Expression and characterization of earthworm Eisenia foetida lumbrokinase-3 in Pichia pastoris |
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Authors: | Yuan Xiaoling Cao Chenhua Shan Yajun Zhao Zhenhu Chen Jiapei Cong Yuwen |
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Affiliation: | Department of Pathophysiology, Beijing Institute of Radiation Medicine, Beijing, P. R. China. |
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Abstract: | Lumbrokinase-3 (LK-3, AY438622), first cloned from the earthworm Eisenia foetida in our laboratory, is a component of earthworm fibrinolytic enzymes. In this study, cDNA encoding the LK-3 gene was sub-cloned into yeast pPIC9K expression vector and transformed into the Pichia pastoris GS115 cells by electroporation. High level expression of LK-3 in yeast cells was confirmed with a different induction time. The activity of expressed LK-3 was observed in fibrin plates. In addition, the expressed LK-3 protein could dissolve fibrinogen and bovine serum albumin. The use of this system for the high level production of biological protein is implicated from this study. |
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