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、资源可利用性和细胞应激的表现。最重要的是，代谢功能受到反应网络化学计量学的限制。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，这些结果是在对相当了解的动力学和调控立场的系统进行理性分析的指导下产生的。然而，许多令人感兴趣的系统尚未得到很好的描述，特别是涉及生产非本地增值产品的系统。即使已经开发了广泛的分子生物学工具来引入遗传控制，基因靶点的识别和假定参数的提取仍然是一个悬而未决的问题。考虑到整个代谢网络的特性，系统和组合方法的结合可以帮助完成这项任务，系统和组合方法的结合可以帮助完成这项任务。这些问题应该在大肠杆菌生产异杂番茄红素的背景下进行计算和实验。系统方法将利用通量平衡分析框架，而组合方法将依靠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.