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

Lee, Matthew J., Brown, Ian R., Juodeikis, Rokas, Frank, Stefanie and Warren, Martin J. ORCID: https://orcid.org/0000-0002-6028-6456 (2016) Employing bacterial microcompartment technology to engineer a shell-free enzyme-aggregate for enhanced 1,2-propanediol production in Escherichia coli. Metabolic Engineering, 36. pp. 48-56. ISSN 1096-7176

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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.

Item Type: Article
Additional Information: Funding Information: We recognise the help of Usha Dura, Mirabel Inyang, Mathilda Oxandaburu and Tulshi Patel for the initial cloning of glycerol dehydrogenase, methylglyoxal synthase and 1,2-propanediol oxidoreductase. We thank Kevin Howland for assistance with GC–MS. Financial support was provided by the Biotechnology and Biological Sciences Research Council ( BB/M002969/1 ) and the Leverhulme Trust ( ECF-213-341 ). Publisher Copyright: © 2016 The Authors.
Uncontrolled Keywords: biotechnology,compartmentalisation,metabolic engineering,protein aggregation,synthetic biology,biotechnology,bioengineering,applied microbiology and biotechnology ,/dk/atira/pure/subjectarea/asjc/1300/1305
Faculty \ School: Faculty of Science
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Depositing User: LivePure Connector
Date Deposited: 20 Sep 2022 14:31
Last Modified: 21 Sep 2022 00:22
URI: https://ueaeprints.uea.ac.uk/id/eprint/88489
DOI: 10.1016/j.ymben.2016.02.007

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