Di-(2-ethylhexyl)-phthalate (DEHP) is the phthalate ester frequently utilized as a plasticizer, commonly found in cosmetics, packaging materials, moreover, it has carcinogenic and mutagenic effect on humans. In the current study, we isolated the soil bacterium Rhodococcus aetherivorans PFS1 and to assess its DEHP degradation ability in various environmental conditions. The strain PFS1 was isolated from paddy field soil and identified by the 16S rRNA sequencing analyses. The strain PFS1 was examined for its biodegradation ability of DEHP at various pH, temperature, salt concentration, glucose concentration, and high and low concentration of DEHP. Moreover, the biodegradation of DEHP at a contaminated soil environment by strain PFS1 was assessed. Further, the metabolic pathway of DEHP degradation by PFS1 was analyzed by HPLC-MS analysis. The results showed that the strain PFS1 effectively degraded the DEHP at neutral pH and temperature 30 °C, moreover, expressed excellent DEHP degradation at the high salt concentration (up to 50 g/L). The strain PFS1 was efficiently degraded the different tested phthalate esters (PAEs) up to 90%. Significantly removed the DEHP contamination in soil along with native organisms which are present in soil up to 94.66%, nevertheless, the PFS1 alone degraded the DEHP up to 87.665% in sterilized soil. According to HPLC-MS analysis, DEHP was degraded into phthalate (PA) by PFS1 strain via mono (2-ethylehxyl) phthalate (MEHP), then PA was utilized for cell growth. These results suggest that R. aetherivorans PFS1 has excellent potential to degrade DEHP at various environmental conditions especially in contaminated paddy field soil.