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Nat Chem Biol. 2018 Feb;14(2):142-147. doi: 10.1038/nchembio.2535. Epub 2017 Dec 11.

Engineered synthetic scaffolds for organizing proteins within the bacterial cytoplasm.

Author information

1
Industrial Biotechnology Centre, School of Biosciences, University of Kent, Canterbury, UK.
2
School of Biochemistry, University of Bristol, Medical Sciences Building, University Walk, Bristol, UK.
3
Wolfson Bioimaging Facility, Medical Sciences Building, University Walk, Bristol, UK.
4
School of Chemistry, University of Bristol, Cantock's Close, Bristol, UK.
5
Department of Biochemical Engineering, University College London, Bernard Katz Building, Gordon Street, London, UK.
6
BrisSynBio, Life Sciences Building, Tyndall Avenue, Bristol, UK.

Abstract

We have developed a system for producing a supramolecular scaffold that permeates the entire Escherichia coli cytoplasm. This cytoscaffold is constructed from a three-component system comprising a bacterial microcompartment shell protein and two complementary de novo coiled-coil peptides. We show that other proteins can be targeted to this intracellular filamentous arrangement. Specifically, the enzymes pyruvate decarboxylase and alcohol dehydrogenase have been directed to the filaments, leading to enhanced ethanol production in these engineered bacterial cells compared to those that do not produce the scaffold. This is consistent with improved metabolic efficiency through enzyme colocation. Finally, the shell-protein scaffold can be directed to the inner membrane of the cell, demonstrating how synthetic cellular organization can be coupled with spatial optimization through in-cell protein design. The cytoscaffold has potential in the development of next-generation cell factories, wherein it could be used to organize enzyme pathways and metabolite transporters to enhance metabolic flux.

PMID:
29227472
DOI:
10.1038/nchembio.2535

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