From 9 water samples and 15 soil samples, 42 protease producing bacterial isolates were obtained at 65 degree Celsius. Only 12 isolates could produce clear zone of 2.5 mm or more on BYmedium with 1%. The isolate RW60-6 gave the highest enzyme activity and was identified as Bacillus stearothermophilus, according to Bergey’s Manual of Systematic Bacteriology. The extracellular proteases of the isolate RW60-6 were partially purified by ammonium sulfate fractionation and DEAE Biogel A ion-exchange chromatography. Ion-exchange chromatography resulted in separation of the enzyme preparation into one major (protease U) and one minor (protease B) peak. The two-step purification scheme resulted in 13.16-fold purification and overall recovery of 10.68% of protease U and 4.77-fold purification and 8.28% recovery of protease B. SDS-PAGE assay showed two bands with neutral protease activity, at 33 and 200 kDa of protease U and at 35 and 197 kDa of protease B. Both of partially purified proteases had optimum temperature and pH of 65 degrees Celsius and 7.0 Protease U and B, when together, retained 100% activity at 65 degrees Celsius for 30 min. and 80% activity at pH 7.0-8.5 for 30 min. The enzymes activity was inhibited by metal chelating agents, EDTA. They can thus be classified as neutral-metalloproteases.