Corynebacterium glutamicum was engineered for the first time to produce 3-hydroxypropionic acid (3-HP) from glucose via glycerol pathway. 62.6 g/L of 3-HP was produced from glucose at a yield of 0.51 g/g, the highest titer and yield of 3-HP from sugar reported to date. Downregulation of glyceraldehyde 3-phosphate dehydrogenase and employment of the non-PTS glucose-uptake system are important for increasing the yield of 3-HP. 3-HP overproduction by simultaneous utilization of glucose and xylose was achieved.