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The extracellular polymeric substances of white rot fungi play an important role in the adsorption of heavy metals, but the influence of culture conditions on extracellular polymeric substances production is still unknown. In this paper, we researched on the influence of temperature, incubation time, the rotational speed and the inoculation volume on the yield of extracellular polymeric substances produced by Phanerochaete chrysosporium, a model strain of white rot fungi. The results show that the optimum culture conditions for Phanerochaete chrysosporium to produce extracellular polymeric substances was culturing at 40 °C, incubating for 5 d, rotating at 100 rpm, and inoculating 0.5 ml of spore suspension with concentration of 2.5×106 spores/ml. The highest yield of EPS was 234.65 mg/g when the fungi was cultured at 100 rpm, 40 °C and incubated for 5 days. This study can provide useful information for the follow-up experiments related to extracellular polymeric substances of white rot fungi