AbstractDi(2-ethylhexyl) phthalate (DEHP) is a plasticizer that is widely used to soften plastic products. It is an endocrine-disrupting compound, widely considered an environmental contaminant that is difficult to remove or degrade. The radial growth rate (ur), mycelial biomass, intracellular and extracellular activities of laccase and esterase, intracellular and extracellular contents of protein and glycogen, and the contents of soluble and insoluble glucans were evaluated in colonies of Lentinula edodes, grown in various concentrations of DEHP (0, 750, 1200, and 1500 mg/L) on agar. Glucose and yeast extracts were added to all media. The highest ur was shown in medium lacking DEHP, followed by that shown in the medium containing 1500 mg of DEHP/L. The greatest protein extracellular contents were observed in medium with added DEHP. The largest extracellular glycogen contents and mycelial biomass were found in media containing 1200 and 1500 mg of DEHP/L, respectively. These results suggest that DEHP was degraded and used as a substrate by L. edodes during diauxic growth (glucose metabolized first, followed by DEHP), and laccases were more important than esterases in the metabolism of DEHP. L. edodes can be used to remove DEHP from phthalate-polluted environments.