Immobilization of an Industrial β-Glucosidase from Aspergillus fumigatus and Its Use for Cellobiose Hydrolysis

In this study, several covalent methods of immobilization based on acrylic supports, Schiff bases and epoxides have been applied to a commercial cocktail with a high β-glucosidase activity secreted by Aspergillus fumigatus. This cocktail was preliminary compared to a commercial secretome of Aspergillus niger, which was also subjected to the aforementioned immobilization methods. Due to its higher activity, the cocktail from A. fumigatus immobilized on ReliZyme™ HA403 activated with glutaraldehyde was employed for pNPG and cellobiose hydrolysis in diverse operational conditions and at diverse enzyme loadings, showing a very high activity at high enzyme load. A kinetic model based on the Michaelis–Menten hypothesis, in which double inhibition occurs due to glucose, has been selected upon fitting it to all experimentally retrieved data with the lowest-activity immobilized enzyme. This model was compared to the one previously established for the free form of the enzyme, observing that cellobiose acompetitive inhibition does not exist with the immobilized enzyme acting as the biocatalyst. In addition, stability studies indicated that the immobilized enzyme intrinsically behaves as the free enzyme, as expected for a one-bond low-interaction protein-support immobilization.