Regulatory Tasks of the Phosphoenolpyruvate-Phosphotransferase System of <named-content content-type="genus-species">Pseudomonas putida</named-content> in Central Carbon Metabolism

ABSTRACT Two branches of the phosphoenolpyruvate-phosphotransferase system (PTS) operate in the soil bacterium Pseudomonas putida KT2440. One branch encompasses a complete set of enzymes for fructose intake (PTSFru), while the other (N-related PTS, or PTSNtr) controls various cellular functions unre...

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Autores principales:	Max Chavarría, Roelco J. Kleijn, Uwe Sauer, Katharina Pflüger-Grau, Víctor de Lorenzo
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2012
Materias:	Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/3360aa076889444ab08afb70c211d31d
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spelling	oai:doaj.org-article:3360aa076889444ab08afb70c211d31d2021-11-15T15:39:09ZRegulatory Tasks of the Phosphoenolpyruvate-Phosphotransferase System of <named-content content-type="genus-species">Pseudomonas putida</named-content> in Central Carbon Metabolism10.1128/mBio.00028-122150-7511https://doaj.org/article/3360aa076889444ab08afb70c211d31d2012-05-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.00028-12https://doaj.org/toc/2150-7511ABSTRACT Two branches of the phosphoenolpyruvate-phosphotransferase system (PTS) operate in the soil bacterium Pseudomonas putida KT2440. One branch encompasses a complete set of enzymes for fructose intake (PTSFru), while the other (N-related PTS, or PTSNtr) controls various cellular functions unrelated to the transport of carbohydrates. The potential of these two systems for regulating central carbon catabolism has been investigated by measuring the metabolic fluxes of isogenic strains bearing nonpolar mutations in PTSFru or PTSNtr genes and grown on either fructose (a PTS substrate) or glucose, the transport of which is not governed by the PTS in this bacterium. The flow of carbon from each sugar was distinctly split between the Entner-Doudoroff, pentose phosphate, and Embden-Meyerhof-Parnas pathways in a ratio that was maintained in each of the PTS mutants examined. However, strains lacking PtsN (EIIANtr) displayed significantly higher fluxes in the reactions of the pyruvate shunt, which bypasses malate dehydrogenase in the TCA cycle. This was consistent with the increased activity of the malic enzyme and the pyruvate carboxylase found in the corresponding PTS mutants. Genetic evidence suggested that such a metabolic effect of PtsN required the transfer of high-energy phosphate through the system. The EIIANtr protein of the PTSNtr thus helps adjust central metabolic fluxes to satisfy the anabolic and energetic demands of the overall cell physiology. IMPORTANCE This study demonstrates that EIIANtr influences the biochemical reactions that deliver carbon between the upper and lower central metabolic domains for the consumption of sugars by P. putida. These findings indicate that the EIIANtr protein is a key player for orchestrating the fate of carbon in various physiological destinations in this bacterium. Additionally, these results highlight the importance of the posttranslational regulation of extant enzymatic complexes for increasing the robustness of the corresponding metabolic networks.Max ChavarríaRoelco J. KleijnUwe SauerKatharina Pflüger-GrauVíctor de LorenzoAmerican Society for MicrobiologyarticleMicrobiologyQR1-502ENmBio, Vol 3, Iss 2 (2012)
institution	DOAJ
collection	DOAJ
language	EN
topic	Microbiology QR1-502
spellingShingle	Microbiology QR1-502 Max Chavarría Roelco J. Kleijn Uwe Sauer Katharina Pflüger-Grau Víctor de Lorenzo Regulatory Tasks of the Phosphoenolpyruvate-Phosphotransferase System of <named-content content-type="genus-species">Pseudomonas putida</named-content> in Central Carbon Metabolism
description	ABSTRACT Two branches of the phosphoenolpyruvate-phosphotransferase system (PTS) operate in the soil bacterium Pseudomonas putida KT2440. One branch encompasses a complete set of enzymes for fructose intake (PTSFru), while the other (N-related PTS, or PTSNtr) controls various cellular functions unrelated to the transport of carbohydrates. The potential of these two systems for regulating central carbon catabolism has been investigated by measuring the metabolic fluxes of isogenic strains bearing nonpolar mutations in PTSFru or PTSNtr genes and grown on either fructose (a PTS substrate) or glucose, the transport of which is not governed by the PTS in this bacterium. The flow of carbon from each sugar was distinctly split between the Entner-Doudoroff, pentose phosphate, and Embden-Meyerhof-Parnas pathways in a ratio that was maintained in each of the PTS mutants examined. However, strains lacking PtsN (EIIANtr) displayed significantly higher fluxes in the reactions of the pyruvate shunt, which bypasses malate dehydrogenase in the TCA cycle. This was consistent with the increased activity of the malic enzyme and the pyruvate carboxylase found in the corresponding PTS mutants. Genetic evidence suggested that such a metabolic effect of PtsN required the transfer of high-energy phosphate through the system. The EIIANtr protein of the PTSNtr thus helps adjust central metabolic fluxes to satisfy the anabolic and energetic demands of the overall cell physiology. IMPORTANCE This study demonstrates that EIIANtr influences the biochemical reactions that deliver carbon between the upper and lower central metabolic domains for the consumption of sugars by P. putida. These findings indicate that the EIIANtr protein is a key player for orchestrating the fate of carbon in various physiological destinations in this bacterium. Additionally, these results highlight the importance of the posttranslational regulation of extant enzymatic complexes for increasing the robustness of the corresponding metabolic networks.
format	article
author	Max Chavarría Roelco J. Kleijn Uwe Sauer Katharina Pflüger-Grau Víctor de Lorenzo
author_facet	Max Chavarría Roelco J. Kleijn Uwe Sauer Katharina Pflüger-Grau Víctor de Lorenzo
author_sort	Max Chavarría
title	Regulatory Tasks of the Phosphoenolpyruvate-Phosphotransferase System of <named-content content-type="genus-species">Pseudomonas putida</named-content> in Central Carbon Metabolism
title_short	Regulatory Tasks of the Phosphoenolpyruvate-Phosphotransferase System of <named-content content-type="genus-species">Pseudomonas putida</named-content> in Central Carbon Metabolism
title_full	Regulatory Tasks of the Phosphoenolpyruvate-Phosphotransferase System of <named-content content-type="genus-species">Pseudomonas putida</named-content> in Central Carbon Metabolism
title_fullStr	Regulatory Tasks of the Phosphoenolpyruvate-Phosphotransferase System of <named-content content-type="genus-species">Pseudomonas putida</named-content> in Central Carbon Metabolism
title_full_unstemmed	Regulatory Tasks of the Phosphoenolpyruvate-Phosphotransferase System of <named-content content-type="genus-species">Pseudomonas putida</named-content> in Central Carbon Metabolism
title_sort	regulatory tasks of the phosphoenolpyruvate-phosphotransferase system of <named-content content-type="genus-species">pseudomonas putida</named-content> in central carbon metabolism
publisher	American Society for Microbiology
publishDate	2012
url	https://doaj.org/article/3360aa076889444ab08afb70c211d31d
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Regulatory Tasks of the Phosphoenolpyruvate-Phosphotransferase System of <named-content content-type="genus-species">Pseudomonas putida</named-content> in Central Carbon Metabolism

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