The present investigations dealt with the optimization of the physical parameters for production of alkaline protease by alkaliphilic Bacillus subtilis SH-2 isolated from slaughter house soil of Warangal, Telangana State, India. Primary screening of four different samples revealed one potent isolate. Morphological and Biochemical characterization followed by Molecular signature of 16s rRNA homology confirmed that the isolate SH-2 belongs to Bacillus subtilis. Bacillus subtilis SH-2 was screened on four different reported mediums (M1213, M660, Horikoshi and Halophilic Bacillus medium) under shake culture conditions. Maximum alkaline protease production (500 EU/ml) obtained on M1213 and Horikoshi mediums. Further optimization of physical parameters by OVAT method revealed that mean generation time (41.18 min), 4% level inoculum, incubation time 72 hrs, pH 10, temperature 350C and agitation 150 rpm are ideal for enzyme production. OVAT method resulted in 2.2 fold increased production of alkaline protease production (1100 EU/ml)
