Biological denitrification of high nitrate-containing wastewater was examined in a sulfur-packed column using a smaller amount of methanol than required stoichiometrically for heterotrophic denitrification. In the absence of methanol, the observed nitrate removal efficiency was only about 40%, and remained at 400 mg NO3--N/l, which was due to an alkalinity deficiency of the pH buffer and of CO2 as a carbon source. Complete denitrification was achieved by adding approximately 1.4 g methanol/g nitrate-nitrogen (NO3--N) to a sulfur-packed reactor. As the methanol concentration increased, the overall nitrate removal efficiency increased. As influent methanol concentrations increased from 285 to 570, 855, and 1, 140 mg/l, the value of Delta mg alkalinity as CaCO3 consumed/Delta mg NO3--N removed increased from -1.94 to -0.84, 0.24, and 0.96, and Delta mg SO42- produced/Delta mg NO3--N removed decreased from 4.42 to 3.57, 2.58, and 1.26, respectively. These results imply the co-occurrence of simultaneous autotrophic and heterotrophic denitrification. Sulfur-utilizing autotrophic denitrification in the presence of a small amount of methanol is very effective at decreasing both sulfate production and alkalinity consumption. Most of methanol added was removed completely in the effluent. A small amount of nitrite accumulated in the mixotrophic column, which was less than 20 mg NO2--N/l, while under heterotrophic denitrification conditions, nitrite accumulated steadily and increased to 60 mg NO2--N/l with increasing column height.