The objective of the present study was to study the optimization conditions for the production of xanthan by Xanthomonas campestris from pre-treated sugarcane molasses. In the study, the optimization was carried out for different parameters including pH, temperature, and incubation time for the pre-treated sugarcane molasses. The age of inoculums and time of culture growth (6, 12, 18 and 24 hrs), size of inoculums (2%, 5%, 7.5% and 10%), pH (6.6, 6.8, 7.0 and 7.2) and temperature (25°C, 28°C, 30°C, 32°C and 37°C) were studied. It was observed that the xanthan production was maximal with 7.5% (v/v) inoculums, pH. 7 at 30°C for 48 hrs. The study suggested that cane molasses is an appropriate agro-industrial substrate for xanthan gum fermentations, and further scale-up study is needed for gum production in the stirred fermenter.