Improving lycopene production in Escherichia coli by engineering metabolic control
Improving lycopene production in Escherichia coli by engineering metabolic control
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DOI：
10.1038/75398
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发表时间：
2000-05-01
影响因子：
46.9
通讯作者：
Liao, JC
中科院分区：
文献类型：
--
作者：
Farmer, WR;Liao, JC
代谢工程已经在多种宿主中产生外源代谢物方面取得了令人鼓舞的成功。然而，工程代谢途径的常见策略集中在扩增所需的酶和解除细胞控制。因此，不受控制或失调的代谢途径导致代谢失衡和次优生产力。在这里，我们已经证明了代谢工程的第二阶段的努力，通过设计和工程的调控电路，以控制基因表达，响应细胞内代谢状态。具体来说，我们招募并改变了大肠杆菌中的一个全局调节系统，即Ntr调节子，以控制工程番茄红素生物合成途径。人工工程调节子，通过感测细胞内代谢物乙酰磷酸，由过量的糖酵解通量刺激，控制番茄红素合成中两个关键酶的表达，以响应通量动力学。这种细胞内控制回路显着提高番茄红素的生产，同时减少代谢失衡造成的负面影响。虽然我们证明了这种策略的代谢产物的生产，它可以扩展到其他领域的基因表达必须密切控制的细胞内生理，如基因治疗。
Metabolic engineering has achieved encouraging success in producing foreign metabolites in a variety of hosts. However, common strategies for engineering metabolic pathways focus on amplifying the desired enzymes and deregulating cellular controls. As a result, uncontrolled or deregulated metabolic pathways lead to metabolic imbalance and suboptimal productivity. Here we have demonstrated the second stage of metabolic engineering effort by designing and engineering a regulatory circuit to control gene expression in response to intracellular metabolic states. Specifically, we recruited and altered one of the global regulatory systems in Escherichia coli, the Ntr regulon, to control the engineered lycopene biosynthesis pathway. The artificially engineered regulon, stimulated by excess glycolytic flux through sensing of an intracellular metabolite, acetyl phosphate, controls the expression of two key enzymes in lycopene synthesis in response to flux dynamics. This intracellular control loop significantly enhanced lycopene production while reducing the negative impact caused by metabolic imbalance. Although we demonstrated this strategy for metabolite production, it can be extended into other fields where gene expression must be closely controlled by intracellular physiology, such as gene therapy.