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http://dx.doi.org/10.5352/JLS.2005.15.4.513

Cloning and Characterization of a Gene for Fibrinolytic Enzyme from Bacillus subtilis BB-1 Isolated from Black Bean Chung-kuk  

Lee Young-Hoon (Department of Microbiological Engineering, Jinju National University)
Lee Sung-Ho (Environmental Biotechnology National Core Research center, Gyeongsang National University)
Jeon Ju-Mi (Environmental Biotechnology National Core Research center, Gyeongsang National University)
Kim Hong-Chul (Department of Microbiological Engineering, Jinju National University)
Cho Yong-Un (Department of Microbiological Engineering, Jinju National University)
Park Ki-Hoon (Department of Applied Life Science, Gyeongsang National University)
Choi Young-Ju (Department of Food and Nutrition, Silla University)
Gal Sang-Wan (Department of Microbiological Engineering, Jinju National University)
Publication Information
Journal of Life Science / v.15, no.4, 2005 , pp. 513-521 More about this Journal
Abstract
A bacterium producing five fibrinolytic isozymes was isolated from black bean chung kuk. The bacterium was identified as Bacillus subtilis BB-1 by 16s rDNA sequence homology search. A gene out of five fibrinolytic genes in the Bacillus subtilis BB-1 was cloned by shot-gun method. A Cla I DNA fragment of B. subtilis BB-1 chromosome was cloned in to pBluescript II SK(-) and showed the fibrinolytic activity to bacterial cells. The Cla I DNA fragment was sequenced and the sequences did not show homology with gene for protease or fibrinolytic enzyme genes in other organisms. The Cla I DNA fragment was reduced to 2,142 bp by activity-guided PCR cloning method. The optimum pH and temperature of the enzyme were 5.0 and $35^{\circ}C$, respectively. Substrate specificity of the fibrinolytic enzyme was detected in skim milk, casein, gelatin and blood agar plates. The activity of the enzyme was not detected with these substrates. Taken together, this enzyme is a new fibrinolytic enzyme and may be used to prevent thrombosis and arteriosclerosis.
Keywords
Clung-kuk; Bacillus subtilis BB-1 (KFCC 1134P); fibrinolytic enzyme; fibrinolytic isozymes; deletion mutation;
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