Applied Biochemistry and Biotechnology, Vol.175, No.6, 2872-2882, 2015
Impact of Enzyme Loading on the Efficacy and Recovery of Cellulolytic Enzymes Immobilized on Enzymogel Nanoparticles
Cellulase and beta-glucosidase were adsorbed on a polyacrylic acid polymer brush grafted on silica nanoparticles to produce enzymogels as a form of enzyme immobilization. Enzyme loading on the enzymogels was increased to a saturation level of approximately 110 mu g (protein) mg(-1) (particle) for each enzyme. Enzymogels with varied enzyme loadings were then used to determine the impact on hydrolysis rate and enzyme recovery. Soluble sugar concentrations during the hydrolysis of filter paper and Solka-Floc with the enzymogels were 45 and 53 %, respectively, of concentrations when using free cellulase. beta-Glucosidase enzymogels showed lower performance; hydrolyzate glucose concentrations were just 38 % of those using free enzymes. Increasing enzyme loading on the enzymogels did not reduce net efficacy for cellulase and improved efficacy for beta-glucosidase. The use of free cellulases and cellulase enzymogels resulted in hydrolyzates with different proportions of cellobiose and glucose, suggesting differential attachment or efficacy of endoglucanases, exoglucanases, and beta-glucosidases present in cellulase mixtures. When loading beta-glucosidase individually, higher enzyme loadings on the enzymogels produced higher hydrolyzate glucose concentrations. Approximately 96 % of cellulase and 66 % of beta-glucosidase were recovered on the enzymogels, while enzyme loading level did not impact recovery for either enzyme.