Bacterial cellulose (BC) producing bacterial strains were isolated from bio-extract (BE). Nine isolates that can produce BC in Hestrin–Schramm medium (HS medium) were identified. The BC production of these isolates was then investigated using agricultural waste as a raw material. The agricultural waste (banana, papaya, dragon fruit, and mango peels) was used as a carbon source for BC production. After incubation, the highest dry weight of BC reached 0.93±0.27 g/L, and 4.07±0.27 g/L was obtained from isolate BE073 in a medium containing mango and dragon fruit peels because the raw materials state is appropriate for bacterial growth. In a medium with papaya peel, the highest dry weight of BC was obtained from isolate BE052 at about 1.08±0.05 g/L. None of the strains was able to grow with the banana medium. However, all the isolate strains could grow and produce BC in the HS medium. The maximum dry weights of BC of 4.31±0.45 g/L, 4.23±0.13 g/L, and 4.21±0.25 g/L were obtained from isolates BE123, BE052, and BE073, respectively, and Acetobacter xylinum produced BC at 2.39±0.11 g/L. The structure and physical properties of BC produced from bacterial isolates using agricultural waste were characterized. It was similar to BC produced from HS medium and production from the reference strain A. xylinum . This study demonstrates the ability for BC production of bacterial strains isolated from bio-extract. It is also demonstrated that agricultural waste is a suitable and alternative carbon source for raw material in BC production.