Extremophiles

, Volume 4, Issue 3, pp 157–164

Activity and stability of hyperthermophilic enzymes: a comparative study on two archaeal β-glycosidases

Authors

  • Jeroen Pouwels
    • Laboratory of Microbiology, Wageningen Agricultural University, Wageningen, The Netherlands
  • Marco Moracci
    • Institute of Protein Biochemistry and Enzymology, Consiglio Nazionale delle Ricerche, Via Marconi 10, 80125 Naples, Italy Tel. +39 081 5932411; Fax +39 081 2396525 e-mail: rossi@dafne.ibpe.na.cnr.it
  • Beatrice Cobucci-Ponzano
    • Institute of Protein Biochemistry and Enzymology, Consiglio Nazionale delle Ricerche, Via Marconi 10, 80125 Naples, Italy Tel. +39 081 5932411; Fax +39 081 2396525 e-mail: rossi@dafne.ibpe.na.cnr.it
  • Giuseppe Perugino
    • Institute of Protein Biochemistry and Enzymology, Consiglio Nazionale delle Ricerche, Via Marconi 10, 80125 Naples, Italy Tel. +39 081 5932411; Fax +39 081 2396525 e-mail: rossi@dafne.ibpe.na.cnr.it
  • John van der Oost
    • Laboratory of Microbiology, Wageningen Agricultural University, Wageningen, The Netherlands
  • Thijs Kaper
    • Laboratory of Microbiology, Wageningen Agricultural University, Wageningen, The Netherlands
  • J.H.G. Lebbink
    • Laboratory of Microbiology, Wageningen Agricultural University, Wageningen, The Netherlands
  • W.M. de Vos
    • Laboratory of Microbiology, Wageningen Agricultural University, Wageningen, The Netherlands
  • Maria Ciaramella
    • Institute of Protein Biochemistry and Enzymology, Consiglio Nazionale delle Ricerche, Via Marconi 10, 80125 Naples, Italy Tel. +39 081 5932411; Fax +39 081 2396525 e-mail: rossi@dafne.ibpe.na.cnr.it
  • M. Rossi
    • Institute of Protein Biochemistry and Enzymology, Consiglio Nazionale delle Ricerche, Via Marconi 10, 80125 Naples, Italy Tel. +39 081 5932411; Fax +39 081 2396525 e-mail: rossi@dafne.ibpe.na.cnr.it
ORIGINAL PAPER

DOI: 10.1007/s007920070030

Cite this article as:
Pouwels, J., Moracci, M., Cobucci-Ponzano, B. et al. Extremophiles (2000) 4: 157. doi:10.1007/s007920070030

Abstract

Sβgly 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 Sβgly 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.

Key words Thermophilic enzymes β-Glycosidase Thermostability Sulfolobus solfataricus Pyrococcus furiosus

Copyright information

© Springer-Verlag Tokyo 2000