An adapted strain of the flocculating yeast S. cerevisiae NRRL Y-265 was obtained through adaptation by cell recycle batch fermentation. The exposure to increased levels of lignocellulosic inhibitors in fermentation media simulating lignocellulosic hydrolysates generated a tolerant strain after 39 days of successive batch culture, with ethanol productivity and yield increased by 50–60% in the presence of moderately high levels of inhibitors. When used in batch fermentation in the presence of inhibitors derived from steam-exploded eucalyptus, the adapted Y-265A3 strain showed higher ethanol productivity and growth rate, and higher transformation rates for furfural, HMF and syringaldehyde, when compared to the non-adapted strain, which can be considered an important advantage for bioethanol production.