Immobilization of laccase on itaconic acid grafted and Cu(II) ion chelated chitosan membrane for bioremediation of hazardous materials

Background: Chitosan membranes were formed through a phase inversion technique and then cross-linked with epichlorohydrin (CHX). Heterogeneous graft copolymerization of itaconic acid (IA) onto membrane was carried out with different monomer concentrations (CHX-g-p(IA)). The membrane properties such as equilibrium swelling ratio, porosity, and contact angle were measured, together with analysis by scanning electron microscopy (SEM), energy dispersive analysis of X-rays (EDAX), atomic force microscopy (AFM), and Fourier transform infrared (FTIR) spectroscopy. Results: The Cu(II) ion incorporated membranes (i.e. CHX-g-p(IA)-Cu(II)) were used for reversible immobilization of laccase using CHX-g-p(IA) membrane as a control system. Maximum laccase adsorption capacities of the CHX-g-p(IA) and CHX-g-p(IA)-Cu(II) membranes (with 9.7% grafting yield) were found to be 6.3 and 17.6 mg mL -1 membrane at pH 4.0 and 6.0, respectively. The K m value for immobilized laccase on CHX-g-p(IA)-Cu(II) (4.16 × 10 -2 mmol L -1) was 2.11-fold higher than that of free enzyme (1.97 × 10 -2 mmol L -1). Finally, the immobilized laccase was used in a batch system for degradation of three different dyes (Reactive Black 5, RB5; Cibacron Blue F3GA, CB; and Methyl Orange, MO). The immobilized laccase on CHX-g-p(IA)-Cu(II) membrane was more effective for removal of MO dye than removal of CB and RB5 dyes. CONCLUSION: Flexibility of the enzyme immobilized grafted polymer chains is expected to provide easy reaction conditions without diffusion limitation for substrate dye molecules and their products. The support described, prepared from green chemicals, can be used for the immobilization of industrially important enzymes. © 2012 Society of Chemical Industry