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The success of xanthan biosynthesis depends on several factors, most importantly the genetic potential of the production microorganism and cultivation media composition. Cultivation media composition affects the yield and quality of the desired product as well as production costs. This is why many studies focus on finding cheap alternative raw materials, especially carbon sources, to replace commercially used glucose and sucrose. In addition to the Xanthomonas campestris ATCC 13951 which is the primary industrial production microorganism, other Xanthomonas strains can produce xanthan as well. Under the same conditions, different strains produce different amounts of the biopolymer of varying quality. The aim of this paper is to compare producibility of phytopathogenic X. campestris strains, isolated from the environment with the reference X. campestris ATCC 13951 strain and to estimate the possibility of xanthan production using alternative glycerol-based media than the synthetic glucose-based media. Submerged cultivation on the medium based on glucose or glycerol (2.0 %w/v) was performed using the reference strain and eight isolated X. campestris strains. In order to assess the success of biosynthesis, xanthan yield and rheological properties were determined. Strains isolated from the environment produced yields between 2.98 g/L and 12.17 g/L on the glucose-based medium and 1.68 g/L and 6.31 g/L on the glycerol-based medium. Additionally, X. campestris ATCC 13951 provided the highest yield when using glucose (13.24 g/L), as well as glycerol-based medium (7.44 g/L). The obtained …