Programmable biomolecular switches for rewiring flux in Escherichia coli
Programmable biomolecular switches for rewiring flux in Escherichia coli
复制标题
用于在大肠杆菌中重新布线通量的可编程生物分子开关
DOI：
10.1038/s41467-019-11793-7
复制
发表时间：
2019-08-21
影响因子：
16.6
通讯作者：
Liu, Liming
中科院分区：
文献类型：
--
作者：
Gao, Cong;Hou, Jianshen;Liu, Liming
合成生物学的目标是开发可编程工具来执行复杂的功能，如重新分配工业微生物中的代谢流量。然而，蛋白质级电路的发展受到可设计、正交和可组合工具的限制。在这里，我们借助工程病毒蛋白水解酶和蛋白分解信号，构建了两套可控的蛋白质单元，可以合理地配置为三个工具。利用基于蛋白水解酶的动态调节电路对代谢流进行微调，在不加诱导剂的最低限度培养条件下，获得了12.63g L-1莽草酸滴度。此外，在多种碳源下，基于蛋白酶的逆转器介导的通量重分配可以缓解碳分解代谢的抑制。通过在大肠杆菌中使用基于酶的振荡器来协调反应速度，我们获得了D-木酸的产率7.12g L-1h(-1)和效价199.44 g L-1。这些结果突出了可编程蛋白质开关在代谢工程中的适用性，以生产有价值的化学品。
Synthetic biology aims to develop programmable tools to perform complex functions such as redistributing metabolic flux in industrial microorganisms. However, development of protein-level circuits is limited by availability of designable, orthogonal, and composable tools. Here, with the aid of engineered viral proteases and proteolytic signals, we build two sets of controllable protein units, which can be rationally configured to three tools. Using a protease-based dynamic regulation circuit to fine-tune metabolic flow, we achieve 12.63 g L-1 shikimate titer in minimal medium without inducer. In addition, the carbon catabolite repression is alleviated by protease-based inverter-mediated flux redistribution under multiple carbon sources. By coordinating reaction rate using a protease-based oscillator in E. coil, we achieve D-xylonate productivity of 7.12 g L-1 h(-1) with a titer of 199.44 g L-1 . These results highlight the applicability of programmable protein switches to metabolic engineering for valuable chemicals production.