Feather waste is a significant byproduct of raising hens and poses a threat to the environment due to its abundance and resistance to degradation. Keratinase enzyme is the natural solution for degrading such kind of the wastes. The objective of this study was to isolate and characterize keratinolytic bacteria from poultry waste in Chennai. Soil samples from five locations were collected, and bacterial strains were isolated and characterized through various biochemical tests. Keratinolytic bacteria were screened by growth on skim milk agar and secondary screening on feather meal agar. Keratinase enzyme activity and feather degradation at different pH levels were assessed. Molecular identification of keratinase species was performed by extracting total DNA, partially amplifying 16S rRNA genes, and conducting sequence analysis