Engineered synthetic scaffolds for organizing proteins within bacterial cytoplasms

Matthew J. Lee, Judith Mantell, Lorna Hodgson, Dominic Alibhai, Jordan M. Fletcher, Ian R. Brown, Stefanie Frank, Wei Feng Xue, Paul Verkade, Derek N. Woolfson, Martin J. Warren

Research output: Contribution to journalArticle (Academic Journal)peer-review

55 Citations (Scopus)
359 Downloads (Pure)

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 with 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 for the development of next-generation cell factories, where it could be used to organize enzyme pathways and metabolite transporters to enhance metabolic flux.
Original languageEnglish
Pages (from-to)142-147
Number of pages6
JournalNature Chemical Biology
Volume14
Issue number2
Early online date11 Dec 2017
DOIs
Publication statusPublished - 1 Feb 2018

Structured keywords

  • BrisSynBio
  • Bristol BioDesign Institute

Keywords

  • synthetic biology
  • bacteria
  • metabolic engineering
  • protein design

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