Glycerol production from alcoholic fermentation of glucose, a by-product of cane sugar molasses, and wheat-milling residues (akalona) was studied. Experiments were run for the three different substrates under the same operating conditions. The highest glycerol production was obtained from molasses (24.35%), followed by glucose (22.92%) after 48 h. Maximum glycerol production was obtained from akalona hydrolyzate (17.47%) after 58 h. Stirring increased the glycerol production by about 18.5%. The effect of calcium on glycerol production was studied. The time courses of the fermentation processes are presented.