Employing bacterial microcompartment technology to engineer a shell-free enzyme-aggregate for enhanced 1,2-propanediol production in Escherichia coli

Matthew J. Lee, Ian R. Brown, Rokas Juodeikis, Stefanie Frank, Martin J. Warren

Research output: Contribution to journalArticlepeer-review

56 Citations (Scopus)

Abstract

Bacterial microcompartments (BMCs) enhance the breakdown of metabolites such as 1,2-propanediol (1,2-PD) to propionic acid. The encapsulation of proteins within the BMC is mediated by the presence of targeting sequences. In an attempt to redesign the Pdu BMC into a 1,2-PD synthesising factory using glycerol as the starting material we added N-terminal targeting peptides to glycerol dehydrogenase, dihydroxyacetone kinase, methylglyoxal synthase and 1,2-propanediol oxidoreductase to allow their inclusion into an empty BMC. 1,2-PD producing strains containing the fused enzymes exhibit a 245% increase in product formation in comparison to un-tagged enzymes, irrespective of the presence of BMCs. Tagging of enzymes with targeting peptides results in the formation of dense protein aggregates within the cell that are shown by immuno-labelling to contain the vast majority of tagged proteins. It can therefore be concluded that these protein inclusions are metabolically active and facilitate the significant increase in product formation.

Original languageEnglish
Pages (from-to)48-56
Number of pages9
JournalMetabolic Engineering
Volume36
DOIs
Publication statusPublished - 1 Jul 2016

Keywords

  • Biotechnology
  • Compartmentalisation
  • Metabolic engineering
  • Protein aggregation
  • Synthetic biology

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