Generally, high- and low-performance nongenetic variants and young and aged cells co-existed in culture at all growth phases. In this regard, individually and dynamically regulating the metabolic flux of single cells based on their cellular state is highly useful for improving the performance of populations. However, balancing the trade-offs between biomass formation and compound over-production requires sophisticated dynamic regulation networks (DRNs) which can be challenging. Here, we developed a growth coupled NCOMB (Naringenin-Coumaric acid-Malonyl-CoA-Balanced) DRN with systematic optimization of (2S)-naringenin and p-coumaric acid-responsive regulation pathways for real-time control of intracellular supply of malonyl-CoA. In doing so, the acyl carrier protein was used as a new critical node for fine-tuning malonyl-CoA consumption instead of fatty acid synthase. Following directed evolution of the NCOMB DRN, we obtained a strain with cumulative 8.4-fold improvement in (2S)-naringenin production with balanced cell growth, demonstrating the high efficiency of this system for improving pathway production.

中文翻译：

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平衡丙二酰-CoA节点的生长耦合动态调控网络的开发，以增强大肠杆菌中的（2S）-柚皮素合成

通常，高性能和低性能的非遗传变异以及年轻和衰老的细胞在所有生长阶段都共存于培养物中。在这方面，基于单个细胞的细胞状态单独动态地调节其代谢通量对于改善群体的性能非常有用。但是，要在生物质形成与化合物过量生产之间权衡取舍，就需要复杂的动态调节网络（DRN），这可能会带来挑战。在这里，我们开发了生长耦合NCOMB（萘林宁-香豆酸-丙二酸-CoA-平衡）DRN，具有系统优化的（2S）-柚皮素和对香豆酸响应调节途径，可实时控制细胞内丙二酸- CoA。在这样做，酰基载体蛋白代替脂肪酸合酶被用作微调丙二酰辅酶A消耗的新关键节点。遵循NCOMB DRN的定向进化，我们获得了一种菌株，该菌株在（2S）-柚皮素生产中的累积累积提高了8.4倍，并且细胞生长平衡，证明了该系统在改善途径生产方面的高效性。