The wild, edible mushroom Rugiboletus extremiorientalis has an excellent taste and also medicinal effects, and is very popular in China. In this study, hot water extraction is used to isolate polysaccharides from R. extremiorientalis fruiting bodies, and response surface methodology is used to optimize the extraction conditions in terms of temperature, liquid–solid ratio, and time. The crude extract (REP) is purified in turn by DEAE‐52 chromatography and Sephadex G‐100 chromatography, resulting in three fractions (REP‐1, REP‐2, and REP‐3) with recovery rates of 16.7%, 57.65%, and 6.63%, respectively. In vitro antioxidant activity is highest for REP‐2, and the subsequent research is therefore focused on that fraction. High‐performance liquid chromatography reveals that REP‐2 is comprised of fucose, arabinose, galactose, glucose, xylose, and mannose. Fourier‐transform infrared spectroscopy of REP‐2 indicates the presence of functional groups typical for polysaccharides. High‐performance gel filtration chromatography indicates the REP‐2 has a molecular weight of 342.4 kDa. The results of this study provide a foundation for future research concerning the antioxidant activity of polysaccharides in R. extremiorientalis.