Many studies have focused on using crude glycerol from biodiesel to obtain valuable products, but few of these studies have focused on obtaining polysaccharides. A mutant strain of Xanthomonas campestris CCTCC M2015714 that could use glycerol to produce high-transparency and low-viscosity xanthan gum was obtained by adaptive evolution, and the yield of xanthan gum reached 11.0 g/L. We found that transcriptional levels of genes related to glycerol metabolism (glpF, glpK, glpD, and fbp) in the mutant strain were all higher than those from the parent strain. Using 5 g/L sucrose or glucose as starter substrate, cell growth time decreased from 36 h to 24 h and xanthan gum yield increased. Moreover, the mutant strain can tolerate high titer glycerol, and its activity was not affected by the impurities in crude glycerol. All these results proved that crude glycerol from biodiesel industries can be used for xanthan gum production. (C) 2016 Elsevier Ltd. All rights reserved.