Entwicklung von systematischen und kombinatorischen Methoden zum Metabolic Engineering von Escherichia coli zur Produktion von aromatischen Aminosäuren

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Metabolic engineering aims to alter cellular phenotype through introduction of genetic controls and cellular alterations. On the physiological level, the phenotype is a manifestation of gene expression levels, metabolic demand, resource availability, and cellular stresses. Above all, metabolic function is constrained by the stoichiometry of the reaction network. During the past decade, metabolic engineering has produced an impressive portfolio of results that were guided by rational analysis of rather well understood systems of a kinetic and regulatory standpoint. However, many systems of interest have not been well characterized, particularly systems involving the production of non-native, value-added products. Even with extensive molecular biological tools which have been developed to introduce genetic control, the identification of gene targets and extraction of putative parameters remains an open problem. A combination of systematic and combinatorial approaches can aid to accomplish this task, of systematic and coombinatorial approaches can aid to accomplish this task, considering the properties of the overall metabolic network. These issues should be addressed computationally and experimentally in the context of heterolougos lycopene production in Escherichia coli. The systematic method will make use of the Flux Balance Analysis framework while the combinatorial method will rely on random transposon mutagenesis and promoter engineering.