Substrate-selective uptake is a novel approach for the simultaneous consumption of sugars found in biomass hydrolysates. Of the five strains studied, E. coli W showed generally the fastest growth on carbohydrates. The triple-knockout strain W glk, ptsG manZ (KD777) consumed 7 g/L L-arabinose or D-xylose within 5.5 h, and subsequently consumed glucose only slowly. The presence of glucose did affect the utilization of D-galactose by KD777. Similar results were obtained with W glk, ptsG manZ crr (KD915). Two-sugar carbon-limited chemostats with KD777 at a growth rate of 0.25 h-1 showed that including glucose in the feed slowed xylose, arabinose, galactose consumption by 8%, 20%, and 45% respectively. However, glucose was not consumed in the presence of xylose.