Bacterial poly [(R)-3-hydroxybutyrate-co-(R)-3-hydroxyhexanoate] (PHBH) highly oriented films were prepared by the combination of roll and uniaxial drawing processes. The change in the higher-order structure of PHBH films was investigated by wide-angle X-ray diffraction (WAXD) and small angle X-ray scattering (SAXS). Extended films, which show superior mechanical properties and high ductility, have complicated structures. By roll extension, both deformed and undeformed spherulites co-exist, the former inclined to the direction perpendicular to the film surface. The latter were destroyed by further uniaxial extension. The tie-molecules between uniaxially oriented lamellae were extended and transformed to the beta-form with a planar zig-zag conformation. Three kinds of structures, c-axis parallel to the uniaxial drawing direction, c-axis inclining to the normal vector of the film surface, and the beta-form between lamellae, are intermingled in the film. (C) 2008 Published by Elsevier Ltd.