This research explores various immobilized enzyme support materials, including the novel nylon-6 fiber membrane (NFM), observing the increase of surface area and what effect that has on enzyme binding potential. This study also manipulates incubation and reaction conditions and observes what affect that has on activity and stability of β-galactosidase comparing various solid support materials and free enzyme. Nylon-6 fiber membranes were created using the process of electrospinning and were compared with other materials as solid support materials for enzyme binding. The other materials included polyvinylidene fluoride 5 kD nanofiltration dairy membranes, nylon-6 pellets, silica glass beads, and free—dissolved—enzyme. Scanning electron microscopy images exposed the nylon-6 fiber membrane’s large amount of surface area which coordinated with greater enzyme activity as compared to the relatively flatter surfaces of the other solid support materials. Enzyme activity was measured spectrophotometrically with the color-changing substrate ortho-Nitrophenyl-β-galactoside. NFM had greater maximum enzyme binding potential than the other solid supports. Across pH conditions ranging from 3.5 to 6.0., enzyme activity was maintained on the membrane immobilized samples whereas free enzyme did not maintain activity. Altering storage temperature (4, 22, and 50 °C) affected enzyme stability, the ability of the enzyme to maintain activity over time, of free and polyvinylidene fluoride membrane samples. However, nylon-6 fiber membrane samples maintained stability across the varying storage temperatures. Increasing the immobilization solution enzyme concentration above maximum enzyme binding capacity had no significant effect on enzyme stability for membrane immobilized samples. Although, both had lower mean stability than free enzyme by approximately 74% percent. With further development, β-galactosidase immobilized on nylon-6 fiber membranes, or other membranes, could be used in continuous processing in the dairy industry for a combination of filtration and lactose hydrolysis—creating products reduced in lactose and increased in sweetness with no “added sugars” requirement for a nutrition label and no enzyme listed as final product ingredient.
College and Department
BYU ScholarsArchive Citation
Hutchins, Deborah Ann, "Activity, Stability, and Binding Capacity of β-Galactosidase Immobilized on Electrospun Nylon-6 Fiber Membrane" (2020). Theses and Dissertations. 9173.
nylon-6, membrane, immobilization, β-galactosidase, stability