Enhancement of stability and activity of RSD amylase from Paenibacillus lactis OPSA3 for biotechnological applications by covalent immobilization on green silver nanoparticles

Abstract

The key challenge to the biotechnological applications of amylases is achieving high activity and stability under extreme pH, temperature and often high levels of enzyme denaturants. This study immobilized a novel raw starch-digesting (RSD) amylase from Paenibacillus lactis OPSA3 on glutaraldehyde-activated silver nanoparticles. Effects of time, glutaraldehyde concentration, pH, temperature, and enzyme concentration on immobilization were studied, and the immobilized enzymes were characterized. pH 9.0 was optimum for the enzyme immobilization. The maximum immobilization efficiency of 82.23 ± 7.99 % was achieved at 25 °C for 120 min. After immobilization, the optimum pH and temperature changed from 9.0 to 11.0 and 60 to 70, respectively. Immobilization reduced the amylase's activation energy (KJ/mol) from the initial 58.862 to 45.449 following immobilization. The Km of the amylase decreased after immobilization, while the Vmax increased. The immobilized amylase showed significantly greater storage and thermal stability than the free amylase. At 80, enzyme half-life (min) and D value (min) increased from 12.33 to 179.11 and 40.94 to 594.98, respectively. The immobilized amylase (80–88 %) had more stability to the effects of the studied surfactants than the free enzyme. It also showed improved stability in the presence of commercial detergents compared to the free enzyme. The amylase's enhanced kinetic parameters and stability following successful immobilization on silver nanoparticles indicate its potential for application in the range of biotechnological processes where alkaline- and temperature-stable amylases are employed.