Characterization of maltotriose transporters from the Saccharomyces eubayanus subgenome of the hybrid Saccharomyces pastorianus lager brewing yeast strain Weihenstephan 34/70

The genome from the Saccharomyces pastorianus industrial lager brewing strain Weihenstephan 34/70, a natural Saccharomyces cerevisiae/Saccharomyces eubayanus hybrid, indicated the presence of two different maltotriose transporter genes: a new gene in the S. eubayanus subgenome with 81% of homology to the AGT1 permease from S. cerevisiae, and an amplification of the S. eubayanus MTY1 maltotriose permease previously identified in S. pastorianus yeasts. To characterize these S. eubayanus transporter genes, we used a S. cerevisiae strain deleted in the AGT1 permease and introduced the desired permease gene(s) into this locus through homologous recombination. Our results indicate that both the MTY1 and AGT1 genes from the S. eubayanus subgenome encode functional maltotriose transporters that allow fermentation of this sugar by yeast cells, despite their apparent differences in the kinetics of maltotriose‐H+ symport activity. The presence of two maltotriose transporters in the S. eubayanus subgenome not only highlights the importance of sugar transport for efficient maltotriose utilization by industrial yeasts, but these new genes can be used in breeding and/or selection programs aimed at increasing yeast fitness for the efficient fermentation of brewer's wort. Significance and Impact of the Study:The Saccharomyces pastorianus lager yeast usually ferments maltose and maltotriose efficiently, and transport of these α‐glucosides into the brewer's yeast cell is thought to be rate limiting in the fermentation of these sugars from wort. Our results revealed a new maltotriose transporter gene in the Saccharomyces eubayanus subgenome of the industrial lager brewing strain Weihenstephan 34/70 that allows efficient maltotriose fermentation by yeast cells. The characterization of maltotriose transporters from industrial yeasts opens new opportunities to increase yeast fitness for fermentation of brewer's wort, preventing incomplete or sluggish maltotriose fermentations.