Antioxidant activities of 80% methanol, water, and 70% ethanol extracts of Lentinula edodes GNA01 were compared and estimated. The yield of Lentinula edodes GNA01 was identified to be in the following order: water>70% ethanol>80% methanol, but there was no significant difference between 80% methanol and 70% ethanol extracts. The highest total phenolic content (TPC) and total flavonoid content (TFC) were found in water extract, and TPC of 80% methanol extract was higher than that of 70% ethanol extract and TFC of 70% ethanol extract was higher than that of 80% methanol extract. Water extract exhibited the strongest DPPH, ABTS radicals, and nitrite scavenging activities, Fe2+ chelating ability, and FRAP among the three extracts. In addition, antioxidant activity of 80% methanol extract was higher than that of 70% ethanol extract in most of the experiments. As a result, antioxidant activity of Lentinula edodes GNA01 showed a difference according to extraction solvent and concentration; nevertheless, water extract exhibiting high polarity had the strongest antioxidant effect. Consequently, water extract from Lentinula edodes GNA01 is anticipated to be useful for the development of a high value-added functional product