To explore Candida guilliermondii for the production of long-chain dicarboxylic acids (DCA), we performed metabolic pathway engineering aiming to prevent DCA consumption during beta-oxidation, but also to increase its production via the omega-oxidation pathway. We identified the major beta- and omega-oxidation pathway genes in C. guilliermondii and performed first steps in the strain improvement. A double pox disruption mutant was created that slowed growth with oleic acid but showed accelerated DCA degradation. Increase in DCA production was achieved by homologous overexpression of a plasmid borne cytochrome P450 monooxygenase gene. C. guilliermondii is a promising biocatalyst for DCA production but further insight into its fatty acid metabolism is necessary.