Incomplete and/or sluggish maltotriose fermentation causes both quality and economic problems in the ale-brewing industry. Although it has been proposed previously that the sugar uptake must be responsible for these undesirable phenotypes, there have been conflicting reports on whether all the known ␣-glucoside transporters in Saccharomyces cerevisiae (MALx1, AGT1, and MPH2 and MPH3 transporters) allow efficient maltotriose utilization by yeast cells. We characterized the kinetics of yeast cell growth, sugar consumption, and ethanol production during maltose or maltotriose utilization by several S. cerevisiae yeast strains (both MAL constitutive and MAL inducible) and by their isogenic counterparts with specific deletions of the AGT1 gene. Our results clearly showed that yeast strains carrying functional permeases encoded by the MAL21, MAL31, and/or MAL41 gene in their plasma membranes were unable to utilize maltotriose. While both highand low-affinity transport activities were responsible for maltose uptake from the medium, in the case of maltotriose, the only low-affinity (K m , 36 ؎ 2 mM) transport activity was mediated by the AGT1 permease. In conclusion, the AGT1 transporter is required for efficient maltotriose fermentation by S. cerevisiae yeasts, highlighting the importance of this permease for breeding and/or selection programs aimed at improving sluggish maltotriose fermentations.Several important industrial applications of the yeast Saccharomyces cerevisiae, such as brewing, baking, and the production of distilled beverages, rely on the efficient fermentation of starch hydrolysates rich in glucose and in the ␣-glucosides maltose and maltotriose. In brewer's wort, for example, the most abundant fermentable sugar is maltose (50 to 60%), followed by maltotriose (15 to 20%) and glucose (10 to 15%). Of these sugars, glucose is preferentially and rapidly utilized by the yeast cells, but process efficiency also requires the complete fermentation of both maltose and maltotriose. After glucose exhaustion, maltose is easily fermented by the majority of S. cerevisiae strains, and not only is maltotriose the least preferred sugar for uptake by these yeast cells, but many yeasts may not use it at all (23, 37, 44). The difficulty that some strains have in consuming maltotriose leads to one of the problems experienced by many breweries, namely, a high content of residual sugar in the finished beer, low ethanol yields, and atypical beer flavor profiles. Therefore, the rate of uptake and fermentation of maltotriose is one of the major determinants of fermentation efficiency and product quality. Due to its relevance for beer fermentation, maltotriose utilization has been studied mostly with the two classes of yeast strains used for brewing, S. cerevisiae ale strains and S. pastorianus lager strains. These studies have revealed that maltotriose utilization by ale strains is significantly slower than that by lager yeasts and that, consequently, residual maltotriose is more common at the end of ale fermentations (22,23,44...

A synthetic pathway for (d)-xylose assimilation was stoichiometrically evaluated and implemented in Escherichia coli strains. The pathway proceeds via isomerization of (d)-xylose to (d)-xylulose, phosphorylation of (d)-xylulose to obtain (d)-xylulose-1-phosphate (X1P), and aldolytic cleavage of the latter to yield glycolaldehyde and DHAP. Stoichiometric analyses showed that this pathway provides access to ethylene glycol with a theoretical molar yield of 1. Alternatively, both glycolaldehyde and DHAP can be converted to glycolic acid with a theoretical yield that is 20% higher than for the exclusive production of this acid via the glyoxylate shunt. Simultaneous expression of xylulose-1 kinase and X1P aldolase activities, provided by human ketohexokinase-C and human aldolase-B, respectively, restored growth of a (d)-xylulose-5-kinase mutant on xylose. This strain produced ethylene glycol as the major metabolic endproduct. Metabolic engineering provided strains that assimilated the entire C2 fraction into the central metabolism or that produced 4.3 g/L glycolic acid at a molar yield of 0.9 in shake flasks.