清酒酵母の醸造特性に関する遺伝子の解析

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The sake yeast Kyokai no. 7 is a pantothenic acid auxotroph at 35 and this phenotype is used to distinguish Kyokai no. 7 from other sake yeasts. We found that ECM31 of a Saccharomyces cerevisiae laboratory strain could complement the pantothenic acid auxotrophy of this yeast. Disruption of ECM31 in the laboratory strain resulted in pantothenic acid auxotrophy. ECM31 can be used as a selective marker in the transformation of Kyokai no. 7. Almost all sake yeasts form a thick foam layer on sake mash during the fermentation process. We cloned a gene from a foam-forming sake yeast that confers foaming ability on a non-foaming mutant. This gene was named AWAI and encodes a GPI anchor protein that is localized to the cell wall. An AWAI disruptant showed a non-foaming phenotype in sake mash, indicating that Awalp is required for the foam-forming phenotype. Several ethanol-tolerant mutants have been bred from industrial sake yeasts. We compared expression profiles between the ethanol-tolerant mutant and its parent sake yeast using DNA microarray. We found that several stress-induced genes were highly expressed even in the unstressed condition. We also found that the ethanol-tolerant mutant also exhibited resistance to other stresses, including heat, high osmolarity, and oxidative stress in addition to ethanol tolerance. The mechanism of ethanol tolerance in yeast Saccharomyces cerevisiae is thought to be regulated by many genes. To identify some of them, we screened for ethanol-sensitive mutants among a collection of mutants obtained using transposon mutagenesis. By this method, we identified the genes BEM2, PAT1, ROM2, VPS34, and ADA2, whose disruption resulted in ethanol sensitivity. We also found that the integrity of the cell wall plays an important role in ethanol tolerance in S. cerevisiae.
社団法人日本生物工学会の論文
2002-02-25

清酒酵母の醸造特性に関する遺伝子の解析

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