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Synthesis of dextrans with controlled amounts of α-1,2 linkages using the transglucosidase GBD–CD2

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Abstract

GBD–CD2 is an α-1,2 transglucosidase engineered from DSR-E, a glucansucrase naturally produced by Leuconostoc mesenteroides NRRL B-1299. This enzyme catalyses from sucrose, the α-1,2 transglucosylation of glucosyl moieties onto α-1,6 dextran chains. Steady-state kinetic studies showed that hydrolysis and transglucosylation reactions occurred at the early stage of the reaction in the presence of 70 kDa dextran as acceptor and sucrose. The transglucosylation reaction catalysed by GBD–CD2 follows a Ping Pong Bi Bi mechanism with a high k cat value of 970 s−1. The amount of the synthesised α-1,2 side chains was found to be directly dependent on the initial molar ratio [Sucrose]/[Dextran]. Dextrans with controlled α-1,2 linkage contents ranging from 13% to 40% were synthesised. The procedure resulted in the production of dextrans with the highest content of α-1,2 linkages ever reported.

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Acknowledgements

Part of this work was carried out at MetaSys—the Metabolomics & Fluxomics Center at the Laboratoire d’Ingénierie des Systèmes Biologiques et des Procédés (LISBP, Toulouse, France)—which is supported by the Région Midi-Pyrénées (France) and the European Regional Development Fund (ERDF). We also gratefully acknowledge Pr. Philippe Besse, Sandrine Laguerre, Dr. Gabrielle Potocki-Véronèse and Pierre Escalier for their assistance. Yoann Brison is supported by a grant from the Région Midi-Pyrénées, France.

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Author notes

  1. Emeline Fabre

    Present address: Unité de Glycobiologie Structurale et Fonctionnelle, CNRS UMR 8576, IFR 147, Université Lille1, Sciences et Technologies, 59655, Villeneuve d’Ascq cedex, France

Authors and Affiliations

  1. Université de Toulouse; INSA, UPS, INP; LISBP, F-31077, Toulouse, France

    Yoann Brison, Claire Moulis, Jean-Charles Portais, Pierre Monsan & Magali Remaud-Siméon

  2. CNRS, UMR5504, F-31400, Toulouse, France

    Yoann Brison, Claire Moulis, Jean-Charles Portais, Pierre Monsan & Magali Remaud-Siméon

  3. INRA, UMR792 Ingénierie des Systémes Biologiques et des Procédés, F-31400, Toulouse, France

    Yoann Brison, Claire Moulis, Jean-Charles Portais, Pierre Monsan & Magali Remaud-Siméon

Authors
  1. Yoann Brison
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  2. Emeline Fabre
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  3. Claire Moulis
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Correspondence to Magali Remaud-Siméon.

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Yoann Brison and Emeline Fabre contributed equally to this work.

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Brison, Y., Fabre, E., Moulis, C. et al. Synthesis of dextrans with controlled amounts of α-1,2 linkages using the transglucosidase GBD–CD2. Appl Microbiol Biotechnol 86, 545–554 (2010). https://doi.org/10.1007/s00253-009-2241-z

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  • DOI: https://doi.org/10.1007/s00253-009-2241-z

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