Accelerating the identification of novel secondary metabolites in bioenergy plant root exudates using microcrystal electron diffraction (MicroED)
EMSL Project ID
60664
Abstract
Small molecule metabolites drive inter- and intraspecies communication and dependencies in diverse biological systems, yet a large proportion of these important chemical compounds remain uncharacterized in plants and microbes. We need advanced analytical capabilities that will support rapid discovery and structural elucidation of metabolites from biological samples that may be limited in quantity and high in complexity. Approximately 90% of the metabolites in root exudate profiles are unknown compounds, despite the importance of root exudate composition in plant-microbe interactions. To fill this gap, we will implement a novel cryo-electron microscopy (cryoEM) method ideal for sub-angstrom structural characterization of small molecules, known as microcrystal electron diffraction (MicroED), which has not yet been extensively used to study metabolites from complex biological systems. We will use MicroED to identify novel secondary metabolites from sorghum root exudates with the goal of annotating highly abundant unknown compounds previously observed in sorghum metabolomics datasets. Using crude root exudates from sorghum, this project will develop purification strategies in conjunction with crystallization screening to identify optimized conditions for generating quality metabolite crystals for MicroED analysis. This optimized sample preparation process will then be integrated with the existing data collection and data analysis pipelines for MicroED at PNNL to facilitate rapid natural product discovery.
Project Details
Start Date
2022-12-16
End Date
2023-09-30
Status
Closed
Released Data Link
Team
Principal Investigator
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