BACKGROUND: The crude glycerol from biodiesel production represents an abundant and inexpensive source which can be used as raw material for lactic acid production. The first aim of this investigation was to select a strain suitable for producing lactic acid from glycerol with a high concentration and productivity. The second aim was to obtain the optimum fermentation conditions, as a basis for large-scale lactate production in the future. RESULTS: Eight bacterial strains, which could aerobically convert glycerol to lactic acid, were screened from soil samples. One of the strains, AC-521, which synthesized lactic acid with a higher concentration, was identified based on its 16S rDNA sequences and physiological characteristics. These results indicated that this strain was a member of Escherichia coli. The optimal fermentation conditions for Escherichia coli AC-521 were 42 degrees C, pH 6.5, 0.85 min(-1) (K(L)a). CONCLUSION: Escherichia coli AC-521 suitable for producing lactic acid from glycerol with high concentration and productivity was identified. After 88 h of fed-batch fermentation, both the lactic acid concentration and glycerol consumption reached maximum, giving 85.8 g L-1 of lactic acid with a productivity of 0.97 g L-1 h(-1) and a yield of 0.9 mol mol(-1) glycerol. (C) 2009 Society of Chemical Industry