Escherichia coil Lin43 is a strain which has some mutations in glycerol kinase (GlpK) and the repressor for the glycerol 3-phosphate regulon (GlpR). When exposed to glycerol, it quickly accumulates lethal levels of methylglyoxal, which is a precursor of acetol; acetol is important for the manufacture of polyols, acrolein, dyes, and skin tanning agents. This work reports the engineering of E. coil Lin 43 for the conversion of glycerol into acetol. First, the glyoxalase system was interrupted by deleting the gloA gene, which increased the acetol yield by 32%. In addition, the aldehyde reductase YqhD was overexpressed which led to an increase of acetol production by 11.4-fold. Acetol production was optimized by varying the cell density, glycerol concentration, supplemental carbon source, pH and temperature. Under the optimal conditions (OD600 = 20, 20 g/L glycerol, 2 g/L succinate, pH 7.0, and 28 degrees C), we obtained 5.4 g/L acetol in 21 h. (C) 2013 Elsevier Ltd. All rights reserved.