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  Modulation of the Major Paths of Carbon in Photorespiratory Mutants of Synechocystis

Huege, J., Goetze, J., Schwarz, D., Bauwe, H., Hagemann, M., & Kopka, J. (2011). Modulation of the Major Paths of Carbon in Photorespiratory Mutants of Synechocystis. PLoS One, 6(1), e16278. doi:10.1371/journal.pone.0016278.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-0014-21BD-1 Version Permalink: http://hdl.handle.net/11858/00-001M-0000-0018-05AE-9
Genre: Journal Article

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Huege, J.1, Author              
Goetze, J.2, Author
Schwarz, D.2, Author
Bauwe, H.2, Author
Hagemann, M.2, Author
Kopka, J.1, Author              
Affiliations:
1Applied Metabolome Analysis, Department Willmitzer, Max Planck Institute of Molecular Plant Physiology, Max Planck Society, ou_1753338              
2External Organizations, ou_persistent22              

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Free keywords: metabolic flux analysis sp strain pcc-6803 mass-spectrometry gas-chromatography wild-type cycle cyanobacteria microorganisms purification arabidopsis
 Abstract: Background: 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. Methodology/Principal Findings: The C-13 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 CO2 (5%, HC) or limited CO2 (0.035%, LC) conditions were pulse-labelled under very high (2% w/w) C-13-NaHCO3 (VHC) conditions followed by treatment with ambient C-12 at HC and LC conditions, respectively. The C-13 enrichment, relative changes in pool size, and C-13 flux of selected metabolites were evaluated. We demonstrate two major paths of CO2 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, C-13 labelling of glycolate was observed under conditions thought to suppress photorespiration. Using the glycolate-accumulating Delta glcD1 mutant, we demonstrate enhanced C-13 partitioning into the glycolate pool under conditions favouring photorespiration and enhanced C-13 partitioning into the glycine pool of the glycine-accumulating Delta gcvT mutant. Under LC conditions, the photorespiratory mutants Delta glcD1 and Delta gcvT showed enhanced activity of the additional carbon-fixing PEP carboxylase pathway. Conclusions/Significance: With our approach of non-steady-state C-13 labelling and analysis of metabolite pool sizes with respective C-13 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.

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Language(s): eng - English
 Dates: 2011-01-212011
 Publication Status: Published in print
 Pages: -
 Publishing info: -
 Table of Contents: -
 Rev. Type: -
 Identifiers: ISI: ISI:000286522300029
DOI: 10.1371/journal.pone.0016278
ISSN: 1932-6203
URI: ://000286522300029http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3025020/pdf/pone.0016278.pdf?tool=pmcentrez
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Title: PLoS One
Source Genre: Journal
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Publ. Info: San Francisco, CA : Public Library of Science
Pages: - Volume / Issue: 6 (1) Sequence Number: - Start / End Page: e16278 Identifier: ISSN: 1932-6203
CoNE: https://pure.mpg.de/cone/journals/resource/1000000000277850