Organophosphorus insecticides (OPs) have been widely used to control agricultural pests, which has raised concerns about OPs residues in crops and the environment. The research group had previously isolated a highly efficient OPs degradation strain , Cupriavidus nantongensis X1^T. In this study, the degradation characteristics of chiral organophosphorus pesticide profenofos by strain X1^T under different substrate concentrations, bacterial counts, pH and metal ions were measured. The results showed that strain X1^T could preferentially degrade S-profenofos with high toxicity and low insecticidal activity. With the increase of substrate concentration, the degradation rate of profenofos by strain X1^T gradually slowed down the optimal pH of profenofos degradation by strain X1^T was 7.5; the valence state of metal ions was an important factor to affect the degradation of profenofos, of which Cr³⁺ and Fe³⁺ could inhibit the degradation of profenofos, while Ca²⁺, Mn²⁺, Mg²⁺, Co²⁺, Ni²⁺ could promote the degradation of S-profenofos. Meanwhile, there was no significant difference between the degradation rate of S-profenofos and R-profenofos by the crude enzyme in strain X1^T and purified enzyme opdB. The results provide a theoretical basis for bioremediation of profenofos residuals.