The profenofos (PFF), an organophosphate (OP) insecticide, is extensively use in agriculture to control lepidopteran pests and its indiscriminate use in agriculture causes accumulation of the pesticide and its residue in the ecosystem. Interestingly, the microorganisms play a crucial role in detoxification of pesticides and other xenobiotics compounds. One PFF degrading novel bacterium, Pseudomonas aeruginosa strain BRD1 was isolated through enrichment culture technique from an agricultural soil of Baharpur, Burdwan, Purba Bardhaman, West Bengal, India. The bacterium is rod shaped, motile and Gram-negative in nature and shown optimal growth in 100 ppm PFF at 32 °C. The isolate BRD1 was identified as Pseudomonas aeruginosa strain BRD1 using 16S rRNA gene based molecular phylogenetic approach, phenotypic characterization and whole genome analyses. Quantitative biodegradation analysis by High-performance liquid chromatography (HPLC) chromatograms reveals that the strain can degrade 76% of PFF after 96 hrs incubation. The HPLC and Electrospray Ionization Mass Spectrometry (ESI-MS) investigation indicates that the strain degrades the PFF via the formation of intermediate metabolites, 4-bromo-2-chlorophenol (BCP), 3-methoxyphenol, 2-chloro-4-benzoquinone, O-ethyl S-propyl phosphorothioic acid (EPPA), 3,4-dimethyl benzoic acid and phosphoric acid but, BCP was produced as a major hydrolytic intermediate. The genome of the bacterium possesses several OP and PFF pesticide degrading genes like bphD, glpD, estB_1, estB_2, pdeB, pdeN, pdeG, gstB_2, yfcF, gstB_3, cpg2, lap, pepN and encodes several OP degrading enzyme groups like phosphodiesterase, cytochrome p450, glutathione S-transferase, metal dependent hydrolase, metallophosphoesterase, amidohydrolase. The current study elucidates that strain BRD1 efficiently can remove PFF within a short time.