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Functional Enzyme Discovery by LC-MS Metabolomics in Yeast
Ort / Verlag
ProQuest Dissertations & Theses
Erscheinungsjahr
2015
Quelle
ProQuest Dissertations & Theses A&I
Beschreibungen/Notizen
In pursuit of novel enzymes, I have carried out a systematic analysis of gene function in the budding yeast, Saccharomyces cerevisiae. I used LC-MS metabolomics to profile metabolite levels for 120 prototrophic mutants carrying deletions in uncharacterized candidate enzyme-encoding genes. Methodology I developed in this study was successful in capturing metabolomic phenotypes for 26 candidate enzyme deletion mutants. Furthermore, I explored the function of a set of mutants deleted for genes not previously known to have a role in metabolism. I showed these mutants have altered levels of NAD+ biosynthetic intermediates. New phenotypes, here discovered, will lead to dissection of biochemical functions of uncharacterized genes and determination of their contributions to metabolism. I have also uncovered biologically-derived compounds that are uncharacterized and may be new metabolites. Overall, my work underscores LC-MS metabolomics as a powerful tool for functional genomics in its ability capture the metabolic functions of uncharacterized genes.