Abstract
In this work, we propose a facile technique to dually-immobilize alpha-amylase and horseradish peroxidase (HRP) as two different enzyme models via entrapment within two distinct polymeric electrospun fibers by simple mixing steps and compare their properties with both individually immobilized forms and with the free counterparts. The immobilization was verified using Fourier transform infrared spectroscopy (FTIR) and Field emission scanning electron microscope (FESEM). The immobilization efficiencies for the dual-immobilized HRP and alpha-amylase were 89% and 85%, respectively. The retained catalytic activities of the dual-immobilized HRP and alpha-amylase enzymes were observed to be 79% and 80.2% after 10 cycles, respectively. After storage for 12 weeks, the dual-immobilized enzymes still retained nearly 90% activities similar to the individually immobilized ones. This immobilization procedure did not appear to exert either negative or back inhibitory effects upon both enzymes with respect to the different enzymatic assay procedures. This approach demonstrates that two or more type of enzymes could be mixed with different polymers individually and undergoes electrospinning simultaneously. We believe that this approach is expected to considerably promote and extend the application of multienzyme systems and worth further investigation for potential enzyme mediated cascade reactions. (C) 2020 Elsevier B.V. All rights reserved.