Screening gelatin hydrogel and calcium alginate beads as scaffold approach for immobilization of horseradish peroxidase enzyme: Comparative operational and thermal stability study
Abstract
Immobilization of enzymes is a highly beneficial technique where it augments industrial process economics by allowing enzyme re-use and improving process productivity and robustness. Horseradish peroxidase (HRP) is a highly available enzyme of wide industrial applications. Calcium alginate (Ca-Alg-HRP) and gelatin (Gel-HRP) mixed gel were screened for the immobilization of (HRP) hydrogen peroxide enzyme for improvement of its industrial applications. The immobilization procedure onto Ca alginate beads was accomplished by gelation method where calcium ions were used as cross-linking agent. For gelatin hydrogel, glutraldehyde was added to enzyme/gelatin mixture to complete cross-linking. The effect of immobilization on the pH and temperature profile, thermal and operational stability and reusability of biocatalyst activity was assessed. Immobilization caused a change in pH/activity profile of HRP with a marginal shift in optimum pH from 7.0 to 7.5 for free and immobilized HRP, respectively. Whereas the temperature/activity profile was improved upon immobilization. Both thermal and operational stability were enhanced as a result of immobilization. The half-life of immobilized HRP at both 50°C and 70°C was found to be higher than the free enzyme with a reported half life of 1034 min and stabilization factor of (61.5%) . Reusability of the immobilized HRP was also studied and the results showed that both methods allowed for consecutive runs with residual catalytic activity of 80% after five successive cycles. The suggested immobilization supports proved to provide protective effect for HRP with improved thermal and operational stability and allowed re-use and recovery of the enzyme.
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