Aim of this study was to isolate keratinolytic actinomycetes from poultry waste. Four soil samples were collected from three different poultry farm of Gwalior, Guna and Bhind district of MP, India. Soil suspension were prepared by serial dilution method, inoculated into two different media actinomycetes isolation agar and starch casein agar and incubated at 28°C for 7-14 days. Thirty five isolate were purified and obtained after screening on basis of their different morphology and colour of aerial and substrate mycelium. After gram staining all isolate were found gram positive. Biochemical characterization of gram positive isolate was also performed. Out of Thirty five isolates, twenty nine showed keratinolytic activity in primary screening on Skim milk agar plate, in which clear zone around the colonies were selected responsible for degradation of chicken feathers. Secondary screenings of twenty nine isolates were performed in modified liquid basal medium supplemented with feather for keratinolytic activity. Twenty seven isolates out of twenty nine, showed keratinolytic activity and able to efficiently utilized feather as source of carbon and nitrogen. All isolates were preserved on ISP1 broth (Tryptone – yeast extract Broth) and glycerol stock for storage. The study showed that isolates from poultry soil from 3 districts were able to degrade the keratin waste (feather) by their keratinolytic activity of keratinase enzyme.