A method combining a Plackett-Burman design(PBD), the steepest ascent method (SA), and a Box-Behnken design (BBD) was developed to optimize succinicacid production from cane molasses by Actinobacillussuccinogenes GXAS137. The important parameters were(g/L): total sugars of cane molasses (85 g/L), yeast extract(8.84 g/L), and MgCO3 (63.1 g/L). Verification experimentsindicated that the maximal succinic acid productionreached 57.43±0.86 g/L, which agreed with the predictedvalue (57.12 g/L). In addition, batch and fed-batchfermentations were carried out in a 1.3 L stirred bioreactor. Compared with a batch fermentation that produced 57.96g/L of succinic acid at 60 h, a fed-batch fermentation,performed to minimize the inhibition effect of the substrate,produced 64.34 g/L of succinic acid at 60 h. The combinedmethod is powerful for selection of optimized conditionsfor succinic acid production from cane molasses.