Iron deficiency anemia has been a widespread disease. As an effective and stable iron supplement, the physiochemical properties of the polysaccharide iron complex have been widely studied. In this study, we characterized a novel polysaccharide-iron(III) complex extracted in an edible fungal species Auricularia auricular (AAPS-iron(III)). The highest iron content (28.40%) in the AAPS-iron(III) complex was obtained under the optimized preparation conditions including an AAPS to FeCl3∙ 6H2O ratio of 2:3 (w/w), a pH value of 8.0 in solution, a reaction temperature of 50°C, and a reaction time of 3 h. The physical and chemical properties of the AAPS-iron(III) complex were characterized by qualitative and quantitative analyses using scanning electron microscope, particle size distribution, thermogravimetric analyzer, Fourier transform infrared spectroscopy, circular dichroism, and 1H nuclear magnetic resonance. Result showed that, although the iron was bound to the polysaccharide, it was released under artificial gastrointestinal conditions. The AAPS-iron(III) complex exhibited high stability (under 50-256°C) and water solubility. The AAPS-iron(III) complex also showed high antioxidant activity in vitro, demonstrating an additional health benefit over other typical nonantioxidant iron nutritional supplements. Furthermore, the AAPS-iron(III) complex showed high efficiency on the treatment of the iron deficiency anemia in the model rats. Therefore, the AAPS-iron(III) complex can be used as a nutritional fortifier to supply iron in industrial processing and to assist the treatment of iron deficiency anemia.