| Summary: | <i>Hyphozyma roseonigra</i> is a dimorphic yeast used as a biocatalyst to convert sclareol, a plant diterpenoid to ambradiol. The latter is an intermediate in the synthesis of ambrafuran, a high-value chemical in the fragrance industry. Unfortunately, little is known about the underlying biochemistry of this microorganism. In this study, the integration of multi-platform-based metabolomics was used to better comprehend <i>H. roseonigra</i> from a biochemical perspective. The focus on metabolomic changes during growth and development was accomplished using untargeted LC–MS and NMR analyses. Cell suspensions were grown in batch culture over a 14-day period, and cells from the early-, log-, and stationary phases were harvested every second day using platform-compatible extraction procedures. Following chemometric analysis of LC–MS and NMR data acquired from both intra- and extracellular extracts, the identified discriminatory ions annotated from the endo- and exometabolomes (metabo-fingerprinting and metabo-footprinting) were found to fall predominantly in the primary metabolism class. Pathway mapping and feature-based network correlation analysis assisted in gaining insights into the active metabolic pathways during growth and development and did not flag terpene synthesis. This study provides novel insights into the basic metabolic capabilities of <i>H. roseonigra</i> and suggests that sclareol is metabolized as the detoxification of a hydrophobic xenobiotic compound.
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