In this study, a genetically engineered Escherichia coli strain, HBUT-D (Delta pflB Delta pta Delta frdABCD Delta adhE Delta ald Delta cscR), was initially evaluated on a laboratory scale (7 L) in a glucose (130 g L-1) mineral salts medium for D-lactic acid fermentation using 6 N KOH, Ca(OH)(2) or NH4OH as the neutralizing agent. Fermentations neutralized by Ca(OH)(2) achieved a volumetric productivity of 6.35 g L-1 h(-1), tripling that achieved by KOH (1.71 g L-1 h(-1)) and NH4OH (1.5 g L-1 h(-1)). The facilitative effect of Ca(OH)(2) neutralization was then demonstrated on a pilot scale (6 ton vessel, 130 kg glucose ton(-1)), resulting in a volumetric productivity of 6 kg ton(-1) h(-1), a titer of 126 kg ton(-1), a yield of 97%, and an optical purity of 99.5%. These results demonstrated that E. coli HBUT-D is a promising strain for large scale D-lactic acid fermentation using mineral salts medium and Ca(OH)(2) for neutralization. (C) 2014 Elsevier Ltd. All rights reserved.