Proteases are one of the most studied microbial group of enzymes for theirimportance at commercial, industrial, pharmaceutical and also diagnostic fields. The aim of this work was the isolation of proteolytic bacteria from soil and characterization of their proteolytic activity. Two effective protease producing bacterial isolates have been obtained from soil from Sadat City and evaluated its extracellular protease production properties by exhibiting a clear zone on media containing skimmed milk casein as a substrate for the protease enzyme. The strains have been identified according to basic biochemical tests using Bergey's Manual of Systematic Bacteriology as Bacillus sp. Scanning electron microscopy revealed morphology of Cells of Bacillus colonies tended to be longer than in liquid culture and irregular in shape and to give the appearance of branching. The 16s rRNA gene pcr and sequencing were used for species identification of the 2 isolates. And by analysis of the sequence results using the BLASTn, it revealed that one isolate (Ash1) was identified as bacillus cereus and the other (Ash2) was bacillus subtilis. The enzyme production reached its maximum level at pH 8.0 at 37^oC incubation temperature, under aerated environment, suggesting the enzyme produced is alkaline in nature. The Bacillus cereus strain Ash1 and B. subtilis Ash2 showed maximum growth at 37^oC with alkaline protease production at pH 8.0 and at 150 rpm. This study reveals the importance of genus bacillus in protease production among the bacterial population in soil, although the narrow scale this study was applied on, but it has to be studied on larger scales in Egypt, an issue that will provide many beneficial information for various fields particularly the industrial sectors.