African Journal of
Biotechnology

  • Abbreviation: Afr. J. Biotechnol.
  • Language: English
  • ISSN: 1684-5315
  • DOI: 10.5897/AJB
  • Start Year: 2002
  • Published Articles: 12488

Full Length Research Paper

High-density fermentation and functional characterization of a recombinant echistatin mutant

Tao Yang1, Bo Niu1,2, Niuliang Cheng1, Jun Xie1 and Lijun Yang1*
  1Department of Biochemistry and Molecular Biology; Key Laboratory of Cellular Physiology (Shanxi Medical University), Ministry of Education, Taiyuan, Shanxi 030001, China. 2Capital Institute of Paediatrics, Beijing 100020, China.
Email: [email protected]

  •  Accepted: 12 May 2009
  •  Published: 06 July 2009

Abstract

 

A disintegrin echistatin mutant, R24K-echistatin, was obtained by overlap extension polymerase chain reaction. It was efficiently expressed as a soluble and fusion protein in Escherichia coli at optimized fermentation conditions. The recombinant bacteria had the highest wet weight of 120 g/l in 2´YT medium with IPTG induction. The R24K-echistatin was purified by chitin affinity chromatography and had a yield of 13 mg/l with DTT-cleavage. The purified R24K-echistatin inhibited the aggregation of platelet in vitro with IC50 of 21 nM which is better efficient than echistatin. This work lays the foundation for further research on specific anti-thrombus disintegrin agents.

 

Key words: Disintegrin, echistatin, Escherichia coli, fermentation, mutant.

Copyright © 2024 Author(s) retain the copyright of this article.

This article is published under the terms of the Creative Commons Attribution License 4.0

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