The aim of this present study was to isolate the keratinase producing bacteria from poultry farm and slaughter house. In that, feather dumping soil (100 samples) was collected from several area includes the Coimbatore, Pollachi, and Erode. Among these samples, the eighteen isolate were subjected for primary screening using Milk casein agar plate. From these isolates, the Bacillus subtilis showing a highest keratinase activity in secondary screening using Casein agar medium & it was confirmed by 16s rRNA molecular sequence. The specific organism subjected to keratinase activity assay and identified by the spectrometric using the keratin azure as a substrate.The enzyme were undergone with the standard Purification process, its includes the ammonium sulphate precipitation, DEAE Sephadex K-50, Biogel G-200.The molecular mass of specific protein is 40KDa. There are enomorous types of azo dyes were classified and connected to this three types of azo dyes were used, which include Kerria Lacca, Rubia cardifolia, Accacia catechu. The newly isolated Bacillus subtilis can able to decolour the Rubia cardifolia effectively. According to the wet lab analyses , the OD value is 7.31 (Rubia cardifolia). In dry lab analysis, the “G” score value about -4.01; which indicates that there is an higher indication profile of keratinase with alizarin.

Keywords : Soil sample, Bacillus subtilis, Azo dye, alizarin.