Applied Biochemistry and Biotechnology

, Volume 160, Issue 7, pp 2129–2147

Immobilization of Naringinase in PVA–Alginate Matrix Using an Innovative Technique

  • Mário A. P. Nunes
  • Hélder Vila-Real
  • Pedro C. B. Fernandes
  • Maria H. L. Ribeiro
Article

DOI: 10.1007/s12010-009-8733-6

Cite this article as:
Nunes, M.A.P., Vila-Real, H., Fernandes, P.C.B. et al. Appl Biochem Biotechnol (2010) 160: 2129. doi:10.1007/s12010-009-8733-6

Abstract

A synthetic polymer, polyvinyl alcohol (PVA), a cheap and nontoxic synthetic polymer to organism, has been ascribed for biocatalyst immobilization. In this work PVA–alginate beads were developed with thermal, mechanical, and chemical stability to high temperatures (<80 °C). The combination of alginate and bead treatment with sodium sulfate not only prevented agglomeration but produced beads of high gel strength and conferred enzyme protection from inactivation by boric acid. Naringinase from Penicillium decumbens was immobilized in PVA (10%)–alginate beads with three different sizes (1–3 mm), at three different alginate concentrations (0.2–1.0%), and these features were investigated in terms of swelling ratio within the beads, enzyme activity, and immobilization yield during hydrolysis of naringin. The pH and temperature optimum were 4.0 and 70 °C for the PVA–alginate-immobilized naringinase. The highest naringinase activity yield in PVA (10%)–alginate (1%) beads of 2 mm was 80%, at pH 4.0 and 70 °C. The Michaelis constant (KMapp) and the maximum reaction velocity (Vmaxapp) were evaluated for both free (KMapp = 0.233 mM; Vmaxapp = 0.13 mM min−1) and immobilized naringinase (KMapp = 0.349 mM; Vmaxapp = 0.08 mM min−1). The residual activity of the immobilized enzyme was followed in eight consecutive batch runs with a retention activity of 70%. After 6 weeks, upon storage in acetate buffer pH 4 at 4 °C, the immobilized biocatalyst retained 90% of the initial activity. These promising results are illustrative of the potential of this immobilization strategy for the system evaluated and suggest that its application may be effectively performed for the entrapment of other biocatalysts.

Keywords

NaringinNaringinase entrapmentPoly(vinyl alcohol)PVADNS micromethodMicroassayHydrolysisBoric acidAlginate

Copyright information

© Humana Press 2009

Authors and Affiliations

  • Mário A. P. Nunes
    • 1
  • Hélder Vila-Real
    • 1
  • Pedro C. B. Fernandes
    • 2
  • Maria H. L. Ribeiro
    • 1
  1. 1.Faculdade de Farmácia, Research Institute for Medicines and Pharmaceutical Sciences (i-Med-UL)University of LisbonLisbonPortugal
  2. 2.IBB—Institute for Biotechnology and Bioengineering, Center for Biological and Chemical EngineeringInstituto Superior TécnicoLisbonPortugal