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Isolation of a Copper‐resistant Sake Yeast Mutant with Improved Flavour Compound Production
Authors:Kiyoo Hirooka  Akira Ogita  Ken‐Ichi Fujita  Yoshihiro Yamamoto  Toshio Tanaka
Affiliation:1. Kyoto Municipal Institute of Industrial Technology and Culture, 91 Chudoji Awata‐cho, Shimogyo‐ku, Kyoto 600–8815, Japan.;2. Research Center for Urban Health and Sports, Osaka City University, Osaka, Japan, 3–3–138 Sugimoto, Sumiyoshi‐ku, Osaka 558–8585, Japan.;3. Department of Bio and Geosciences, Graduate School of Science, Osaka City University, 3–3–138 Sugimoto, Sumiyoshi‐ku, Osaka 558–8585, Japan.
Abstract:The sake (traditional Japanese alcoholic beverage) yeast mutant A1 was previously isolated as a strain resistant to an isoprenoid analog. This strain is used for industrial sake brewing because of its increased production of isoamyl acetate. In this study, a physiological event was identified which was closely related to the elevation of alcohol acetyltransferase (AATase) activity in strain A1. This finding was applied for the isolation of another mutant with an improved capacity for flavour compound production. Strain A1 revealed an additional phenotype showing resistance to Cu2+, as seen from its growth and isoamyl acetate production, even in a medium with the copper ion at 6 mM. Mutant strains were successfully isolated with increased isoamyl acetate production capacity from sake yeast strain 2NF on the basis of a Cu2+‐resistant phenotype at a high yield. Among them, strain Cu7 was characterized by its ability to produce isoamyl acetate at the highest concentration under condition where isoamyl alcohol (its precursor) was accumulated to the lowest extent. Such a phenotype of strain Cu7 is applicable for the practical production of an alcoholic beverage of excellent quality in terms of flavour.
Keywords:alcohol acetyltransferase  Cu2+ resistance  E/A ratio  isoamyl acetate  sake yeast
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