Determination of some adsorption and kinetic parameters of α-amylase onto Cu-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-attached poly(2-hydroxyethyl methacrylate) (Cu-PHEMA) beads, (2 µm size) were synthesized, then composite cryogel column was prepared by composing beads and PHEMA cryogels. After the preparation of Cu-PHEMA beads embedded cryogel column (Cu-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. V and K of α-amylase were detected as 1.149 U/mg protein, and 11.6 × 10 mM, respectively. α-Amylase was utilized 35 times repeatedly without losing the productivity.