Dataset Information

The proteome of Synechocystis sp. PCC 6803 cells grown under light-activated heterotrophic growth

ABSTRACT: Cyanobacteria are photoautotrophic prokaryotes with a plant-like photosynthetic machinery. Besides being able to grow photoautotrophically, some cyanobacteria are also capable to grow photoheterotrophically, where they use reduced organic compounds as carbon source, or even completely heterotrophically by using reduced organic compounds as carbon and energy source. The well characterized cyanobacterium Synechocystis sp. PCC 6803 can grow in darkness under light-activated heterotrophic growth (LAHG) conditions by using glucose as carbon and energy source. In the present work, we combined pre-fractioning of Synechocystis cellular membranes with a global proteome and lipidome analysis, to shift the analytical focus towards the rearrangement of the internal thylakoid membrane system observed in Synechocystis cells under LAHG conditions.

INSTRUMENT(S): Thermo Scientific instrument model

ORGANISM(S): Synechocystis Sp. Pcc 6803

TISSUE(S): Photosynthetic Cell

SUBMITTER: Nicole Plohnke

LAB HEAD: Sascha Rexroth

PROVIDER: PXD000980 | Pride | 2015-03-03

REPOSITORIES: Pride

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Publications

The proteome and lipidome of Synechocystis sp. PCC 6803 cells grown under light-activated heterotrophic conditions.

Plohnke Nicole N Seidel Tobias T Kahmann Uwe U Rögner Matthias M Schneider Dirk D Rexroth Sascha S

Molecular & cellular proteomics : MCP 20150105 3

Cyanobacteria are photoautotrophic prokaryotes with a plant-like photosynthetic machinery. Because of their short generation times, the ease of their genetic manipulation, and the limited size of their genome and proteome, cyanobacteria are popular model organisms for photosynthetic research. Although the principal mechanisms of photosynthesis are well-known, much less is known about the biogenesis of the thylakoid membrane, hosting the components of the photosynthetic, and respiratory electron ...[more]

PMID: 25561504

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Project description:Cyanobacteria are phototrophic prokaryotes that can convert inorganic carbon as CO2 into organic carbon compounds at the expense of light energy. In addition, they need only a few inorganic nutrients and can be cultivated in high densities using non-arable land and seawater. This features qualified cyanobacteria as attractive organisms for the production of third generation biofuels as part of the development of future CO2-neutral energy production. Synechocystis sp. PCC 6803 represents one of the most widely used cyanobacterial model strains. On the basis of its available genome sequence and genetic tools, many strains of Synechocystis have been generated that produce different biotechnological products. Efficient isoprene production is an attractive goal, since this compound represents not only an energy-rich biofuel but is also used as chemical feedstock. Here, we report on our attempts to generate isoprene-producing strains of Synechocystis. The cDNA of a codon-optimized plant isoprene synthase (IspS) was cloned under the control of different Synechocystis promoters, which ensure strong constitutive or light-regulated ispS expression. The expression of the ispS gene was quantified by qPCR, whereas the amount of isoprene was quantified using GC-MS. Incubation of our strains at different salt conditions had marked impact on the isoprene production rates. Under low salt conditions, a good correlation was found between ispS expression and isoprene production rate. However, the cultivation of isoprene production strains under salt-supplemented conditions decreased isoprene production despite the fact that ispS expression was salt-stimulated. The characterization of the metabolome of isoprene producing strains indicated that isoprene production might be limited by insufficient precursor levels. Our isoprene production rates under low salt conditions were 2 - 6.5times higher compared to the previous report of Lindberg et al. (2010). These results can be used to guide future attempts establishing the isoprene production with cyanobacterial host systems.

Project description:Montagud2010 - Genome-scale metabolic network of Synechocystis sp. PCC6803 (iSyn669) This model is described in the article: Reconstruction and analysis of genome-scale metabolic model of a photosynthetic bacterium. Montagud A, Navarro E, Fernández de Córdoba P, Urchueguía JF, Patil KR. BMC Syst Biol 2010; 4: 156 Abstract: BACKGROUND: Synechocystis sp. PCC6803 is a cyanobacterium considered as a candidate photo-biological production platform--an attractive cell factory capable of using CO2 and light as carbon and energy source, respectively. In order to enable efficient use of metabolic potential of Synechocystis sp. PCC6803, it is of importance to develop tools for uncovering stoichiometric and regulatory principles in the Synechocystis metabolic network. RESULTS: We report the most comprehensive metabolic model of Synechocystis sp. PCC6803 available, iSyn669, which includes 882 reactions, associated with 669 genes, and 790 metabolites. The model includes a detailed biomass equation which encompasses elementary building blocks that are needed for cell growth, as well as a detailed stoichiometric representation of photosynthesis. We demonstrate applicability of iSyn669 for stoichiometric analysis by simulating three physiologically relevant growth conditions of Synechocystis sp. PCC6803, and through in silico metabolic engineering simulations that allowed identification of a set of gene knock-out candidates towards enhanced succinate production. Gene essentiality and hydrogen production potential have also been assessed. Furthermore, iSyn669 was used as a transcriptomic data integration scaffold and thereby we found metabolic hot-spots around which gene regulation is dominant during light-shifting growth regimes. CONCLUSIONS: iSyn669 provides a platform for facilitating the development of cyanobacteria as microbial cell factories. This model is hosted on BioModels Database and identified by: MODEL1507180008. To cite BioModels Database, please use: BioModels Database: An enhanced, curated and annotated resource for published quantitative kinetic models. To the extent possible under law, all copyright and related or neighbouring rights to this encoded model have been dedicated to the public domain worldwide. Please refer to CC0 Public Domain Dedication for more information.

Dataset Information

The proteome of Synechocystis sp. PCC 6803 cells grown under light-activated heterotrophic growth

Publications

The proteome and lipidome of Synechocystis sp. PCC 6803 cells grown under light-activated heterotrophic conditions.

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