Abstract
Engineering subcellular organization in microbes shows great promise in addressing bottlenecks in metabolic engineering efforts; however, rules guiding selection of an organization strategy or platform are lacking. Here, we study compartment morphology as a factor in mediating encapsulated pathway performance. Using the 1,2-propanediol utilization microcompartment (Pdu MCP) system from Salmonella enterica serovar Typhimurium LT2, we find that we can shift the morphology of this protein nanoreactor from polyhedral to tubular by removing vertex protein PduN. Analysis of the metabolic function between these Pdu microtubes (MTs) shows that they provide a diffusional barrier capable of shielding the cytosol from a toxic pathway intermediate, similar to native MCPs. However, kinetic modeling suggests that the different surface area to volume ratios of MCP and MT structures alters encapsulated pathway performance. Finally, we report a microscopy-based assay that permits rapid assessment of Pdu MT formation to enable future engineering efforts on these structures.
| Original language | English (US) |
|---|---|
| Article number | 3746 |
| Journal | Nature communications |
| Volume | 13 |
| Issue number | 1 |
| DOIs | |
| State | Published - Dec 2022 |
Funding
The authors thank and acknowledge members of the Mangan, Olvera de la Cruz, and Tullman-Ercek groups for insightful discussions around this work. The authors specifically acknowledge Dr. Chris Jakobson for plasmids used in this study. This work was funded in part by the Army Research Office (grant W911NF-19-1-0298 to D.T.E. and M.C.J.) and the Department of Energy (grant DE-SC0019337 to D.T.E. and N.M.M. and grant DE-FG02-08ER46539 to M.O.d.l.C). N.W.K. was funded by the National Science Foundation Graduate Research Fellowship Program (grant DGE-1842165), and by the National Institutes of Health Training Grant (T32GM008449) via the Northwestern University Biotechnology Training Program. M.O.d.l.C. thanks the computational support of the Sherman Fairchild Foundation. This work made use of the BioCryo facility of Northwestern University’s NUANCE Center, which has received support from the SHyNE Resource (NSF ECCS-2025633), the IIN, and Northwestern’s MRSEC program (NSF DMR-1720139). Molecular graphics and analyses performed with UCSF Chimera, developed by the Resource for Biocomputing, Visualization, and Informatics at the University of California, San Francisco, with support from NIH P41-GM103311. The authors thank and acknowledge members of the Mangan, Olvera de la Cruz, and Tullman-Ercek groups for insightful discussions around this work. The authors specifically acknowledge Dr. Chris Jakobson for plasmids used in this study. This work was funded in part by the Army Research Office (grant W911NF-19-1-0298 to D.T.E. and M.C.J.) and the Department of Energy (grant DE-SC0019337 to D.T.E. and N.M.M. and grant DE-FG02-08ER46539 to M.O.d.l.C). N.W.K. was funded by the National Science Foundation Graduate Research Fellowship Program (grant DGE-1842165), and by the National Institutes of Health Training Grant (T32GM008449) via the Northwestern University Biotechnology Training Program. M.O.d.l.C. thanks the computational support of the Sherman Fairchild Foundation. This work made use of the BioCryo facility of Northwestern University’s NUANCE Center, which has received support from the SHyNE Resource (NSF ECCS-2025633), the IIN, and Northwestern’s MRSEC program (NSF DMR-1720139). Molecular graphics and analyses performed with UCSF Chimera, developed by the Resource for Biocomputing, Visualization, and Informatics at the University of California, San Francisco, with support from NIH P41-GM103311.
ASJC Scopus subject areas
- General Chemistry
- General Biochemistry, Genetics and Molecular Biology
- General Physics and Astronomy