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Alkalophilic coryneform bacteria TU-19 isolated from soil extracellularly produced at least three proteases (Protease Ⅰ, Ⅱ, and Ⅲ). Investigating the cultural conditions related to the enzyme production of this bacterial cell, the optimum pH and temperature were 10.0 and 30℃, respectively. In order to purify these enzymes from the 2 day culture broth ammonium sulfate fractionation, gel filtration and QAE-Sephadex column chromatography were performed step by step. And then these three proteases were purified to near homogeneity by judging from SDS-PAGE pattern, and had the molecular weights of 120, 80, and 45 kilodaltons, respectively. The optimum pH and temperature for the enzyme activity of Protease Ⅰ and Ⅱ were 10.5 and 45℃, respectively, and Protease Ⅱ were 11.0 and 50℃. And the enzymes were completely inhibited by PMSF suggesting serine protease, but not affected by pCMB. 1,10-phenanthroline, IAA, and EDTA.
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