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Published on: March 2, 2009
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High-throughput Saccharomyces cerevisiae cultivation method for credentialing-based untargeted metabolomics
Lorenzo Favilli1, Corey M Griffith2, Emma L Schymanski2
1Luxembourg Centre for Systems Biomedicine (LCSB), University of Luxembourg, Avenue du Swing 6, Belvaux, L-4367, Luxembourg. lorenzo.favilli@uni.lu.
Analytical and Bioanalytical Chemistry
|May 22, 2023
Summary
This study introduces a high-throughput method for yeast metabolomics using stable isotope labeling to accurately identify biological metabolites. This approach enhances the understanding of cellular metabolism and aids in discovering new metabolic functions.
Area of Science:
- Metabolomics
- Systems Biology
- Yeast Genetics
Background:
- Understanding cellular metabolism is crucial for biology and disease research.
- Many genes in model organisms like Saccharomyces cerevisiae remain uncharacterized.
- Untargeted high-resolution mass spectrometry detects many features, but distinguishing biological signals from noise is challenging.
Purpose of the Study:
- To develop a high-throughput, stable isotope labeling (SIL)-based approach for untargeted metabolomics in yeast.
- To improve the credentialing of biologically relevant metabolites detected by mass spectrometry.
- To enable efficient molecular phenotypic screening and complete metabolic networks.
Main Methods:
- Developed a SIL-based approach for high-throughput untargeted metabolomics in S. cerevisiae.
- Utilized deep-48 well format cultivation and metabolite extraction.
- Employed HILIC and RP liquid chromatography coupled to Orbitrap Q Exactive HF mass spectrometry.
- Applied open-source software (MS-DIAL, MetFrag, SIRIUS CSI:FingerID, MetaboAnalyst) for data analysis and peak annotation and verification engine (PAVE) tool.
Main Results:
- Credentialed and analyzed 3-7% of approximately 37,000 detected features.
- Successfully annotated 198 metabolites using MS2 database matching.
- Observed comparable metabolic profiles between deep-well plates and shake flasks.
- Confirmed expected metabolic changes in the sdh1Δ yeast strain.
Conclusions:
- The developed approach enables high-throughput yeast cultivation and credentialing-based untargeted metabolomics.
- This method efficiently performs molecular phenotypic screens.
- It provides a means to help complete metabolic networks in well-characterized organisms.
Keywords:
High-throughput sample generationLiquid chromatographyMetabolomicsSaccharomyces cerevisiaeStable isotope labellingUntargeted high-resolution mass spectrometryMore Related Videos
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