<jats:title>ABSTRACT</jats:title> <jats:p> The oleaginous yeast <jats:named-content content-type="genus-species">Yarrowia lipolytica</jats:named-content> is an industrially important host for production of organic acids, oleochemicals, lipids, and proteins with broad biotechnological applications. Albeit known for decades, the unique native metabolism of <jats:named-content content-type="genus-species">Y. lipolytica</jats:named-content> for using complex fermentable sugars, which are abundant in lignocellulosic biomass, is poorly understood. In this study, we activated and elucidated the native sugar metabolism in <jats:named-content content-type="genus-species">Y. lipolytica</jats:named-content> for cell growth on xylose and cellobiose as well as their mixtures with glucose through comprehensive metabolic and transcriptomic analyses. We identified 7 putative glucose-specific transporters, 16 putative xylose-specific transporters, and 4 putative cellobiose-specific transporters that are transcriptionally upregulated for growth on respective single sugars. <jats:named-content content-type="genus-species">Y. lipolytica</jats:named-content> is capable of using xylose as a carbon source, but xylose dehydrogenase is the key bottleneck of xylose assimilation and is transcriptionally repressed by glucose. <jats:named-content content-type="genus-species">Y. lipolytica</jats:named-content> has a set of 5 extracellular and 6 intracellular β-glucosidases and is capable of assimilating cellobiose via extra- and intracellular mechanisms, the latter being dominant for growth on cellobiose as a sole carbon source. Strikingly, <jats:named-content content-type="genus-species">Y. lipolytica</jats:named-content> exhibited enhanced sugar utilization for growth in mixed sugars, with strong carbon catabolite activation for growth on the mixture of xylose and cellobiose and with mild carbon catabolite repression of glucose on xylose and cellobiose. The results of this study shed light on fundamental understanding of the complex native sugar metabolism of <jats:named-content content-type="genus-species">Y. lipolytica</jats:named-content> and will help guide inverse metabolic engineering of <jats:named-content content-type="genus-species">Y. lipolytica</jats:named-content> for enhanced conversion of biomass-derived fermentable sugars to chemicals and fuels. </jats:p>