Lumbrokinase derived from earthworms, Lumbricus rubellus is known to have fibrinolytic enzymes that have potential as therapeutic drugs due to its ability to dissolve fibrin. The current study is aimed to purify the Lumbrokinase from L. rubellus and identify its protein component. Water extract of local earthworm Lumbricus rubellus revealed several proteins. Therefore, to identify its protein component, purification through HiPrep DEAE fast flow and proteomic analysis were conducted prior to identifications. A combination of two-dimension gel electrophoresis (2DE) and electrospray ionization mass spectrometry analysis was used to identify the purified fractions. The purified fractions contain five protein bands, namely F25-1, F25-2, F85-1, F85-2, and F85-3, which displayed strong fibrinogenolytic activity. F25 fractions showed fibrinogenolytic activity of 974.85 U/mg, while F85 fractions showed higher activity of 1,484.11 U/mg. Fractions F85-1, F85-2, and F85-3 showed molecular weights of 42.6 kDa, 27.03 kDa, and 14 kDa, respectively and were identified as Lumbrokinase iso-enzymes. This preliminary study indicates that the F25 and F85 fractions are similar to published fibrinolytic protease-1 and lumbrokinase, respectively, in terms of their amino acid sequence.

中文翻译：

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从 Lumbricus rubellus 中提取的强效纤溶酶的纯化和蛋白质组学分析

来自蚯蚓的蚓激酶，已知 Lumbricus rubellus 具有纤维蛋白溶解酶，由于其溶解纤维蛋白的能力，具有作为治疗药物的潜力。目前的研究旨在从风疹杆菌中纯化出蚓激酶并鉴定其蛋白质成分。当地蚯蚓 Lumbricus rubellus 的水提取物揭示了几种蛋白质。因此，为了鉴定其蛋白质成分，在鉴定之前通过 HiPrep DEAE 快速流动和蛋白质组学分析进行了纯化。二维凝胶电泳 (2DE) 和电喷雾电离质谱分析的组合用于鉴定纯化的级分。纯化的级分包含五个蛋白质条带，即 F25-1、F25-2、F85-1、F85-2 和 F85-3，它们显示出很强的纤维蛋白原溶解活性。F25 组分显示纤维蛋白原溶解活性为 974.85 U/mg，而 F85 组分显示更高的活性，为 1,484.11 U/mg。组分 F85-1、F85-2 和 F85-3 的分子量分别为 42.6 kDa、27.03 kDa 和 14 kDa，被鉴定为蚓激酶同工酶。这项初步研究表明，F25 和 F85 组分在氨基酸序列方面分别与已发表的纤溶蛋白酶 1 和蚓激酶相似。