In this study, a genetically engineered Escherichia coli strain, HBUT-D (ΔpflB Δpta ΔfrdABCD ΔadhE Δald ΔcscR), was initially evaluated on a laboratory scale (7L) in a glucose (130gL−1) mineral salts medium for d-lactic acid fermentation using 6N KOH, Ca(OH)2 or NH4OH as the neutralizing agent. Fermentations neutralized by Ca(OH) 2 achieved a volumetric productivity of 6.35gL−1h−1, tripling that achieved by KOH (1.71gL−1h−1) and NH4OH (1.5gL−1h−1). The facilitative effect of Ca(OH)2 neutralization was then demonstrated on a pilot scale (6ton vessel, 130kgglucoseton−1), resulting in a volumetric productivity of 6kgton−1h−1, a titer of 126kgton−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.