Lipases have bio-imprinted with common substrate-interfaces and interesterification activities compared with amphiphile bio-imprinted counterparts. Bio-imprinting has yielded a 3.5- to 4.5-fold activity enhancement. Solvent-free medium was equally effective as hexane medium. Water addition erased the bio-imprinting effect. Bio-imprinting caused rate acceleration in the interesterification reaction and increased thermostability of the enzyme.