Bacterial cellulose pellicle produced by Gluconacetobacter xylinus (G. xylinus) is one of the best biobased materials having a unique supernetwork structure with remarkable physiochemical properties for a wide range of medical and tissue-engineering applications. It is still necessary to modify them to obtain materials suitable for biomedical use with satisfactory mechanical strength, biodegradability, and bioactivity. The aim of this research was to develop a gene-transformation route for the production of bacterial cellulose/Curdlan (beta-1,3-glucan) nanocomposites by separate but simultaneous in vivo synthesis of cellulose and Curdlan. Modification of the cellulose-nanofiber-producing system of G. xylinus enabled Curdlan to be synthesized simultaneously with cellulose nanofibers in vivo, resulting in biopreparation of nanocomposites. The obtained Curdlan/cellulose composites were characterized, and their properties were compared with those of normal bacterial cellulose pellicles, indicating that Curdlan mixed with the cellulose nanofibers at the nanoscale without disruption of the nanofiber network structure in the pellicle.