In this work, we demonstrate the activity of enzyme invertase immobilized in the pores of nanoporous anodized 3 μm thick aluminum oxide (AA). The porous anodic alumina has uniform nanosized pores with an interpore distance of p = 100 nm, with pore diameters on the order of 60-65 nm. The pores trap the enzyme and continuous monitoring of the activity is carried out in a flow cell where the substrate is made to flow and the product is detected. The activity of the immobilized enzyme has been determined for the different concentrations of sucrose and for pH ranging from 3 to 6.5. Maximum activity was found for pH 4.5. Adsorption of the enzyme followed by its interaction with the substrate have been analyzed using confocal laser scanning microscopy (CLSM) and surface plasmon spectroscopy (SPR) and the results obtained show excellent correlation. SPR results show a biphasic kinetics for the adsorption of the enzyme as well as its interaction with the substrate with rates of adsorption for the enzyme at k = 2.9 × 10(5) M(-1) s(-1) and 1.17 × 10(5) M(-1) s(-1). The rate of interaction of the substrate with the invertase is initially rapid with k = 4.49 × 10(5) M(-1) s(-1) followed by a slower rate 1.43 × 10(4) M(-1) s(-1).
Stability of collagenous matrixes such as skin and leather with respect to changes in their dimensions on heating has long been correlated with degree and type of cross links formed and short-range ordering in angstrom unit scales. Macroscopic dimensional changes may be expected to involve alterations in the long-range order as well as supramolecular assemblies in skin and leather. This study relates thermal shrinkage of skin matrixes with alterations observed in micro-, meso-, and macroporic structures. Changes in the pore structure of skin associated with thermal shrinkage have been studied using nitrogen adsorption and mercury intrusion porosimetry measurements. A comparison of results obtained using both techniques has been made. These results indicate that although the percentage porosity of the matrix decreases, the BET specific surface area increases on shrinkage. An insight into the changes in the pore systems of skin induced by thermal shrinkage has been gained.
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