Isolation and identification of a fibrinolytic enzyme producing bacterium and its analysis of the fibrinolytic components

Abstract:

[Objective] To isolate and identify a fibrinolytic enzyme producing bacterium, and to analyze the component and characteristic of fibrinolytic enzyme. [Methods] Plasmin-producing bacterium was isolated by casein medium and AGAR-fibrin double-layer tablets from sea mud, soil, etc. Then its enzyme activity was measured by urokinase standard curve. The morphological, biochemical and physiological characteristics and 16S rDNA gene were analyzed to identify the taxonomic position of strain CNY16. The composition of the extracellular plasmin system was analyzed by SDS-PAGE and fiber protease zymogram. [Results] A strain named CNY16 with fibrinolytic activity was isolated and identified as Bacillus safensis. The results of SDS-PAGE and fiber protease zymogram showed that the extracellular fibrinolytic enzyme system contained at least two fibrinolytic enzymes with molecular masses of approximately 33 kD and 23 kD, respectively. The plasmin effectively dissolved fibrin clots, but did not hydrolyze blood cells. [Conclusion] CNY16 was a novel plasmin-producing bacterium, the fibrinolytic enzyme produced by strain CNY16 hold the valuable property in high activity and good stability. The strain might provide a new source for novel fibrinolytic enzymes.

Key words:Fibrinolytic enzyme, Bacillus safensis, Identification, Fiber protease zymogra