Enhancement of biochemical aspects of lipase adsorbed on halloysite nanotubes and entrapped in a polyvinyl alcohol/alginate hydrogel: strategies to reuse the most stable lipase

Abstract

Entrapment of halloysite nanotubes (HNTs) loaded with enzyme, into a polymer matrix (PVA/Alg), is a way to produce an environment surrounding the adsorbed enzyme molecules which improves the enzyme properties such as storage and operational stability. Hence, in this study, we optimised the factors affecting lipase adsorption onto halloysite nanotubes including halloysite amounts (5, 42.5 and 80 mg), lipase concentrations (30, 90 and 150 µg/ml), temperatures (5, 20 and 35 °C) and adsorption times (30, 165 and 300 min). The optimal conditions were determined as an halloysite amount of 50 to 80 mg, a lipase concentration of 30 to 57 μg/ml, an adsorption temperature of 20 °C and an adsorption time of 165 min, which resulted in a specific activity and adsorption efficiency of 15,000 (U/g protein) and 70%, respectively. Then, lipase adsorbed under optimal conditions was entrapped in a PVA/Alg hydrogel. The formation mechanism of immobilized lipase was investigated by FESEM and FTIR. Subsequent entrapment of adsorbed lipase improved the lipase storage and operational stability. K_m, V_max, K_cat and K_cat/K_m values showed an increase in the entrapped HNT-lipase performance in comparison with the free and adsorbed lipase.

Keywords: Alginate hydrogel; Halloysite nanotubes; Immobilization; Lipase reusability; Poly vinyl alcohol.