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A novel metabolic network leads to enhanced citrate biogenesis in Pseudomonas fluorescens exposed to aluminum toxicity

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Abstract

Aluminum (Al), an environmental toxin, is known to have a negative impact on various biological systems. However, some microbes have devised intricate mechanisms to combat the toxic influence of this trivalent metal. In this study, Pseudomonas fluorescens grown in malate invoked a unique metabolic shift to promote the synthesis of citrate, a metabolite involved in the sequestration of Al. Electrophoretic and spectrophotometric assays revealed several malate-metabolizing enzymes including malate dehydrogenase (MDH) and malic enzyme (ME) displayed increases in activity and expression in the Al-treated cells. Whereas pyruvate dehydrogenase (PDH) also showed increased activity and expression in the Al-stressed cultures, phosphoenolpyruvate carboxykinase (PEPCK) displayed a marked diminution in the Al-treated cells. The upregulation of citrate synthase (CS) coupled with the diminished activities of aconitase (ACN) and NAD-isocitrate dehydrogenase (NAD-ICDH) appeared to be instrumental in the accumulation of citrate. HPLC experiments revealed high levels of citrate in the Al-stressed cultures. Thus, an Al-enriched environment provoked a metabolic shift in P. fluorescens dedicated to the conversion of malate to citrate.

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Abbreviations

ME:

Malic enzyme

MDH:

Malate dehydrogenase

PEPCK:

Phosphoenol pyruvate carboxykinase

ACN:

Aconitase

PDH:

Pyruvate dehydrogenase

CS:

Citrate synthase

NAD-ICDH:

NAD-dependent isocitrate dehydrogenase

Al:

Aluminum

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Acknowledgments

This work was funded by Ontario Center of Excellence and the Northern Ontario Heritage Fund.

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Authors and Affiliations

  1. Department of Chemistry and Biochemistry, Laurentian University, Sudbury, ON, Canada, P3E 2C6

    Ryan J. Mailloux, Joseph Lemire & Vasu Appanna

  2. Department of Chemical Enzymology, Moscow State University, 119992, Moscow, Russia

    Sergey Kalyuzhnyi

Authors
  1. Ryan J. Mailloux
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  2. Joseph Lemire
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  3. Sergey Kalyuzhnyi
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  4. Vasu Appanna
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Corresponding author

Correspondence to Vasu Appanna.

Additional information

Communicated by G. Antranikian.

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Mailloux, R.J., Lemire, J., Kalyuzhnyi, S. et al. A novel metabolic network leads to enhanced citrate biogenesis in Pseudomonas fluorescens exposed to aluminum toxicity. Extremophiles 12, 451–459 (2008). https://doi.org/10.1007/s00792-008-0150-1

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  • DOI: https://doi.org/10.1007/s00792-008-0150-1

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