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Rapid regulations of metabolic reactions in Escherichia coli via light-responsive enzyme redistribution
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  • Zikang Huang,
  • Lize Sun,
  • Genzhe Lu,
  • Hongrui Liu,
  • Zihan Zhai,
  • Site Feng,
  • Ji Gao,
  • Chunyu Chen,
  • Chuheng Qing,
  • Meng Fang,
  • Bowen Chen,
  • Jiale Fu,
  • Xuan Wang,
  • George Chen
Zikang Huang
Tsinghua University

Corresponding Author:huangzk17@tsinghua.org.cn

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Hongrui Liu
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Zihan Zhai
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Chunyu Chen
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Chuheng Qing
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Bowen Chen
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George Chen
School of Life Sciences, Tsinghua University
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Protein-based condensates have been proposed to accelerate biochemical reactions by enriching reactants and enzymes simultaneously. Here, we engineered those condensates into a Photo-Activated Switch in E. coli (PhASE) to regulate enzymatic reactions via tuning the spatial correlation of enzymes and substrates. In this system, scaffold proteins undergo liquid-liquid phase separation (LLPS) to form light-responsive compartments. Tethered with a light-responsive protein, enzymes of interest (EOIs) can be recruited by those compartments from cytosol within only a few seconds after a pulse of light induction and fully released in 15 minutes. Furthermore, we managed to enrich small molecular substrates simultaneously with enzymes in the compartments and achieved the acceleration of luciferin and catechol oxidation by 2.3 folds and 1.6 folds, respectively. We also developed a quantitative model to guide the further optimization of this de-mixed regulatory system. Our tool can thus be used to study the rapid redistribution of proteins, and reversibly regulate enzymatic reactions in E. coli.
15 Mar 2022Submitted to Biotechnology Journal
16 Mar 2022Submission Checks Completed
16 Mar 2022Assigned to Editor
23 Mar 2022Reviewer(s) Assigned
11 Apr 2022Editorial Decision: Revise Major
15 May 20221st Revision Received
16 May 2022Submission Checks Completed
16 May 2022Assigned to Editor
16 May 2022Reviewer(s) Assigned
25 May 2022Editorial Decision: Accept
Sep 2022Published in Biotechnology Journal volume 17 issue 9 on pages 2200129. 10.1002/biot.202200129