Hydrogel nanoparticles can be prepared by using linoleic acid (LA) modified carboxymethyl chitosan (CMCS) after sonication. Bromelain can be loaded onto nanoparticles of LA-CMCS. Factors affecting the activity of the immobilized enzyme, including temperature, storage etc., were investigated in this study. The results showed that the stability of bromelain for heat and storage was improved after immobilization on nanoparticles. The Michaelis constant (Km) of the immobilized enzyme, was smaller than that of free enzyme which indicated that the immobilization could promote the stability of the enzyme and strengthen the affinity of the enzyme for the substrate.