Abstract
Bacterial membrane vesicles (MVs) mediate diverse microbial processes and are emerging as powerful biomedical tools, but MV population heterogeneity remains an open question. Here, we separate, enumerate, and characterize two MV populations from the soil bacterium Pseudomonas putida during growth with or without lignin-derived carbon, a major carbon source from plant cells in the rhizosphere. Small MVs (MV-S, diameter ~100 nm) were produced from all cultures, whereas large MVs (MV-L, diameter ~300 nm) were observed during the late stationary phase of lignin cultivations. MV-S contained selectively packaged proteins with diverse physiological functions, whereas the MV-L proteome was smaller and largely enriched in outer membrane proteins. Interestingly, enzymes known to mediate the catabolism of lignin-derived aromatic compounds were enriched in MV-S. Overall, this study highlights the need for careful consideration of MV populations in microbial systems.
| Original language | English |
|---|---|
| Journal | Applied and Environmental Microbiology |
| Volume | 91 |
| Issue number | 10 |
| DOIs | |
| State | Published - Oct 2025 |
Funding
This work was authored in part by the National Renewable Energy Laboratory for the U.S. Department of Energy (DOE) under contract no. DE-AC36-08GO28308. This work was also authored in part by Oak Ridge National Laboratory, managed by UT-Battelle, LLC for the US DOE under contract no. DE-AC05-00OR22725. This work was supported by the Center for Bioenergy Innovation (CBI), U.S. Department of Energy, Office of Science, Biological and Environmental Research Program, under award number ERKP886 and U.S. Department of Energy (DOE) DE-SC0022181. We thank Sean Woodworth for analytical assistance. The views expressed in the article do not necessarily represent the views of the DOE or the U.S. Government. The U.S. Government retains and the publisher, by accepting the article for publication, acknowledges that the U.S. Government retains a nonexclusive, paid-up, irrevocable, worldwide license to publish or reproduce the published form of this work, or allow others to do so, for U.S. Government purposes. A.Z.W., R.J.G., D.S., R.L.H., and G.T.B. conceived and designed the study. A.Z.W. conducted microbial cultivation and M.V. enrichment experiments, C.P. and S.K.R.W. conducted the AF4 experiments and analysis, and A.Z.W., R.J.G., M.J.K., D.L.C., P.E.A., and R.L.H. conducted the proteomics and lipidomics experiments and analyses. A.Z.W., R.J.G., M.J.K., D.L.C., and R.J.W. conducted formal analyses and data visualization, and L.A., R.L.H., and G.T.B. acquired funding. A.Z.W. wrote the manuscript with input from all authors. Biological and Environmental Research DE-SC0022181 Ludmilla Aristilde Biological and Environmental Research ERKP886 Gregg T. Beckham This work was supported by the Center for Bioenergy Innovation (CBI), U.S. Department of Energy, of Science, Biological and Environmental Research Program, under award number ERKP886 and U.S. Department of Energy (DOE) DE-SC0022181. We thank Sean Woodworth for analytical assistance. This work was authored in part by the National Renewable Energy Laboratory for the U.S. Department of Energy (DOE) under contract no. DE-AC36-08GO28308. This work was also authored in part by Oak Ridge National Laboratory, managed by UT-Battelle, LLC for the US DOE under contract no. DE-AC05-00OR22725.
Keywords
- biological lignin valorization
- extracellular vesicles
- lipidomics
- outer membrane vesicles
- proteomics