Abstract
Immobilization of enzymes is one of the protein engineering methods used to improve their thermal and long-term stabilities. Immobilized pectinase has become an essential biocatalyst for optimization in the food processing industry. Herein, nanostructured magnetic nanoparticles were prepared in situ for use as supports to immobilize pectinase. The structural, morphological, optical and magnetic features and the chemical compositions of the nanoparticles were characterized. Nanoparticle agglomeration and low porosity were observed due to the synthetic conditions. These nanoparticles exhibited superparamagnetic behavior, which is desirable for biotechnological applications. The maximum retention rate for the enzyme was observed at pH 4.5 with a value of 1179.3 U/mgNP (units per milligram of nanoparticle), which was equivalent to a 65.6% efficiency. The free and immobilized pectinase were affected by the pH and temperature. The long-term instability caused 40% and 32% decreases in the specific activities of the free and immobilized pectinase, respectively. The effects of immobilization were analyzed with kinetic and thermodynamic studies. These results indicated a significant affinity for the substrate, a decreased reaction rate, and improved thermal stability of the immobilized pectinase. The reusability of the immobilized pectinase was preserved effectively during cycling, with only a 21.2% decrease in activity observed from the first to the last use. Therefore, alternative magnetic nanoparticles are presented for immobilizing and maintaining the thermostability of pectinase.