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    IMAC application of extracellular polymeric substances doped composite membranes for α-amylase immobilization and kinetic studies
    (Elsevier, 2024) Bani, Nurullah; Inanan, Tulden; Acet, Omur; Odabasi, Mehmet
    Immobilized metal affinity chromatography (IMAC), extensively utilized technique in affinity chromatography, has proven to be highly effective in purifying alpha-amylase, a crucial enzyme in industrial applications. Here, Poly (2-hydroxyethyl methacrylate) (PHEMA) based composite membranes were fabricated using the radical copolymerization technique in a petri dish. Extracellular polymeric materials (EPS) as the metal binding agent. The attachment of Cu 2+ ions to the membranes occurred through the utilization of EPSs (Cu 2+ -EPS). The membranes having Cu 2+ -EPS (Cu 2+ -EPS-PM) were subjected to various characterization techniques, including scanning electron microscopy (SEM), attenuated total reflectance-fourier transform infrared spectroscopy (ATR-FTIR), swelling tests, and surface area measurements. The investigation also encompassed the examination of various factors, including pH, initial alpha-amylase concentration, temperature, and ionic strength, that exert an influence on the adsorption of alpha-amylase from an aqueous medium. The Cu 2+ -EPS-PMs demonstrated maximum adsorption capacity with a value of 54.2 mg/g polymer (pH 5.0 sodium acetate buffer, temperature: 25 degrees C, initial alpha-amylase concentration of 2 mg/mL). Additionally, it has been observed that the enzyme is able to be used in adsorption and desorption cycles on Cu 2+ -EPS-PM many times. The investigation also focused on examining the impact of pH, temperature, storage, and operational stability over the activities of both free and immobilized alpha-amylase. The outcomes showed that immobilization had positive effects on activity.