Cloning of fibrinolytic enzyme gene from Bacillus subtilis isolated from Cheonggukjang and its expression in protease-deficient Bacillus subtilis strains |
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Authors: | Jeong Seon-Ju Kwon Gun-Hee Chun Jiyeon Kim Jong Sang Park Cheon-Seok Kwon Dae Young Kim Jeong Hwan |
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Institution: | Division of Applied Life Science, Graduate School, Gyeongsang National University, Jinju 660-701, Korea. |
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Abstract: | Bacillus subtilis CH3-5 was isolated from cheonggukjang prepared according to traditional methods. CH3-5 secreted at least four different fibrinolytic proteases (63, 47, 29, and 20 kDa) into the culture medium. A fibrinolytic enzyme gene, aprE2, encoding a 29 kDa enzyme was cloned from the genomic DNA of CH3-5, and the DNA sequence determined. aprE2 was overexpressed in heterologous B. subtilis strains deficient in extracellular proteases using a E. coli-Bacillus shuttle vector. A 29 kDa AprE2 band was observed and AprE2 seemed to exhibit higher activities towards fibrin rather than casein. |
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