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Optimization of nutrient supplements i.e., yeast extract (1, 3 and 5 g·L−1), dried spent yeast (DSY: 4, 12 and 20 g·L−1) and osmoprotectant (glycine: 1, 3 and 5 g·L−1) to improve the efficiency of ethanol production from a synthetic medium under very high gravity (VHG) fermentation by Saccharomyces cerevisiae NP 01 was performed using a statistical method, an L9 (34) orthogonal array design. The synthetic medium contained 280 g·L−1 of sucrose as a sole carbon source. When the fermentation was carried out at 30 °C, the ethanol concentration (P), yield (Yp/s) and productivity (Qp) without supplementation were 95.3 g·L−1, 0.49 g·g−1 and 1.70 g·L−1·h−1, respectively. According to the orthogonal results, the order of influence on the P and Qp values were yeast extract > glycine > DSY, and the optimum nutrient concentrations were yeast extract, 3; DSY, 4 and glycine, 5 g·L−1, respectively. The verification experiment using these parameters found that the P, Yp/s and Qp values were 119.9 g·L−1, 0.49 g g−1 and 2.14 g·L−1·h−1, respectively. These values were not different from those of the synthetic medium supplemented with 9 g·L−1 of yeast extract, indicating that DSY could be used to replace some amount of yeast extract. When sweet sorghum juice cv. KKU40 containing 280 g·L−1 of total sugar supplemented with the three nutrients at the optimum concentrations was used as the ethanol production medium, the P value (120.0 g·L−1) was not changed, but the Qp value was increased to 2.50 g·L−1·h−1

High Level Ethanol Production by Nitrogen and Osmoprotectant Supplementation under Very High Gravity Fermentation Conditions