Purification and Characterization of a Novel Fibrinolytic Enzyme from Cipangopaludina Cahayensis

BACKGROUND: Cipangopaludina cahayensis contains active fibrinolytic proteins and has been considered a potential anti-cancer agent. However, its anti-cancer characteristics and functions have yet to be elucidated OBJECTIVES: To study the fibrinolytic activity and anticancer activity of crude protein extracts from Cipangopaludina cahayensis. MATERIALS AND METHODS: Crude proteases were separated and extracted from the Cipangopaludina cahayensis through homogenization, desalting, ammonium sulfate fractionation, dialysis, and ion exchange chromatography. The fibrinolytic activity of extracted proteins was assessed using the fiber plate method. Total protein concentrations of the crude proteases were determined via BCA assay. Molecular weights (MWs) were determined through SDS-PAGE electrophoresis. RESULTS: The crude extract had a MW of ~ 50 kDa, and the highest protein concentration was 3.026 mg.mL(-1). The optimum pH for fibrinolytic activity was 7.0. Cell culture assays demonstrated that the addition of the crude enzyme extracts to the human ovary cancer cell line Ovcar-3 resulted in significant growth defects. CONCLUSIONS: Our data showed that crude proteins purified from Cipangopaludina cahayensis are novel fibrinolytic proteases and have potential anti-cancer propertie