L-Dopa synthesis using tyrosinase immobilized on magnetic beads

Tuncagil S., Kayahan S. K. , BAYRAMOĞLU G. , Arica M. Y. , TOPPARE L. K.

JOURNAL OF MOLECULAR CATALYSIS B-ENZYMATIC, vol.58, pp.187-193, 2009 (Journal Indexed in SCI) identifier identifier

  • Publication Type: Article / Article
  • Volume: 58
  • Publication Date: 2009
  • Doi Number: 10.1016/j.molcatb.2008.12.014
  • Page Numbers: pp.187-193


Magnetic beads were prepared via Suspension polymerization of glycidyl methacrylate (GMA) and methyl methacrylate (MMA) in the presence of ferric ions. Following polymerization, thermal co-precipitation of the Fe(III) ions in the beads with Fe(II) ions under alkaline condition resulted in encapsulation of Fe3O4 nano-crystals within the polymer matrix. The magnetic beads were activated with glutaraldehyde, and tyrosinase enzyme was covalently immobilized on the support via reaction of amino groups under mild conditions. The immobilized enzyme was used for the synthesis of L-Dopa (1-3,4-dihydroxy phenylalanine) which is a precursor of dopamine. The immobilized enzyme was characterized by temperature, pH, operational and storage stability experiments. Kinetic parameters, maximum velocity of the enzyme (V-max) and Michaelis-Menten constant (K-m) values were determined as 1.05 U/mg protein and 1.0 mM for 50-75 mu m and 2.00 U/mg protein and 4.0 mM for 75-150 mu m beads fractions. respectively. Efficiency factor and catalytic efficiency were found to be 1.39 and 0.91 for 75-150 mu m beads and 0.73 and 0.75 for 50-75 mu m beads fractions, respectively. The catalytic efficiency of the soluble tyrosinase was 0.37. The amounts of immobilized protein were on the 50-75 mu m and 75-150 mu m fractions were 2.7 and 2.8 mg protein/g magnetic beads, respectively. (C) 2008 Elsevier B.V. All rights reserved.