PHB nanofibers are synthesized by electros pinning of a PHB solution prepared using HFIP as the solvent. The nanofibrous scaffold supports rapid cell growth with normal morphology and attains a viability of 87% after 48 h. Kanamycin sulphate-loaded PHB nanofiber mats are synthesized, with the antibiotic on the surface and sandwiched within the nanofiber mats: their antimicrobial property is proved by the good zone of inhibition tested against Staphylococcus aureus. The drug shows more than 95% release within 8 h. These results indicate that nanofibers loaded with the antibiotic have potential applications as a template for tissue engineering and as a drug carrier.