To enhance the protease production and decrease cost, corn flour and soy peptone were screened as cheap raw materials for the production of extracellular proteases by Bacillus strains. Their compositions in the medium suitable for enzyme production of Bacil- lus sp. B001 were optimized using statistical experi- ment designs. Under the optimized conditions, the protease production of Bacillus sp. B001 was stable at the stationary stage and reached to 63,200 U/mL, ap- proximately 1.84-fold increase compared with that using the original medium. These improvements could be attributed to the release of the catabolite repression by crude materials corn flour and soy peptone which contained low level of available nutri- ents. Additionally, a highly pure protease which dis- played excellent stability and compatibility with high salinity, commercial laundry detergents, and organic solvents, was rapidly obtained by two-step procedure involving ammonium sulphate precipitation and an- ion exchange from the fermentation cultures of B001 in the optimized medium. When the culture method applied to other Bacillus strains, their protease yields were all remarkably increased approximately 2.9 to 8.5 folds. In conclusion, a low-cost, easy-purified, and effective producing strategy using the cheap raw ma- terials was developed here, representing a potential application for protease production in various Indus- trial processes.