Song Tuo, Diao Xiaozhen, Cheng Jun, Man Yang, Chen Boyu, Zhang Haixing, Wu Wenhui
Department of Marine Biopharmacology, College of Food Science and Technology, Shanghai Ocean University, Shanghai 201306, China.
Solvo Biotherapeutics (Shanghai) Co., Ltd., Block 6, No.999 Huanke Road, Pudong New District, Shanghai 201210, China.
Bioengineering (Basel). 2024 Oct 15;11(10):1030. doi: 10.3390/bioengineering11101030.
As an essential thrombolytic agent, the tissue plasminogen activator receives increasing attention due to its longer half-life, lower immunogenicity, and easier administration, which are superior to other thrombolytic agents. In this study, the isolated and purified plasminogen activator from the sandworm () was expressed in . () to investigate its potential for simplifying the development process. The sandworm plasminogen activator was previously successfully cloned and expressed in . with low yield and activity in the culture supernatant. This low yield and activity prompted us to optimize its DNA sequence. Furthermore, to raise the efficiency in the separation of the target protein, the protein's solubility was enhanced by fusing it with maltose-binding protein (MBP) tags. Eventually, the fibrinolytic activity was successfully restored after digestion with tobacco etch virus (TEV) protease. This study provides an innovative method of efficiently expressing and purifying plasminogen activators from sandworm in . and broadens its applications in therapeutic treatment of cardiovascular diseases, including thrombosis, stroke, and coronary atherosclerotic heart disease.
作为一种重要的溶栓剂，组织型纤溶酶原激活剂因其半衰期长、免疫原性低且给药方便等优点，优于其他溶栓剂，因而受到越来越多的关注。在本研究中，从沙蚕（）中分离纯化的纤溶酶原激活剂在中表达（），以研究其简化开发过程的潜力。沙蚕纤溶酶原激活剂先前已成功克隆并在中表达，但培养上清液中的产量和活性较低。这种低产量和活性促使我们优化其DNA序列。此外，为了提高目标蛋白的分离效率，通过将其与麦芽糖结合蛋白（MBP）标签融合来增强蛋白的溶解性。最终，用烟草蚀纹病毒（TEV）蛋白酶消化后成功恢复了纤溶活性。本研究提供了一种在中高效表达和纯化沙蚕纤溶酶原激活剂的创新方法，并拓宽了其在心血管疾病治疗中的应用，包括血栓形成、中风和冠状动脉粥样硬化性心脏病。
