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NanoSIMS visualization of P. putida nanotubes during utilization of 13C-lignin-derived aromatics


EMSL Project ID
51008

Abstract

Specific Aim: Image spatial 13C enrichment into Pseudomonas putida intercellular nanotubes and/or associated outer membrane vesicles during static cultivations on lignin-derived aromatic compounds.
Resource Request:
One week of NanoSIMS imaging and analysis is requested. NanoSIMS is uniquely equipped to address the central hypothesis of this work: Nanotubes and/or associated membrane vesicles are involved in carbon assimilation from aromatic compounds by the soil bacterium Pseudomonas putida KT2440 (P. putida). NanoSIMS is the only imaging mass spectrometry platform with spatial resolution suitable for analyzing 13C enrichment into nanotubes. Here, we propose to leverage the methodology developed at NREL for preparing well preserved, high vacuum compatible, conductive nanotubes with EMSL’s NanoSIMS capability and expertise. Results from this study will provide spatiotemporal resolution and a novel mechanism for the assimilation of lignin-derived compounds by soil bacteria. This research is in-line with DOE BER’s mission to understand the underlying mechanisms of carbon conversion and energy flow among plants and soil bacteria, towards improving a plant based bio-economy.

Project Details

Project type
Limited Scope
Start Date
2019-09-23
End Date
2019-11-23
Status
Closed

Team

Principal Investigator

Bryon Donohoe
Institution
National Renewable Energy Laboratory

Co-Investigator(s)

Davinia Salvachua Rodriguez
Institution
National Renewable Energy Laboratory

Team Members

Allison Werner
Institution
National Renewable Energy Laboratory

Gregg Beckham
Institution
National Renewable Energy Laboratory