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Aspartate metabolism and pyruvate homeostasis triggered by oxidative stress in Pseudomonas fluorescens: a functional metabolomic study

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Abstract

There is mounting evidence that metabolic reprogramming is critical for the survival of organisms exposed to changing and stressed environments. Using the soil microbe Pseudomonas fluorescens as a model system, we demonstrate that the metabolic networks aimed at the conversion of aspartate into pyruvate are enhanced in the presence of hydrogen peroxide (H2O2). The metabolites pyruvate, oxaloacetate and acetate were increased in the treated cultures as measured by HPLC. Enzymes such as aspartate transaminase and phosphoenolpyruvate carboxylase (PEPC) that mediate the conversion of aspartate to phosphoenolpyruvate (PEP) were up-regulated. This high-energy phosphate was readily converted into ATP, a process facilitated by the increased activity of pyruvate orthophosphate dikinase (PPDK) and phosphoenolpyruvate synthase (PEPS) as oxidative phosphorylation was severely compromised. The ensuing formation of pyruvate readily detoxified reactive oxygen species with the concomitant formation of acetate. This H2O2-induced metabolic reconfiguration not only helps generate the antioxidants necessary to thwart oxidative stress but also powers the formation of energy.

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Abbreviations

AL:

Aspartate lyase

AT:

Aspartate transaminase

AK:

Adenylate kinase

CFE:

Cell free extract

G6PDH:

Glucose 6-phosphate dehydrogenase

H2O2 :

Hydrogen peroxide

ICDH:

Isocitrate dehydrogenase

MDH:

Malate dehydrogenase

ME:

Malic enzyme

OP:

Oxidative phosphorylation

PEPC:

Phosphoenolpyruvate carboxylase

PEPS:

Phosphoenolpyruvate synthase

PPDK:

Pyruvate orthophosphate dikinase

ROS:

Reactive oxygen species

NADPH:

Reduced nicotinamide adenine dinucleotide phosphate

TCA:

Tricarboxylic acid

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Acknowledgment

This study was funded by Laurentian University and the Northern Ontario Heritage Fund. Azhar Alhasawi is a recipient of funding from the Ministry of Higher Education of Saudi Arabia.

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Authors and Affiliations

  1. Faculty of Science and Engineering, Laurentian University, Sudbury, ON, P3E2C6, Canada

    Azhar Alhasawi, Martine Leblanc, Nishma D. Appanna, Christopher Auger & Vasu D. Appanna

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  1. Azhar Alhasawi
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  2. Martine Leblanc
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  3. Nishma D. Appanna
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  4. Christopher Auger
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  5. Vasu D. Appanna
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Correspondence to Vasu D. Appanna.

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Alhasawi, A., Leblanc, M., Appanna, N.D. et al. Aspartate metabolism and pyruvate homeostasis triggered by oxidative stress in Pseudomonas fluorescens: a functional metabolomic study. Metabolomics 11, 1792–1801 (2015). https://doi.org/10.1007/s11306-015-0841-4

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