For the production of riboflavin, strain development of Ashbya gossypii NRRL Y-1056 was attempted by NTG(N-methyl-N`-nitro-N-nitrosoguanidine) treatment. The optimum composition of culture medium and other culture conditions for the production of riboflavin by selected mutant Ashbya gossypii JAG-13 were determined. The optimum composition of medium was 9% of corn oil, 3% of gellatone, 4% of CSL, 0.3% of glycine, 0.2% of S770. The optimum culture temperature and initial pH of medium was 28℃ and 6.5, respectively. oxygen was essential for the production of riboflavin, but excess oxygen inhibit the production of riboflavin. When Ashbya gossypii JAG-13 was cultured under above conditions for 12 days with a bioreactor, 6.9 ㎎/㎖ of riboflavin was produced.
Saccharomyces diastaticus KCTC 1804 균주와 Saccharomyces cerevisiae KOY-1 균주를 이용하여 전분분해활성과 알코올 발효능을 동시에 가지는 효모융합체를 개발하였다. Saccharomyces cerevisiae KOY-1 균주의 단수체 유도를 통하여 Saccharomyces cerevisiae KH-12 균주를 획득하였다. EMS를 사용한 돌연변이 유발을 통하여 Saccharomyces cerevisiae KH-12 균주의 Met 영양요구성 변이주를 획득하였다. Lyticase 처리로 Saccharomyces cerevisiae KOY-1 Met?), Saccharomyces diastaticus KCTC 1804(Trp?) 균주의 원형질체 형성률은 각각 90.5%, 97.7%로 관찰되었다. 원형질체 융합은 polyethylene glycol 4,000을 이용하여 실시하였고, 이때 원형질체 융합율은 1.79 × 10-4으로 관찰되었다. 원형질체융합과정을 통해 총 1,000주의 융합체를 획득하였고, 획득한 융합체의 전분분해능과 알코올발효능을 비교하여 최종적으로 전분배지 YPS24에서 알코올생성능이 가장 우수한 효모융합체인 FA 776을 최종 선발하였다. 최종 선발된 효모융합체 FA 776은 10세대 계대배양 후의 revertent 복귀율이 4.64%로 유전적 안정성을 확인하였으며, 효모융합체의 total DNA 함량을 비교한 결과, Saccharomyces cerevisiae 균주로부터 분리된 단수체 효모 KH-12는 26.56 fg/cll, Saccharomyces diastaticus KCTC 1804 균주는 25.40 fg/cell이었고, 이들을 사용해서 얻어진 융합체 FA 776의 DNA 함량은 42.67 fg/cell이었다. 전분배지 YPS24 배지에서 60시간 발효하였을때 Saccharomyces cerevisiae KOY-1 균주는 알코올 생성하지 못하였지만 효모융합체 FA 776은 13 mg/mL의 알코올을 생성하였다. 이는 전분이용성 효모인 Saccharomyces diastaticus KCTC 1804 균주에 비해 1.86배나 향상된 알코올 발효능을 나타냈다.
The objectives of this study were to evaluate the physico-chemical and sensory characteristics of a Korean traditional alcoholic beverage (yakju) prepared using different nuruk (Korean-style koji) concentrations and yeasts such as the fusant FA776 and Saccharomyces cerevisiae KOY-1, respectively. The fusant FA776, which has alcohol-fermenting and starch-utilizing properties, was formed by Saccharomyces cerevisiae KOY-1 and Saccharomyces diastaticus KCTC1804. The fermentation trial was conducted in a 5 L lab-scale jar at 25℃. The maximum alcohol production of the K-100 and F-50 reached levels of 135.0 mg/mL and 119.4 mg/mL, respectively. The pH values were in a range of 4.3-4.5. Total acidity was in a range of 0.47-0.60%. Organic acids and amino acids were analyzed in order to evaluate variations in its composition and content via HPLC analysis. Organic acids including lactic acid, citric acid, malic acid, and pyruvic acid, and 16 kinds of amino acids, including aspartic acid, were detected in all treatments. K-100 showed the highest amino acid contents, whereas F-50 exhibited the lowest amino acid contents. Volatile flavor components such as phenylethyl alcohol, isoamyl alcohol, 2-methylthiophane, isobutyl alcohol, and ethyl succinate were detected as a major component in all treatments, as determined via gas chromatography. The results of our sensory evaluation demonstrated that Yakju fermented by the FA776 fusant yielded more favorable results than S. cerevisiae KOY-1.
