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Supplementary data for
Branching pattern of gluco-oligosaccharides and 1.5 kDa dextran

grafted by the -1,2 branching sucrase GBD-CD2


Yoann Brison1,2,3, Sandrine Laguerre1,2,3, François Lefoulon4, Sandrine Morel1,2,3, Nelly Monties1,2,3, Gabrielle Potocki-Véronèse1,2,3, Pierre Monsan1,2,3, Magali Remaud-Simeon1,2,3
1INSA, UPS, INP, LISBP, Université de Toulouse, 135 avenue de Rangueil, 31077 Toulouse, France

2UMR 5504, CNRS, 31400 Toulouse, France

3UMR 792, Ingénierie des Systèmes Biologiques et des Procédés, INRA, 31400 Toulouse, France

4Technologie Servier, 25/27 rue Eugène Vignat 45000 Orléans, France

To whom correspondence may be addressed: Magali Remaud-Simeon, Laboratoire d’Ingénierie des Systèmes Biologiques et des Procédés, Institut National des Sciences Appliquées, 135 avenue de Rangueil, 31077 Toulouse Cedex, France; Tel: +33 561 559 446; Fax: +33 561 559 400; E-mail remaud@insa-toulouse.fr


Supplementary figure for the materials and methods section


Fig. S1b



Fig. S1c

Reducing end


Fig. S1. Examples of data matrices corresponding to the modelling of branching scenarios. 1a, Data matrix corresponding to the dextran model. 1b, Representation of isomaltopentaose branching for the simulated scenario T. 1c, Representation of isomaltopentaose branching for the simulated scenario A (see Table 1).

Supplementary figures and tables for the result section
1D and 2D NMR spectra and additional LC-MS chromatograms

Fig. S2. 1H NMR spectra of anomeric regions of gluco-oligosaccharides 1, 2 and 3 (see Fig. 2).

The NMR peaks are designated by the proton position on the residue and followed by the letter of the D-Glcp unit. For example, H1B -(1→4) indicates that the anomeric proton of D-Glcp unit B has the -configuration and is linked to the C4 of D-Glcp unit A.

Fig. S3. HMBC spectrum of tetrasaccharide 1 recorded in deuterium oxide at 298 K.

The code H-1/C-6 indicates a long-range coupling between C-6 of a D-Glcp unit and H-1 of a D-Glcp unit.



Fig. S4. HMQC spectrum of pentasaccharide 2 recorded in deuterium oxide at 300.3 K.

The code H-1 / C-1 indicates couplings between C-1 of a D-Glcp unit and H-1 of the same D-Glcp unit.



Fig. S5. HMBC spectrum of pentasaccharide 2 recorded in deuterium oxide at 298 K.

The code H-1/C-2, C-4/C-6 indicates a long-range coupling between C-2, C-4 or C-6 of a D-Glcp unit and H-1 of another D-Glcp unit.

Fig. S6. HMBC spectrum of hexasaccharide 3 recorded in deuterium oxide at 298 K.

The code H-1/C-2, C-4/C-6 indicates a long-range coupling between C-2, C-4 or C-6 of a D-Glcp unit and H-1 of another D-Glcp unit.

Fig. S7. LC-MS analyses of the products obtained after hydrolysis by Aspergillus niger amyloglucosidase of -(1→2) glucosylated pentasaccharide 4 (-D-Glcp-(1→6)--D-Glcp-(1→6)--D-Glcp-(1→6)--D-Glcp-(1→4)-D-Glcp).

Numerals refer to the structures drawn in Fig. 4. Time value over each chromatogram indicates the reaction time. (♦), (*) before and after the Aspergillus niger amyloglucosidase hydrolysis, respectively.

Fig. S8. 1H NMR spectra of anomeric regions of gluco-oligosaccharides 4, 6a or 6b and 7.



The NMR peaks are designated by the proton position on the D-Glcp unit and followed by the letter of the unit. For example, H1B -(1→4) indicates that the anomeric proton of D-Glcp unit B has the -configuration and is linked to the C4 of unit A.


Fig. S9. Percentage of glucose released from -(1→2) branched dextrans after the action of Aspergillus niger amyloglucosidase. The hydrolysis ratio is expressed as the molar ratio of the released D-Glcp units versus the D-Glcp units contained in the branched dextrans.




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