This review presents the recent advances in the achievement of organized proteo-lipidic nanostructures based on Langmuir-Blodgett technology and their potential applications in the nanobioscience area. By using the self-assembled properties of amphiphilic biomolecules at the air-water interface, the Langmuir-Blodgett (LB) technique offers the possibility to prepare ultrathin layers suitable for biomolecule immobilization at the molecular level. This review will provide a general overview of the enzyme association with preformed Langmuir-Blodgett films in connection with their potential applications in biosensing device developments, and then introduce the design of a new functionalised biomimetic nanostructure with oriented recognition site. The potential applications of such an organized proteo-lipidic nanostructure for biocatalysis investigations of an immobilised enzyme in a biomimetic situation and for the development of bioelectronic devices are finally discussed.