title: Immobilization of alcohol dehydrogenase onto metal-chelated cryogels.
abstract: In this presented work, poly(HEMA-GMA) cryogel was synthesized and used for the immobilization of alcohol dehydrogenase. For this, synthesized cryogels were functionalized with iminodiacetic acid and chelated with Zn(2+). This metal-chelated cryogels were used for the alcohol dehydrogenase immobilization and their kinetic parameters were compared with free enzyme. Optimum pH was found to be 7.0 for both immobilized and free enzyme preparations, while temperature optima for free and immobilized alcohol dehydrogenase was 25 °C. Kinetic constants such as K(m), V(max), and k(cat) for free and immobilized form of alcohol dehydrogenase were also investigated. k(cat) value of free enzyme was found to be 3743.9 min(-1), while k(cat) for immobilized enzyme was 3165.7 min(-1). Thermal stability of the free and immobilized alcohol dehydrogenase was studied and stability of the immobilized enzyme was found to be higher than free form. Also, operational stability and reusability profile of the immobilized alcohol dehydrogenase were investigated. Finally, storage stability of the free and immobilized alcohol dehydrogenase was studied, and at the end of the 60 days storage, it was demonstrated that, immobilized alcohol dehydrogenase was exhibited high stability than that of free enzyme.
authors.affiliation: a Chemistry Department , Adnan Menderes University , Aydın , Turkey.
authors.forename: Deniz Aktaş
authors.initials: DA
authors.lastname: Uygun
authors.forename: Begüm
authors.initials: B
authors.lastname: Akduman
authors.forename: Murat
authors.initials: M
authors.lastname: Uygun
authors.forename: Sinan
authors.initials: S
authors.lastname: Akgöl
authors.forename: Adil
authors.initials: A
authors.lastname: Denizli
mesh_terms: D000426:Alcohol Dehydrogenase / Q000737:chemistry / Q000378:metabolism*; D002614:Chelating Agents / Q000737:chemistry*; D059985:Cryogels; D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000737:chemistry / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007098:Imino Acids / Q000737:chemistry*; D007700:Kinetics; D008422:Materials Testing; D008855:Microscopy, Electron, Scanning; D011102:Polyhydroxyethyl Methacrylate / Q000031:analogs & derivatives* / Q000737:chemistry; D016062:Porosity; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000378:metabolism*; D013499:Surface Properties; D013696:Temperature; D015032:Zinc / Q000737:chemistry*
chemical_list: D002614:Chelating Agents; D059985:Cryogels; D004800:Enzymes, Immobilized; D007098:Imino Acids; D029701:Saccharomyces cerevisiae Proteins; C515720:poly(hydroxyethyl methacrylate-co-glycerol methacrylate); D011102:Polyhydroxyethyl Methacrylate; C516801:ADH1 protein, S cerevisiae; D000426:Alcohol Dehydrogenase; D015032:Zinc; C008109:iminodiacetic acid
keywords: IMAC; alcohol dehydrogenase; cryogel; immobilization