Xu Zhe-rong, Yang Yun-mei, Gui Qi-feng, Zhang Li-na, Hu Lin
Department of Geriatrics, First Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou 310003, China.
Protein Expr Purif. 2010 Feb;69(2):198-203. doi: 10.1016/j.pep.2009.08.013. Epub 2009 Aug 28.
Lumbrokinase (LK) is an important fibrinolytic enzyme derived from earthworms. It has been found that LK is composed of a group of isoenzymes. To construct and express the mature peptide of LK PI239 in Escherichia coli, we amplified and optimized the gene of LK which was then cloned into the prokaryotic expression vector pET-22b(-). The recombinant LK (rLK) protein was expressed as inclusion bodies and we have developed a purification process of rLK from these inclusion bodies. A step-down urea concentration strategy was applied to the rLK renaturation process. The purified and renatured rLK apparently ameliorated the conditions of the model thrombosis rats used, and may be developed into a therapeutic agent for thrombotic-associated diseases.
蚓激酶（LK）是一种从蚯蚓中提取的重要纤溶酶。已发现LK由一组同工酶组成。为了在大肠杆菌中构建并表达LK PI239的成熟肽，我们对LK基因进行了扩增和优化，然后将其克隆到原核表达载体pET-22b(-)中。重组LK（rLK）蛋白以包涵体形式表达，我们开发了从这些包涵体中纯化rLK的工艺。在rLK复性过程中采用了逐步降低尿素浓度的策略。纯化并复性后的rLK明显改善了所使用的模型血栓大鼠的状况，有望开发成为治疗血栓相关疾病的药物。
