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Xanthan is one of the most important biopolymers characterised by a high commercial value. Since the availability of essential nutrients influences its biosynthesis, the aim of this study was the optimisation of carbon, organic and inorganic nitrogen content in glycerol-based medium employing response surface methodology. The cultivation of strain Xanthomonas campestris ATCC 13951 was carried out under appropriate conditions on media with glycerol, peptone and ammonium-nitrate formulated according to Box-Behnken design (3³), while the desirability function approach was used for determination of optimal nutrient levels. The final model predicts that the maximal amount of xanthan (12.95 g/L) is produced when the initial contents of glycerol, peptone and ammonium-nitrate in the medium are 32.96 g/L, 0.55 g/L and 0.73 g/L, respectively. To minimize the residual nutrient content and therefore the …