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Plant Cell Reports

, Volume 38, Issue 12, pp 1485–1499 | Cite as

Production of bacteriophage-encoded endolysin, LysP11, in Nicotiana benthamiana and its activity as a potent antimicrobial agent against Erysipelothrix rhusiopathiae

  • Md Reyazul Islam
  • Namil Son
  • Junho Lee
  • Dong Wook Lee
  • Eun-Ju Sohn
  • Inhwan HwangEmail author
Original Article

Abstract

Key message

We produced a biologically active phage-encoded endolysin, LysP11, in N. benthamiana. Plant-produced LysP11 exhibited robust antimicrobial activity against E. rhusiopathiae, and C-terminal domain of LysP11 bound specifically to E. rhusiopathiae.

Abstract

Bacterial resistance to antibiotics, a serious issue in terms of global public health, is one of the leading causes of death today. Thus, new antimicrobial agents are needed to combat pathogens. Recent research suggests that bacteriophages and endolysins derived from bacteriophages are potential alternatives to traditional antibiotics. Here, we examined the antimicrobial activity of LysP11, which is encoded by Propionibacterium phage P1.1 and comprises an N-terminal amidase-2 domain and a C-terminal domain with no homology to other bacteriophage endolysins. LysP11 was produced in Nicotiana benthamiana (N. benthamiana) using an Agrobacterium-mediated transient expression strategy. LysP11 was purified on microcrystalline cellulose-binding resin after attachment of the Clostridium thermocellum-derived family 3 cellulose-binding domain as an affinity tag. The affinity tag was removed using the small ubiquitin-related modifier (SUMO) domain and SUMO-specific protease. Plant-produced LysP11 showed strong antimicrobial activity toward Erysipelothrix rhusiopathiae (E. rhusiopathiae), mediated via lysis of the cell wall. Lytic activity was optimal at pH 8.0–9.0 (37 °C) and increased at higher concentrations of NaCl up to 400 mM. Furthermore, the C-terminal domain of LysP11 bound specifically to the E. rhusiopathiae cell wall. Based on these results, we propose that LysP11 is a potential candidate antimicrobial agent against E. rhusiopathiae.

Keywords

Plant-based expression systems Nicotiana benthamiana Endolysin LysP11 Cellulose-binding domain Erysipelothrix rhusiopathiae 

Notes

Acknowledgment

This work was supported by grants from the Ministry of Trade, Industry and Energy (MOTIE, Korea) under Industrial Technology Innovation Program (no. 10063301), Cooperative Research Program for Agriculture Science and Technology Development (no. PJ010953012019), Republic of Korea. Junho Lee and Dong Wook Lee supported by grants from the Next-Generation BioGreen 21 Program (nos. PJ01365001 and SSAC: PJ01335801 respectively), Rural Development Administration, Republic of Korea.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no competing interests to report.

Supplementary material

299_2019_2459_MOESM1_ESM.docx (4 mb)
Supplementary material 1 (DOCX 4106 kb)

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Md Reyazul Islam
    • 1
  • Namil Son
    • 1
  • Junho Lee
    • 1
  • Dong Wook Lee
    • 1
  • Eun-Ju Sohn
    • 3
  • Inhwan Hwang
    • 1
    • 2
    Email author
  1. 1.Division of Integrative Biosciences and BiotechnologyPohang University of Science and TechnologyPohangSouth Korea
  2. 2.Department of Life SciencesPohang University of Science and TechnologyPohangSouth Korea
  3. 3.BioApplications Inc.PohangSouth Korea

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