Background: Bioremediation of groundwater and soil contamination is more economical than physicochemical remediation. The present study focused on the bioremediation capability of two bacterial species (Klebsiella planticola and Enterobacter cloacae) from the family Enterobacteriaceae. These bacteria have been identified as new species with capability of degrading methyl tertiary-butyl ether (MTBE). In order to enhance their degradation capability, selected concentrations and retention time were investigated. Methods: The bacteria were cultured on the nutrient agar (NA) medium at room temperature. pH of the medium was adjusted to 7. The medium was autoclaved at 121°C for 15 minutes and incubated for 24 hours at 35°C. After 24 hours, the mixture was inoculated into 50 mL of Luria Bertani (LB) liquid medium containing 50 and 150 ppm MTBE. The cultures were incubated for 2 and 5 days at 35°C and shacked on a shaker at 150 rpm. Cell concentrations of the bacteria in pure culture were determined from the optical density at 600 nm using a UV-VIS spectrophotometer. Then, the culture was centrifuged at 3800 rpm for 20 minutes. In the next step, the MTBE concentration in the supernatant was measured by gas chromatography/mass spectrometry (GC/MS, Agilent Technologies, 5975, US10304411, 5.02.07). Results:The results showed that both strains are able to grow in the presence of 50 and 150 ppm MTBE. In the best conditions, when cell density was 3×108 CFU/mL during 5 days, the highest rate of MTBE degradation for K. planticola and E. cloacae, was 43% and 40%, respectively. It was also revealed that Escherichia coli can degrade 50 and 150 ppm MTBE about 19.8% and 13.65%, respectively. Conclusion: It seems that E. coli can be a good candidate for MTBE degradation at high concentrations for a time longer than that in the present study. It was also found that the species have high performance at 50 ppm than 150 ppm. So, these bacteria can remove MTBE from the environment.