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In mid to completion of moromi mashing, due to high ethanol concentration yeast cells cease growing, become inactive, and eventually lysed, resulting in unfavorable increase of amino acids and color . To solve the problem we attempted to isolate sake yeast strains which could grow in the medium containing more than 10% ethanol. Cell of Saccharomyces cerevisiae Kyokai No.7 were treated with ethyl methanesulfonte. Colonies appeared on the yeast extractpolypepton-dextrose agar medium (YEPD) were replicated onto 0.6% sodium acetate agar medium for nitrogen starvation . Through the primary screening on the YEPD agar medium added with 12% ethanol as a final concentration and the secondary screening in the same medium, 6 mutants showed abundant growth comparing with poor growth by the parental strain. In a small-scale sake brewing test with the 6 mutants, mutant strain 7-68 was selected. A pilot-scale sake brewing experiment with strain 7-68 and the parental strain was carried out. Moromi fermentation progressed similarly for the both strains. Activities of yeast cells measured by the methylene blue staining method showed 20% active for strain 7-68 and 32% active for strain Kyokai No.7 at the completion of the moromi mashing. It was also observed that amino acidity was 1.4 a in the resulted sake made with strain 7-68 and 1.7 me with the parental strain. The result suggests that the mutant obtained by the above procedures would be helpful in solving the problem.

Isolation of Sake Yeast Able to Grow in the Medium Containing More Than Ten Percent Ethanol