Being a lignocellulose-rich biomass, pineapple leaves waste (PL) could be a potential raw material for the production of biofuel, biochemicals, and other value-added products. The main aim of this study was to investigate the potential of pineapple leaves in the sustainable production of bioethanol via stepwise saccharification and fermentation. For this purpose, PL was subjected to hydrothermal pretreatment in a high-pressure reactor at 150 °C for 20 min without any catalyst, resulting in a maximum reducing sugar yield of 38.1 g/L in the liquid fraction after solid-liquid separation of the pretreated hydrolysate. Inhibitors (phenolics, furans) and oligomers production were also monitored during the pretreatment in the liquid fraction of pretreated PL. Enzymatic hydrolysis (EH) of both pretreated biomass slurry and cellulose-rich solid fraction maintained at a solid loading (dry basis) of 5% wt. was performed at 50 °C and 150 rpm using commercial cellulase at an enzyme dose of 10 FPU/gds. EH resulted in a glucose yield of 13.7 and 18.4 g/L from pretreated slurry and solid fractions, respectively. Fermentation of the sugar syrup obtained by EH of pretreated slurry and the solid fraction was performed at 30 °C for 72 h using Saccharomyces cerevisiae WLP300, resulting in significant ethanol production with more than 91% fermentation efficiency. This study reveals the potential of pineapple leaves waste for biorefinery application, and the role of inhibitors in the overall efficiency of the process when using whole biomass slurry as a substrate.