Bio-imprinting has been introduced as a technique of interfacial activation of lipase for anhydrous reaction applications. In this study, air-water (bubble) interfaces were compared to amphiphile and substrate interfaces in microbial lipase bio-imprinting. Results indicated that the bubble interface is equally effective on lipase interesterification activity and produces a 4-4.5-fold increase compared with the enzymes as supplied. Interesterification activity can be explained in terms of effects upon the accessibility of the lipase active site. This technique provides an easier, cheaper and product-friendly way of lipase bio-imprinting.