Polyhydroxyalkanoates (PHAs) are biodegradable polymers, which have recently attracted much attention as a promising substitute for petroleum-based plastics. For circumventing the financial barrier of PHA production, non-edible Cerbera odollam oil was proposed as a novel PHA feedstock. The objective of this study was to investigate the feasibility of C. odollam oil for PHA production using Cupriavidus necator and Pseudomonas resinovorans. The PHA production performance of P. resinovorans on different oil conditions (i.e., crude, hydrolyzed, and emulsified oil) was also compared. Direct fermentation of crude C. odollam oil by P. resinovorans showed the most promising results with a biomass accumulation of 1.32 g/L and a maximum PHA concentration of 0.25 g/L (corresponding to 18.8 polymer content). The polymer was composed of 3-hydroxybutyrate (3HB), 3-hydroxyhexanoate (3HHx), 3-hydroxyoctanoate (3HO), and 3-hydroxydecanoate (3HD). For the first time, C. odollam oil has been proven to be a feasible feedstock for an economical production of PHAs. © 2023