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

  • T Yang
  • B Niu
  • N Cheng
  • J Xie
  • L Yang
Keywords: Disintegrin, echistatin, Escherichia coli, fermentation, mutant.

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.
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eISSN: 1684-5315