Applied Microbiology and Biotechnology

, Volume 102, Issue 8, pp 3711–3721 | Cite as

Functional identification of the prnABCD operon and its regulation in Serratia plymuthica

  • Xiaoguang Liu
  • Xiaoli Yu
  • Yang Yang
  • Stephan Heeb
  • Shao Gao
  • Kok Gan Chan
  • Miguel Cámara
  • Kexiang Gao
Applied genetics and molecular biotechnology

Abstract

The antibiotic pyrrolnitrin (PRN) is a tryptophan-derived secondary metabolite that plays an important role in the biocontrol of plant diseases due to its broad-spectrum of antimicrobial activities. The PRN biosynthetic gene cluster remains to be characterised in Serratia plymuthica, though it is highly conserved in PRN-producing bacteria. To better understand PRN biosynthesis and its regulation in Serratia, the prnABCD operon from S. plymuthica G3 was cloned, sequenced and expressed in Escherichia coli DH5α. Furthermore, an engineered strain prnind which is a conditional mutant of G3 prnABCD under the control of the Ptac promoter was constructed. This mutant was able to overproduce PRN with isopropylthiogalactoside (IPTG) induction by overexpressing prnABCD, whilst behaving as a conditional mutant of G3 prnABCD in the absence of IPTG. These results confirmed that prnABCD is responsible for PRN biosynthesis in strain G3. Further experiments involving lux-/dsRed-based promoter fusions, combined with site-directed mutagenesis of the putative σS extended -10 region in the prnA promoter, and liquid chromatography-mass spectrometry (LC-MS) analysis extended our previous knowledge about G3, revealing that quorum sensing (QS) regulates PRN biosynthesis through cross talk with RpoS, which may directly activated prnABCD transcription. These findings suggest that PRN in S. plymuthica G3 is produced in a tightly controlled manner, and has diverse functions, such as modulation of cell motility, in addition to antimicrobial activities. Meanwhile, the construction of inducible mutants could be a powerful tool to improve PRN production, beyond its potential use for the investigation of the biological function of PRN.

Keywords

Serratia plymuthica G3 The prnABCD operon A conditional mutant Overexpressing Pyrrolnitrin biosynthesis and regulation 

Notes

Acknowledgements

We would like to thank N. Halliday for kind assistance with LC-MS analysis.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical statement

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

Supplementary material

253_2018_8857_MOESM1_ESM.pdf (277 kb)
ESM 1 (PDF 276 kb)

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Xiaoguang Liu
    • 1
  • Xiaoli Yu
    • 1
  • Yang Yang
    • 1
  • Stephan Heeb
    • 2
  • Shao Gao
    • 3
  • Kok Gan Chan
    • 4
  • Miguel Cámara
    • 2
  • Kexiang Gao
    • 5
  1. 1.Institute of Life SciencesJiangsu UniversityZhenjiangChina
  2. 2.Centre for Biomolecular Sciences, School of Life SciencesThe University of NottinghamNottinghamUK
  3. 3.Department of Architecture and Built Environment, Faculty of EngineeringThe University of NottinghamNottinghamUK
  4. 4.Division of Genetics and Molecular Biology, Institute of Biological Sciences, Faculty of ScienceUniversity of MalayaKuala LumpurMalaysia
  5. 5.Department of Plant PathologyShandong Agricultural UniversityTai’anChina

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