Production of a novel bioflocculant by a fed-batch culture of Citrobacter sp. TKF04 was investigated using acetic acid as a sole carbon source. Synthesis of the bioflocculant was favored by dissolved O-2 tension at 20% of air saturation and C/N ratio (mol acetic acid/mol ammonium) of 10:1 in the feed solution. Under optimal conditions, 4.6 g crude bioflocculant per liter broth was produced, whose flocculating activity was 22 300 units. This activity was 9 times higher than that of the control (only acetic acid was supplied).