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

代谢工程旨在通过引入遗传控制和细胞改变来改变细胞表型。在生理水平上，表型是基因表达水平、代谢需求、资源可获得性和细胞压力的表现。最重要的是，代谢功能受制于反应网络的化学计量比。在过去的十年中，代谢工程产生了令人印象深刻的结果组合，这些结果是由对动力学和监管立场的相当好的理解的系统的理性分析指导的。然而，许多感兴趣的系统没有得到很好的特征，特别是涉及生产非本地增值产品的系统。即使已经开发了大量的分子生物学工具来引入基因控制，但识别基因靶标和提取假定参数仍然是一个悬而未决的问题。考虑到整个代谢网络的特性，系统和组合方法的组合可以帮助完成这一任务，系统和组合方法的组合可以帮助完成这一任务。这些问题应该在大肠杆菌中异色番茄红素生产的背景下通过计算和实验来解决。系统化方法将利用通量平衡分析框架，而组合方法将依赖于随机转座子突变和启动子工程。

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.