Acetobacter xylinum KJ1 efficiently producing bacterial cellulose(BC) In shaking culture was isolated from a rotten grape. The strain was used to investigate optimum operating conditions for increasing BC production and factorial design model was employed for the optimization. The results of experiments were statistically analyzed by SAS program. Reciprocal effects of each factors(carbon source concentration, shaking speeds(rpm), oxygen pressure, and CSL concentration) and culture condition of BC production were examined by getting regression equation of the dependent variable. Comparisons between experimental results and predicted results about BC concentration were done in total 24 experiments by combination of each factors using SAS program, and the correlation coefficients of BC concentration and BC yield were 0.91 and 0.81, respectively. The agitated cultures were peformed in various operation conditions of factors which affected considerably to BC production in jar fermentor. The results showed that BC concentration was 11.67 g/L in 80 hours cultivation under the condition of carbon source concentration : shaking speeds(rpm) : oxygen pressure : CSL concentration : 4% : 460 rpm : 0.28 : 6%. On the other hand BC yield was 0.42 g/g in 80 hours cultivation under the condition of carbon source concentration : shaking speeds(rpm) : oxygen pressure : CSL concentration : 4% : 564 rpm : 0.21 : 2%. The BC production could be enhanced up to more than 2.4 times by factorial design. The result of a verifying experiment under the optimal conditions determined by the factorial design to the BC production showed that the model was appropriate by obtaining BC concentration of 11.47 g/L in the optimum condition.