Screening of High-yield Fibrinolytic Enzyme-producing Strains from Traditional Fermented Douchi (Honghe, Yunnan) and Analysis of the Gene Encoding the Fibrinolytic Enzyme
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Abstract:
High-yield fibrinolytic enzyme-producing bacterial strains were isolated and screened from traditionally fermented douchi from Honghe, by a two-step screening method, in which skim milk agar and fibrin agar were used. The bacterial gene encoding the fibrinolytic enzyme was cloned and analyzed to provide bacterial reference strains and a theoretical basis to develop a new version of functional douchi. Using these methods, a high-yield fibrinolytic enzyme-producing strain Bacillus subtilis LC-2-1 was successfully isolated, which secreted a kind of douchi fibrinolytic enzyme. The molecular mass of the enzyme was 27.4 kDa that contained 275 amino acids encoded by 825-bp. Compared to other douchi fibrinolytic enzymes and nattokinase, the enzyme produced by B. subtilis LC-2-1 showed significant differences, and the amino acid sequence homology was 85.1%. Additionally, the enzyme had high fibrinolytic activity, which was up to 79.84 U/mL. Thus, these results could provide a bacterial reference strain and theoretical basis to develop a new version of fermented douchi with thrombolytic activity.
