Construction of a Novel Chimeric Dextransucrase Fused to the Carbohydrate-Binding Module CBM2a

Abstract

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.