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

本研究的目的是研究基于代谢和基因工程的风味化合物的微生物生产。主要目的是研究大肠杆菌在辅酶辅酶a存在扰动时中央代谢途径的反应。研究的具体目标是：(1)通过干扰辅酶CoA合成或使用途径的突变菌株，研究CoA代谢改变后大肠杆菌代谢通量的重新分布；(2)在确定的生物反应器实验下，研究这些基因操作对细胞产生风味化合物能力的影响；(3)对乙醇乙酰转移酶进行定向进化，改变其特异性，生成不同模式的酯。本研究开发的框架可应用于与生物基工业产品相关的生物过程应用中代谢功能的改善。

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.