A key step in any process that converts lignocellulosic biomass to biofuels is the efficient fermentation of all sugars found in the hydrolysate. In this work we performed fed-batch experiments using Scheffersomyces stipitis to investigate the efficiency of ethanol production by this strain, during consumption of pentoses sugars. To better predict how this yeast behaves in fed-batch culture and to optimize the system performance, the impact of xylose concentration (100-200 g.L-1) in feed stream on sugar consumption and ethanol production was investigated. Fed-batch mode cultivation has been utilized to avoid overflow metabolism as well as to ensure an accurate balanced feeding of nutrient substrate into reactor. The operation in fed-batch mode with an optimized feeding rate and cell growth rate resulted in higher titer, yield and productivity of ethanol compared with batch cultures. The maximum ethanol concentration achieved was 46.4 g.L-1, obtained for xylose concentration of 200 g.L-1 in the feed stream. The maximum ethanol yield (0.43 g.g-1) and productivity (0.58 h-1) was 1.1 and 2.3-fold than those in batch mode. The fed-batch strategy developed in this work was capable to achieve higher efficiency in second generation ethanol production by Scheffersomyces stipitis, and these results might be useful to guide future improvements in industrial scale.