Formulation and Engineering of Biomaterials

Assembly of horseradish peroxidase within supported cationic bilayers

  1. Ana Carolina Leão-Silva1,
  2. Alliny F. Naves2,
  3. Edla M. A. Pereira1,2,
  4. Denise F. S. Petri2,
  5. Ana M. Carmona-Ribeiro1,*

Article first published online: 25 MAY 2011

DOI: 10.1002/btpr.640

Issue

Biotechnology Progress

Biotechnology Progress

Volume 27, Issue 5, pages 1433–1441, September/October 2011

Additional Information

How to Cite

Leão-Silva, A. C., Naves, A. F., Pereira, E. M. A., Petri, D. F. S. and Carmona-Ribeiro, A. M. (2011), Assembly of horseradish peroxidase within supported cationic bilayers. Biotechnol Progress, 27: 1433–1441. doi: 10.1002/btpr.640

Author Information

  1. 1

    Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, CP 26077, CEP 05513-970, São Paulo SP, Brazil

  2. 2

    Departamento de Química Fundamental, Instituto de Química, Universidade de São Paulo, CP 26077, CEP 05513-970, São Paulo SP, Brazil

*Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, CP 26077, CEP 05513-970, São Paulo SP, Brazil

Publication History

  1. Issue published online: 10 OCT 2011
  2. Article first published online: 25 MAY 2011
  3. Accepted manuscript online: 2 MAY 2011 07:01AM EST
  4. Manuscript Revised: 19 APR 2011
  5. Manuscript Received: 30 JAN 2011

Funded by

  • FAPESP and CNPq

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Keywords:

  • dioctadecyldimethylammonium bromide;
  • horseradish peroxidase;
  • supported cationic bilayers;
  • particles;
  • catalytic activity

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 H2O2 in presence of free or immobilized enzyme. HRP incorporation (3.0 mg/m2) 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. © 2011 American Institute of Chemical Engineers Biotechnol. Prog., 2011

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