清酒酵母からのウラシル要求性変異株の単離と変異株による清酒醸造

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Uracil auxotrophic mutants were isolated from sake yeast strains (K-6,K-7,K-9,K-10) of Saccharomyces cerevisiae, which were though to be diploids. The isolation was made by a positive selection using a medium on which only ura3 or ura5 mutants were able to grow. The method was much more effective and simpler than the conventional replica plating technique. Mild mutagenesis with ethyl methanesulfonate (EMS) was done to avoid defects in important genes for the sake brewing at the survival rate of 66-97%. EMS-treated yeast cells (2-4×10^6) were spread on each of four FOA (5-fluoroorotic acid) plates and colonies were isolated after incubation for 1 week at 30℃. (K-6,46 colonies; K-7,13 colonies; K-9,18 colonies; K-10,105 colonies.) Most (60-80%) of the isolated colonies were uracil auxotrophic mutants, as expected. Some of the uracil mutants were transformed with pCP2-4-10(URA3,SMR1) to examine whether they were ura3 or ura5 mutants. The result showed that all of these uracil mutants carried ura3 mutations since their uracil requirements were complemented by pCP2-4-10. Brewing profiles of sake using the ura3 mutants were inferior to those by their parent strains, but those by the transformants with YIp5(URA3) were at the same level as parent strains, showing that the inferiority was closely related with the ura3 mutation. The ura3 mutation is useful as a genetic maker for breeding of sake yeasts by cell fusion and recombinant DNA techniques.
社団法人日本生物工学会の論文
1990-09-25

清酒酵母からのウラシル要求性変異株の単離と変異株による清酒醸造

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