*Article* **Optimization of Yeast, Sugar and Nutrient Concentrations for High Ethanol Production Rate Using Industrial Sugar Beet Molasses and Response Surface Methodology**

**Jean-Baptiste Beigbeder , Julia Maria de Medeiros Dantas and Jean-Michel Lavoie \***

Biomass Technology Laboratory, Department of Chemical Engineering and Biotechnology Engineering, Université de Sherbrooke, 2500 Boul. de l'Université, Sherbrooke, QC J1K 2R1, Canada; jean-baptiste.beigbeder@usherbrooke.ca (J.-B.B.); Julia.M.M.Dantas@USherbrooke.ca (J.M.d.M.D.) **\*** Correspondence: Jean-Michel.Lavoie2@USherbrooke.ca

**Abstract:** Among the various agro-industrial by-products, sugar beet molasses produced by sugar refineries appear as a potential feedstock for ethanol production through yeast fermentation. A response surface methodology (RSM) was developed to better understand the effect of three process parameters (concentration of nutrient, yeast and initial sugar) on the ethanol productivity using diluted sugar beet molasses and *Saccharomyces cerevisiae* yeast. The first set of experiments performed at lab-scale indicated that the addition of 4 g/L of nutrient combined with a minimum of 0.2 g/L of yeast as well as a sugar concentration lower than 225 g/L was required to achieve high ethanol productivities (>15 g/L/d). The optimization allowed to considerably reduce the amount of yeast initially introduced in the fermentation substrate while still maximizing both ethanol productivity and yield process responses. Finally, scale-up assays were carried out in 7.5 and 100 L bioreactors using the optimal conditions: 150 g/L of initial sugar concentration, 0.27 g/L of yeast and 4 g/L of nutrient. Within 48 h of incubation, up to 65 g/L of ethanol were produced for both scales, corresponding to an average ethanol yield and sugar utilization rate of 82% and 85%, respectively. The results obtained in this study highlight the use of sugar beet molasses as a low-cost food residue for the sustainable production of bioethanol.

**Keywords:** RSM; bioethanol; yeast fermentation; sugar beet molasses; industrial by-product; scale-up
