The influence of nanofiller, plasticizer and hydroxyvalerate (HV) contents on the properties of poly(hydroxybutyrate-co-hydroxyvalerate) (PHBV)/clay nano-biocomposites is studied. Analyses by wide-angle X-ray diffraction and differential scanning calorimetry showed that both the organo-modified montmorillonite clay and acetyl tributyl citrate plasticizer facilitate the crystallization process of PHBV. Despite its nucleating effect, the nanoclay hampers the spherulite growth and leads to a reduction in the copolymer crystallinity, while the addition of plasticizer induces the formation of thicker spherulites. For all studied systems, it was shown that the variation of mechanical properties of these nano-biocomposites is strongly influenced by their changes in crystallinity. (C) 2013 Elsevier Ltd. All rights reserved.