Ms. Yue Wang
Liquid-liquid phase separation (LLPS) forms biomolecular condensates or coacervates in cells. Metabolic enzymes can form phase-separated subcellular compartments that enrich enzymes, cofactors, and substrates. Here, we report the construction of synthetic multienzyme condensates that catalyze the biosynthesis of a terpene, 𝛼-farnesene in the prokaryote E. coli. RGGRGG derived from LAF-1 was used as the scaffold protein to form the condensates via LLPS. Multienzyme condensates were then formed by assembling two enzymes Idi and IspA through an RIAD/RIDD interaction. Multienzyme condensates constructed inside E. coli cells compartmentalized the cytosolic space into regions of high and low enzyme density and led to a significant enhancement of 𝛼-farnesene production. This work manifests LLPS-driven compartmentalization of the cytosolic space of prokaryotic cells, condensation of a biosynthetic pathway, and enhancement of of the biosynthesis of 𝛼-farnesene.
Formation of RGG protein condensate in E. coli
LLPS-mediated protein clustering in E. coli cells
Compartmentalizing biosynthetic enzymes to increase 𝛼-farnesene production in E. coli
 P. A. Srere, Trends Biochem. Sci. 1985, 10, 109-110.
 S. Boeynaems, S. Alberti, N. L. Fawzi, T. Mittag, M. Polymenidou, F. Rousseau, J. Schymkowitz, J. Shorter, B. Wolozin, L. Van Den Bosch, Trends Cell Biol. 2018, 28, 420-435.
 M. Saito, D. Hess, J. Eglinger, A. W. Fritsch, M. Kreysing, B. T. Weinert, C. Choudhary, P. Matthias, Nat. Chem. Biol. 2019, 15, 51-61.
 S. F. Banani, H. O. Lee, A. A. Hyman, M. K. Rosen, Nat. Rev. Mol. Cell Biol. 2017, 18, 285-298.
 B. R. Sabari, A. Dall’Agnese, A. Boija, I. A. Klein, E. L. Coffey, K. Shrinivas, B. J. Abraham, N. M. Hannett, A. V. Zamudio, J. C. Manteiga, Science 2018, 361, eear3958.
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