One of the challenges in biofuel production from lignocellulosic wastes is to improve its conversion to solvents; therefore, new strategies to enhance xylose uptake are required due to be the secondary abundant sugar. In this context, a novel fermentation strategy integrating a co-culture of Clostridium acetobutylicum and Saccharomyces cerevisiae with pH control was developed. Initially, two different buffers, ammonium acetate and calcium carbonate, were tested under pHmin > 4.8 by fermenting 60 g L−1 of glucose with the C. acetobutylicum monoculture. Ammonium acetate was selected for fermenting media as butanol production was increased from 9.8 to 10.9 g L−1 over the calcium carbonate test. Comparing with the spontaneous acetone-butanol-ethanol (ABE) fermentation with C. acetobutylicum when no xylose consumption was observed, xylose consumption was efficiently increased by controlling pHmin > 4.8. The xylose consumption was > 47% either by using a 45:15 g L−1 glucose:xylose mixture or with rice straw (RS) hydrolysate. Clostridium monoculture using RS hydrolysate and pHmin > 4.8 produced a butanol (ABE) concentration of 6.5 (9.5) g L−1. While it increased to 7.0 (13.1) g L−1 when the co-culture with S. cerevisiae was used using same pH regulation strategy mainly due to ethanol increase up to 2.7 g L−1. Moreover, the xylose uptake doubled to 94% due to amino-acid secretion by yeast. Overall, this combined strategy was a very effective method for promoting sugar consumption and ABE solvent production from lignocellulosic waste.