Abstract: Background and Objectives: Keratins comprise largest complex family of cytoskeletal filament proteins found in human hairs, chicken feathers and wool. Biotechnical applications might augment the human efforts to degrade these keratins which pose a problem to environment. Present study was aimed to enumerate the potential microbial sources for production and purification of keratinase followed by characterization. Materials and Methods: Poultry waste samples (n = 20) collected and used for the isolation of organism through serial dilution. Keratin broth media employed to assess the activity using azocasein as substrate by spectrophotometry. Biochemical and morphological identifications done and confirmed as Serratia sp. by 16S rRNA sequencing. To purify the enzyme, ammonium sulphate precipitation, dialysis, ion exchange and gel filtration chromatography were carried out followed by SDS-PAGE based molecular weight determination. Results: Out of 12 organisms, one showed increased keratinase activity. The activity and yield were increased 6.4 and 16.25%, respectively followed by purification. Enzyme activity was maximum at pH 7 and temperature 37°C with keratin azure as substrate. Molecular weight of purified enzyme was shown to have 50 kDa as determined by SDS-PAGE. Conclusion: It is revealed that Serratia sp. could be a potential microbial source can be harnessed for alleviation of environmental pollution and feather waste utilization.