Characterization of substrate and product ...
Type de document :
Article dans une revue scientifique
PMID :
URL permanente :
Titre :
Characterization of substrate and product specificity of the purified recombinant glycogen branching enzyme of Rhodothermus obamensis
Auteur(s) :
Roussel, Xavier [Auteur]
Lancelon-Pin, Christine [Auteur]
Viksø-Nielsen, Anders [Auteur]
Rolland-Sabaté, Agnès [Auteur]
Grimaud, Florent [Auteur]
Potocki-Véronèse, Gabrielle [Auteur]
Buléon, Alain [Auteur]
Putaux, Jean-Luc [Auteur]
D'hulst, Christophe [Auteur]
Unité de Glycobiologie Structurale et Fonctionnelle (UGSF) - UMR 8576
Lancelon-Pin, Christine [Auteur]
Viksø-Nielsen, Anders [Auteur]
Rolland-Sabaté, Agnès [Auteur]
Grimaud, Florent [Auteur]
Potocki-Véronèse, Gabrielle [Auteur]
Buléon, Alain [Auteur]
Putaux, Jean-Luc [Auteur]
D'hulst, Christophe [Auteur]
Unité de Glycobiologie Structurale et Fonctionnelle (UGSF) - UMR 8576
Titre de la revue :
Biochimica et biophysica acta
Nom court de la revue :
Biochim. Biophys. Acta
Numéro :
1830
Pagination :
2167-2177
Date de publication :
2013-01
ISSN :
0006-3002
Mot(s)-clé(s) en anglais :
Substrate Specificity
1,4-alpha-Glucan Branching Enzyme
Catalysis
Rhodothermus
Bacterial Proteins
Enzyme Stability
1,4-alpha-Glucan Branching Enzyme
Catalysis
Rhodothermus
Bacterial Proteins
Enzyme Stability
Discipline(s) HAL :
Chimie/Chimie théorique et/ou physique
Résumé en anglais : [en]
BACKGROUND: Glycogen and starch branching enzymes catalyze the formation of α(1→6) linkages in storage polysaccharides by rearrangement of preexisting α-glucans. This reaction occurs through the cleavage of α(1→4) linkage ...
Lire la suite >BACKGROUND: Glycogen and starch branching enzymes catalyze the formation of α(1→6) linkages in storage polysaccharides by rearrangement of preexisting α-glucans. This reaction occurs through the cleavage of α(1→4) linkage and transfer in α(1→6) of the fragment in non-reducing position. These enzymes define major elements that control the structure of both glycogen and starch. METHODS: The kinetic parameters of the branching enzyme of Rhodothermus obamensis (RoBE) were established after in vitro incubation with different branched or unbranched α-glucans of controlled structure. RESULTS: A minimal chain length of ten glucosyl units was required for the donor substrate to be recognized by RoBE that essentially produces branches of DP 3-8. We show that RoBE preferentially creates new branches by intermolecular mechanism. Branched glucans define better substrates for the enzyme leading to the formation of hyper-branched particles of 30-70nm in diameter (dextrins). Interestingly, RoBE catalyzes an additional α-4-glucanotransferase activity not described so far for a member of the GH13 family. CONCLUSIONS: RoBE is able to transfer α(1→4)-linked-glucan in C4 position (instead of C6 position for the branching activity) of a glucan to create new α(1→4) linkages yielding to the elongation of linear chains subsequently used for further branching. This result is a novel case for the thin border that exists between enzymes of the GH13 family. GENERAL SIGNIFICANCE: This work reveals the original catalytic properties of the thermostable branching enzyme of R. obamensis. It defines new approach to produce highly branched α-glucan particles in vitro.Lire moins >
Lire la suite >BACKGROUND: Glycogen and starch branching enzymes catalyze the formation of α(1→6) linkages in storage polysaccharides by rearrangement of preexisting α-glucans. This reaction occurs through the cleavage of α(1→4) linkage and transfer in α(1→6) of the fragment in non-reducing position. These enzymes define major elements that control the structure of both glycogen and starch. METHODS: The kinetic parameters of the branching enzyme of Rhodothermus obamensis (RoBE) were established after in vitro incubation with different branched or unbranched α-glucans of controlled structure. RESULTS: A minimal chain length of ten glucosyl units was required for the donor substrate to be recognized by RoBE that essentially produces branches of DP 3-8. We show that RoBE preferentially creates new branches by intermolecular mechanism. Branched glucans define better substrates for the enzyme leading to the formation of hyper-branched particles of 30-70nm in diameter (dextrins). Interestingly, RoBE catalyzes an additional α-4-glucanotransferase activity not described so far for a member of the GH13 family. CONCLUSIONS: RoBE is able to transfer α(1→4)-linked-glucan in C4 position (instead of C6 position for the branching activity) of a glucan to create new α(1→4) linkages yielding to the elongation of linear chains subsequently used for further branching. This result is a novel case for the thin border that exists between enzymes of the GH13 family. GENERAL SIGNIFICANCE: This work reveals the original catalytic properties of the thermostable branching enzyme of R. obamensis. It defines new approach to produce highly branched α-glucan particles in vitro.Lire moins >
Langue :
Anglais
Établissement(s) :
CNRS
Université de Lille
Université de Lille
Équipe(s) de recherche :
Plant Storage Polysaccharides
Date de dépôt :
2020-02-12T15:11:46Z