The effects of agitation and aeration in the production of gluconic acid by Aspergillus niger from a glucose medium were investigated. Experiments were conducted at aeration rates of 5.0 and 10.0 L/min. Four different agitation speeds were investigated for each aeration rate. Gluconic acid concentration and biomass concentration were analyzed, and the rate of consumption of substrate by A. niger was noted. The main purpose of this work was to find the optimal conditions of agitation and aeration for the growth of A. niger and production of gluconic acid in submerged culture in a batch fermenter at a bench-top scale. The oxygen-transfer rates at different agitation and aeration rates were calculated. The gluconic acid concentration and rate of growth of A. niger increased with increase in the agitation and aeration rates.