Optimal Immobilization of β-Galactosidase onto κ-Carrageenan Gel Beads Using Response Surface Methodology and Its Applications

beta-Galactosidase (beta-gal) was immobilized by covalent binding on novel kappa-carrageenan gel beads activated by two- step method; the gel beads were soaked in polyethyleneimine followed by glutaraldehyde. 2(2) full-factorial central composite experiment designs were employed to optimize the conditions for the maximum enzyme loading efficiency. 11.443 U of enzyme/g gel beads was achieved by soaking 40 units of enzyme with the gel beads for eight hours. Immobilization process increased the pH from 4.5 to 5.5 and operational temperature from 50 to 55 degrees C compared to the free enzyme. The apparent K-m after immobilization was 61.6 mM compared to 22.9 mM for free enzyme. Maximum velocity V-max was 131.2 mu mol.min(-1) while it was 177.1 mu mol.min(-1) for free enzyme. The full conversion experiment showed that the immobilized enzyme form is active as that of the free enzyme as both of them reached their maximum100% relative hydrolysis at 4 h. The reusability test proved the durability of the kappa-carrageenan beads loaded with beta-galactosidase for 20 cycles with retention of 60% of the immobilized enzyme activity to be more convenient for industrial uses.