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Web End = Biotechnol Lett (2016) 38:251258 DOI 10.1007/s10529-015-1979-x
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Web End = Rapid extraction and purication of lumbrokinasefrom Lumbricus rubellus using a hollow ber membrane and size exclusion chromatography
Tingming Fu . Fengyun Yang . Huaming Zhu . Huaxu Zhu . Liwei Guo
Received: 5 August 2015 / Accepted: 12 October 2015 / Published online: 19 October 2015 Springer Science+Business Media Dordrecht 2015
AbstractObjectives To develop a purication process using hollow ber membrane separation combined with size exclusion chromatography for the extraction of lumbrokinase from earthworms (Lumbricus rubellus). Results To extract the protein, the earthworms were rst homogenized for 10 min, to produce ultrane particles. Polyether sulfone hollow ber membranes with MW cut offs of 50 and 6 kDa were used for initial purication of the crude extract. Further purication was carried out on a Sephadex G-75 column, and yielded three fractions of high purity protein. One of these fractions showed brinolytic activity. Conclusions Three samples of high purity protein were obtained and one protein (LK1) showed strong brinolytic activity. The method has higher purication efciency in comparison with existing methods.
Keywords Earthworms Hollow ber membrane
Fibrinolytic activity Lumbrokinase Lumbricus
rubellus
Introduction
Thrombosis is a major cause of morbidity and mortality worldwide. It plays an important role in the pathogenesis of numerous cardiovascular disorders, such as acute coronary syndrome (e.g. unstable angina and myocardial infarction), sudden cardiac death, peripheral arterial occlusion, ischemic stroke, deep vein thrombosis, and pulmonary embolism (Ruppert et al. 2010). Treatment of venous thrombosis currently relies on the use of anti-platelet drugs, anticoagulants, and thrombolytic agents. Urokinase and streptokinase are the most commonly used thrombolytic drugs, but cannot be administered orally (Prasad et al. 2006).
A novel thrombolytic agent, lumbrokinase, has been isolated from some earthworm species and thoroughly characterized as a serine protease isoenzyme (Wang et al. 2005). Compared with urokinase and streptokinase, lumbrokinase has higher thermal stability and alkali resistance. More importantly, it can be administered orally (Lee et al. 2007).
Purication of lumbrokinase requires multiple steps (Choi and Sa 2001), with a...