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Engineered synthetic scaffolds for organizing proteins within the bacterial cytoplasm

Lee, Matthew J., Mantell, Judith, Hodgson, Lorna, Alibhai, Dominic, Fletcher, Jordan M, Brown, I.R., Frank, Stefanie, Xue, Wei-Feng, Verkade, Paul, Woolfson, Derek N, and others. (2017) Engineered synthetic scaffolds for organizing proteins within the bacterial cytoplasm. Nature Chemical Biology, 14 . pp. 142-147. ISSN 1552-4450. E-ISSN 1552-4469. (doi:10.1038/nchembio.2535) (KAR id:65739)

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.

Item Type: Article
DOI/Identification number: 10.1038/nchembio.2535
Divisions: Divisions > Division of Natural Sciences > Biosciences
Depositing User: Martin Warren
Date Deposited: 16 Jan 2018 12:21 UTC
Last Modified: 09 Dec 2022 07:13 UTC
Resource URI: https://kar.kent.ac.uk/id/eprint/65739 (The current URI for this page, for reference purposes)

University of Kent Author Information

Brown, I.R..

Creator's ORCID:
CReDIT Contributor Roles:

Xue, Wei-Feng.

Creator's ORCID: https://orcid.org/0000-0002-6504-0404
CReDIT Contributor Roles:

Warren, Martin J..

Creator's ORCID: https://orcid.org/0000-0002-6028-6456
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