Gao, CongHou, JianshenXu, PengGuo, LiangChen, XiulaiHu, GuipengYe, ChaoEdwards, HarleyChen, JianChen, WeiLiu, Liming2019-11-042019-11-042019-08-21Gao, Cong; Hou, Jianshen; Xu, Peng; Guo, Liang; Chen, Xiulai; Hu, Guipeng; Ye, Chao; Edwards, Harley; Chen, Jian; Chen, Wei; Liu, Liming; Programmable biomolecular switches for rewiring flux in Escherichia coli; Nature Communication 10, 3751 (2019); doi:10.1038/s41467-019-11793-7https://doi.org/10.1038/s41467-019-11793-7http://hdl.handle.net/11603/16025Synthetic 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.12 pagesen-USThis item is likely protected under Title 17 of the U.S. Copyright Law. Unless on a Creative Commons license, for uses protected by Copyright Law, contact the copyright holder or the author.Attribution 4.0 International (CC BY 4.0)Applied microbiologyGenetic circuit engineeringMetabolic engineeringSynthetic biologyText