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Adaptation of Bacillus subtilis carbon core metabolism to simultaneous nutrient limitation and osmotic challenge


ABSTRACT: Here, we investigated for the first time the systems-wide response of B. subtilis to different simultaneous stresses, i.e. nutrient limitation and high osmolarity. To address the anticipated complexity of the cellular response networks, we combined chemostat experiments under conditions of carbon limitation, salt stress and osmoprotection with multi-omics analyses at the transcriptome, proteome, metabolome and fluxome levels. Our results indicate that the flux through central carbon and energy metabolism is very robust under all conditions studied. The key to achieve this robustness is the adjustment of the biocatalytic machinery to compensate for solvent-induced impairment of enzymatic activities during osmotic stress. The accumulation of the exogenously provided osmoprotectant glycine betaine helps the cell to rescue enzyme activities in the presence of high salt. A major effort of the cell during osmotic stress is the production of the compatible solute proline. This is achieved by the concerted adjustment of multiple reactions around the 2-oxoglutarate node, which drives metabolism towards the proline precursor glutamate. The fine-tuning of the transcriptional and metabolic networks involves functional modules that overarch the individual pathways. We applied transcriptomic, mass spectrometry-based protein, metabolite and 13C-metabolic flux analysis techniques to B. subtilis cells grown under well-controlled conditions in a glucose-limited chemostat at a growth rate of 0.1 h-1 under i) reference conditions, ii) in the presence of 1.2 M NaCl, and iii) in the presence of 1.2 M NaCl and 1 mM GB. Microarray hybridizations were performed with RNA from three biological replicates. The individual samples were labeled with Cy5; a reference pool containing equal amounts of RNA from all 9 samples was labeled with Cy3.

ORGANISM(S): Bacillus subtilis

SUBMITTER: Ulrike MM-CM-$der 

PROVIDER: E-GEOD-53333 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Adaptation of Bacillus subtilis carbon core metabolism to simultaneous nutrient limitation and osmotic challenge: a multi-omics perspective.

Kohlstedt Michael M   Sappa Praveen K PK   Meyer Hanna H   Maaß Sandra S   Zaprasis Adrienne A   Hoffmann Tamara T   Becker Judith J   Steil Leif L   Hecker Michael M   van Dijl Jan Maarten JM   Lalk Michael M   Mäder Ulrike U   Stülke Jörg J   Bremer Erhard E   Völker Uwe U   Wittmann Christoph C  

Environmental microbiology 20140331 6


The Gram-positive bacterium Bacillus subtilis encounters nutrient limitations and osmotic stress in its natural soil ecosystem. To ensure survival and sustain growth, highly integrated adaptive responses are required. Here, we investigated the system-wide response of B. subtilis to different, simultaneously imposed stresses. To address the anticipated complexity of the cellular response networks, we combined chemostat experiments under conditions of carbon limitation, salt stress and osmoprotect  ...[more]

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