Stabilizing of catechol 1,2 dioxygenase on the surface of magnetic nanoparticles and investigating the enzyme activity

Catechol 1,2 dioxygenase is one of the important enzymes in the removal and analysis of phenolic compounds from the environment. In the process of biodegradation, the nonstabilized enzymes are inactivated as affected by environmental cues. One of the strategies to cope with the problem is to stabilize the enzyme. In the present study, catechol 1, 2 dioxygenase was stabilized on the surface of magnetic nanoparticles (MNPs). Firstly, the MNPs were synthesized by tetraethoxysilane and 3-aminopropyltriethoxysilane containing modified hydroxyl and amine groups, respectively. To investigate the structure and the size of synthesized MNPs, ScanningElectron Microscope (SEM) and dynamical light scattering (DLS) were used. The enzyme was then stabilized on MNPs by EDC and NHS linkers at concentrations of 5 μg/mL. The concentration of MNPs, catechol 1, 2 dioxygenase, EDC and NHS linkers, the reaction temperature, and the time required for determining the enzyme activity were also optimized. Finally, the enzyme activity was determined under optimum conditions at 260 nm wavelength by spectrophotometry. The results of the SEM and DLS showed that the size of the synthesized MNPs was 23-35nm. The results of comparing the activity of enzyme showed that the stabilized enzyme had an approximately 1.5 times more than a non-stabilized enzyme

کلیدواژه ها:

Catechol 1 ، 2 dioxygenase ، Enzyme stabilization ، MNPs