Alkaline Protease from Bacillus subtilis IAS 01 (KF:761633) was successively purified by 70% ammonium sulfate precipitation, ion exchange chromatography on DEAE-cellulose and gel filtration chromatography on Sephadex G-100 with 10.45 % recovery and 11.63 fold increase in specific activity. The molecular weight of the purified protease was determined as 14.3 kDa under SDS-PAGE, respectively. The characteristic of 70% (NH4)2SO4 precipitated protease was active at 55°C and pH 12.0. Among the tested mono Na+ (76.42±1.41%), divalent ion CaCl2 (130.88±1.48%) represent the highest activity and highly stability towards non-ionic, ionic surfactants, commercial detergent and 1mM EDTA inhibited by 94.2% of its original activity indicated alkaline type of metalloprotease. In addition wash performance was exhibited that the 70% ammonium sulfate precipitate metalloprotease could successfully remove the dried blood stain from hospitalized bet cloth within 30 min than commercial detergents. Considering its promising properties, Bacillus subtilis IAS 01 enzymatic preparation may be considered a potential nominee for bio-detergent in future use, respectively.
