In the present investigation, the effects of crude glycerol concentration, spore inoculum concentration, yeast extract concentration and shaking frequency on seed morphology of Aspergillus niger PJR1 on malic acid production were investigated and dispersed fungal mycelium with higher biomass (20.25 +/- 0.91 g/L) was obtained when A. niger PJR1 grow on crude glycerol. Dry cell weight under dispersed fermentation was 21.28% higher than usual pellet fermentation. The optimal crude glycerol, nitrogen source and nitrogen source concentration were found to be 160 g/L, yeast extract and 1.5 g/L, respectively. Batch fermentation in a shake flask culture containing 160 g/L crude glycerol resulted in the yield of malic acid 83.23 +/- 1.86 g/L, after 192 h at 25 degrees C. Results revealed that morphological control of A. niger is an efficient method for increased malic acid production when crude glycerol derived from biodiesel production is used as feedstock.