Construction of a Novel Chimeric Dextransucrase Fused to the Carbohydrate-Binding Module CBM2a
2021
Fraga Vidal, Reinaldo | Arísticas Ribalta, Roberto, Carlos | Martínez Valdés, Lisandra, Teresa | Lafargue Gámez, Meinardo | Montes Alvarez, Amanda | Rubio Sánchez, Arianne | Dubreucq, Eric | Moreau, Benoît | Instituto Cubano de Investigaciones de los Derivados de la Caña de Azúcar (ICIDCA) | Ingénierie des Agro-polymères et Technologies Émergentes (UMR IATE) ; Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad)-Université de Montpellier (UM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Institut Agro - Montpellier SupAgro ; Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)-Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro) | Haute Ecole Condorcet | Development Research Project financed by the Development Cooperation Program of ARES (Académie de Recherche et d’Enseigment Supérieur) from Belgium (2017–2022)
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Afficher plus [+] Moins [-]anglais. Lactic acid bacteria (LAB) have the potential to produce homoexopolysaccharides (HoPS). Their health benefits and physicochemical properties have been the subject of extensive research. The HoPS functional properties are determined by molecular weight, the type of glycosidic linkages, degrees of branching and chemical composition. The dextransucrases (DSases) produce a kind of HoPS (dextrans), which are among the first biopolymers produced at industrial scale with applications in medicine and biotechnology. The glycodiversification opens additional applications for DSases. Therefore, the design and characterization of new DSases is of prime importance. Previously, we described the isolation and characterization of a novel extracellular dextransucrase (DSR-F) encoding gene. In this study, from DSR-F, we design a novel chimeric dextransucrase DSR-F-∆SP-∆GBD-CBM2a, where DSR-F-∆SP-∆GBD (APY repeats and a CW repeat deleted) was fused to the carbohydrate-binding module (CBM2a) of the β-1-4 exoglucanase/xylanase Cex (Xyn10A) of Cellulomonas fimi ATCC 484. This dextransucrase variant is active and the specificity is not altered. The DSR-F-∆SP-∆GBD-CBM2a was purified by cellulose affinity chromatography for the first time. This research showed that hybrids and chimeric biocatalyst DSases with novel binding capacity to cellulose can be designed to purify and immobilize using renewable lignocellulosic materials as supports.
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