title: Stability and kinetic behavior of immobilized laccase from Myceliophthora thermophila in the presence of the ionic liquid 1-ethyl-3-methylimidazolium ethylsulfate.
abstract: The use of ionic liquids (ILs) as reaction media for enzymatic reactions has increased their potential because they can improve enzyme activity and stability. Kinetic and stability properties of immobilized commercial laccase from Myceliophthora thermophila in the water-soluble IL 1-ethyl-3-methylimidazolium ethylsulfate ([emim][EtSO4 ]) have been studied and compared with free laccase. Laccase immobilization was carried out by covalent binding on glyoxyl-agarose beads. The immobilization yield was 100%, and the activity was totally recovered. The Michaelis-Menten model fitted well to the kinetic data of enzymatic oxidation of a model substrate in the presence of the IL [emim][EtSO4 ]. When concentration of the IL was augmented, the values of Vmax for free and immobilized laccases showed an increase and slight decrease, respectively. The laccase-glyoxyl-agarose derivative improved the laccase stability in comparison with the free laccase regarding the enzymatic inactivation in [emim][EtSO4 ]. The stability of both free and immobilized laccase was slightly affected by small amounts of IL (<50%). A high concentration of the IL (75%) produced a large inactivation of free laccase. However, immobilization prevented deactivation beyond 50%. Free and immobilized laccase showed a first-order thermal inactivation profile between 55 and 70°C in the presence of the IL [emim][EtSO4 ]. Finally, thermal stability was scarcely affected by the presence of the IL.
authors.affiliation: Dept. of Chemical Engineering, University of Vigo, Lagoas Marcosende, 36310, Vigo, Spain.
authors.forename: María
authors.initials: M
authors.lastname: Fernández-Fernández
authors.forename: Diego
authors.initials: D
authors.lastname: Moldes
authors.forename: Alberto
authors.initials: A
authors.lastname: Domínguez
authors.forename: M Ángeles
authors.initials: MÁ
authors.lastname: Sanromán
authors.forename: Ana Paula M
authors.initials: AP
authors.lastname: Tavares
authors.forename: Oscar
authors.initials: O
authors.lastname: Rodríguez
authors.forename: Eugénia A
authors.initials: EA
authors.lastname: Macedo
mesh_terms: D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000737:chemistry*; D007093:Imidazoles / Q000737:chemistry; D052578:Ionic Liquids / Q000737:chemistry; D007700:Kinetics; D042845:Laccase / Q000737:chemistry* / Q000378:metabolism; D020037:Sordariales / Q000201:enzymology*; D014867:Water / Q000737:chemistry
chemical_list: D004800:Enzymes, Immobilized; D007093:Imidazoles; D052578:Ionic Liquids; D014867:Water; D042845:Laccase; C518739:1-ethyl-3-methylimidazolium
keywords: 1-ethyl-3-methylimidazolium ethylsulfate; agarose; immobilization; ionic liquid; laccase; stability