Abstract Background: A single-step ethanol production is the combination of raw cassava starch hydrolysis and fermentation. For the development of raw starch consolidated bioprocessing (CBP) technologies, this research work was to investigate the optimum conditions and technical procedures for the production of ethanol from raw cassava starch in a single step. This resulted high yields and productivities of all the experiments from the laboratory, the pilot, through the industrial scales. The yields of ethanol concentration are comparable with those in the commercial industries that use molasses and hydrolyzed starch as the raw materials. Results: Before single-step ethanol production, the studies of raw cassava starch hydrolysis by a granular starch hydrolyzing enzyme, StargenTM002, were carefully conducted. It successfully converted 80.19% (w/v) of raw cassava starch to glucose at a concentration of 176.41 g/L with a productivity of 2.45 g/L/h when the raw starch was pretreated at 60 °C for 1 h with 0.10% (v/w dry starch basis) of Distillase ASP before hydrolysis. A single-step ethanol production at 34 °C in a 5-L fermenter showed that S. cerevisiae (Fali, active dry yeast) produced the maximum ethanol concentration, p of 81.86 g/L (10.43% v/v) with a yield coefficient, Y p/s of 0.41 g/g, a productivity or production rate, r p of 1.14 g/L/h with an efficiency, Ef of 71.44%. The scale-up experiments of the single-step ethanol production using this method, from the 5-L fermenter to the 200-L fermenter and further to the 3,000-L industrial fermenter were successfully achieved with essentially good results. The p, Y p/s , r p , and Ef values of the 200-L scale were 80.85 g/L (10.23% v/v), 0.41 g/g, 1.12 g/L/h and 72.47% , respectively ; of the 3,000-L scale were 70.74 g/L (9.01% v/v), 0.34 g/g, 0.98 g/L/h and 59.82% , respectively. Because of using raw starch, the major by-products of all the three scales were very low; glycerol lactic acid and acetic acid, in ranges of 0.94-1.14%, 0.046-0.052%, 0-0.059% (w/v), respectively, where are less than those values in the industries. Conclusions: This single-step ethanol production using a combination of raw cassava starch hydrolysis and fermentation of the three fermentation scales here is practicable and feasible for the scale-up of industrial production of ethanol from raw starch.