Techniques using immobilization of enzymes in various types of support have increased the interest due to the various advantages provided by immobilized enzymes against the use of free enzymes, mainly by the decrease in the operational cost. Functionalized magnetic nanoparticles have shown great application potentials in processes because of the facility of catalyst recovery from the reactional medium. In this study, magnetic nanoparticles (MNP) modified with L-Histidine have been synthesized by a two-step process and the lipase from Thermomyces lanuginosus (TLL) was then immobilized. Results revealed that the particles are magnetite and the crystallite size calculated from XRD analysis reveal a single structure natural from iron oxide particles. The FT-IR measurements revealed that the L-Histidine was bonded to the surface of the particles via the carboxyl groups and the zeta potentials showed good stability. Superparamagnetic behavior was proved by measurements on VSM. The biocatalysts with L-Histidine showed a final activity of 74% of the initial activity. The particles fabricated in this study have great potential on the immobilization of TLL and other lipases.