Applied Microbiology and Biotechnology

, Volume 98, Issue 16, pp 6973–6982

Expression and purification of cyto-insectotoxin (Cit1a) using silkworm larvae targeting for an antimicrobial therapeutic agent

Authors

  • M. P. Ali
    • Laboratory of Biotechnology, Integrated Bioscience Section, Graduate School of Science and TechnologyShizuoka University
  • Katsuhiko Yoshimatsu
    • Research Institute of Green Science and TechnologyShizuoka University
  • Tomohiro Suzuki
    • Research Institute of Green Science and TechnologyShizuoka University
  • Tatsuya Kato
    • Research Institute of Green Science and TechnologyShizuoka University
    • Laboratory of Biotechnology, Integrated Bioscience Section, Graduate School of Science and TechnologyShizuoka University
    • Research Institute of Green Science and TechnologyShizuoka University
Biotechnological products and process engineering

DOI: 10.1007/s00253-014-5728-1

Cite this article as:
Ali, M.P., Yoshimatsu, K., Suzuki, T. et al. Appl Microbiol Biotechnol (2014) 98: 6973. doi:10.1007/s00253-014-5728-1

Abstract

Antimicrobial peptides (AMPs), both synthetic and from natural sources, have raised interest recently as potential alternatives to antibiotics. Cyto-insectotoxin (Cit1a) is a 69-amino-acid antimicrobial peptide isolated from the venom of the central Asian spider Lachesana tarabaevi. The synthetic gene Cit1a fused with the enhanced green fluorescent protein (EGFP) gene was expressed as the EGFP-Cit1a fusion protein using a cysteine protease-deleted Bombyx mori nucleopolyhedrovirus (BmNPV-CP) bacmid in silkworm larva and pupa. The antimicrobial effect of the purified protein was assayed using disk diffusion and broth microdilution methods. The minimum inhibitory concentration of EGFP-Cit1a was also measured against several bacterial strains and showed similar antimicrobial activity to that of the synthetic Cit1a reported earlier. The EGFP-Cit1a fusion protein showed antibiotic activity toward gram-positive and gram-negative bacteria at the micromolar concentration level. These results show that active Cit1a can be produced and purified in silkworm, although this peptide is insecticidal. This study demonstrates the potential of active Cit1a purified from silkworms to use as an antimicrobial agent.

Keywords

Cyto-insectotoxin (Cit1a)Antimicrobial peptidesSilkwormBombyx mori nucleopolyhedrovirus

Copyright information

© Springer-Verlag Berlin Heidelberg 2014