Agricultural wastes and other non-food sources can be used to produce biofuels. Despite multiple attempts using engineered yeast strains expressing exogenous genes, the native Saccharomyces cerevisiae produces low amount of second generations of biofuels. Here, we focused on Znf1, a non-fermentable carbon transcription factor and the suppressor protein Bud21 to overcome this challenge. Several mutants of engineered S. cerevisiae strains were engineered to enhance production of biofuels and xylose-derived compounds such as xylitol. This study demonstrates Znf1's novel transcriptional regulatory control of xylose and offer an initial step toward a more sustainable production of advanced biofuels from xylose.