Ectoine is widely produced by various bacteria as a natural cell protectant against environment stress, e.g., osmotic and temperature stress. Its protective properties therefore exhibit high commercial value, especially in agriculture, medicine, cosmetics, and biotechnology. Here, we successfully constructed an engineered Escherichia coli for the heterologous production of ectoine. Firstly, the ectABC genes from Halomonas elongata were introduced into E. coli MG1655 to produce ectoine without high osmolarity. Subsequently, lysA gene was deleted to weaken the competitive L-lysine biosynthesis pathway and ectoine bioconversion was further optimized, leading to an increase of ectoine titer by 16.85-fold. Finally, at the low cell density of 5 OD600/mL in Erlenmeyer flask, the concentration of extracellular ectoine was increased to 3.05 mg/mL. At the high cell density of 15 OD600/mL, 12.7 g/L of ectoine was achieved in 24 h and the overall yield is 1.27 g/g glycerol and sodium aspartate. Our study herein provides a feasible and valuable biosynthesis pathway of ectoine with a potential for large-scale industrial production using simple and cheap feedstocks.