Abstract—Amylases are starch degrading enzyme with wide industrial applications. Microbial production of amylase is more simplified and economical than other sources. In this study, amylase producing bacteria were isolated from hot springs of Munger (Bihar, India). Water samples were collected from three different kunds, viz., Sita kund, Rishi kund and Bhimbandh kund with temperature 40-50°C and pH 5.2-6.3. Amongst amylase positive isolates, RK6 was selected for amylase characterization, exhibiting enzyme activity at highest optimum temperature of 80°C. On the basis of 16S rRNA gene sequence, it was identified as Bacillus subtilis RK6 (KX247637). Amylase of RK6 was characterized with optimum pH of 8.0 and was thermoactive, retaining its activity upto 90°C and stability at 60-90°C after preheating for 30 min. Vmax and Km for enzyme activity were 60.56 U/ml and 1.86 mg/ml respectively, showing an appreciable affinity for substrate. The enzyme was activated in the presence of Ca2+, Mg2+, and Fe2+, while that of Zn2+ and Cu2+ resulted in its inhibition. The present finding indicates potential of thermo-alkaliphilic amylase of B. subtilis RK6 for various biotechnological applications.