![]() KEY POINTS: An increase in malate level of GID4 mutant resulted from the accumulation of Mdh2. Our results suggest that high-malate-producing disruptants adapt their metabolism to produce malate in excess via the regulation of protein expression in glucose assimilation and ethanol fermentation. The high-malate-producing disruptants showed the activation of several glycolytic enzymes and a reduction in enzymes involved in the conversion of pyruvate to ethanol. Comparison of protein abundance among the GID disruptants revealed variations in protein expression profiles, including in glycolysis and tricarboxylic acid cycle–related enzymes. The disruptants of GID1, 2, 3, 4, 5, 8, and 9 exhibited high malate production. ![]() Next, we investigated the effects of GID protein–coding genes (GID1–GID9) on organic acid production and enzyme expression profiles in yeast. Furthermore, target proteome analysis indicated that an increase in malate production resulted from the accumulation of Mdh2 in gid4 disruptant due to the loss of GID complex–mediated degradation. The evaluation of disruptants and mutants of gluconeogenic enzymes revealed that cytosolic malate dehydrogenase (Mdh2) participated in the malate production. In this study, the mechanism by which this mutation led to high malate production in yeast cells was investigated. Gid4 is a component of the GID (glucose-induced degradation-deficient) complex and stimulates the catabolic degradation of gluconeogenic enzymes. We previously isolated a high-malate-producing strain and found that a missense mutation in GID4 was responsible for the high-malate-producing phenotype. Since malate contributes to the pleasant flavor of sake (a Japanese alcoholic beverage), various methods for breeding high-malate-producing yeast have been developed. 4971-4983 ISSN: 0175-7598 Subject: Saccharomyces cerevisiae, acetates, biotechnology, ethanol, ethanol fermentation, flavor, glucose, glycolysis, malate dehydrogenase, malates, missense mutation, mutants, phenotype, protein synthesis, proteome, pyruvic acid, sake, succinic acid, yeasts Abstract: During alcohol fermentation, Saccharomyces cerevisiae produces organic acids, including succinate, acetate, and malate. Effects of mutations of GID protein–coding genes on malate production and enzyme expression profiles in Saccharomyces cerevisiae Author: Hiroaki Negoro, Kengo Matsumura, Fumio Matsuda, Hiroshi Shimizu, Yoji Hata, Hiroki Ishida Source: Applied microbiology and biotechnology 2020 v.104 no.11 pp.
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