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

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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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Acknowledgments

This study was financially supported by the 863 (Hi-tech research and development program of China) program under contract No. 2008AA10Z336.

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Authors and Affiliations

  1. Key Laboratory of Industrial Microbiology, Ministry of Education, Tianjin, People’s Republic of China

    Xiao-Lan Jing, Xue-Gang Luo & Wen-Jing Tian

  2. College of Bioengineering, Tianjin University of Science and Technology, Tianjin, People’s Republic of China

    Xiao-Lan Jing, Xue-Gang Luo, Li-Hui Lv, Yong Jiang, Nan Wang & Tong-Cun Zhang

  3. Tianjin Key Laboratory of Industrial Microbiology, No. 29, 13th Street, Economic-Technological Development Area, 300457, Tianjin, People’s Republic of China

    Tong-Cun Zhang

  4. School of Medicine, Wuhan University of Science and Technology, 430080, Wuhan, Hubei Province, People’s Republic of China

    Tong-Cun Zhang

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  1. Xiao-Lan Jing
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  2. Xue-Gang Luo
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  3. Wen-Jing Tian
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  4. Li-Hui Lv
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  5. Yong Jiang
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  6. Nan Wang
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  7. Tong-Cun Zhang
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Corresponding author

Correspondence to Tong-Cun Zhang.

Additional information

Xiao-Lan Jing and Xue-Gang Luo contributed equally to the project.

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Jing, XL., Luo, XG., Tian, WJ. et al. High-Level Expression of the Antimicrobial Peptide Plectasin in Escherichia coli . Curr Microbiol 61, 197–202 (2010). https://doi.org/10.1007/s00284-010-9596-3

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  • DOI: https://doi.org/10.1007/s00284-010-9596-3

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