Evaluating enzyme stabilizations in calcium carbonate: Comparing in situ and crosslinking mediated immobilization

Enzyme immobilization using inorganic materials has been shown to preserve enzyme activity improving and improve their practical applications in biocatalytic process designs. Proper immobilization methods have been used to obtain high recycling and storage stability. In this study, we compared the activity and stability of in situ or crosslink-immobilized enzymes in a CaCO3 biomineral carrier. More than 30% of the initial enzyme activity was preserved for both the systems after 180 days upon 15 activity measurements at room temperature, confirming the improved stability of these enzyme systems (100 mM phosphate buffer, pH 8.0): however, differences in enzyme loading, activity, and characteristics were observed for each of these methods. Each system exhibited efficacy of 80% and 20%, respectively. Based on the same amount of immobilized enzyme (0.2 mg), the specific activities of hydrolysis of p-nitrophenyl butyrate substrate at room temperature of in situ immobilized carboxyl esterase (CE) and crosslinked CE were 11.37 and 7.63 mM min(-1) mg(-1), respectively (100 mM phosphate buffer, pH 8.0). Moreover, based on the kinetic behavior, in situ immobilized CE exhibited improved catalytic efficiency (Vmax Km(-1)) of the enzyme, exhibiting 4-fold higher activity and efficiency values than those of the CE immobilized in CaCO3. This is the first study to describe the stabilization of enzymes in CaCO3 and compare the enzyme kinetics and efficiencies between in situ immobilization and crosslinking in CaCO3 carriers. (C) 2021 Elsevier B.V. All rights reserved.

Automated summary

Enzyme immobilization using inorganic materials, such as CaCO3, has been shown to improve enzyme stability and activity. In this study, in situ and crosslink-immobilized enzymes in a CaCO3 carrier were compared, with both methods preserving over 30% of initial enzyme activity after 180 days. However, differences in enzyme loading, activity, and characteristics were observed between the two methods.