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中国生物工程杂志

China Biotechnology
China Biotechnology  2010, Vol. 30 Issue (08): 47-51    DOI: Q819
    
Purification and Characterization of a Novel Fibrinolytic Enzyme from a marine animal,Urechis unicinctus
WANG Dian-liang1,2**,JIANG He-zuo1,WANG Rui-ling1,LIU Wan-shun2,ZHANG Yan-mei1,SUN Jin-wei1
1.Central Laboratory, the Second Artillery General Hospital, PLA, Beijing 100088,China
2.Department of Marine Bioengineering, Ocean University of China, Qingdao 266003, China
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Abstract  

Urechis unicinctus living in the inter-tidal and hypotidal zone of the lower coastal mud-sand areas is a member of the group of worms classified as the Phylum Echiura since 1940, but increasingly thought to represent a clade within the polychaete annelids. A systematically analysis revealed that the highest thrombolytic activity was found in the haemocoelomic fluid and gut. Therefore the isolation of  fibrinolytic enzyme  (UFE)was undergone from the two tissues by centrifugation, filtration, ammonium sulfate precipitation, ultrafiltration, and chromatographic steps, including Q. Sepharose Fast Flow, Sephadex G-75, and Sephacry S-100. UFE was finally purified approximately 11 fold with a yield of 29.82%. UFE appeared to be homogenous by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) with a low molecular weight of about 10.38 kDa.

Functional studies showed that using urokinase (UK) as a positive control and phosphate buffer saline (PBS) as a negative control, UFE mainly hydrolyzed fibrin directly, but it also exihibited some UK activity. UFE appeared to degrade fibrin by transformation of plasminogen into plasmin, which was  similar to lumbrokinase from different earthworms, whereas UK did  not directly hydrolyze fibrin  without the  presence of active plasminogen. These suggest that UFE  is a potent thrombolytic agent with less immunogenicity and strong fibrinolysis for treatment of thrombosis.



Key wordsUrechis unicincts      Marine worm      Fibrinolytic enzyme      Thrombolytic agent      Thrombotic disease     
Received: 18 March 2010      Published: 25 August 2010
Cite this article:

WANG Dian-Liang, JIANG Ge-Zuo, WANG Rui-Ling, LIU Mo-Shun, ZHANG Yan-Mei, SUN Jin-Wei. Purification and Characterization of a Novel Fibrinolytic Enzyme from a marine animal,Urechis unicinctus. China Biotechnology, 2010, 30(08): 47-51.

URL:

https://manu60.magtech.com.cn/biotech/Q819     OR     https://manu60.magtech.com.cn/biotech/Y2010/V30/I08/47

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