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Edible mushrooms are an important source of nutraceuticals and for the discovery of bioactive metabolites as pharmaceuticals. In this work, the OSMAC (One Strain, Many Active Compounds) approach was used to isolate two new compounds (<b>1</b> and <b>2</b>) along with seven known compounds (<b>3</b>&#8722;<b>9</b>) from a mycelial culture of a unique North American edible mushroom <i>Hericium</i> sp. The fruiting body was collected in Marine on St. Croix, Minnesota (USA), and mycelial cultures were grown on four different solid and liquid media. Extracts from the mycelial cultures were screened for antimicrobial activity and only the extract from the Cheerios substrate culture exhibited antifungal activity. Bioassay guided fractionation and HPLC analysis were used to isolate nine pure compounds and the structures of the known compounds were established by analysis of the NMR and mass spectrometry data and comparison to published reports. Compound <b>1</b> is a new erinacerin alkaloid and <b>2</b> is an aldehyde derivative of 4-hydroxy chroman. Four chlorinated orcinol derivatives (<b>3</b>&#8722;<b>6</b>), a pyran (<b>7</b>), erinaceolactone (<b>8</b>), and erinacine (<b>9</b>) were identified. Compound <b>4</b> showed antifungal activity against <i>C. albicans</i> and <i>C. neoformans</i> (MIC = 31.3&#8722;62.5 &#956;g/mL, respectively). Compound <b>4</b> also inhibited biofilm formation of <i>C. albicans</i> and <i>C. neoformans</i> at 7.8 &#956;g/mL. These results suggest that mycelial cultures of edible fungi may provide useful, bioactive compounds.