Programmable biomolecular switches for rewiring flux in Escherichia coli

Cong Gao, Jianshen Hou, Peng Xu, Liang Guo, Xiulai Chen, Guipeng Hu, Chao Ye, Harley Edwards, Jian Chen, Wei Chen and Liming Liu ()
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Cong Gao: Jiangnan University
Jianshen Hou: Jiangnan University
Peng Xu: University of Maryland Baltimore County
Liang Guo: Jiangnan University
Xiulai Chen: Jiangnan University
Guipeng Hu: Jiangnan University
Chao Ye: Jiangnan University
Harley Edwards: University of Maryland Baltimore County
Jian Chen: Jiangnan University
Wei Chen: Jiangnan University
Liming Liu: Jiangnan University

Nature Communications, 2019, vol. 10, issue 1, 1-12

Abstract: Abstract 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. coli, 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.

Date: 2019
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DOI: 10.1038/s41467-019-11793-7

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