Is the acid/base catalytic residue mutation in beta-D-mannosidase DtMan from Dictyoglomus thermophilum sufficient enough to provide thioglycoligase activity? - Université d'Orléans Accéder directement au contenu
Article Dans Une Revue Biochimie Année : 2017

Is the acid/base catalytic residue mutation in beta-D-mannosidase DtMan from Dictyoglomus thermophilum sufficient enough to provide thioglycoligase activity?

Résumé

Glycoside hydrolases can be turned into thioglycoligase by mutation of the acid/base catalytic carboxylate residue. These mutants have proven valuable to generate S-glycosides, however, few examples in literature have described efficient thioglycoligase activity, and even fewer the underlying molecular mechanism. DtMan, a GH2 family beta-D-mannosidase from the thermophilic Dictyoglomus thermophilum was cloned and expressed in E. coll. The recombinant protein is highly specific for beta-D-mannosides, and exhibits efficient catalysis constants coupled to thermostability. However, seven variants bearing mutated acid/base residue could not be turned into efficient thioligases. Crystal structure of DtMan Glu425Cys mutant and molecular modeling calculations have demonstrated that unlike other GH2 thioligase reported, active site accessibility of thiol acceptor may be impaired by entrance loop rigidity. This structural feature may explain why DtMan mutants do not exhibit thioglycoligase activity.
Fichier non déposé

Dates et versions

hal-01608341 , version 1 (03-10-2017)

Identifiants

Citer

Laure Guillotin, Nicolas Richet, Pierre Lafite, Richard Daniellou. Is the acid/base catalytic residue mutation in beta-D-mannosidase DtMan from Dictyoglomus thermophilum sufficient enough to provide thioglycoligase activity?. Biochimie, 2017, 137, pp.190-196. ⟨10.1016/j.biochi.2017.03.020⟩. ⟨hal-01608341⟩
72 Consultations
0 Téléchargements

Altmetric

Partager

Gmail Facebook X LinkedIn More