Extracellular maltotriose hydrolysis by Saccharomyces cerevisiae cells lacking the AGT1 permease

In brewing, maltotriose is the least preferred sugar for uptake by Saccharomyces cerevisiae cells. Although the AGT1 permease is required for efficient maltotriose fermentation, we have described a new phenotype in some agt1 strains of which the cells do not grow on maltotriose during the first 3-4days of incubation, but after that, they start to grow on the sugar aerobically. Aiming to characterize this new phenotype, we performed microarray gene expression analysis which indicated upregulation of high-affinity glucose transporters (HXT4, HXT6 and HXT7) and -glucosidases (MAL12 and IMA5) during this delayed cellular growth. Since these results suggested that this phenotype might be due to extracellular hydrolysis of maltotriose, we attempted to detect glucose in the media during growth. When an hxt-null agt1 strain was grown on maltotriose, it also showed the delayed growth on this carbon source, and glucose accumulated in the medium during maltotriose consumption. Considering that the poorly characterized -glucosidase encoded by IMA5 was among the overexpressed genes, we deleted this gene from an agt1 strain that showed delayed growth on maltotriose. The ima5 agt1 strain showed no maltotriose utilization even after 200h of incubation, suggesting that IMA5 is likely responsible for the extracellular maltotriose hydrolysis. Significance and Impact of the StudyMaltotriose is the second most abundant sugar present in brewing. However, many yeast strains have difficulties to consume maltotriose, mainly because of its low uptake rate by the yeast cells when compared to glucose and maltose uptake. The AGT1 permease is required for efficient maltotriose fermentation, but some strains deleted in this gene are still able to grow on maltotriose after an extensive lag phase. This manuscript shows that such delayed growth on maltotriose is a consequence of extracellular hydrolysis of the sugar. Our results also indicate that the IMA5-encoded -glucosidase is likely the enzyme responsible for this phenotype.