Regulator LdhR and D-lactate dehydrogenase LdhA of Burkholderia multivorans play roles in carbon overflow and in planktonic cellular aggregate formation

Inês N. Silva, Marcelo J. Ramires, Lisa A. Azevedo, Ana R. Guerreiro, Andreia C. Tavares, Jörg D. Becker, Leonilde M. Moreira

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Abstract

LysR-type transcriptional regulators (LTTRs) are the most commonly found regulators in Burkholderia cepacia complex, comprising opportunistic pathogens causing chronic respiratory infections in cystic fibrosis (CF) patients. Despite LTTRs being global regulators of pathogenicity in several types of bacteria, few have been characterized in Burkholderia. Here, we show that gene ldhR of B. multivorans encoding an LTTR is cotranscribed with ldhA encoding a D-lactate dehydrogenase and evaluate their implication in virulence traits such as exopolysaccharide (EPS) synthesis and biofilm formation. A comparison of the wild type (WT) and its isogenic ΔldhR mutant grown in medium with 2% D-glucose revealed a negative impact on EPS biosynthesis and on cell viability in the presence of LdhR. The loss of viability in WT cells was caused by intracellular acidification as a consequence of the cumulative secretion of organic acids, including D-lactate, which was absent from the ΔldhR mutant supernatant. Furthermore, LdhR is implicated in the formation of planktonic cellular aggregates. WT cell aggregates reached 1,000 μm in size after 24 h in liquid cultures, in contrast to ΔldhR mutant aggregates that never grew more than 60 μm. The overexpression of D-lactate dehydrogenase LdhA in the ΔldhR mutant partially restored the formed aggregate size, suggesting a role for fermentation inside aggregates. Similar results were obtained for surface-attached biofilms, with WT cells producing more biofilm. A systematic evaluation of planktonic aggregates in Burkholderia CF clinical isolates showed aggregates in 40 of 74. As CF patients' lung environments are microaerophilic and bacteria are found as free aggregates/biofilms, LdhR and LdhA might have central roles in adapting to this environment.

Original languageEnglish
Article numbere01343-17
JournalApplied and Environmental Microbiology
Volume83
Issue number19
DOIs
Publication statusPublished - 1 Jan 2017

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Keywords

  • Biofilms
  • Burkholderia multivorans
  • Cystic fibrosis
  • D-lactate dehydrogenase
  • Exopolysaccharide
  • LysR family transcriptional regulator
  • Planktonic cellular aggregates

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