EAGER: Synthetic Control of Metabolism by Dynamic Metabolons

该项目解决了代谢工程中的一个具有挑战性的问题，即以有效和动态控制的方式将细胞中的化学反应引导到代谢途径中，但不影响细胞生长。这两个合作者在代谢流量分析和蛋白质工程方面提供互补的专业知识，以追求高度创新的设计概念来实现这一目标。在这个项目上接受培训的学生将在预期显著提高生物制造能力的研究背景下接受多学科培训。其目标是开发合成的动态代谢物，使碳通量能够根据需要进行重定向。最终结果将是为最佳产品形成提供微生物代谢的动态优化的能力。这将通过合成代谢物的组装和分解之间的动态转换来实现，以调节整体输出函数。具体地说，连续的代谢酶将被连接到PUF模块上，该模块也能够结合特定的核糖开关。这些核糖开关被设计成对特定的代谢或合成信号做出反应。可行性将通过设计两种多酶动态代谢物来证明，其中一种将甲醇转化为己酮糖-6-磷酸，这是一种有价值的化学品和燃料生产的前体。第二种代谢蛋白包括将乙酰辅酶A转化为甲氧戊酸的酶。最终目标是将所获得的知识结合起来，设计用于增强脂肪酸合成的动态代谢物。

This project addresses a challenging problem in metabolic engineering, namely the channelling of chemical reactions in the cell into metabolic pathways in an efficient and dynamically controlled manner, but without affecting cellular growth. The two collaborators offer complementary expertise in the analysis of metabolic flux and protein engineering in order to pursue a highly innovative design concept to achieve this goal. The students trained on this project will be exposed to multi-disciplinary training in the context of research that is anticipated to significantly advance biomanufacturing capabilities. The goal is to develop synthetic dynamic metabolons that will allow carbon flux to be redirected as desired. The end result will be the ability to provide dynamic optimization of microbial metabolism for optimal product formation. This will be achieved by dynamic shifts between the assembly and disassembly of synthetic metabolons in order to modulate the overall output function. Specifically, successive metabolic enzymes will be coupled onto Pumilio (PUF) modules, which are also capable of binding specific riboswitches. These riboswitches are designed to respond to specific metabolic or synthetic signals. Feasibility will be demonstrated by designing two multi-enzyme dynamic metabolons, one that converts methanol into hexulose-6-phosphate, which is a valuable precursor for chemical and fuel production. The second metabolon includes enzymes that convert acetyl-CoA into mevalonate. The ultimate goal is to combine the knowledge gained toward the design of dynamic metabolons for enhanced fatty acid synthesis.