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Bacterial cellulose (BC) has shown potential as a separation material. Herein, the performance of BC in pressure-driven separation is investigated as a function of incubation conditions and post-culture treatment. The pure water flux of never-dried BC (NDBC), was found to be 9 to 16 times higher than that for dried BC (DBC), in a pressure range of 0.25 to 2.5 bar. The difference in pressure response of NDBC and DBC was observed both in cross-flow filtration and capillary flow porometry experiments. DBC and NDBC were permeable to polymers