Isolation, purification and characterization of a surfactants-, laundry detergents- and organic solvents-resistant alkaline protease from Bacillus sp. HR-08.

摘要: Bacillus sp. HR-08 screened from soil samples of Iran, is capable producing proteolytic enzymes. 16S rDNA analysis showed that this strain closely related to subtilis, licheniformis, pumilus, mojavensis, and atrophaeus. The zymogram the crude extract revealed presence five extracellular proteases. One proteases was purified in three steps procedure involving ammonium sulfate precipitation, DEAE-Sepharose ionic exchange Sephacryl S-200 gel filtration chromatography. molecular mass enzyme on SDS-PAGE estimated be 29 kDa. protease exhibited maximum activity at pH 10.0 60 degrees C inhibited by PMSF but it not affected cysteine inhibitors, suggesting a serine alkaline protease. Irreversible thermoinactivation examined 50, 60, 70 10 mM CaCl(2). Results retains more than 80% 50% its initial after incubation for 30 min C, respectively. This had good stability H(2)O(2), nonionic surfactant, local detergents enhanced 20% dimethyl sulfoxide (DMSO), formamide (DMF) isopropanol. retained 90% pre-incubation 1 h room temperature these solvents. Also, activation can seen high concentration (50%, v/v) DMF DMSO.