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Download fileConstruction of Recombinant Pdu Metabolosome Shells for Small Molecule Production in Corynebacterium glutamicum
journal contribution
posted on 2017-08-21, 00:00 authored by Isabel Huber, David J. Palmer, Kira N. Ludwig, Ian R. Brown, Martin J. Warren, Julia FrunzkeBacterial microcompartments have
significant potential in the area
of industrial biotechnology for the production of small molecules,
especially involving metabolic pathways with toxic or volatile intermediates. Corynebacterium glutamicum is an established industrial
workhorse for the production of amino acids and has been investigated
for the production of diamines, dicarboxylic acids, polymers and biobased
fuels. Herein, we describe components for the establishment of bacterial
microcompartments as production chambers in C. glutamicum. Within this study, we optimized genetic clusters for the expression
of the shell components of the Citrobacter freundii propanediol utilization (Pdu) bacterial compartment, thereby facilitating
heterologous compartment production in C. glutamicum. Upon induction, transmission electron microscopy images of thin
sections from these strains revealed microcompartment-like structures
within the cytosol. Furthermore, we demonstrate that it is possible
to target eYFP to the empty microcompartments through C-terminal fusions
with synthetic scaffold interaction partners (PDZ, SH3 and GBD) as
well as with a non-native C-terminal targeting peptide from AdhDH
(Klebsiella pneumonia). Thus, we show that it is
possible to target proteins to compartments where N-terminal targeting
is not possible. The overproduction of PduA alone leads to the construction
of filamentous structures within the cytosol and eYFP molecules are
localized to these structures when they are N-terminally fused to
the P18 and D18 encapsulation peptides from PduP and PduD, respectively.
In the future, these nanotube-like structures might be used as scaffolds
for directed cellular organization and pathway enhancement.
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cytosolCorynebacterium glutamicumGBDmicrocompartmentacidCitrobacter freundii propanediol utilization. glutamicumtransmission electron microscopy imagesSmall Molecule ProductioncomponentRecombinant Pdu Metabolosome ShellseYFPPDZscaffold interaction partnersSHheterologous compartment productionD 18 encapsulation peptidespathwaymoleculeC-terminalC . glutamicum