Abstract

Proteobacteria are known to communicate via signaling molecules and this process is known as quorum sensing. The most commonly studied quorum sensing molecules are N-acylhomoserine lactones (AHLs) that consists of a homoserine lactone moiety and an N-acyl side chain with various chain lengths and degrees of saturation at the C-3 position. We have isolated a bacterium, RB-44, from a site which was formally a landfill dumping ground. Using matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry analysis, this isolate was identified as a Pandoraea sp.which was then screened for AHL production using biosensors which indicated its quorum sensing properties. To identify the AHL profile of Pandoraea sp. RB-44, we used high resolution tandem mass spectrometry confirming that this isolate produced N-octanoylhomoserine lactone (C8-HSL). To the best of our knowledge, this is the first report that showed quorum sensing activity exhibited by Pandoraea sp. Our data add Pandoraea sp. to the growing number of bacteria that possess QS systems.

Highlights

  • Often reported as non-fermentative Gram-negative bacilli or misidentified as Burkholderia cepacia complex (Bcc) or Ralstonia spp. in clinical microbiology laboratory [1,2,3], Pandoraea sp. is a taxonomically distinct β-subclass of proteobacteria where genera of Bukholderia and Ralstonia are the closest neighbors [4]

  • Most Gram negative bacteria use quorum sensing (QS) to establish communications between neighboring cells through secretion and detection of diffusible signaling molecule known as autoinducers [13,14,15]

  • The most widely studied QS molecules are N-acyl-homoserine lactones (AHLs) which are synthesized by AHL synthase that will diffuse and bind to its cognate receptor where this AHL-LuxR complex will be activated to regulate

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Summary

Introduction

Often reported as non-fermentative Gram-negative bacilli or misidentified as Burkholderia cepacia complex (Bcc) or Ralstonia spp. in clinical microbiology laboratory [1,2,3], Pandoraea sp. is a taxonomically distinct β-subclass of proteobacteria where genera of Bukholderia and Ralstonia are the closest neighbors [4]. Often reported as non-fermentative Gram-negative bacilli or misidentified as Burkholderia cepacia complex (Bcc) or Ralstonia spp. in clinical microbiology laboratory [1,2,3], Pandoraea sp. Despite all the documentation on this species, little is known about the mechanism and regulation of its pathogenicity and bio-catalytic activities. Virulence factor and bio-catalytic activities is through characterizing its AHL production expression [20]. We used matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry analysis for strain identification and high resolution quadrupole mass spectrometry to confirm its AHL profile. To our knowledge, this is the first report of AHL production by Pandoraea sp

Soil Sampling
Enrichment and Isolation
Bacteria Strains and Culture Conditions
AHL Extraction
Strain Identification Using 16S rDNA Amplification
Sampling and Screening for AHL Producing Bacteria
Strain Identification and MALDI-TOF Score-Oriented Dendrogram
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