Applied Biochemistry and Biotechnology

, Volume 84, Issue 1, pp 791–800

l-DOPA production by immobilized tyrosinase

  • Gabriela M. J. Carvalho
  • Tito Lívio M. Alves
  • Denise M. G. Freire
Article

DOI: 10.1385/ABAB:84-86:1-9:791

Cite this article as:
Carvalho, G.M.J., Alves, T.L.M. & Freire, D.M.G. Appl Biochem Biotechnol (2000) 84: 791. doi:10.1385/ABAB:84-86:1-9:791

Abstract

The production of l-DOPA using l-tyrosine as substrate, the enzyme tyrosinase (EC 1.14.18.1) as biocatalyst, and l-ascorbate as reducing agent for the o-quinones produced by the enzymatic oxidation of the substrates was studied. Tyrosinase immobilization was investigated on different supports and chemical agents: chitin flakes activated with hexamethylenediamine and glutaraldehyde as crosslinking agent, chitosan gel beads, chitosan gel beads in the presence of glutaraldehyde, chitosan gel beads in the presence of polyvinyl pyrrolidone, and chitosan flakes using glutaraldehyde as crosslinking agent. The last support was considered the best using as performance indexes the following set of immobilization parameters: efficiency (90.52%), yield (11.65%), retention (12.87%), and instability factor (0.00). The conditions of immobilization on chitosan flakes were optimized using a two-level full factorial experimental design. The independent variables were enzyme-support contact time (t), glutaraldehyde concentration (G), and the amount of enzyme units initially offered (UC). The response variable was the total units of enzymatic activity shown by the immobilized enzyme (UIMO). The optimal conditions were t=24 h, G=2% (v/v), and UC=163.7 U. Under these conditions the total units of enzymatic activity shown by the immobilized enzyme (UIMO) was 23.3 U and the rate of l-DOPA production rate was 53.97 mg/(L·h).

Index Entries

Tyrosinase enzymatic production l-DOPA immobilized enzyme 

Copyright information

© Humana Press Inc 2000

Authors and Affiliations

  • Gabriela M. J. Carvalho
    • 1
  • Tito Lívio M. Alves
    • 1
  • Denise M. G. Freire
    • 2
  1. 1.Chemical Engineering Program-COPPEFederal University of Rio de Janeiro, Centro de Ciências da SaúdeRio de JaneiroBrazil
  2. 2.Faculty of PharmacyFederal University of Rio de Janeiro, Centro de Ciências da SaúdeRio de JaneiroBrazil

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