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In this study the effect of different nitrogen source substrates on the amount of chitosan production by Aspergillus niger was investigated. A. niger PTCC 5012 from the Persian Type Culture Collection (PTCC) was grown on soy bean, corn seed and canola residues at 30 °C for specified cultivation days under sterilized conditions. Chitosan was extracted from the fungal mycelia using hot alkaline and acid treatment. The results were shown that soy bean residue at moisture of 37% and 8.40.26 % of nitrogen content produced the highest amount of chitosan (17.0530.95 g/kg of dry substrate), after 12 days of incubation. Corn seed residue produced very low amount of chitosan (1.90.4 % of nitrogen content). The chitosan was analyzed by FTIR spectroscopy and its spectrum was recorded.