Here, we investigated if and how the genomes of Asian S. eubayanus strains could have contributed to maltotriose utilization by generating chromosome-level genome assemblies using Oxford Nanopore Technology (ONT) long-read sequencing. Since the Asian strains were unable to utilize maltotriose, we functionally characterized the assembled MAL genes and identified genetic determinants limiting maltotriose utilization. Finally, we generated hybrids between an Asian S. eubayanus strain and a maltotriose-deficient ale S. cerevisiae strain to investigate the heredity of maltotriose utilization. We analyzed the implications of our results for the role and origin of SeAGT1 in S. pastorianus and for the potential of hybridization to enable maltotriose consumption in novel S. pastorianus hybrids.