Assembly of horseradish peroxidase within supported cationic bilayers |
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Authors: | Leão-Silva Ana Carolina Naves Alliny F Pereira Edla M A Petri Denise F S Carmona-Ribeiro Ana M |
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Institution: | 1. Departamento de Bioquímica, Instituto de Química, Universidade de S?o Paulo, CP 26077, CEP 05513‐970, S?o Paulo SP, Brazil;2. Departamento de Química Fundamental, Instituto de Química, Universidade de S?o Paulo, CP 26077, CEP 05513‐970, S?o Paulo SP, Brazil |
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Abstract: | The interaction between horseradish peroxidase (HRP) and dioctadecyldimethylammonium bromide (DODAB) bilayers supported on polystyrene microspheres (PSS) or on flat silicon wafers was evaluated from the following techniques: (1) dynamic light-scattering for determining size distributions, zeta-potentials and polydispersities for dispersions; (2) spectrophotometric determination of HRP concentration in supernatants of centrifuged mixtures; (3) in situ ellipsometry for mean thickness of deposited layers on wafers; (4) kinetics of product appearance for oxidation of 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid by H(2) O(2) in presence of free or immobilized enzyme. HRP incorporation (3.0 mg/m(2)) did not alter mean diameter and zeta-potential of PSS/DODAB particles but reduced enzyme activity by 50%, though activity persisted after several rinsing steps. In situ ellipsometry could not detect any HRP layer on top of the DODAB bilayer. HRP insertion in the bilayer core explained all results for both systems. Useful biotechnological applications are anticipated for such assemblies. |
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Keywords: | dioctadecyldimethylammonium bromide horseradish peroxidase supported cationic bilayers particles catalytic activity |
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