the opportunistic pathogen Malassezia pachydermatis causes bloodstream infections in preterm infants or individuals with immunodeficiency disorders and has been associated with a broad spectrum of diseases in animals such as seborrheic dermatitis, external otitis and fungemia. The current approaches to treat these infections are failing as a consequence of their adverse effects, changes in susceptibility and antifungal resistance. Thus, the identification of novel therapeutic targets against M. pachydermatis infections are highly relevant. Here, Gene Essentiality Analysis and Flux Variability Analysis was applied to a previously reported M. pachydermatis metabolic network to identify enzymes that, when absent, negatively affect biomass production. Three novel therapeutic targets (i.e., homoserine dehydrogenase (MpHSD), homocitrate synthase (MpHCS) and saccharopine dehydrogenase (MpSDH)) were identified that are absent in humans. Notably, L-lysine was shown to be an inhibitor of the enzymatic activity of MpHCS and MpSDH at concentrations of 1 mM and 75 mM, respectively, while L-threonine (1 mM) inhibited MpHSD. Interestingly, L-lysine was also shown to inhibit M. pachydermatis growth during in vitro assays with reference strains and canine isolates, while it had a negligible cytotoxic activity on HEKa cells. Together, our findings form the bases for the development of novel treatments against M. pachydermatis infections. The yeast M. pachydermatis is part of the skin microbiota of domestic and wild animals and behaves as an opportunistic pathogen causing external otitis and seborrheic dermatitis in dogs and cats. Particular conditions such as the presence of lipid-rich microenvironments, a local imbalance of the natural microbiota and altered immune states favor these infections 1. Dermatologic infections caused by M. pachydermatis often exhibit a chronic (recurrent) course and their treatment can be complicated due to the ability of this yeast to form biofilms 1. In addition, M. pachydermatis causes bloodstream infections in preterm infants or in individuals with immunodeficiency disorders. These infections are related to contamination of medical devices such as catheters, the transmission through medical staff and the administration of lipids through intravenous way 2,3. Recently, several factors contributing to M. pachydermatis virulence have been determined, which include the production of proteinases, phospholipases, hyaluronidases, and chondroitin-sulfatases 4. Currently, five classes of antifungal agents are used orally, topically or intravenously for the treatment of fungal infections. The first class is formed by the azoles (ketoconazole, itraconazole, clotrimazole, miconazole, and