In the present investigation, six isolates of Bacillus subtilis were isolated from soil of different areas in Nagpur district for production of amylase. All isolates were aerobic with straight rods, motile, endospore forming, gram positive, strongly catalase positive and indicative of Bacillus subtilis. The maximum clear zone surrounding the bacterial growth was observed in Isolate No. B.S-1 and B.S-3 indicating starch hydrolysis. The agro wastes used as substrates were Wheat Bran, Mustard Oilseed Cake and Gram Husk respectively. Crude enzyme obtained by using Wheat Bran as a substrate, showed maximum results for enzyme activity as compared to Mustard Oilseed Cake and Gram Husk. The results of the assay showed that bacterial isolate B.S-3 showed the highest enzymatic activity of 177mg/ml/min with wheat bran as substrate. Bacillus isolate B.S-3 and B.S-1 showed highest enzyme activity in presence of Wheat bran (177mg/ml/min, 151mg/ml/min) followed by Gram husk (126mg/ml/min, 71mg/ml/min) and low yield with Mustard oilseed cake by both isolates (91mg/ml/min).
