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Metabolomics

, Volume 11, Issue 6, pp 1792–1801 | Cite as

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

  • Azhar Alhasawi
  • Martine Leblanc
  • Nishma D. Appanna
  • Christopher Auger
  • Vasu D. AppannaEmail author
Original Article

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.

Keywords

Pyruvate Antioxidant Metabolic networks Pyruvate synthase Energy 

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

Notes

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.

Compliance with ethical standards

Conflict of Interest

All authors declare that they have no conflict of interest.

Research involving Human and Animal Rights

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

Supplementary material

11306_2015_841_MOESM1_ESM.tif (2.2 mb)
Supplementary material 1 (TIFF 2296 kb)

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Azhar Alhasawi
    • 1
  • Martine Leblanc
    • 1
  • Nishma D. Appanna
    • 1
  • Christopher Auger
    • 1
  • Vasu D. Appanna
    • 1
    Email author
  1. 1.Faculty of Science and EngineeringLaurentian UniversitySudburyCanada

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