Engineered synthetic scaffolds for organizing proteins within the bacterial cytoplasm

Lee, Matthew J, Mantell, Judith, Hodgson, Lorna, Alibhai, Dominic, Fletcher, Jordan M, Brown, Ian R, Frank, Stefanie, Xue, Wei-Feng, Verkade, Paul, Woolfson, Derek N and Warren, Martin J ORCID: (2018) Engineered synthetic scaffolds for organizing proteins within the bacterial cytoplasm. Nature Chemical Biology, 14 (2). pp. 142-147. ISSN 1552-4450

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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.

Item Type: Article
Faculty \ School: Faculty of Science
UEA Research Groups: Faculty of Medicine and Health Sciences > Research Centres > Norwich Institute for Healthy Aging
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Depositing User: LivePure Connector
Date Deposited: 24 Sep 2020 00:04
Last Modified: 21 Apr 2023 00:45
DOI: 10.1038/nchembio.2535

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