Microbial proteases are relevant biocatalysts with diverse applications. Production of protein hydrolysates is recently focused, since they might display biological activities. Therefore, the extracellular protease from Bacillus sp. CL18 was partially purified through ammonium sulfate precipitation (25-50% saturation) and gel filtration chromatography, with a 60.7-fold purification (40,593 U/mg protein) and 21.3% recovery. The partially purified protease (PPP) was characterized as a serine protease, with optimal activity at 51-59 degrees C and pH 7.4-8.8 and low thermal stability. Thermal inactivation followed first-order kinetics. PPP depended on Ca2+ for higher thermal stability, depicted by increases in half-lives (t(1/2)), activation energy (E-a), and free energy (Delta G(#)) for kinetic inactivation. PPP preferentially hydrolyzed casein > soy protein isolate (SPI) >>> keratinous materials. SPI hydrolysis by PPP was further investigated, and the obtained hydrolysates exhibited increased in vitro bioactivities. Hydrolysates displayed antioxidant capacities through the scavenging of synthetic organic radicals and Fe3+-reducing ability. In addition, hydrolysates inhibited the activities of dipeptidyl peptidase IV (DPP IV) and angiotensin-converting enzyme (ACE), suggesting antidiabetic and antihypertensive potentials, respectively. From its biochemical properties, PPP might be used to produce protein hydrolysates with multifunctional bioactivities. Both PPP and SPI hydrolysates can find applications in food biotechnology.