Transient Expression of Lumbrokinase (PI239) in Tobacco (Nicotiana tabacum) Using a Geminivirus-Based Single Replicon System Dissolves Fibrin and Blood Clots

文献类型： 外文期刊

第一作者： Dickey, Alexia

作者： Dickey, Alexia;Tull, Lauren;Breedlove, Drew;Wang, Kevin Yueju;Wang, Nan;Liu, Dehu;Cooper, Edwin;Mason, Hugh;Mason, Hugh

作者机构：

期刊名称：EVIDENCE-BASED COMPLEMENTARY AND ALTERNATIVE MEDICINE （ 影响因子：2.629； 五年影响因子：2.845 ）

ISSN： 1741-427X

年卷期： 2017 年

页码：

收录情况： SCI

摘要： Lumbrokinases, a group of fibrinolytic enzymes extracted from earthworm, have been widely used to prevent and treat various cardiovascular diseases. They specifically target fibrin to effectively degrade thrombi without major side effects. Plant expression systems are becoming potential alternative expression platforms for producing pharmaceutical proteins. In this work, a lumbrokinase (PI239) was produced from a plant system. Both wild-type (WT) and plant codon-optimized (OP) PI239 gene sequences were synthesized and cloned into a geminivirus-based single-vector DNA replicon system. Both vectors were independently expressed in tobacco (Nicotiana tabacum) leaves transiently by agroinfiltration. Overexpressed PI239 resulted in sudden tissue necrosis 3 days after infiltration. Remaining proteins were purified through His-tag affinity chromatography and analyzed with SDS-PAGE and Western blot methods. Purified PI239 successfully degraded artificial fibrin with relative activity of 13,400 U/mg when compared with commercial lumbrokinase product. In vitro tests demonstrated that plant-derived PI239 dissolved human blood clots and that the plant expression system is capable of producing functional PI239.

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