Title :

Characterization of hyperbranched glycopolymers produced in vitro using enzymes

Author(s) :

Rolland-Sabaté, Agnès [Auteur]
Guilois, Sophie [Auteur]
Grimaud, Florent [Auteur]
Lancelon-Pin, Christine [Auteur]
Roussel, Xavier [Auteur]
Laguerre, Sandrine [Auteur]
Viksø-Nielsen, Anders [Auteur]
Putaux, Jean-Luc [Auteur]
D'hulst, Christophe [Auteur]
Unité de Glycobiologie Structurale et Fonctionnelle (UGSF) - UMR 8576
Potocki-Véronèse, Gabrielle [Auteur]
Buléon, Alain [Auteur]

Journal title :

Analytical and bioanalytical chemistry

Abbreviated title :

Anal Bioanal Chem

Volume number :

406

Pages :

1607-1618

Publication date :

2014-02

ISSN :

1618-2650

English keyword(s) :

Fractionation, Field Flow
Amylases
Molecular Weight
Sucrose
Glycosyltransferase
Glycogen
Light
Glucans
Bacteria
Molecular Structure
Scattering, Radiation

HAL domain(s) :

Chimie/Chimie théorique et/ou physique

English abstract : [en]

Asymmetrical flow field flow fractionation (AF4) has proven to be a very powerful and quantitative method for the determination of the macromolecular structure of high molar mass branched biopolymers, when coupled with ...
Show more >Asymmetrical flow field flow fractionation (AF4) has proven to be a very powerful and quantitative method for the determination of the macromolecular structure of high molar mass branched biopolymers, when coupled with multi-angle laser light scattering (MALLS). This work describes a detailed investigation of the macromolecular structure of native glycogens and hyperbranched α-glucans (HBPs), with average molar mass ranging from 2 × 10(6) to 4.3 × 10(7) g mol(-1), which are not well fractionated by means of classical size-exclusion chromatography. HBPs were enzymatically produced from sucrose by the tandem action of an amylosucrase and a branching enzyme mimicking in vitro the elongation and branching steps involved in glycogen biosynthesis. Size and molar mass distributions were studied by AF4, coupled with online quasi-elastic light scattering (QELS) and transmission electron microscopy. AF4-MALLS-QELS has shown a remarkable agreement between hydrodynamic radii obtained by online QELS and by AF4 theory in normal mode with constant cross flow. Molar mass, size, and dispersity were shown to significantly increase with initial sucrose concentration, and to decrease when the branching enzyme activity increases. Several populations with different size range were observed: the amount of small size molecules decreasing with increasing sucrose concentration. The spherical and dense global conformation thus highlighted was partly similar to native glycogens. A more detailed study of HBPs synthesized from low and high initial sucrose concentrations was performed using complementary enzymatic hydrolysis of external chains and chromatography. It emphasized a more homogeneous branching pattern than native glycogens with a denser core and shorter external chains.Show less >

Language :

Anglais

Administrative institution(s) :

CNRS
Université de Lille

Collections :

• Unité de Glycobiologie Structurale et Fonctionnelle (UGSF) - UMR 8576

Research team(s) :

Plant Storage Polysaccharides

Submission date :

2020-02-12T15:11:44Z