‘Phase‐Separated Multienzyme Compartmentalization for Terpene Biosynthesis in a Prokaryote'

Abstract

Liquid–liquidphase separation(LLPS) forms biomolecular condensates or coacervatesin cells. Metabolic enzymescan form phase-separated subcellular compartments that enrich enzymes,cofactors, and substrates. Herein, 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 by LLPS. Multienzyme condensates were then formed by assembling two enzymes Idi and IspA through an RIAD/RIDD interaction. Multienzyme condensates constructed inside E.colicells compartmentalized the cytosolic space into regions of high and low enzyme density and led to a significant enhancement of α-farnesene production. This work demonstrates LLPS-driven compartmentalization of the cytosolic space of prokaryotic cells, condensation of a biosynthetic pathway,and enhancement of the biosynthesis of α-farnesene.

Publication
Angewandte Chemie International Edition, (2022)