Current Microbiology

, Volume 61, Issue 3, pp 197–202

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

  • Xiao-Lan Jing
  • Xue-Gang Luo
  • Wen-Jing Tian
  • Li-Hui Lv
  • Yong Jiang
  • Nan Wang
  • Tong-Cun Zhang
Article

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.

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Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • Xiao-Lan Jing
    • 1
    • 2
  • Xue-Gang Luo
    • 1
    • 2
  • Wen-Jing Tian
    • 1
  • Li-Hui Lv
    • 2
  • Yong Jiang
    • 2
  • Nan Wang
    • 2
  • Tong-Cun Zhang
    • 2
    • 3
    • 4
  1. 1.Key Laboratory of Industrial Microbiology, Ministry of EducationTianjinPeople’s Republic of China
  2. 2.College of BioengineeringTianjin University of Science and TechnologyTianjinPeople’s Republic of China
  3. 3.Tianjin Key Laboratory of Industrial MicrobiologyTianjinPeople’s Republic of China
  4. 4.School of MedicineWuhan University of Science and TechnologyWuhan, Hubei ProvincePeople’s Republic of China

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