Smitha K V, Pradeep B V
Department of Microbiology, SAFI Institute of Advanced Study, Malappuram, Kerala, 673 633, India.
Department of Microbiology, Karpagam Academy of Higher Education, Coimbatore, Tamil Nadu, 641021, India.
Curr Microbiol. 2025 Mar 4;82(4):166. doi: 10.1007/s00284-025-04141-0.
Thrombosis, the major cause of heart disease, is on the rise owing to the current lifestyle habits of the population. Current treatments using thrombolytics, although successful, are plagued by side effects and costs. Fibrinolytic enzymes derived from microbial sources are ideal substitutes for chemical thrombolytics. Bacillus altitudinis S-CSR 0020, a soil bacterium, was shown to efficiently produce fibrinolytic enzyme. In this study, we further purified the enzyme using a combination of ammonium sulfate precipitation and ion-exchange chromatography to achieve a purification fold of 7.8 and a yield of 7%. SDS-PAGE and zymogram analysis revealed that the enzyme was 95 kDa in size and exhibited high stability across a broad pH range (6-11) and high temperature (30-60 ℃) even after 24 h incubation, along with an optimum pH and temperature of 9 and 40 °C, respectively. Further characterization revealed that the fibrinolytic activity was maximum at 4% fibrin concentration, and 3% ferric chloride and 5% β-mercaptoethanol enhanced enzyme activity. Functional analysis revealed that the purified enzyme efficiently removed preformed clots in the capillary and microfuge tubes and exhibited anticoagulant activity. No adverse effects were observed in the goat hepatic portal vein upon application of the fibrinolytic enzyme. This enzyme also displayed antibacterial activity against Staphylococcus aureus. The anticoagulant, clot-degrading potential, and non-toxic nature of the purified fibrinolytic enzyme highlight its potential application in the treatment of thrombosis.
血栓形成是心脏病的主要病因，由于当前人群的生活方式习惯，其发病率正在上升。目前使用溶栓剂的治疗方法虽然取得了成功，但却受到副作用和成本的困扰。源自微生物的纤维蛋白溶解酶是化学溶栓剂的理想替代品。土壤细菌高海拔芽孢杆菌S-CSR 0020被证明能有效产生纤维蛋白溶解酶。在本研究中，我们使用硫酸铵沉淀和离子交换色谱相结合的方法进一步纯化该酶，纯化倍数达到7.8，产率为7%。SDS-PAGE和酶谱分析表明，该酶大小为95 kDa，即使在24小时孵育后，在较宽的pH范围（6-11）和高温（30-60℃）下仍表现出高稳定性，最佳pH和温度分别为9和40℃。进一步表征表明，纤维蛋白溶解活性在纤维蛋白浓度为4%时最高，3%的氯化铁和5%的β-巯基乙醇可增强酶活性。功能分析表明，纯化后的酶能有效去除毛细管和微量离心管中预先形成的血栓，并具有抗凝活性。在山羊肝门静脉中应用纤维蛋白溶解酶未观察到不良反应。该酶还对金黄色葡萄球菌表现出抗菌活性。纯化后的纤维蛋白溶解酶的抗凝、降解血栓潜力和无毒性质突出了其在血栓治疗中的潜在应用价值。
