The properties of PHB may be affected by the carbon source used in its production and this may affect nanofibres made from this polymer by electrospinning. In this study, P(3-HB) was produced from glucose, rapeseed oil, and olive oil by Ralstonia eutropha H16. Cell growth and polymer production were higher in olive or rapeseed oil supplemented media compared to glucose supplemented media. FT-IR, ¹H-, ¹³C-NMR, and ESI/MSn confirmed that the synthesized polymers were P(3-HB). SEM micrograph showed the formation of nanofibres from P(3-HB) samples with the fibre diameters dependent on the source of the carbon used in polymer synthesis and the concentration of the polymer in the electrospinning solution. GPC showed that P(3-HB) from glucose (G-PHB) had a higher molecular weight (7.35x10 gmol¯¹) compared to P(3-HB) from rapeseed (R-PHB) and olive (O-PHB) oil. Differential scanning calorimetry (DSC) showed that the crystallinity of the electrospun polymers reduces with decreasing polymer concentration with R-PHB having lower crystallinity at all concentrations used. These observation shows that more PHB yield can be obtained using either rapeseed or olive oil compared to glucose with glucose producing polymers of higher molecular weight. It also show that electrospinning could be used to reduce the crystallinity of PHB fibres.