International Journal of Industrial Entomology and Biomaterials

Korean Society of Sericultural Science (한국잠사학회)
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Expression of Active Antibacterial Bumblebee Abaecin in Escherichia coli Cells

  • Kim, Seong-Ryul (Department of Agricultural Biology, NIAST, Rural Development Administration) ;
  • Hwang, Jae-Sam (Department of Agricultural Biology, NIAST, Rural Development Administration) ;
  • Yoon, Hyung-Joo (Department of Agricultural Biology, NIAST, Rural Development Administration) ;
  • Park, Kwan-Ho (Department of Agricultural Biology, NIAST, Rural Development Administration) ;
  • Hong, Mee-Yeon (College of Agriculture & Life Sciences, Chonnam National University) ;
  • Kim, Kee-Young (Department of Agricultural Biology, NIAST, Rural Development Administration) ;
  • Jin, Byung-Rae (College of Natural Resources and Life Science, Dong-A University) ;
  • Kim, Ik-Soo (College of Agriculture & Life Sciences, Chonnam National University)
  • Published : 2008.09.30
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Abstract

We previously isolated and cloned a cDNA of abaecin from the Bombus ignitus. In an effort to produce a large amount of soluble abaecin at low cost, we successfully expressed the peptide in Escherichia coli that are highly sensitive to its mature form. For this, we fused the peptide encoding 39 amino acids of mature B. ignitus abaecin to the thioredoxin gene together with a C-terminal 6xHis tag. An enterokinase cleavage site was introduced between the 6xHis tag and mature abaecin to allow final release of the recombinant peptide. A high yield of 9.6 mg soluble fusion protein from 200 ml of bacterial culture was purified by $Ni^{2+}$-charged His-Bind resin affinity column, and 1.4 mg of pure active recombinant abaecin was readily obtained by enterokinase cleavage, followed by affinity chromatograph. The molecular mass of recombinant abaecin peptide was determined by Tricin-SDS-PAGE analysis. The recombinant abaecin exhibited antibacterial activity against Gram-negative bacteria.

Keywords

References

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