A large amount of wastewater containing various organic compounds is generated in the paper-making industry. Therefore, the re-utilization of wastewater from paper-making is of great importance. In this study, acetic acid and formic acid, the dominant inhibitors existing in the paper mill wastewater, were used to produce succinate by metabolically engineered Escherichia coli strains, HB04(pTrc99a-gltA) and HB04(pTrc99a-gltA, pBAD33-Trc-fdh). The distilled paper mill wastewater could be utilized and transformed directly to succinate efficiently. However, the utilization of acetate was inhibited dramatically using raw paper mill wastewater. After adsorption by activate carbon, the acetate consumption and succinate production were improved significantly. In the resting cells experiment with high cell density (50 OD600), acetate was consumed completely and the titer of succinate reached 65.44 mM in 6 h. These results showed that the metabolically engineered E. coli strains had great potential to utilize acetic acid in the paper mill wastewater for succinate biosynthesis.