특징을 파악하고자, 6개의 하우스키핑(housekeeping) 유전자 서열을 이용하여 SGD (Saccharomyces Genome Database, https://www.yeastgenome.org/)의 얼터너티브 레퍼런스 스트레인(alternative reference strain)들과 비교하여 멀티 로코스 시퀀스 타이핑(multi locus sequence typing) 분석을 실시하였다. To determine the genetic characteristics of the S. cerevisiae strain NIBRFGC000501771, six housekeeping gene sequences were used to determine the genetic characteristics of Saccharomyces Genome Database (SGD, https://www.yeastgenome.org/). ) A multi locus sequence typing analysis was performed in comparison with alternative reference strains.
구체적으로 NIBRFGC000501771 균주로부터 DNA를 추출하고, 잘 보존된 6종 마커 유전자를 증폭한 후 염기서열을 확보하였다. 6개 마커 유전자((ACC1, ADP1, GLN4, MET4, NUP116, RPN2)의 프라이머는 하기 [표 8]과 같다. Specifically, DNA was extracted from strain NIBRFGC000501771, six well-preserved marker genes were amplified, and base sequences were obtained. Primers for six marker genes ((ACC1, ADP1, GLN4, MET4, NUP116, RPN2) are shown in [Table 8] below.
[표 8][Table 8]
비교군으로는 효모 유전체 데이터베이스인 SGD(Saccharomyces Genome Database)에 등록된 얼터너티브 레퍼런스 스트레인(alternative reference strain)을 이용하였다. 분자계통수(Phylogenetic tree)는 Mega X 프로그램에서 설정된 기본값으로 네이버-조이닝(Neighbor-joining) 방법을 이용하여 작성하였다. 이때 유전적 거리는 타무라-네이(Tamura-Nei) 방법에 따라 계산하였다.As a comparison group, an alternative reference strain registered in the Saccharomyces Genome Database (SGD), a yeast genome database, was used. The molecular phylogenetic tree was created using the Neighbor-joining method with default values set in the Mega X program. At this time, the genetic distance was calculated according to the Tamura-Nei method.
균주의 유연관계 분석 결과를 정리한 도 5를 참조하면, NIBRFGC000501771 균주는 미국의 캘리포니아의 와인통 부산물에서 분리된 BC187 균주 및 칠레 와인 생산 균주인 L1528 균주와 가장 가까운 유연관계를 형성하는 것으로 나타났다. 또한, 일본에서 분리된 사케 균주인 Kyokai7 균주와는 유전적으로 다소 거리가 있는 것으로 확인되었다.Referring to Figure 5, which summarizes the results of the strain relationship analysis, the NIBRFGC000501771 strain was found to form the closest affinity with the BC187 strain isolated from wine barrel by-products in California, USA, and the L1528 strain, a Chilean wine production strain. In addition, it was confirmed to be somewhat genetically distant from the Kyokai7 strain, a sake strain isolated in Japan.
상기와 같이 생리적, 유전적 특징 분석을 완료한 NIBRFGC000501771 균주는 2022년 2월 17일자로 국립농업과학원 농업유전자원센터(KACC)에 사카로마이세스 세레비지애 NIBRFGC000501771 균주로 기탁하여 수탁번호 KACC 83061BP를 부여받았다. The NIBRFGC000501771 strain, which completed the analysis of physiological and genetic characteristics as described above, was deposited as Saccharomyces cerevisiae NIBRFGC000501771 strain at the Agricultural Genetic Resources Center (KACC) of the National Academy of Agricultural Sciences on February 17, 2022, and assigned the accession number KACC 83061BP. It was granted.
이상의 설명으로부터, 본 발명이 속하는 기술분야의 당업자는 본 발명이 그 기술적 사상이나 필수적 특징을 변경하지 않고서 다른 구체적인 형태로 실시될 수 있다는 것을 이해할 수 있을 것이다. 이와 관련하여, 이상에서 기술한 실시예들은 모든 면에서 예시적인 것이며 한정적인 것이 아닌 것으로서 이해해야만 한다. 본 발명의 범위는 상기 상세한 설명보다는 후술하는 특허 청구범위의 의미 및 범위 그리고 그 등가 개념으로부터 도출되는 모든 변경 또는 변형된 형태가 본 발명의 범위에 포함되는 것으로 해석되어야 한다.From the above description, those skilled in the art to which the present invention pertains will understand that the present invention can be implemented in other specific forms without changing its technical idea or essential features. In this regard, the embodiments described above should be understood in all respects as illustrative and not restrictive. The scope of the present invention should be construed as including the meaning and scope of the patent claims described below rather than the detailed description above, and all changes or modified forms derived from the equivalent concept thereof are included in the scope of the present invention.
Claims (8)
상기 균주로 발효 완료된 술덧의 최종 에틸알코올 도수는 14.0%(v/v) 내지 20.0%(v/v)인 것을 특징으로 하는, 드라이 타입의 약주 제조용 사카로마이세스 세레비지애 신균주 NIBRFGC000501771(기탁번호 KACC 83061BP).According to paragraph 1,
Saccharomyces cerevisiae new strain NIBRFGC000501771 (deposited) for producing dry type herbal liquor, characterized in that the final ethyl alcohol content of the fermented liquor with the above strain is 14.0% (v/v) to 20.0% (v/v) Number KACC 83061BP).
상기 균주는 효모의 응집성이 높은 것을 특징으로 하는, 드라이 타입의 약주 제조용 사카로마이세스 세레비지애 신균주 NIBRFGC000501771(기탁번호 KACC 83061BP).According to paragraph 1,
The strain is a new strain of Saccharomyces cerevisiae for the production of dry type medicinal liquor, NIBRFGC000501771 (accession number KACC 83061BP), which is characterized by high yeast aggregation.
상기 균주는 향기 성분을 고함량으로 생성시키는 것을 특징으로 하는, 드라이 타입의 약주 제조용 사카로마이세스 세레비지애 신균주 NIBRFGC000501771(기탁번호 KACC 83061BP).According to paragraph 1,
The strain is a new Saccharomyces cerevisiae strain NIBRFGC000501771 (accession number KACC 83061BP) for the production of dry type herbal liquor, which is characterized by producing a high content of aroma components.
상기 향기 성분은 메틸 아세테이트(methyl acetate), 에틸 아세테이트(ethyl acetate), 이소-부틸 아세테이트(iso-buthyl acetete), 에틸 부틸레이트(ethyl butylate), 1-프로판올(1-propanol), 이소부탄올(isobutanol), 이소펜틸 아세테이트(isopentyl acetate), 1-부탄올(1-butanol), 이소아밀 알코올(iso-amyl alcohol), 에틸 헥사노에이트(Ethyl hexanoate), 1-헥사놀(1-hexanol), 에틸 옥타노에이트(ethyl octanoate), 푸르푸랄(furfural), 페닐 아세테이트(phenyl acetate), 에틸페닐 아세테이트(ethylphenyl acetate), 및 에틸 도데카노에이트(ethyl dodecanoate)인 것을 특징으로 하는, 드라이 타입의 약주 제조용 사카로마이세스 세레비지애 신균주 NIBRFGC000501771(기탁번호 KACC 83061BP).According to paragraph 4,
The fragrance ingredients include methyl acetate, ethyl acetate, iso-butyl acetate, ethyl butylate, 1-propanol, and isobutanol. ), isopentyl acetate, 1-butanol, iso-amyl alcohol, Ethyl hexanoate, 1-hexanol, ethyl octa Dry type saccharide for manufacturing herbal liquor, characterized in that it contains ethyl octanoate, furfural, phenyl acetate, ethylphenyl acetate, and ethyl dodecanoate. Myces cerevisiae new strain NIBRFGC000501771 (accession number KACC 83061BP).
상기 밑술에 팽화미, 정제효소제 및 정제수를 첨가하고 10일 내지 14일 동안 발효하여 담금하는 단계;를 포함하는,
드라이 타입의 약주 제조 방법.Preparing base liquor by entering purified water and inoculating the new strain of Saccharomyces cerevisiae NIBRFGC000501771 (accession number KACC 83061BP) and fermenting for 2 to 5 days; and
Including the step of adding puffed rice, purified enzyme, and purified water to the base liquor and fermenting and soaking for 10 to 14 days.
Dry type herbal liquor manufacturing method.
상기 향기 성분은 메틸 아세테이트(methyl acetate), 에틸 아세테이트(ethyl acetate), 이소-부틸 아세테이트(iso-buthyl acetete), 에틸 부틸레이트(ethyl butylate), 1-프로판올(1-propanol), 이소부탄올(isobutanol), 이소펜틸 아세테이트(isopentyl acetate), 1-부탄올(1-butanol), 이소아밀 알코올(iso-amyl alcohol), 에틸 헥사노에이트(Ethyl hexanoate), 1-헥사놀(1-hexanol), 에틸 옥타노에이트(ethyl octanoate), 푸르푸랄(furfural), 페닐 아세테이트(phenyl acetate), 에틸페닐 아세테이트(ethylphenyl acetate), 및 에틸 도데카노에이트(ethyl dodecanoate)인 것을 특징으로 하는, 드라이 타입의 약주.In clause 7,
The fragrance ingredients include methyl acetate, ethyl acetate, iso-butyl acetate, ethyl butylate, 1-propanol, and isobutanol. ), isopentyl acetate, 1-butanol, iso-amyl alcohol, Ethyl hexanoate, 1-hexanol, ethyl octa A dry type herbal liquor characterized by ethyl octanoate, furfural, phenyl acetate, ethylphenyl acetate, and ethyl dodecanoate.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR19980049302A (en) | 1996-12-19 | 1998-09-15 | 백운화 | New homegrown medicinal yeast strain Saccharomyces cerevises TD03 and manufacturing method of medicinal herbs using the same |
| KR20080053629A (en) | 2006-12-11 | 2008-06-16 | 주식회사 두산 | Method for preparing rice wine using saccharomyces cerevisiae t084 increasing aromatic compounds in alcoholic liquor and oriental hub |
| KR20170137237A (en) | 2016-06-02 | 2017-12-13 | 숭실대학교산학협력단 | Saccharomycopsis fibuligera KJJ81 strain with high saccharification and fermentation capability and the method for preparing makgeolli |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR19980049302A (en) | 1996-12-19 | 1998-09-15 | 백운화 | New homegrown medicinal yeast strain Saccharomyces cerevises TD03 and manufacturing method of medicinal herbs using the same |
| KR20080053629A (en) | 2006-12-11 | 2008-06-16 | 주식회사 두산 | Method for preparing rice wine using saccharomyces cerevisiae t084 increasing aromatic compounds in alcoholic liquor and oriental hub |
| KR20170137237A (en) | 2016-06-02 | 2017-12-13 | 숭실대학교산학협력단 | Saccharomycopsis fibuligera KJJ81 strain with high saccharification and fermentation capability and the method for preparing makgeolli |
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