Abstract: The pilot fermentation experiment of bacterial cellulose( BC) is a necessary stage for its industrial production.It has a very important value and significance. The pilot fermentation experiment were systematically researched using 20 L- 50 L- 100 L laboratory stirred tank fermentation system. The growth curve of Komagataeibacter nataicola seeds at 20 L fermentor were determined. The effects of different seed culture series on 100 L mechanical fermentation tank were studied. To improve the level of dissolved oxygen,stirring speed-dissolved oxygen( DO) linkage control method were used during fermentation and cellulose production.The results showed that suitable inoculation period was in the middle and late stage of logarithmic phase( 30 ~ 48 h). The fermentation period using grade three seed was 62 h and 14 h shorter than grade one and two,respectively. To shorten the fermentation process delay time,the bacteria concentration in the fermentation tank was more than 2.0× 107 CFU / m L after inoculation with bacteria. Stirring speed- DO linkage control could ensure DO maintained at around 30% and increased BC yield to 2.8 g / L.This research was beneficial with shortening the fermentation cycle,reducing the production cost,improving the yield of cellulose,and laying foundations for the industrial production.The pilot experiment study results of BC had a significant improvement in the production process.