We developed a highly enantioselective separation system for the optically active compounds, (S)-ibuprofen and l-phenylalanine, from their racemic mixtures by employing a supported liquid membrane (SLM) encapsulating a surfactant-lipase complex (or a surfactant-alpha-chymotrypsin complex). In the present system, enzymes encapsulated in the liquid-membrane phase effectively drove the enantioselective transport of optically active compounds through the SLM. This novel SLM allowed high enantioselectivity (ee over 91%) in the optical resolution of racemic ibuprofen and phenylalanine.