Codon optimization increases human kallistatin expression in escherichia coli

Zhiyu Dai, Yifei Chen, Weiwei Qi, Lijun Huang, Yang Zhang, Ti Zhou, Xia Yang, Guoquan Gao*

*Corresponding author for this work

Research output: Contribution to journalArticle

5 Scopus citations

Abstract

A unique serpin, kallistatin, displays vasodilatory, antiangiogenic, anti-inflammatory, and antioxidant activity. Difficulty and low efficacy of obtaining recombinant kallistatin limit the wide investigation of its biological and pathological function. The present study employed a codon optimization algorithm to redesign the kallistatin gene and achieved a high yield of recombinant kallistatin protein. The kallistatin codons were redesigned for a more suitable Escherichia coli host without altering amino acids. Base composition and GC% content were compared between synthetic optimized kallistatin (opti-kallistatin) and wild-type kallistatin (wt-kallistatin). Both opti-kallistatin and wt-kallistatin were purified using Ni-NTA His-binding resins through fast protein liquid chromatography (FPLC). The identity and purity of kallistatin were confirmed by Coomassie blue staining, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), and Western blot analysis. The output of opti-kallistatin protein was ∼2-fold increase (2.09±0.23mg/L) compared to wt-kallistatin (1.05±0.2mg/L). These results suggest that more common codon optimization in the E. coli host significantly increases the yield of heterologous human protein yields. This approach will remarkably facilitate the further investigation of kallistatin in vitro and in vivo.

Original languageEnglish (US)
Pages (from-to)123-136
Number of pages14
JournalPreparative Biochemistry and Biotechnology
Volume43
Issue number1
DOIs
StatePublished - Jan 1 2013

Keywords

  • Kallistatin
  • angiogenesis
  • codon optimization
  • human KBP
  • protein purification
  • serpin

ASJC Scopus subject areas

  • Biotechnology
  • Biochemistry

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