The second-generation bioethanol production in which lignocellulosic material is used as feedstock faces some difficulties. Lignocellulosic materials have to be pretreated prior to fermentation. In the pretreatment stages several inhibitory compounds, which can negatively affect the metabolic and physiologic activity of the microorganism used, Saccharomyces cerevisiae, are released. Moreover, wild strains of Saccharomyces cerevisiae cannot co-utilize the hexose and pentose saccharides present in the lignocellulosic substrate. In this study, reverse membrane bioreactor (rMBR) was applied to address the difficulties faced in the secondgeneration ethanol production. Semi-synthetic medium and pretreated wheat straw slurry containing different level of glucose, xylose and inhibitor concentrations were fermented in rMBR using genetically-modified xylose-consuming S. cerevisiae. The diffusion rate of different substrates and metabolites during fermentation were measured and analyzed. The results showed that the application of rMBR facilitated simultaneous utilization of hexose and pentose sugars and enhanced the cell tolerance of the inhibitor present in the medium.