title: Affinity and stability modifications of immobilized alcohol dehydrogenase through multipoint copolymerization.
abstract: Yeast alcohol dehydrogenase (alcohol:NAD+ oxidoreductase, EC 1.1.1.1), a potentially useful enzyme for cofactor regeneration processes, was covalently immobilized in a multipoint fashion by activation with acryloyl chloride and subsequent copolymerization in a polyacrylamide gel. Several properties such as the activity and stability were systematically studied for the free enzyme, the acryloate-enzyme and the immobilized enzyme. The activation energy was significantly lowered upon immobilization. The thermal stability of the immobilized enzyme was, however, greatly increased. But its maximum activity was observed at a lower temperature. These results suggest an important effect of the diffusional restrictions and of the mode of activation and immobilization on the activity and the stability of the enzyme.
authors.affiliation: Unité de Biochimie Cellulaire, Facultés Universitaires de Namur, Belgium.
authors.forename: V
authors.initials: V
authors.lastname: Bille
authors.forename: D
authors.initials: D
authors.lastname: Plainchamp
authors.forename: J
authors.initials: J
authors.lastname: Remacle
mesh_terms: D000179:Acrylates; D000215:Acylation; D000426:Alcohol Dehydrogenase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D001665:Binding Sites; D055598:Chemical Phenomena; D002621:Chemistry; D004058:Diffusion; D004355:Drug Stability; D004789:Enzyme Activation; D004800:Enzymes, Immobilized / Q000378:metabolism*; D007700:Kinetics; D009243:NAD / Q000378:metabolism; D011108:Polymers; D011489:Protein Denaturation; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D013816:Thermodynamics
chemical_list: D000179:Acrylates; D004800:Enzymes, Immobilized; D011108:Polymers; D009243:NAD; C026200:acryloyl chloride; D000426:Alcohol Dehydrogenase