Abstract

Trichosporon cutaneum is a soil yeast that utilizes a variety of aromatic compounds as sole carbon and energy sources. T. cutaneum is perhaps the best-characterized aromatic-degrading fungus from which further information about biodegradation can be obtained. In this thesis, the global effects on the T. cutaneum proteome associated with growth on phenol have been examined. In order to identify all the enzymes involved in the phenol degradation pathway and to find other proteins associated with the degradation process that are not part of the catabolic pathway, proteomic analysis combine high-resolution two-dimensional gel electrophoresis (2-DE) and mass spectrometry. The optimized sample preparation methods and experimental conditions provide a great advantage in quantitative analysis and potential high-throughout applications. Three mass spectrometry methods, matrix-assisted laser desorption/ionization quadrupole time-of-flight mass spectrometry (MALDI-QTOF-MS), nano-electrospray mass spectrometry (nanoESI-MS/MS) and capillary liquid chromatography mass spectrometry (CapLC-MS/MS), have been successfully applied to studying proteins of T. cutaneum strain ATCC 58094 differentially expressed in response to growth on phenol. Cross-species identification provides a powerful tool in protein identification with an unknown genome and was applied to the mass spectral data obtained. Except for phenol hydroxylase and cis, cis -muconate cyclase, the other identified 24 proteins were not previously known to be associated with phenol degradation in T. cutaneum