Mushroom tyrosinase was covalently immobilized on a poly(acrylic acid)-type, weakly acidic cation-exchange resin (Daiaion WK10, Mitsubishi Chemical Corp., Tokyo, Japan) with 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride salt as a water-soluble carbodiimide. Ion-exchange resins immobilized with tyrosinase were packed in one column, and crosslinked chitosan beads prepared with epichlorohydrin were packed in another column. The enzymatic activity was modified by covalent immobilization, and the immobilized tyrosinase had a high activity in the temperature range of 30-45 degrees C and in the pH range of 7-10. When solutions of various alkylphenols were circulated through the two columns packed wish tyrosinase-immobilized ion-exchange resins and crosslinked chitosan beads at 45 degrees C and pH 7 (the optimum conditions determined for p-cresol), alkylphenols were effectively removed through quinone oxidation with immobilized tyrosinase and Subsequent quinone adsorption on chitosan beads. The use of chemically crosslinked chitosan beads in place of commercially available chitosan beads was effective in removing alkylphenols from aqueous solutions in shorter treatment times. The removal efficiency increased with an increase in the amount of crosslinked chitosan beads packed in the column because the rate of quinone adsorption became higher than the rate of enzymatic quinone generation. The activity of tyrosinase was iteratively used by covalent immobilization on ion-exchange resins. One of the most important findings obtained in this study is the fact that linear and branched alkylphenols suspected of weak endocrine-disrupting effects were effectively removed from aqueous solutions. (C) 2009 Wiley Periodicals, Inc. J Appl Polym Sci 115: 137-145, 2010