The foremost aim of the present study was to enhance butanol production in an extractive fermentation study in presence of non-ionic surfactant using immobilized cells. Earlier studies had shown improved butanol production with non-ionic surfactant and immobilized cells independently. Therefore, in the present work, the combined effect of extractive fermentation and immobilized cells on butanol production was studied. Different matrices (brick, bamboo, cotton fiber, flannel cloth, and polyurethane foam) were tested for immobilization of Clostridium sporogenes. Immobilized biomass thus obtained was used in an extractive fermentation study with non-ionic surfactant L62. Biomass immobilized on polyurethane foam (PF) doubled the butanol production in presence of 6% (v/v) L62 with respect to control (free cells without surfactant). Further, the effect of different carbon sources was also studied to check the suitability of different industrial wastes containing different carbon sources. Glucose was found to be the best carbon source.