The effect of medium composition on the production of bacterial cellulose (BC) by Gluconacetobacter hansenii PJK was investigated. The addition of yeast extract and peptone in the medium increased the production yield (Y_ps/) of BC. The amount of BC produced by G. hansenii PJK was constant if the initial pH of the medium was in the range 4.5 to 6.0. Strains from the supernatant of the culture medium produced more BC than those from inside the BC. BC production was dependent on glucose metabolism, and the addition of fructose or lactate as a carbon source converted cells to Cel^- mutants. Cel^- mutants produced by the addition of fructose or lactate to the medium caused 73% or 30% decreases in BC production, respectively. The addition of succinate, which is one of the constituents of the TCA cycle, did not affect the production of BC.