Modulation of the major paths of carbon in photorespiratory mutants of synechocystis.

<h4>Background</h4>Recent studies using transcript and metabolite profiles of wild-type and gene deletion mutants revealed that photorespiratory pathways are essential for the growth of Synechocystis sp. PCC 6803 under atmospheric conditions. Pool size changes of primary metabolites, suc...

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Autores principales:	Jan Huege, Jan Goetze, Doreen Schwarz, Hermann Bauwe, Martin Hagemann, Joachim Kopka
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2011
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/cf47e77620e74fc7831590322e0bc377
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spelling	oai:doaj.org-article:cf47e77620e74fc7831590322e0bc3772021-11-18T07:00:06ZModulation of the major paths of carbon in photorespiratory mutants of synechocystis.1932-620310.1371/journal.pone.0016278https://doaj.org/article/cf47e77620e74fc7831590322e0bc3772011-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/21283704/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203<h4>Background</h4>Recent studies using transcript and metabolite profiles of wild-type and gene deletion mutants revealed that photorespiratory pathways are essential for the growth of Synechocystis sp. PCC 6803 under atmospheric conditions. Pool size changes of primary metabolites, such as glycine and glycolate, indicated a link to photorespiration.<h4>Methodology/principal findings</h4>The (13)C labelling kinetics of primary metabolites were analysed in photoautotrophically grown cultures of Synechocystis sp. PCC 6803 by gas chromatography-mass spectrometry (GC-MS) to demonstrate the link with photorespiration. Cells pre-acclimated to high CO(2) (5%, HC) or limited CO(2) (0.035%, LC) conditions were pulse-labelled under very high (2% w/w) (13)C-NaHCO(3) (VHC) conditions followed by treatment with ambient (12)C at HC and LC conditions, respectively. The (13)C enrichment, relative changes in pool size, and (13)C flux of selected metabolites were evaluated. We demonstrate two major paths of CO(2) assimilation via Rubisco in Synechocystis, i.e., from 3PGA via PEP to aspartate, malate and citrate or, to a lesser extent, from 3PGA via glucose-6-phosphate to sucrose. The results reveal evidence of carbon channelling from 3PGA to the PEP pool. Furthermore, (13)C labelling of glycolate was observed under conditions thought to suppress photorespiration. Using the glycolate-accumulating ΔglcD1 mutant, we demonstrate enhanced (13)C partitioning into the glycolate pool under conditions favouring photorespiration and enhanced (13)C partitioning into the glycine pool of the glycine-accumulating ΔgcvT mutant. Under LC conditions, the photorespiratory mutants ΔglcD1 and ΔgcvT showed enhanced activity of the additional carbon-fixing PEP carboxylase pathway.<h4>Conclusions/significance</h4>With our approach of non-steady-state (13)C labelling and analysis of metabolite pool sizes with respective (13)C enrichments, we identify the use and modulation of major pathways of carbon assimilation in Synechocystis in the presence of high and low inorganic carbon supplies.Jan HuegeJan GoetzeDoreen SchwarzHermann BauweMartin HagemannJoachim KopkaPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 6, Iss 1, p e16278 (2011)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Jan Huege Jan Goetze Doreen Schwarz Hermann Bauwe Martin Hagemann Joachim Kopka Modulation of the major paths of carbon in photorespiratory mutants of synechocystis.
description	<h4>Background</h4>Recent studies using transcript and metabolite profiles of wild-type and gene deletion mutants revealed that photorespiratory pathways are essential for the growth of Synechocystis sp. PCC 6803 under atmospheric conditions. Pool size changes of primary metabolites, such as glycine and glycolate, indicated a link to photorespiration.<h4>Methodology/principal findings</h4>The (13)C labelling kinetics of primary metabolites were analysed in photoautotrophically grown cultures of Synechocystis sp. PCC 6803 by gas chromatography-mass spectrometry (GC-MS) to demonstrate the link with photorespiration. Cells pre-acclimated to high CO(2) (5%, HC) or limited CO(2) (0.035%, LC) conditions were pulse-labelled under very high (2% w/w) (13)C-NaHCO(3) (VHC) conditions followed by treatment with ambient (12)C at HC and LC conditions, respectively. The (13)C enrichment, relative changes in pool size, and (13)C flux of selected metabolites were evaluated. We demonstrate two major paths of CO(2) assimilation via Rubisco in Synechocystis, i.e., from 3PGA via PEP to aspartate, malate and citrate or, to a lesser extent, from 3PGA via glucose-6-phosphate to sucrose. The results reveal evidence of carbon channelling from 3PGA to the PEP pool. Furthermore, (13)C labelling of glycolate was observed under conditions thought to suppress photorespiration. Using the glycolate-accumulating ΔglcD1 mutant, we demonstrate enhanced (13)C partitioning into the glycolate pool under conditions favouring photorespiration and enhanced (13)C partitioning into the glycine pool of the glycine-accumulating ΔgcvT mutant. Under LC conditions, the photorespiratory mutants ΔglcD1 and ΔgcvT showed enhanced activity of the additional carbon-fixing PEP carboxylase pathway.<h4>Conclusions/significance</h4>With our approach of non-steady-state (13)C labelling and analysis of metabolite pool sizes with respective (13)C enrichments, we identify the use and modulation of major pathways of carbon assimilation in Synechocystis in the presence of high and low inorganic carbon supplies.
format	article
author	Jan Huege Jan Goetze Doreen Schwarz Hermann Bauwe Martin Hagemann Joachim Kopka
author_facet	Jan Huege Jan Goetze Doreen Schwarz Hermann Bauwe Martin Hagemann Joachim Kopka
author_sort	Jan Huege
title	Modulation of the major paths of carbon in photorespiratory mutants of synechocystis.
title_short	Modulation of the major paths of carbon in photorespiratory mutants of synechocystis.
title_full	Modulation of the major paths of carbon in photorespiratory mutants of synechocystis.
title_fullStr	Modulation of the major paths of carbon in photorespiratory mutants of synechocystis.
title_full_unstemmed	Modulation of the major paths of carbon in photorespiratory mutants of synechocystis.
title_sort	modulation of the major paths of carbon in photorespiratory mutants of synechocystis.
publisher	Public Library of Science (PLoS)
publishDate	2011
url	https://doaj.org/article/cf47e77620e74fc7831590322e0bc377
work_keys_str_mv	AT janhuege modulationofthemajorpathsofcarboninphotorespiratorymutantsofsynechocystis AT jangoetze modulationofthemajorpathsofcarboninphotorespiratorymutantsofsynechocystis AT doreenschwarz modulationofthemajorpathsofcarboninphotorespiratorymutantsofsynechocystis AT hermannbauwe modulationofthemajorpathsofcarboninphotorespiratorymutantsofsynechocystis AT martinhagemann modulationofthemajorpathsofcarboninphotorespiratorymutantsofsynechocystis AT joachimkopka modulationofthemajorpathsofcarboninphotorespiratorymutantsofsynechocystis
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Modulation of the major paths of carbon in photorespiratory mutants of synechocystis.

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