In this study, optimal operating parameters of 2,3-Butanediol production using Klebsiell aoxytoca under submerged culture conditions are determined by using Taguchi method. The effect of different factors including medium composition, pH, temperature, mixing intensity and inoculum size on 2,3-butanediol production was analyzed using the Taguchi method in three levels. Based on these analyses the optimum concentrations of glucose, acetic acid and succinic acid were found to be 6, 0.5 and 1.0 (%w/v) respectively. Furthermore, optimum values for temperature, inoculumsize, pH and the shaking speed were determined as 37ï°C, 8 (g/L), 6.1 and 150 rpmrespectively. The optimized conditions showed an enhanced 2,3-Butanediol yield of 40%[from0.539 to 0.897 (g/g)]. The optimal combinations of factors obtained from the proposed DOE methodology was further validated by conducting fermentation experiments and the obtained results revealed an enhanced 2,3-Butanediol yield of 27.63%.