The economic production of bioethanol as a sustainable liquid fuel is particularly needed and attractive. Giant reed as a low-cost and renewable biomass can be utilized as a sustainable feedstock for bioethanol development. The current research focuses on optimizing the fermentation parameters to increase ethanol concentration while lowering production costs. In this work, the giant reed was hydrolyzed thermochemically using HCl; cellulose and hemicellulose fractions were maximally converted at optimized hydrolysis conditions (5% HCl, 30 min, and 120 °C), resulting in a high sugar concentration (≈ 55 g/L), which were fermented by Candida tropicalis Y-26 for bioethanol production (≈ 15 g/L). Taguchi design was used to optimize the fermentation parameters (temperatures, pH, incubation period, and nitrogen sources). Under optimum fermentation conditions (25 °C; 24 h.; pH 5.5; and ammonium nitrate as a nitrogen source), the ethanol concentration at flask level accomplished ≈ 21 g/L, while its scale-up to bioreactor level contributed ≈ 25 g/L (equivalent to 250 kg ethanol/ton biomass) with ≈ 67% increase than the fermentation under unoptimized conditions. Overall, these findings proved that optimizing the fermentation parameters by Taguchi design and scaling up at a bioreactor could improve bioethanol production from giant reed biomass.