Preparation and characterization of nanoparticles containing trypsin based on hydrophobically modified chitosan

Trypsin was immobilized on linolenic acid modified chitosan using glutaraldehyde (GA) as cross-linker, which was confirmed by Fourier transform infrared (FTIR) spectra. The chitosan nanoparticles containing trypsin (TR) can be prepared after the sonication of immobilized trypsin. The GA concentration affected both the enzyme activity of the nanoparticle and particle size. Results indicated that the activity of trypsin immobilized onto linolenic acid modified chitosan nanoparticles increased with increasing concentration of GA up to 0.07% (v/v) and then decreased with increasing amount of GA. On the other hand, particle size increased (from 523 to 1372 nm) with the increasing concentration of GA (from 0.03 to 0.1% v/v). The enzyme catalytic characteristics of nanoparticle solution were also studied. The results showed that the kinetic constant value (K_m) of TR immobilized on nanoparticle (71.9 mg/mL) was higher than that of pure TR (50.2 mg/mL). However, the thermal stability and optimum temperature of TR immobilized on nanoparticles improved, which make it more attractive in the application aspect.