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Efficient ethanol production using dilute acid pretreated rice straw was investigated in multiple-fed simultaneous saccharification and co-fermentation (mf-SSCF) for producing high titer ethanol at a low cost. The mf-SSCF was performed at very high solid loading (29.5% w/v) by multiple feeding of dilute acid pretreated rice straw without any detoxification, solid-liquid separation, and sterilization. After 72 h, an ethanol titer of 68.72 g L⁻¹ was attained with higher ethanol yield (0.42 g g⁻¹) and productivity (0.95 g L⁻¹ h⁻¹). Only magnesium sulfate (6 g L⁻¹) was added as nutrient component and aqueous ammonia was used for pH adjustment of pretreated slurry to 5, which also provides ammonium ions as a nitrogen source for yeast during ethanol fermentation. Almost no water was added in the mf-SSCF process; therefore, higher ethanol titer (8.7 % v/v) was achieved, which further reduce distillation, water input, and …