A white rot fungus Ceriporiopsis subvermispora is known as one of the best bio-pulping microorganisms. This fungus secretes hydrophobic metabolites such as fatty acids and alk(en)ylitaconic acids (ceripiric acids). These metabolites play important roles in the selective lignin-degrading system. However, much less attention has been directed toward the roles of hydrophilic metabolites in the selective ligninolysis. In the present study, we focused on amphipathic metabolites which interact with hydrophobic substances like lipid-related metabolites and lignin fragments. Amphipathic metabolites were screened by surface activity measurements, and purified by open column chromatography and high performance liquid chromatography (HPLC). In combination with liquid chromatography electrospray ionization mass spectrometry (LC/ESI-MS), we found that this fungus produced several series of amphipathic metabolites. The purified amphipathic metabolites lowered surface tension of water from 72.8mN/m to 37.3mN/m at the concentration of 0.1%, and solubilized water-insoluble metabolites, ceriporic acids. These results lead to the conclusion that the amphipathic metabolites were surface active agents (surfactants) directly interacting with hydrophobic substances.