Sorbitol production by Zymomonas mobilis in sugar cane molasses was investigated by batch fermentation. The effects of the mean sugar concentration, temperature, agitation, and culture time were studied simultaneously by factorial analysis. Maximum sorbitol production was determined using a second-order central composite design and analyzed by the superficial response method. Response-surface methodology analysis showed that the best conditions for sorbitol production were 300g/l total reducing sugars (TRS) in the culture medium, 36h at 35 o C fermentation time.