Curdlan is a water insoluble polysaccharide composed exclusively of β-(1, 3) linked glucose residues. Agrobacterium sp. is known to produce extracellular curdlan under nitrogen-limited conditions. The purpose of this study was to investigate the effects of pH, amounts of ammonium, sucrose and trace elements, and time the addition of sucrose, ammonium and uracil on the production of curdlan in a shake flask and to further scale-up the process to a 5 L fermentor. A maximum of 48.7 g L-1 of curdlan was obtained in a shake flask when 150 and 1.4 g L-1 of sucrose and ammonium were used at the initial pH of 6.5. The production was enhanced to 57 g L-1 by adding one third of sucrose and 1 g L-1 of uracil at the 48 th h in a fed batch mode. The process was scaled up to a 5 L bioreactor in a batch mode where the oxygen transfer rate was higher (0.192 mg L-1s-1) when compared to that in the shake flask (0.096 mg L-1s-1). Curdlan production was 58 g L-1 in the bioreactor, which was higher than the shake flask under batch conditions (48.7 g L-1). The viscosity average molecular weight of the curdlan produced was found to be 1.4 × 105. Copyright © 2010 The Berkeley Electronic Press. All rights reserved.