초록 ▼

Ⅳ. 연구개발결과
○ 탁주 제조용 곰팡이 및 효모 종균 선발 및 보존
- 독소 안전성, 효소활성, 알코올 발효능, 향기 등을 가진 종균 선정
· 곰팡이 2종(A. luchuensis 74-5, A. oryzae 75-2), 효모(S. cerevisiae Y204, Y283)
- 선발 종균의 발효 최적조건 설정 : 탄소원, 질소원, 온도 및 배양시간 등
- 선발 종균 보존 및 등록 : 계대, 글리세롤 보존 및 KACC 등록
○ 탁주용 고체 및 액체종균제조기술의 최적조건 구명
- 고체종균(곰팡이,

Ⅳ. 연구개발결과
○ 탁주 제조용 곰팡이 및 효모 종균 선발 및 보존
- 독소 안전성, 효소활성, 알코올 발효능, 향기 등을 가진 종균 선정
· 곰팡이 2종(A. luchuensis 74-5, A. oryzae 75-2), 효모(S. cerevisiae Y204, Y283)
- 선발 종균의 발효 최적조건 설정 : 탄소원, 질소원, 온도 및 배양시간 등
- 선발 종균 보존 및 등록 : 계대, 글리세롤 보존 및 KACC 등록
○ 탁주용 고체 및 액체종균제조기술의 최적조건 구명
- 고체종균(곰팡이, AL, AO) : 액체종국 5%, 제국온도 (AL 40℃, AO 35℃), 제국기간 5일
- 액체종균(효모 Y204, Y283) : YPD 배지, 30℃, 150 rpm, 24 hr
- 종균 보호용 증량제 : 3% skim milk, 5% dextrin, 5% 밀가루, 5% lactomeal
- 고체 및 액체 종균의 세균 등 잡균의 오염 저감화 : 한국(10³) < 일본(10²)
○ 종균 보존 및 현장 보급기술 체계 구축
- 장기 보존기술 개발 : 고체종균 포자수 10⁷ spore/g 이상유지, 효모 액체종균 저장성 확인
- 탁주용 우수 종균 배합 및 시제품화 : 곰팡이 고체종균 6종 및 효모 고체종균 2종
- 탁주의 양조적성기술 개발 : 쌀누룩 10종/효모 2종, 품질특성 및 기호도 조사
- 우수 종균의 현장 보급형 시스템 구축 및 실용화: 곰팡이(장희), 탁주(홍전통도가)
○ 국산 포도주 품질 개선을 위한 토착형 포도주 효모 및 유산균 선정
- 국산 토착형 포도주 효모(S. cerevisiae M12), 캠벨얼리, MBA, 개량머루 포도주 주질 개선용(Han. uvarum S6), 캠벨얼리 포도주의 신맛 조절(Iss. orientalis KMBL 5774)
- 포도주 및 김치 600주 분리 → 말로락틱 발효능 유산균 2종(Lactobacillus plantarum 286, 287)
○ 국산 캠벨얼리 포도주 유래의 유산균(L. plantarum)의 종균제조기술 확립
- 부영제 : 10% skim milk + 10% 당류 첨가 동결건조
- 복원율 : Sorbitol 첨가로 38% → 86% 향상
- L. plantarum 종균 시제품에 의한 포도주의 사과산 젖산화 : 2주, 약 90% 분해
- 발효 초기 유산균 종균 첨가 : 9일 만에 사과산 2.8 mg/mL → 1.97 mg/mL
○ 다양한 부영제를 이용한 효모 종균 저온송풍건조기술 확립
- 효모 건조 시제품 : 밀가루, 누룩, 돼지감자 분말, 락토밀 등
- 건조시간 단축 : 1∼2 hr 완료
- 복원율 : S. cerevisiae 97%, H. uvarum 93%, I. orientalis 79%
○ 효모 및 유산균 종균 시제품 제조 기술 확립
- 우수 유산균 분리 및 특성 관찰 : 유산균 1종 균주 기탁(KACC)
- Ca-alginate bead 제품의 보존성 : 4개월경과, 7.4 Log CFU/mL 이상유지
- 송풍건조 종균 제품의 장기 보존성 확인 : 3개월경과, 9 Log CFU/mL 이상유지
- 동결건조 유산균 종균 제품의 장기 보존성 : 3개월경과, 8.5 Log CFU/mL 이상유지
- 유산균과 와인 효모와의 순차적 혼합발효를 통한 사과산 분해기술 확립(특허출원 1건)
- (주)충무발효와의 공동연구로 시제품 대량생산
○ 종균 생산용 오염 방지기술 개발
- 종균 생산용 제품 오염도 저감화기술 개발 : 배양실 및 생산 설비 오염 방지
- 액상 및 분말(고체) 종균의 안정성 설정 : 2% xylooligo 당, 0.2% NaCl
- 액상 및 분말(고체) 종균의 포장법 설정 : 밀폐형 용기, 은박 파우치 진공 포장법
○ 종균화 개발 균주의 중규모(1톤) 발효식품 제조 및 저장용기 및 포장방법 개발
- 입국용 종균 및 주모용 종효모 사용 → 탁주제조 현장적용 문제없음
- 저장 용기별 종균 제품의 생존율 : 액체종균 분말화 → 곰팡이 생존율 98%
- 포장방법 개선 : 포자비산방지장치, 포자흡착용 자동 선별체, 자동 포장기 등
○ 토착 발효미생물의 종균 상품화를 위한 실용화 기술 개발
- 종균별 대량생산 최적화 배양조건 구축: 유가식(YM배지), 종배양(30℃,36h)⇨본배양
- 개발 종균의 스케일 업 시스템 확립: 건조효모 생산기술(냉풍건조법 확립)
- 주류용 종균의 대규모 산업화 기술 개발 : 고상발효법(곰팡이 종균 생산)
- 종균생산 현장오염 저감법 개발: 과초산, 4급 암모늄계열 살균소독제
- 상품화를 위한 제품 안정성 및 포장법 개발 작업중: 비산포자 발생⇨비산포자 흡착

Abstract ▼

Ⅲ. Results and Recommendation
Section 1. Development of preparing starter technique for Takju
○ Preservation and selection of starter (Molds and Yeast) for Takju
- Starter selection by toxin safety, enzyme activity, alcohol production and aroma
· Selected to molds 2 species (A. luchuensi

Ⅲ. Results and Recommendation
Section 1. Development of preparing starter technique for Takju
○ Preservation and selection of starter (Molds and Yeast) for Takju
- Starter selection by toxin safety, enzyme activity, alcohol production and aroma
· Selected to molds 2 species (A. luchuensis 74-5, A. oryzae 75-2), and Yeast 2 species (S. cerevisiae Y204, Y283)
- Set up optimum fermentation condition of starter : Carbon, nitrogen source, temperature and cultivation times etc.
- Preservation and registration of selected starters : Subculture, glycerol preservation and resister KACC (RDA-GeneBank)
○ Investigation of solid and liquid starter manufacturing technology for Takju
- Solid starter (Mold : A. luchuensis, A. oryzae) : 5% of liquid starter, preparing temperature (AL 40℃, AO 35℃), preparing period : 5 days
- Liquid starter (Yeast : S. cerevisiae Y204, Y283) : YPD broth, 30℃, 150 rpm, 24 hr
- Extending agent for starter safety : 3% of skim milk, 5% of dextrin, 5% of flour, 5% of lactomeal^TM
- Contamination reduction of microbes for solid and liquid starter : Korea(10³) < Japan(10²)
○ Selection starter and set-up optimum condition for Takju
- Selection excellent starter for Takju : Growth rate, enzyme activity, safety verification (Aflatoxin B1)
- Set-up optimum fermentation condition of selection starter : Selection of carbon & nitrogen source, pH, temperature and culture time set-up
- Excellent starter registration of molds & yeast for Takju : 18S rRNA identification, 2 types yeast register KACC
○ Development of starter manufacturing and drying technique
- Solid starter manufacturing technique and optimization : Liquid starter inoculum amount (5%), temp. (AL 40℃, AO 35℃), time (5 days)
- Liquid starter manufacturing technique and optimization : YPD broth, 30℃, 150 rpm, 24 hr
- Selection of an extender & wrapper for starter protection : Solid starter → Skim milk (3%), dextrin (5%)
- Drying technology of starter : Spray drying 〉Freezing drying
○ Development of long period preservation technique, dissemination system buildup and commercialization of starter
- Stater preservation and dissemination system buildup : Mold 2 types and yeast 2 types
- Development of long periods preservation : Solid starter spore count keep over 10⁷ spore/g, yeast liquid starter identify preservation
- Excellent starter mix for Takju and prototyping : Solid starter (mold) 6 types and solid starter (yeast) 2 types
- Development of brewing aptitude technique : Rice nuruk 10 types / yeast 2 types, quality characteristics and preference investigation
- Dissemination system buildup and commercialization of excellent starter : Nuruk (Janghi Co.,), Takju (Hongdoga Co.,)

Section 2. Development of technology for preparation of starters (yeast and lactic acid bacteria) for Wine
○ Selection of Korean indigenous wine yeast and malolactic bacteria to improve wine quality
- S. cerevisiae M12 as a Korean indigenous wine yeast, Han. uvarum S6 to improve Campbell Early and MBA wine quality, Iss. orientalis KMBL 5774 to reduce malic acid content in Campbell Early wine
- Isolation of 600 lactic acid bacterial strains and selection of two malolactic bacterial strains (Lactobacillus plantarum 286, 287)
○ Establishment of an efficient technology for preparation of a dried starter of lactic acid bacteria (L. plantarum) isolated from Campbell Early wine
- Additives for freeze-drying : 10% skim milk + 10% sugar
- Improvement of restoration ratio : 38% → 86% when sorbitol added
- Degradation of malic acid in wine by L. plantarum starter sample : 90% for 2 weeks
- Degradation of malic acid during mixed fermentation of wine by inoculation of lactic acid bacteria starter followed by a wine yeast : malic acid content 2.8 mg/mL → 1.97 mg/mL for 9 days
○ Establishment of air-blast drying of yeasts at a low temperature with various additives
- Preparation of dried yeast starter samples with flour, nuruk, jerusalem artichoke powder, lactomil, etc.
- Reduction of drying time : 1∼2 hr
- Restoration ratio : S. cerevisiae 97%, H. uvarum 93%, I. orientalis 79%
○ Establishment of Korean indigenous starter (yeast and lactic acid bacteria) manufacturing technology
- Isolation and characterization of a lactic acid bacterial strain with a high level of malolactic activity : deposited in KACC
- Preservation of product (Ca-alginate bead) : lapse of 4 month, keep 7.4 Log CFU/mL
- Verification of long period preservation of starter product (air-blast drying) : Lapse of 3 month, keep 7.12 Log CFU/mL
- Preservation of lactic acid bacteria starter (freeze-drying) : Lapse of 3 month, keep 8.5 Log CFU/mL
- Establishment of malic acid degradation technology in wine by sequential inoculation of lactic acid bacteria followed by wine yeast (applied for a patent)
- Mass production of yeast and lactic acid bacteria starters through joint researches with Chung-moo fermentation Co., Ltd

Section 3. Development of commercialization technology of aboriginality fermentation microorganisms
○ Development of contamination reduction method for facility
- Contamination reduction method for factory facility : Fermentation rooms and processing machines
- Stability point setup for liquid and powder (solid) starter : Use of 2% xylooligo saccharide, 0.2% NaCl
- Selection of liquid and powder starter packing method : A method of packing with polyethylene sealed bottle, aluminum paper pouch material
○ Medium scale fermented food production and packing method of starter molds
- Brewery application of starter koji and yeast for Takju → Good field application result to Takju brewing
- Livability of starter molds product according to container type : Powderlike of liquid starter → Livability of koji starter mold is 98%
- Improvement of packing process for starter molds : Fungi spore scatter-proof device, automatic vibrating sieve machine for spore adsorption, automatic packing machine
○ Development of commercialization technology of aboriginality fermentation microorganisms into starter products
- Development of starter manufacturing facility contamination reduction method : Per acetic acid, quaternary ammonium sterilization disinfectant
- Establishment of mass production optimum culture condition from starters : Fed-batch culture (YM broth), seed culture (30℃, 36h) → Main propagation
- Development starter establish scale-up system : Dry yeast production technology (cold wind drying method)
- Production of middle-scale fermentation food : Making Takju by starter molds (A. luchuensis and S. cerevisiae)
- Development of brewing starter large scale commercialization technology : Solid state fermentation method (Mold starter production)
- Packaging method and stability of production progressing for commercialization : Airborne spore occurrence → Adsorption of airborne spore