Novel strategy for protein production using a peptide tag derived from Bacillus thuringiensis Cry4Aa

Tohru Hayakawa, Shinya Sato, Shigehisa Iwamoto, Shigeo Sudo, Yoshiki Sakamoto, Takaaki Yamashita, Motoaki Uchida, Kenji Matsushima, Yohko Kashino, Hiroshi Sakai

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

Numerous proteins cannot be sufficiently prepared by ordinary recombinant DNA techniques because they are unstable or have deleterious effects on the host cell. One idea to prepare such proteins is to produce them as protein inclusions. Here we developed a novel system to effectively prepare proteins by using peptide tags derived from the insecticidal Cry toxin of a soil bacterium, Bacillus thuringiensis. Fusion with this peptide tag, designated 4AaCter, facilitates the formation of protein inclusions of glutathione S-transferase in Escherichia coli without losing the enzyme activity. Application of 4AaCter to the production of syphilis antigens TpN15, TpN17 and TpN47 from Treponema pallidum yielded excellent results, including a dramatic increase in the production level, simplification of the product purification and high reactivity with syphilis antibody. The use of 4AaCter may provide an innovational strategy for the efficient production of proteins.

Original languageEnglish
Pages (from-to)2883-2891
Number of pages9
JournalFEBS Journal
Volume277
Issue number13
DOIs
Publication statusPublished - Jul 1 2010

Keywords

  • 4AaCter peptide tag
  • Bacillus thuringiensis
  • Cry4Aa
  • Escherichia coli
  • TpN syphilis antigen

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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  • Cite this

    Hayakawa, T., Sato, S., Iwamoto, S., Sudo, S., Sakamoto, Y., Yamashita, T., Uchida, M., Matsushima, K., Kashino, Y., & Sakai, H. (2010). Novel strategy for protein production using a peptide tag derived from Bacillus thuringiensis Cry4Aa. FEBS Journal, 277(13), 2883-2891. https://doi.org/10.1111/j.1742-4658.2010.07704.x