Determination of some adsorption and kinetic parameters of α-amylase onto Cu +2 -PHEMA beads embedded column.

In order to investigate the biocatalytic properties of α-amylase on a composite cryogel matrix with immobilized metal affinity chromatography, Cu+2 -attached poly(2-hydroxyethyl methacrylate) (Cu+2 -PHEMA) beads, (2 µm size) were synthesized, then composite cryogel column was prepared by composing beads and PHEMA cryogels. After the preparation of Cu+2 -PHEMA beads embedded cryogel column (Cu+2 -BEC), some experiments were tested. Accordingly, the highest adsorption capacity (676.8 mg/g particles) of cryogels was achieved at acetate buffer of pH 5.0 with initial α-amylase concentration of 4 mg/mL. Immobilized enzyme has more stable pH range, between 6 and 7.5 than, the free one. Immobilization also increased the optimal activity from 25 to temperature range of 25-35 °C. Vmax and Km of α-amylase were detected as 1.149 U/mg protein, and 11.6 × 10-1 mM, respectively. α-Amylase was utilized 35 times repeatedly without losing the productivity.