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.

Influence of Metal Ions and Organic Solutions on Activity and Stability of Beta‐Glucosidase Nanoflowers

“…(Figure 6). In a previous study, the protein‐inorganic (Cu 2+ ) hybrid nanoflowers were synthesized using β‐glucosidase lost about 28 % of its initial activity in cycle 5 [21] . In another study, the reusability of β‐glucosidase‐Zn 3 (PO 4 ) 2 hybrid nanoflower retained residual activity of 98.6 and 93.8 % after five and 10 cycles of reuse, respectively [25] .…”

“…Koca‐Caliskan and coworkers have synthesized a plant extract based inorganic hybrid nanoflower, and for the first time, demonstrated superior α‐glucosidase and α‐amylase enzyme inhibition activity compared to plain extracts [20] . Gulmez synthesized β‐glucosidase hybrid nanoflower by using the fast sonication method to investigate glucose tolerance, anti‐protease activity, recyclability, and levels of total antioxidant and total oxidative capacity [21] . Sonochemical methods, with their advantages of faster reaction rates attributed to localized heating and enhanced mass transfer, contrast with conventional methods that often necessitate longer reaction times.…”

“…[20] Gulmez synthesized β-glucosidase hybrid nanoflower by using the fast sonication method to investigate glucose tolerance, antiprotease activity, recyclability, and levels of total antioxidant and total oxidative capacity. [21] Sonochemical methods, with their advantages of faster reaction rates attributed to localized heating and enhanced mass transfer, contrast with conventional methods that often necessitate longer reaction times. Sonochemical synthesis, well-suited for small-scale reactions, require the use of specialized equipment like an ultrasonic bath or probe.…”

Biotransformed soybean cream as a new nutraceutical for skin care: collagen stimulation in vitro and ex vivo

“…The conversion of soybean's glycosylated isoflavones to aglycones can be conducted in vitro by β-glucosidase enzymes of microorganisms, especially Aspergillus awamori , through a fermentation process ( 12 , 13 ). Since the collagen-stimulating activity of biotransformed soybean extracts has been previously shown by our research group ( 14 ), the present work investigated a cosmetic formulation for this product, determining its estrogenic activity, collagen stimulation, and preliminary characteristics.…”