Abstract
Although nitrosative stress is known to severely impede the ability of living systems to generate adenosine triphosphate (ATP) via oxidative phosphorylation, there is limited information on how microorganisms fulfill their energy needs in order to survive reactive nitrogen species (RNS). In this study we demonstrate an elaborate strategy involving substrate-level phosphorylation that enables the soil microbe Pseudomonas fluorescens to synthesize ATP in a defined medium with fumarate as the sole carbon source. The enhanced activities of such enzymes as phosphoenolpyruvate carboxylase and pyruvate phosphate dikinase coupled with the increased activities of phospho-transfer enzymes like adenylate kinase and nucleoside diphophate kinase provide an effective strategy to produce high energy nucleosides in an O2-independent manner. The alternate ATP producing machinery is fuelled by the precursors derived from fumarate with the aid of fumarase C and fumarate reductase. This metabolic reconfiguration is key to the survival of P. fluorescens and reveals potential targets against RNS-resistant organisms.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Adam RD (2001) Biology of Giardia lamblia. Clin Microbiol Rev 14:447–475
al-Aoukaty A, Appanna VD, Falter H (1992) Gallium toxicity and adaptation in Pseudomonas fluorescnes. FEMS Microbiol Lett 71:265–272
Auger C, Lemire J, Cecchini D, Bignucolo A, Appanna VD (2011) The metabolic reprogramming evoked by nitrosative stress triggers the anaerobic utilization of citrate in Pseudomonas fluorescens. PLoS ONE 6:e28469
Auger C, Appanna V, Castonguay Z, Han S, Appanna VD (2012) A facile electrophoretic technique to monitor phosphoenolpyruvate-dependent kinases. Electrophoresis 33:1095–1101
Auger C, Han S, Appanna VP, Thomas SC, Ulibarri G, Appanna VD (2013) Metabolic reengineering invoked by microbial systems to decontaminate aluminum: implications in bioremediation technologies. Biotechnol Adv 31:266–273
Bignucolo A, Appanna VP, Thomas SC, Auger C, Han S, Omri A, Appanna VD (2013) Hydrogen peroxide stress a metabolic reprogramming in Pseudomonas fluorescens: enhanced production of puyruvate. J Biotechnol 167:309–315
Bradford MB (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254
Charoo NA, Shamsher AAA, Lian LY, Abrahamsson B, Cristofoletti R, Groot DW, Kopp S, Langguth P, Polli J, Shah VP, Dressman J (2014) Biowaiver monograph for immediate-release solid oral dosage forms: bisoprolol fumarate. J Pharma Sci 103:378–391
Chénier D, Bériault R, Mailloux R, Baquie M, Abramia G, Lemire J, Appanna VD (2008) Metabolic adaptation in Pseudomonas fluorescens evoked by aluminum and gallium toxicity: involvement of fumarase C and NADH oxidase. Appl Environ Microbiol 74:3977–3984
Couston V, Besterio S, Biran M, Diolez P, Bouchaud V, Voisin P, Michels PAM, Canioni P, Baltz T, Bringaud F (2003) ATP generation in the Trypanosoma brucei procyclic form cytosolic substrate level phosphorylation is essential, but not oxidative phosphorylation. J Biol Chem 278:49625–49635
Hall JW, Ji Y (2013) Sensing and adapting to anaerobic conditions by Staphylococcus aureus. Adv Appl Microbiol 84:1–25
Han S, Auger C, Castonguay Z, Appanna VP, Thomas SC, Appanna VD (2012) The unravelling of metabolic dysfunctions linked to metal-associated diseases by blue native polyacrylamide gel electrophoresis. Anal Bioanal Chem 405(6):1821-1831
Han S, Auger C, Thomas SC, Beites CL, Appanna VD (2013) Mitochondrial biogenesis and energy production in differentiating stem cells: a functional metabolic study. Cell Reprogram 16:84–90
Hunt KA, Flynn JM, Naranjo B, Shikhare ID, Gralnick JA (2010) Substrate-level phosphorylation is the primary source of energy conservation during anaerobic respiration of Shewanella oneidensis strain MR-1. J Bacteriol 192:3345–3351
Ingram-Smith C, Martin SR, Smith KS (2006) Acetate kinase: not just a bacterial enzyme. Trends Microbiol 14:249–253
Kim D, Yu BJ, Kim JA, Lee Y, Choi S, Kang S (2013) The acetylproteome of Gram-positive model bacterium Bacillus subtilis. Proteomics 13:1726–1736
Lemire J, Auger C, Bignucolo A, Appanna VP, Appanna VD (2012) Metabolic strategies deployed by Pseudomonas fluorescens to combat metal pollutants: biotechnological prospects in current research, technology and education topics in applied microbiology and microbial biotechnology. In: Mendez-vilas A (ed). Formalex Publisher, pp 177–187
Li K, Pidatala RV, Shaik R, Datta R, Ramakrishna W (2014) Integrated metabolomic and proteomic approaches dissect the effect of metal-resistant bacteria on maize biomass and copper uptake. Environ Sci Technol 48:1184–1193
Lushchak OV, Piroddi M, Galli F, Lushchak VI (2014) Aconitase post-translational modification as a key in linkage between Krebs cycle, iron homeostasis, redox signaling, and metabolism of reactive oxygen species. Redox Rep 19:8–15
Ma Y, Guo C, Li H, Peng X (2013) Low abundance of respiratory nitrate reductase is essential for Escherichia coli in resistance to aminoglycoside and cephalosporin. J Proteom 87:78–88
Mailloux RJ, Darwich R, Lemire J, Appanna V (2008) The monitoring of nucleotide diphosphate kinase activity by blue native polyacrylamide gel electrophoresis. Electrophor 29:1484–1489
Mailloux RJ, Lemire J, Appanna VD (2011) Metabolic networks to combat oxidative stress in Pseuodomonas fluorescens. Antonie Van Leeuwenhoek 99:433–442
Miranda KM, Espey MG, Wink DA (2001) A rapid, simple spectrophotometric method for simultaneous detection of nitrate and nitrite. Nitric Oxide 5:62–71
Poole RK (2005) Nitric oxide and nitrosative stress tolerance in bacteria. Biochem Soc Trans 33:176–180
Quijano C, Alvarez B, Gatti RM, Augusto O, Radi R (1997) Pathways of peroxynitrite oxidation of thiol groups. Biochem J 322:167–173
Schagger H, von Jagow G (1991) Blue native electrophoresis for isolation of membrane protein complexes in enzymatically active form. Anal Biochem 199:223–231
Singh R, Chenier D, Beriault R, Mailloux R, Hamel RD, Appanna VD (2005) Blue native polyacrylamide gel electrophoresis and the monitoring of malate- and oxaloacetate-producing enzymes. J Biochem Biophys Methods 64:189–199
Singh R, Lemire J, Mailloux RJ, Chénier D, Hamel R, Appanna VD (2009) An ATP and oxalate generating variant tricarboxylic acid cycle counters aluminum toxicity in Pseudomonas fluorescens. PloS One 4:e7344
Watanabe S, Zimmerman M, Goodwin MB, Sauer U, Barry CE, Boshoff HI (2011) Fumarate reductase activity maintains an energized membrane in anerobic Mycobacterium tuberculosis. PLoS ONE 7:1–15
Zielonka J, Zielonka M, Sikora A, Adamus J, Joseph J, Hardy M, Ouari O, Dranka BP, Kalyanarama B (2012) Global profiling of reactive oxygen and nitrogen species in biological systems: high-throughput real-time analyses. J Biol Chem 287:2984–2995
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Appanna, V.P., Auger, C., Thomas, S.C. et al. Fumarate metabolism and ATP production in Pseudomonas fluorescens exposed to nitrosative stress. Antonie van Leeuwenhoek 106, 431–438 (2014). https://doi.org/10.1007/s10482-014-0211-7
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10482-014-0211-7