title: Enhancement of malate-production and increase in sensitivity to dimethyl succinate by mutation of the VID24 gene in Saccharomyces cerevisiae.
abstract: Malate in sake (a Japanese alcoholic beverage) is an important component for taste that is produced by yeasts during alcoholic fermentation. To date, many researchers have developed methods for breeding high-malate-producing yeasts; however, genes responsible for the high-acidity phenotype are not known. We determined the mutated gene involved in high malate production in yeast, isolated as a sensitive mutant to dimethyl succinate. In the comparative whole genome analysis between high-malate-producing strain and its parent strain, one of the non-synonymous substitutions was identified in the VID24 gene. The mutation of VID24 resulted in enhancement of malate-productivity and sensitivity to dimethyl succinate. The mutation appeared to lead to a deficiency in Vid24p function. Furthermore, disruption of cytoplasmic malate dehydrogenase (Mdh2p) gene in the VID24 mutant inhibited the high-malate-producing phenotype. Vid24p is known as a component of the multisubunit ubiquitin ligase and participates in the degradation of gluconeogenic enzymes such as Mdh2p. We suggest that the enhancement of malate-productivity results from an accumulation of Mdh2p due to the loss of Vid24p function. These findings propose a novel mechanism for the regulation of organic acid production in yeast cells by the component of ubiquitin ligase, Vid24p.
authors.affiliation: Research Institute, Gekkeikan Sake Co. Ltd., 101 Shimotoba-koyanagi-cho, Fushimi-ku, Kyoto 612-8385, Japan. Electronic address: negoro@gekkeikan.co.jp.
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Negoro
authors.affiliation: Research Institute, Gekkeikan Sake Co. Ltd., 101 Shimotoba-koyanagi-cho, Fushimi-ku, Kyoto 612-8385, Japan. Electronic address: a_kotaka@gekkeikan.co.jp.
authors.forename: Atsushi
authors.initials: A
authors.lastname: Kotaka
authors.affiliation: Research Institute, Gekkeikan Sake Co. Ltd., 101 Shimotoba-koyanagi-cho, Fushimi-ku, Kyoto 612-8385, Japan. Electronic address: kengo@gekkeikan.co.jp.
authors.forename: Kengo
authors.initials: K
authors.lastname: Matsumura
authors.affiliation: Research Institute, Gekkeikan Sake Co. Ltd., 101 Shimotoba-koyanagi-cho, Fushimi-ku, Kyoto 612-8385, Japan. Electronic address: h_tsutsumi@gekkeikan.co.jp.
authors.forename: Hiroko
authors.initials: H
authors.lastname: Tsutsumi
authors.affiliation: Research Institute, Gekkeikan Sake Co. Ltd., 101 Shimotoba-koyanagi-cho, Fushimi-ku, Kyoto 612-8385, Japan. Electronic address: y_hata@gekkeikan.co.jp.
authors.forename: Yoji
authors.initials: Y
authors.lastname: Hata
mesh_terms: D000434:Alcoholic Beverages / Q000032:analysis; D003593:Cytoplasm / Q000201:enzymology; D007564:Japan; D008291:Malate Dehydrogenase / Q000235:genetics / Q000378:metabolism; D008293:Malates / Q000378:metabolism*; D009154:Mutation; D012441:Saccharomyces cerevisiae / Q000187:drug effects* / Q000201:enzymology / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics* / Q000378:metabolism*; D013386:Succinates / Q000494:pharmacology*; D044767:Ubiquitin-Protein Ligases / Q000737:chemistry / Q000172:deficiency / Q000235:genetics / Q000378:metabolism; D033921:Vesicular Transport Proteins / Q000172:deficiency / Q000235:genetics* / Q000378:metabolism*
chemical_list: D008293:Malates; D029701:Saccharomyces cerevisiae Proteins; D013386:Succinates; C111452:VID24 protein, S cerevisiae; D033921:Vesicular Transport Proteins; C030298:malic acid; C056451:dimethyl succinate; C000614940:MDH2 protein, S cerevisiae; D008291:Malate Dehydrogenase; D044767:Ubiquitin-Protein Ligases
keywords: Dimethyl succinate; MDH2; Malate; Saccharomyces cerevisiae; VID24