Various challenges facing the industrial production of bio-based lactic acid (LA) such as cost of raw materials and nitrogen sources, as well as contamination risk by mesophilic and neutrophilic producers, should be overcome for the commercial production. This study aimed to investigate the feasibility of corn steep water (CSW) as a raw material for LA production using a newly thermo-alkali-tolerant lactic acid bacterium. The physicochemical characteristics of CSW were investigated. The high carbohydrates, proteins, amino acids, vitamins, essential elements, minerals, and non-protein nitrogenous compounds content confirmed that the CSW is a promising substrate for LA production. Out of 67 bacterial isolates, Enterococcus faecium WH51-1 was selected based on its tolerance to high temperatures and inhibitory compounds (sodium metabisulfate, sodium chloride, sodium acetate, and formic acid). Fermentation factors including sugar concentration, temperature, inoculum size, and neutralizing agents were optimized for LA production. Lactic acid concentration of about 44.6 g/L with a high yield (0.89 ± 0.02 g/g) was obtained using 60 g/L of CSW sugar, inoculum size 10% (v/v), 45 °C, and sodium hydroxide or calcium carbonate as a neutralizing agent. These results demonstrated the potential of strain WH51-1 for LA production using CSW effluent as raw material.