Biodegradable polyurethanes (PUs) were produced from castor oil (CO) and poly (3-hydroxybutyrate) diol (PHBD) using hexamethylene diisocyanate as a crosslinking agent. PHBDs of different molecular weights were synthesized through transesterification of bacterial PHB and ethylene glycol by changing the reaction time. The synthesized PHBDs were characterized in terms of Fourier transform infrared and proton nuclear magnetic resonance spectroscopy. A series of PUs at different NCO/OH and CO/PHBD ratios were prepared. The resulting CO/PHBD-based PUs were then characterized in terms of mechanical and thermal properties. Increasing PHBD content significantly increased the tensile strength of CO/PHBD-based PUs by 300% compared to neat CO-based PU. CO/PHBD-based PUs synthetized from short chain PHBD exhibited higher tensile strength compared to those produced from long chain PHBD. As revealed by scanning electron microscopy analysis, such improvement in stiffness of the resulting PUs is due to the good compatibility between CO and PHBD. Increasing PHBD content also increased the crystallinity of the resulting PUs. In addition, higher degradation rates were obtained for CO/PHBD-based PUs synthetized from long chain PHBD compared to neat CO PU and PUs produced from short chain PHBD.