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For maltose fermentation, budding yeast  Saccharomyces cerevisiae  operates a mechanism that involves transporters ( MALT ), maltases ( MALS ) and regulators ( MALR ) collectively known as  MAL  genes. However, functional relevance of  MAL  genes during  sake  brewing process remains largely elusive, since  sake  yeast is cultured under glucose-rich condition achieved by the co-culture partner  Aspergillus  spp.. Here we isolated an ethyl methane sulfonate (EMS)-mutagenized  sake  yeast strain exhibiting enhanced maltose fermentation compared to the parental strain. The mutant carried a single nucleotide insertion that leads to the extension of the C-terminal region of a previously uncharacterized  MALR  gene  YPR196W-2 , which was renamed as  MAL73 . Introduction of the mutant allele  MAL73L  with extended C-terminal region into the parental or other  sake  yeast strains enhanced the growth rate when fed with maltose as the sole carbon source. In contrast, disruption of endogenous  MAL73  in the  sake  yeasts decreased the maltose fermentation ability of  sake  yeast, confirming that the original  MAL73  functions as a MALR. Importantly, the  MAL73L- expressing strain fermented more maltose in practical condition compared to the parental strain during  sake  brewing process. Our data show that  MAL73 ( L ) is a novel  MALR  gene that regulates maltose fermentation, and has been functionally attenuated in  sake  yeast by single nucleotide deletion during breeding history. Since the  MAL73L- expressing strain showed enhanced ability of maltose fermentation,  MAL73L  might also be a valuable tool for enhancing maltose fermentation in yeast in general.

MAL73, a novel regulator of maltose fermentation, is functionally impaired by single nucleotide polymorphism in sake brewing yeast