Investigations on the Determinants Responsible for Low Molar Mass Dextran Formation by DSR-M Dextransucrase - CNRS - Centre national de la recherche scientifique Accéder directement au contenu
Article Dans Une Revue ACS Catalysis Année : 2017

Investigations on the Determinants Responsible for Low Molar Mass Dextran Formation by DSR-M Dextransucrase

Résumé

Certain enzymes of the GH70 family dextransucrases synthesize very high molar mass dextran polymers, whereas others produce a mixed population of very high and low molar mass products directly from sucrose substrate. Identifying the determinants dictating polymer elongation would allow the tight control of dextran size. To explore this central question, we focus on the recently discovered DSR-M enzyme from Leuconostoc citreum NRRL B-1299, which is the sole enzyme that naturally, exclusively, and very efficiently produces only low molar mass dextrans from sucrose. Extensive biochemical and structural characterization of a truncated form of DSR-M (DSR-MΔ2, displaying the same biochemical behavior as the parental enzyme) and X-ray structural analysis of complexes with sucrose and isomaltotetraose molecules together with accurate monitoring of the resulting polymer formation reveal that DSR-MΔ2 adopts a nonprocessive mechanism attributed to (i) a high propensity to recognize sucrose as a preferred acceptor at the initial stage of catalysis, (ii) an ability to elongate oligodextrans irrespective of their size, and (iii) the presence of a domain V showing a weak ability to bind to the growing dextran chains. In this study, we present the 3D structure with the largest defined domain V reported to date in the GH70 family and map sugar binding pockets on the basis of the structure of the complex obtained with isomaltotetraose. Altogether, these findings give insights into the interplay between the domain V and the catalytic site during polymerization. They open promising strategies for GH70 enzyme engineering aiming at modulating glucan size.
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Dates et versions

hal-01886431 , version 1 (02-12-2021)

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Marion Claverie, Gianluca Cioci, Marlène Vuillemin, Nelly Monties, Pierre Roblin, et al.. Investigations on the Determinants Responsible for Low Molar Mass Dextran Formation by DSR-M Dextransucrase. ACS Catalysis, 2017, 7 (10), pp.7106-7119. ⟨10.1021/acscatal.7b02182⟩. ⟨hal-01886431⟩
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