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Application of Chlorpyrifos on agricultural fields to protect crops against pests results in accumulation of it in soil and other environmental samples. The insecticide transform into3,5,6-Trichloro-2-Pyridinol (TCP) through hydrolysis in soil, which has got antimicrobial property and hence resists its degradation in natural condition. In the current findings, a bacterial isolate capable of mineralizing Chlorpyrifos without accumulation of TCP was isolated from agricultural soil by enrichment method. Based on Morphological, Biochemical Characterization and with Bergey’s Manual comparision, the isolate was identified asStaphylococcus sp. The isolate was found to metabolize chlorpyrifos completely in Mineral salt medium with chlorpyrifos as the sole carbon source. No metabolites of chlorpyrifos were detected in Liquid Chromatography-Mass Spectroscopy (LC-MS) analysis after 7 days of incubation. The novelty of the outcome of the experiment relies onStaphylococcussp.ES-2 in complete mineralization of chlorpyrifos which can be used as a potential bioaugmenting agent in the chlorpyrifos contaminated sites.