The b -glycosidase from the hyperthermophilic Archaeon Pyrococcus horikoshii (Phob-gly) is a monomeric enzyme with wide substrate specificity belonging to family 1 of glycoside hydrolases classification. Inspection of the three-dimensional structure of the enzyme, recently resolved, showed that Phob-gly is membrane bound and that the residues putatively involved in the catalytic activity are Glu155 and Glu324 working as the general acid/base and the nucleophile of the reaction, respectively. We show here that mutation of the latter completely eliminated the activity of the enzyme and that it could be reactivated in the presence of sodium formate. Analysis of the products obtained in the presence of sodium formate buffer pH 4.0 at 758C showed that the Glu324Gly mutant acts as a hyperthermophilic glycosynthase.

Preparation of a glycosynthase from the beta-glycosidase of the hyperthermophilic Archaeon Pyrococcus horikoshii / Perugino, G.; Falcicchio, P.; Corsaro, MARIA MICHELA; Matsui, I.; Parrilli, Michelangelo; Rossi, Mose'; Moracci, Marco. - In: BIOCATALYSIS AND BIOTRANSFORMATION. - ISSN 1024-2422. - STAMPA. - 24:(2006), pp. 23-29.

Preparation of a glycosynthase from the beta-glycosidase of the hyperthermophilic Archaeon Pyrococcus horikoshii.

G. PERUGINO;CORSARO, MARIA MICHELA;PARRILLI, MICHELANGELO;ROSSI, MOSE';MORACCI, Marco
2006

Abstract

The b -glycosidase from the hyperthermophilic Archaeon Pyrococcus horikoshii (Phob-gly) is a monomeric enzyme with wide substrate specificity belonging to family 1 of glycoside hydrolases classification. Inspection of the three-dimensional structure of the enzyme, recently resolved, showed that Phob-gly is membrane bound and that the residues putatively involved in the catalytic activity are Glu155 and Glu324 working as the general acid/base and the nucleophile of the reaction, respectively. We show here that mutation of the latter completely eliminated the activity of the enzyme and that it could be reactivated in the presence of sodium formate. Analysis of the products obtained in the presence of sodium formate buffer pH 4.0 at 758C showed that the Glu324Gly mutant acts as a hyperthermophilic glycosynthase.
2006
Preparation of a glycosynthase from the beta-glycosidase of the hyperthermophilic Archaeon Pyrococcus horikoshii / Perugino, G.; Falcicchio, P.; Corsaro, MARIA MICHELA; Matsui, I.; Parrilli, Michelangelo; Rossi, Mose'; Moracci, Marco. - In: BIOCATALYSIS AND BIOTRANSFORMATION. - ISSN 1024-2422. - STAMPA. - 24:(2006), pp. 23-29.
File in questo prodotto:
Non ci sono file associati a questo prodotto.

I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.

Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11588/204309
Citazioni
  • ???jsp.display-item.citation.pmc??? ND
  • Scopus 11
  • ???jsp.display-item.citation.isi??? 10
social impact