Robust strains are essential towards success of n-butanol production from lignocellulosic feedstock. To find a suitable strain to convert a non-detoxified hemicellulosic hydrolysate of sugarcane bagasse, we first assessed the performance of four wild-type butanol-producing Clostridium strains (C. acetobutylicum DSM 6228, C. beijerinckii DSM 6422, C. saccharobutylicum DSM 13864, and C. saccharoperbutylacetonicum DSM 14923)in batch fermentations containing either xylose or glucose at 30 g L⁻¹ as sole carbon sources. C. saccharoperbutylacetonicum was selected after achieving butanol yields as high as 0.31 g g⁻¹ on glucose and 0.25 g g⁻¹ on xylose. In a 48-h fermentation containing a mixture of sugars (93% xylose and 7% glucose)that mimicked the hydrolysate, C. saccharoperbutylacetonicum delivered the highest butanol concentration (14.5 g L⁻¹)when the initial sugar concentration was 50 g L⁻¹. Moreover, the selected strain achieved the highest butanol yield (0.29 g g⁻¹)on xylose-rich media reported so far. Meanwhile, C. saccharoperbutylacetonicum produced 5.8 g butanol L⁻¹ (0.22 g g⁻¹ butanol yield)when fermenting a non-detoxified sugarcane bagasse hemicellulosic hydrolysate enriched with xylose (30 g total sugars L⁻¹). Although sugars were not exhausted (4.7 g residual sugars L⁻¹)even after 72 h because of the presence of lignocellulose-derived microbial inhibitors, these results show that C. saccharoperbutylacetonicum is a robust wild-type strain. This microorganism with high butanol tolerance and yield on xylose can, therefore, serve as the basis for the development of improved biocatalysts for production of butanol from non-detoxified sugarcane bagasse hemicellulosic hydrolysate.