Isolation, Identification, Characterization of Bacillus  Subtilis Subsp Subtilis., Producing the Keratinase  Enzyme Under Optimization Method

R.Manju; Dr.S.Shanmugam

Volume : III, Issue : XI, November - 2013

Isolation, Identification, Characterization of Bacillus Subtilis Subsp Subtilis., Producing the Keratinase Enzyme Under Optimization Method

R. Manju, Dr. S. Shanmugam

Abstract :

The aim of this study to isolate the keratinase producing bacteria from poultry farm, 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 subsp subtilis., showing a highest keratinase activity in secondary screening using Casein agar medium & its confirmed by 16s rRNA gene sequence. The specific organism subjected to keratinase activity assay and it’s identified by the spectrometric using the keratin azure as a substrate. Thus, this organism was optimized by Response enzyme methodology (RSM) using the production medium. In this, fructose shows a 100% result for the keratinase activity;enzyme activity about 8+/– 1.9; pH 6.5, temperature 370C with incubation of 7 days under OGM and dextrose shows an important role in both biomass production and keratinase activity. The keratinase production was favored in the presence of ammonium tartarate as the nitrogen source. The optimum level of glucose for about 5.61g/l; enzyme activity was around 13.280 U /ml. In same way, the keratinase production was favored in the presence of ammonium tartarate as the nitrogen source; the enzyme activity in yeast extract (nitrogen source) as 9.8+/–1.3. Simultaneously, keratinase production were selected based on the initial screening experiments involving carbon and nitrogen source using glucose, starch, starch and ammonium tartarate, glucose and ammonium tartarate, OMG with ammonium tartarate under the submerged fermentation, among the glucose and ammonium tartarate range from 11.2+/–0.3 (keratinase activity)U/ml.