Optimization of the medium for dextransucrase production by Leuconostoc mesenteroides FT 045B was carried out using response surface methodology. A central composite design was employed to determine maximal dextransucrase activity at optimal values of sugarcane molasses, corn steep liquor and potassium phosphate. The combined effects of these nutrients on dextransucrase activity were used to evaluate agitation speed and aeration rate through a full factorial design. Amodel with satisfactory fit was achieved in both cases. Dextransucrase activity was significantly affected by the independent variables corn steep liquor and K2HPO4 using the central composite design and by the independent variables agitation speed and 24 aeration rate using the full factorial design. Maximal dextransucrase activity of 4.03 U/mL was obtained with sugarcane molasses 40 g/L, K2HPO4 20 24 g/L, corn steep liquor 20 g/Land 3.99 U/mLwith agitation speed 100 rpm and aeration rate 1 vvm.