A total of 58 isolates capable of degrading chlorpyrifos were obtained out of 27 samples collected. Among them 10 isolates showed better efficiencies in degrading chlorpyrifos as compared to others. Isolate CHS23a which showed 38% degradation of chlorpyrifos under unoptimized condition was identified as Pseudomonas putida. After optimization process it was able to show 76% degradation of chlorpyrifos at pH 7, temperature 35°C, pesticide concentration of 2%, 10 ml of inoculum size, shaking speed of 150 rpm and in presence of 200 mg/l glucose and 300 mg/l yeast extract.