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Applied Microbiology and Biotechnology

, Volume 100, Issue 21, pp 9283–9293 | Cite as

Identification of antibacterial peptides from endophytic microbiome

  • M. V. TejesviEmail author
  • P. Picart
  • M. Kajula
  • H. Hautajärvi
  • L. Ruddock
  • H.H. Kristensen
  • A. Tossi
  • H.G. Sahl
  • S. Ek
  • S. Mattila
  • A. M. Pirttilä
Applied microbial and cell physiology

Abstract

Endophytes, microorganisms living inside plant tissues, are promising producers of lead compounds for the pharmaceutical industry. However, the majority of endophytes are unculturable and therefore inaccessible for functional studies. To evaluate genetic resources of endophytes, we analyzed the biodiversity of fungal microbiome of black crowberry (Empetrum nigrum L.) by next-generation sequencing and found that it consists mainly of unknown taxa. We then separated the host and the endophyte genomes and constructed a fosmid expression library from the endophytic DNA. This library was screened for antibacterial activity against Staphylococcus aureus. A unique antibacterial clone was selected for further analysis, and a gene En-AP1 was identified with no similarity to known sequences. The expressed, folded protein En-AP1 was not active against S. aureus, while tryptic digests exhibited antimicrobial activity. Seven out of twelve synthesized peptides, predicted antibacterial in silico, exhibited in vitro activity towards both S. aureus and Escherichia coli. We propose that the En-AP1 protein is degraded in the library host E. coli and antimicrobial fragments are released from the cell, explaining the in vitro antibacterial activity of the clone. This is the first report of a novel gene expressed in vitro derived from an endophytic microbiome, demonstrating the potential of finding novel genes and compounds from unculturable endophytes.

Keywords

Antimicrobial peptide Unculturable Endophyte Microbiome 

Notes

Acknowledgments

The work was funded by the Marie Curie Industry-Academia Partnership and Pathways (IAPP) of the EU 7th Framework Programme, New Antimicrobials (NAM) Project (PIAP-GA-2008-218191) and PIIF-GA-2008-220253.

Authors’ contributions

MVT, PP and AMP designed the study. MVT and PP performed molecular lab experiments. AMP, LR, AT, HHS, and HHK provided reagents, materials and scientific input to the study. MK, HH, SK, and SM performed HPLC and MS experiments. MVT analyzed experiments and prepared figures. MVT, PP, and AMP wrote the manuscript. All authors revised the manuscript. All authors read and approved the final manuscript.

Compliance with ethical standards

Conflict of Interest

The authors declare that they have no conflict of interest.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.

Supplementary material

253_2016_7765_MOESM1_ESM.pdf (391 kb)
ESM 1 (PDF 390 kb)

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Copyright information

© Springer-Verlag Berlin Heidelberg 2016

Authors and Affiliations

  1. 1.Genetics and PhysiologyUniversity of OuluOuluFinland
  2. 2.Pharmaceutical Microbiology Section, Institute for Medical Microbiology, Immunology, and ParasitologyUniversity of BonnBonnGermany
  3. 3.Admescope LtdOuluFinland
  4. 4.Biochemistry and Molecular MedicineUniversity of OuluOuluFinland
  5. 5.Novozymes ASBagsvaerdDenmark
  6. 6.Department of BiochemistryUniversity of TriesteTriesteItaly
  7. 7.Molecular MaterialsUniversity of OuluOuluFinland

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