ME: An Integrated Metabolic Engineering Study of Evolved Alcohol Acetyl Transferase Enzymes in Flavor Compound Formation in E. coli

这项研究的目的是基于代谢和基因工程来研究微生物生产风味化合物。主要目的是研究在辅因子辅酶A存在扰动的情况下，在大肠杆菌中的中心代谢途径的反应。这项研究的具体目标是：(1)通过使用辅因子CoA的合成或使用途径上的扰动突变菌株，研究CoA代谢改变时在大肠杆菌中代谢通量的重新分配；(2)在限定的生物反应器实验下，检查这些遗传操作对细胞产生风味化合物能力的影响；以及(3)进行酒精乙酰转移酶的定向进化，以改变其特异性，以产生不同模式的酯。本研究开发的框架可用于改善与生物基工业产品相关的生物过程应用的代谢功能。

The objective of the proposed research is to study the microbial production of flavor compounds based on metabolic and genetic engineering. The primary objective is to investigate the response of the central metabolic pathway in Escherichia coli in the presence of perturbations in the cofactor CoA. The specific goals of the research are to: (1) study the redistribution of metabolic fluxes in E. coli with alteration of CoA metabolism by the use of mutant strains with perturbations either in the synthesis or in the usage pathways of the cofactor CoA, (2) examine the effect of these genetic manipulations on the cellular capacity to produce flavor compounds under defined bioreactor experiments, and (3) perform directed evolution of the alcohol acetyltransferase to alter its specificity to make a different pattern of esters. The framework developed in this research could be applied to the improvement of metabolic function for bioprocess applications related to biobased industrial products.