title: Activity and stability of hyperthermophilic enzymes: a comparative study on two archaeal beta-glycosidases.
abstract: S beta gly and CelB are well-studied hyperthermophilic glycosyl hydrolases, isolated from the Archaea Sulfolobus solfataricus and Pyrococcus furiosus, respectively. Previous studies revealed that the two enzymes are phylogenetically related; they are very active and stable at high temperatures, and their overall three-dimensional structure is very well conserved. To acquire insight in the molecular determinants of thermostability and thermoactivity of these enzymes, we have performed a detailed comparison, under identical conditions, of enzymological and biochemical parameters of S beta gly and CelB, and we have probed the basis of their stability by perturbations induced by temperature, pH, ionic strength, and detergents. The major result of the present study is that, although the two enzymes are remarkably similar with respect to kinetic parameters, substrate specificity, and reaction mechanism, they are strikingly different in stability to the different physical or chemical perturbations induced. These results provide useful information for the design of further experiments aimed at understanding the structure-function relationships in these enzymes.
authors.affiliation: Laboratory of Microbiology, Wageningen Agricultural University, The Netherlands.
authors.forename: J
authors.initials: J
authors.lastname: Pouwels
authors.forename: M
authors.initials: M
authors.lastname: Moracci
authors.forename: B
authors.initials: B
authors.lastname: Cobucci-Ponzano
authors.forename: G
authors.initials: G
authors.lastname: Perugino
authors.forename: J
authors.initials: J
authors.lastname: van der Oost
authors.forename: T
authors.initials: T
authors.lastname: Kaper
authors.forename: J H
authors.initials: JH
authors.lastname: Lebbink
authors.forename: W M
authors.initials: WM
authors.lastname: de Vos
authors.forename: M
authors.initials: M
authors.lastname: Ciaramella
authors.forename: M
authors.initials: M
authors.lastname: Rossi
mesh_terms: D002480:Cellulase / Q000378:metabolism; D003902:Detergents; D004795:Enzyme Stability; D006026:Glycoside Hydrolases / Q000378:metabolism*; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D020027:Pyrococcus furiosus / Q000201:enzymology*; D012492:Salts; D012967:Sodium Dodecyl Sulfate; D016944:Sulfolobus / Q000201:enzymology*
chemical_list: D003902:Detergents; D012492:Salts; D012967:Sodium Dodecyl Sulfate; D006026:Glycoside Hydrolases; D002480:Cellulase