Current Microbiology

, Volume 61, Issue 3, pp 197–202

High-Level Expression of the Antimicrobial Peptide Plectasin in Escherichia coli

Authors

  • Xiao-Lan Jing
    • Key Laboratory of Industrial Microbiology, Ministry of Education
    • College of BioengineeringTianjin University of Science and Technology
  • Xue-Gang Luo
    • Key Laboratory of Industrial Microbiology, Ministry of Education
    • College of BioengineeringTianjin University of Science and Technology
  • Wen-Jing Tian
    • Key Laboratory of Industrial Microbiology, Ministry of Education
  • Li-Hui Lv
    • College of BioengineeringTianjin University of Science and Technology
  • Yong Jiang
    • College of BioengineeringTianjin University of Science and Technology
  • Nan Wang
    • College of BioengineeringTianjin University of Science and Technology
    • College of BioengineeringTianjin University of Science and Technology
    • Tianjin Key Laboratory of Industrial Microbiology
    • School of MedicineWuhan University of Science and Technology
Article

DOI: 10.1007/s00284-010-9596-3

Cite this article as:
Jing, X., Luo, X., Tian, W. et al. Curr Microbiol (2010) 61: 197. doi:10.1007/s00284-010-9596-3

Abstract

Plectasin is a defensin-like antimicrobial peptide isolated from a fungus, the saprophytic ascomycete Pseudoplectania nigrella. Plectasin showed marked antibacterial activity in vitro against Gram-positive bacteria, especially Streptococcus pneumoniae, including strains resistant to conventional antibiotics. Plectasin could kill the sensitive strain as efficaciously as vancomycin and penicillin and without cytotoxic effects on mammalian cell viability. In order to establish a bacterium-based plectasin production system, in the present study, the coding sequence of plectasin was optimized, and then cloned into pET32a (+) vector and expressed as a thioredoxin (Trx) fusion protein in Escherichia coli. The soluble fusion protein collected from the supernatant of the cell lysate was separated by Ni2+-chelating affinity chromatography. The purified protein was then cleaved by Factor Xa protease to release mature plectasin. Final purification was achieved by Ni2+-chelating chromatography again. The recombinant plectasin exhibited the same antimicrobial activity as reported previously. This is the first study to describe the expression of plectasin in E. coli expression system, and these works might provide a significant foundation for the following production or study of plectasin, and contribute to the development and evolution of novel antimicrobial drugs in clinical applications.

Copyright information

© Springer Science+Business Media, LLC 2010