Developing an effective fibrinolytic drug for treating thrombolysis with minimal undesirable side effects is of great importance. In the current study, an optimum solvent was selected for the extraction of fibrinolytic active components. Furthermore, a strong fibrinolytic enzyme named WPI01 was purified from Whitmania pigra Whitman through various chromatographic steps. WPI01 has a molecular mass of 27044.297 Da, and the N-terminal 8 amino acid sequence was determined as VVGGVEAR. WPI01 was stable within the pH range of 6.0–10.0 and with maximum fibrinolytic activity at 40 °C and a pH of 8.0. At 500 U/mL, WPI01 induced 50.59% blood clot reduction in vitro within 6 h, which was higher than that induced by urokinase at 1000 U/mL. In an analysis of the plasminogen activator activity, WPI01 produced obvious halos on heated and unheated fibrin plates, suggesting that WPI01 may not only act as a plasminogen activator but also degrade fibrin clots directly, and more study is needed to support this. In conclusion, WPI01 is obviously different from known fibrinolytic enzymes in terms of substrate specificity and fibrinolytic mode of action, suggesting that it is a novel fibrinolytic enzyme with potential applications in the treatment and prevention of thrombosis.

开发有效的纤溶药物以最小的不良副作用来治疗血栓溶解是非常重要的。在当前的研究中，选择了一种最佳的溶剂来提取纤维蛋白溶解活性成分。此外，从Whitmania pigra Whitman通过各种色谱步骤纯化了一种名为WPI01的强纤溶酶。WPI01的分子量为27044.297 Da，N端8个氨基酸序列确定为VVGGVEAR。WPI01在6.0-10.0的pH范围内稳定，在40°C和8.0的pH下具有最大的纤溶活性。在500 U / mL下，WPI01在体外可导致50.59％的血块减少6 h内，高于尿激酶1000 U / mL诱导的水平。在对纤溶酶原激活剂活性的分析中，WPI01在加热和未加热的纤维蛋白板上均产生明显的光晕，这表明WPI01不仅可以充当纤溶酶原激活剂，而且可以直接降解纤维蛋白凝块，需要更多的研究来支持这一点。总之，WPI01在底物特异性和纤溶作用方式方面与已知的纤溶酶明显不同，这表明它是一种新型的纤溶酶，在治疗和预防血栓形成方面具有潜在的应用前景。