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http://dx.doi.org/10.18419/opus-2348
Autor(en): | Feuer, Ronny Gottlieb, Katrin Viertel, Gero Klotz, Johannes Schober, Steffen Bossert, Martin Sawodny, Oliver Sprenger, Georg Ederer, Michael |
Titel: | Model-based analysis of an adaptive evolution experiment with Escherichia coli in a pyruvate limited continuous culture with glycerol |
Erscheinungsdatum: | 2012 |
Dokumentart: | Zeitschriftenartikel |
Erschienen in: | EURASIP journal on bioinformatics and systems biology (2012), 2012:14. URL http://dx.doi.org./10.1186/1687-4153-2012-14 |
URI: | http://nbn-resolving.de/urn:nbn:de:bsz:93-opus-98271 http://elib.uni-stuttgart.de/handle/11682/2365 http://dx.doi.org/10.18419/opus-2348 |
Zusammenfassung: | Bacterial strains that were genetically blocked in important metabolic pathways and grown under selective conditions underwent a process of adaptive evolution: certain pathways may have been deregulated and therefore allowed for the circumvention of the given block. A block of endogenous pyruvate synthesis from glycerol was realized by a knockout of pyruvate kinase and phosphoenolpyruvate carboxylase in E. coli. The resulting mutant strain was able to grow on a medium containing glycerol and lactate, which served as an exogenous pyruvate source. Heterologous expression of a pyruvate carboxylase gene from Corynebacterium glutamicum was used for anaplerosis of the TCA cycle. Selective conditions were controlled in a continuous culture with limited lactate feed and an excess of glycerol feed. After 200–300 generations pyruvate-prototrophic mutants were isolated. The genomic analysis of an evolved strain revealed that the genotypic basis for the regained pyruvate-prototrophy was not obvious. A constraint-based model of the metabolism was employed to compute all possible detours around the given metabolic block by solving a hierarchy of linear programming problems. The regulatory network was expected to be responsible for the adaptation process. Hence, a Boolean model of the transcription factor network was connected to the metabolic model. Our model analysis only showed a marginal impact of transcriptional control on the biomass yield on substrate which is a key variable in the selection process. In our experiment, microarray analysis confirmed that transcriptional control probably played a minor role in the deregulation of the alternative pathways for the circumvention of the block. |
Enthalten in den Sammlungen: | 04 Fakultät Energie-, Verfahrens- und Biotechnik |
Dateien zu dieser Ressource:
Datei | Beschreibung | Größe | Format | |
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Feuer2012.pdf | 756,53 kB | Adobe PDF | Öffnen/Anzeigen |
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