Glucose Tolerance, Antiprotease Activity and Total Oxidant/Antioxidant Capacity Studies of β-Glucosidase Hybrid Nanoflower for Industrial Applications.

Abstract

β-Glycosidases, which catalyse the hydrolysis of glycoside bonds, have a wide spectrum of industrial applications. However, the reaction product glucose inhibits the activities of many β-glucosidases. Consequently, the reduced catalytic activities of the enzyme limit the industrial applications of the enzymes. For that reason, the studies dealing with maintaining the activities of the relevant enzymes at high glucose concentrations are a great interest among the researchers. In this context, herein, protein-inorganic hybrid nanoflowers were synthesized using β-glucosidase and copper ion by fast sonication method for 10 min. After characterization of synthesized nanoflowers, pH/temperature studies, glucose tolerance, anti-protease activity, recyclability and total antioxidant and total oxidative capacity levels were estimated. Accordingly, the optimum pHs of free β-glucosidase and hybrid nanoflower (β-GNF) were found to be 6 and 5, respectively, and the optimum temperature values for both hybrid nanoflowers and free enzyme were 40 °C. β-GNF exhibited better activity than free enzyme in low acidic and alkaline environment and at high temperature. The nanoflower retained nearly all (99 %) of its initial activity at all glucose concentrations (0.01, 0.05 and 0.1 mg/mL), especially at pH 5 and 6. Also, β-GNF maintained more than 90 % of initial activity at 0.01 and 0.05 mg/mL glucose at pH 4 and 7. It also displayed about 96 % high residual activity after proteinase K treatment for 3 h at 37 °C, while that of the free β-glucosidase was about 87 %. The reusability studies showed that β-GNF only lost ∼28 % of its initial activities at the end of five cycles. The hybrid nanoflowers at 5 mg/mL concentration exhibited the high total antioxidant capacity. In addition, low total oxidant capacity and oxidative stress index levels were recorded at the same concentration of the hybrid nanoflower. The findings of the present study revealed that β-GNFs may be evaluated as a candidate for various industrial applications due to its high glucose tolerance, anti-protease activity, reusability and resistance to low acidic/alkaline environment and high temperature.