The present investigations were carried out to enhance the alkaline protease production by a mutant strain Bacillus subtilis SH2 isolated from slaughter house soils of Warangal and improved through two-tier mutagenesis first by UV and then HNO2. Initially three efficient mutants with over production of alkaline protease were identified and among them only one stable mutant SHmIIIa was selected for further improvement through popular Response Surface Methodology of the FFCCD. Only X2 agitation, X6 KH2PO4 and interactive effects of X3*X3 inoculum, X4*X5 glucose and peptone have shown a significant improvement. The maximum alkaline protease production was achieved with the medium containing of X1 pH 9.8; X2 agitation 237.5 rpm; X3, inoculum size 4%; X4, glucose 6 g/L; X5, peptone 4g/L and X6, KH2PO4 2 g/L; under batch fermentative conditions with 33.33 fold increase
