Isolation of sake yeast strains possessing various levels of succinate- and/or malate-producing abilities by gene disruption or mutation

Food Technology Research Institute of Nagano Prefecture, 205-1 Nishibanba, Kurita, Nagano City 380-0921, Japan; Department of Applied Biology, Faculty of Textile Science and Technology, Shinshu University, 3-15-1 Tokida, Ueda, Nagano 386-0018, Japan; Gene Research Center, Shinshu University, 3-15-1 Tokida, Ueda, Nagano 386-0018, Japan; Section of Clinical Biochemistry, School of Health Science, Kyorin University, Miyashita, Hachioji, Tokyo 192-0005, Japan
Journal of Bioscience and Bioengineering (Impact Factor: 1.79). 02/1999; DOI: 10.1016/S1389-1723(99)80041-3

ABSTRACT Succinate and malate are the main taste components produced by yeast during sake (Japanese alcohol beverage) fermentation. Sake yeast strains possessing various organic acid productivities were isolated by gene disruption. Sake fermented using the aconitase gene (ACO1) disruptant contained a two-fold higher concentration of malate and a two-fold lower concentration of succinate than that made using the wild-type strain K901. The fumarate reductase gene (OSM1) disruptant produced sake containing a 1.5-fold higher concentration of succinate as compared to the wild-type, whereas the α-ketoglutarate dehydrogenase gene (KGD1) and fumarase gene (FUMI) disruptants gave lower succinate concentrations. The Δkgd1 disruptant exhibited lower succinate productivity in the earlier part of the sake fermentation, while the Δfum1 disruptant showed lower succinate productivity later in the fermentation, indicating that succinate is mainly produced by an oxidative pathway of the TCA cycle in the early phase of sake fermentation and by a reductive pathway in the later phases. Sake yeasts with low succinate productivity and/or high malate productivity was bred by isolating mutants unable to assimilate glycerol as a carbon source. Low malate-producing yeasts were also obtained from phenyl succinate-resistant mutants. The mutation of one of these mutant strains with low succinate productivity was found to occur in the KGD1 gene. These strains possessing various succinate- and/or malate-producing abilities are promising for the production of sake with distinctive tastes.

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