Poly(hydroxybutyrate) (PHB) obtained from sugar cane was dissolved in a blend of chloroform and dimethylformamide (DMF) and electrospun at 40 ºC. By adding DMF to the solution, the electrospinning process for the PHB polymer becomes more stable, allowing complete polymer crystallization during the jet travelling between the tip and the grounded collector. The influence of processing parameters on fiber size and distribution was systematically studied. It was observed that an increase of tip inner diameter promotes a decrease of the fiber average size and a broader distribution. On the other hand, an increase of the electric field and flow rate produces an increase of fiber diameter until a maximum of ~2.0 m, but for electric fields higher than 1.5 kV.cm-1, a decrease of the fiber diameter was observed. Polymer crystalline phase seems to be independent of the processing conditions and a crystallinity degree of 53 % was found. Moreover, thermal degradation of the as-spun membrane occurs in single step degradation with activation energy of 91 kJ/mol. Furthermore, MC-3T3-E1 cell adhesion was not inhibited by the fiber mats preparation, indicating their potential use for biomedical applications. ; This work is funded by FEDER funds through the "Programa Operacional Factores de Competitividade – COMPETE" and by national funds arranged by FCT- Fundação para a Ciência e a Tecnologia, project references NANO/NMed-SD/0156/2007, PTDC/CTM/73030/2006 and PTDC/CTM/69316/2006. The authors also thank support from the COST Action MP1003, 2010 ‘European Scientific Network for Artificial Muscles’. V.S. and D.M.C thanks the FCT for the SFRH/BPD/63148/2009, SFRH/BD/ 82411/2011 grants respectively. C. R. thanks the IINL for a financial support via a PhD. JLGR acknowledge the support of the Spanish Ministry of Science and Innovation through project No. MAT2010-21611-C03-01 (including the FEDER financial support). CIBER-BBN is an initiative funded by the VI National R&D&i Plan 2008-2011, Iniciativa Ingenio 2010, Consolider Program, CIBER Actions and financed by the Instituto de Salud Carlos III with assistance from the European Regional Development Fund.