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Title:
A THROBIN-LIKE ENZYME OF AGKISTRODON ACUTUS
Document Type and Number:
WIPO Patent Application WO/2010/034176
Kind Code:
A1
Abstract:
A thrombin-like enzyme isolated from Agkistrodon acutus venom, comprising an alpha subunit having a defined sequence of SEQ ID NO.1 given in the description and a beta subunit having a defined sequence of SEQ ID NO. 2 given in the description, interconnected by seven disulfide linkages, is provided. The enzyme of present invention is a serine proteinase capable of hydrolyzing alpha chain of human fibrinogen, and has a molecular weight of 29.3~29.5kD and an isoelectric point of 5.5. Methods for purifying such enzyme, comprising removing insoluble substance of the venom by pretreatment, conducting twice of anion-exchange chromatography, collecting active eluting peak, dialysing, ultrafiltrating and desalinating, are also provided. The thrombin-like enzyme obtained from said methods has a specific activity of no less then 180u/mg protein, a HPLC purity of more than 95% and a yield of 0.7%-0.8% based on weight of venom raw material.

Inventors:
SUN DI (CN)
WANG XIJUAN (CN)
Application Number:
PCT/CN2009/000230
Publication Date:
April 01, 2010
Filing Date:
March 04, 2009
Export Citation:
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Assignee:
HEALTHSTAR PHARMACEUTICAL CO L (CN)
SUN DI (CN)
WANG XIJUAN (CN)
International Classes:
A61K9/12; C12N9/64; A61K9/19; A61K9/70; A61K38/43; A61P7/04
Domestic Patent References:
WO2005017139A12005-02-24
Foreign References:
CN101358184A2009-02-04
CN1332241A2002-01-23
CN1332242A2002-01-23
CN1502693A2004-06-09
Other References:
TANI A. ET AL: "Characterization, primary structure and molecular evolution of anticoagulant protein from Agkistrodon actus venom", TOXICON, vol. 40, 31 December 2002 (2002-12-31), pages 803 - 813, XP008145772
CHENG X. ET AL: "Purification, characterization, and cDNA cloning of a new fibrinogenlytic venom protein, Agkisacutacin, from Agkistrodon acutus venom", BIOCHEM. BIOPHYS. RES. COMMUN., vol. 265, 31 December 1999 (1999-12-31), pages 530 - 535, XP008145795
ZHANG YANYU ET AL: "Cloning and Bioinformatics Analysis of a Thrombin-like Enzyme Gene from Agkistrodon Acutus", JOURNAL OF EXPERIMENTAL HEMATOLOGY, vol. 13, no. 4, 31 December 2005 (2005-12-31), pages 542 - 547, XP008145774
LIANQ NINGSHENG ET AL: "Molecular Cloning of the New cDNAfor the Thrombin-Like Enzyme form the Venom of Agkistrodon Acutus", JOURNAL OF GUANGXI MEDICAL UNIVERSITY, vol. 19, no. 1, 28 February 2002 (2002-02-28), pages 27 - 30, XP008145793
Attorney, Agent or Firm:
CN-KNOWHOW INTELLECTUAL PROPERTY AGENT LIMITED (Fortune International CenterNo. 17 Daliushu Rd, Beijing 1, CN)
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