title: Thermostability of glucose oxidase in silica gel obtained by sol-gel method and in solution studied by fluorimetric method.
abstract: The thermostability of glucose oxidase entrapped in silica gel obtained by sol-gel method was studied by thermostimulated fluorescence of FAD at pH 5 and 7 and compared with that of the native enzyme in the solution and at the presence of ethanol. The unfolding temperatures were found to be lower for the enzyme immobilised in gel as compared with the native enzyme but higher as for the enzyme at the presence of ethanol. In gel, the thermal denaturation of glucose oxidase is independent on pH while in solution the enzyme is more stable at pH 5. The investigation the enzyme in different environment by steady-state fluorescence of FAD and tryptophan, synchronous fluorescence and time-resolved fluorescence of tryptophan indicates that the state of the molecule (tertiary structure and molecular dynamics) is different in gel and in solution. The ethanol produced during gel precursor hydrolysis is not the main factor influencing the thermostability of the enzyme but more important are interactions of the protein with the gel lattice.
authors.affiliation: Technical University of Łódź, Department of Biotechnology and Food Science, Institute of General Food Chemistry, Stefanowskiego 4/10, 90-924 Łódź, Poland. malgorzata.przybyt@p.lodz.pl
authors.forename: Małgorzata
authors.initials: M
authors.lastname: Przybyt
authors.forename: Ewa
authors.initials: E
authors.lastname: Miller
authors.forename: Tomasz
authors.initials: T
authors.lastname: Szreder
mesh_terms: D001234:Aspergillus niger / Q000201:enzymology; D004795:Enzyme Stability / Q000187:drug effects; D004800:Enzymes, Immobilized / Q000737:chemistry*; D000431:Ethanol / Q000494:pharmacology; D005470:Fluorometry / Q000379:methods*; D005949:Glucose Oxidase / Q000737:chemistry*; D058767:Protein Unfolding / Q000187:drug effects; D058428:Silica Gel / Q000737:chemistry*; D012996:Solutions; D013050:Spectrometry, Fluorescence; D013696:Temperature
chemical_list: D004800:Enzymes, Immobilized; D012996:Solutions; D000431:Ethanol; D058428:Silica Gel; D005949:Glucose Oxidase