[Background] 1,3-Dimethyl-2-imidazolidinone (DMI) is a nonproton solvent with strong polarity, which remains steadily in waste water from production and application processes and causes potential hazard to people's health. [Objective] Acquisition of DMI degrading microbial strains. [Methods] Isolation and screen of DMI degrading microbial strains from the soil of manufacture plant by DMI tolerance assay. The strains were identified by morphological features and 16S rRNA gene sequence analysis. [Results] Two putative Bacillus velezensis strains tolerant to 5% (V/V) DMI were isolated from the soil samples, which were designated as DT-1 and DT-2 respectively. Both the whole cells and the cell-free extracts could reduce the DMI levels in the reaction systems. The degrading rates of whole cells and cell-free extract of strain DT-1 reached 48% and 68% respectively at 1% (V/V) DMI concentration. [Conclusion] DMI degrading Bacillus strains were obtained, which provided a basis for microbial degradation of DMI in the polluted environments. Moreover, the results in this study expand our knowledge of the biological function of Bacillus.