We optimized the conditions of mixed fermentation of very high gravity ethanol with cassava flour and sugarcane juice. Based on the single factor experiment, we screened the important parameters for very high gravity ethanol fermentation with cassava flour and sugarcane juice by the Plackeet-burman design. Then, we obtained the optimum values of the important parameters by the orthogonal experiments: the mixing ratio of cassava flour to sugarcane juice, 1:5; initial pH of fermentation, 4.0-4.5; the concentrations of urea and MgSO4, 0.25% and 0.04% (W/W), respectively. Finally, we used a gradient temperature control strategy with the optimized conditions, and ethanol concentration of 17.84% (V/V) and fermentation efficiency of 91.82% were achieved, correspondingly.

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