Glycosylation of polyphenolic compounds: design of a self-sufficient biocatalyst by co-immobilization of a glycosyltransferase, a sucrose synthase and the cofactor UDP

Trobo Maseda, LaraRomero Fernandez, MariaGuisan, JoseRocha Martín, Javier2023-12-202023-12-202023Trobo-Maseda, Lara, et al. «Glycosylation of Polyphenolic Compounds: Design of a Self-Sufficient Biocatalyst by Co-Immobilization of a Glycosyltransferase, a Sucrose Synthase and the Cofactor UDP». International Journal of Biological Macromolecules, vol. 250, octubre de 2023, p. 126009. https://doi.org/10.1016/j.ijbiomac.2023.126009.0141-813010.1016/j.ijbiomac.2023.126009https://hdl.handle.net/20.500.14352/91618Glycosyltransferases catalyze the regioselective glycosylation of polyphenolic compounds, increasing their solubility without altering their antioxidant properties. Leloir-type glycosyltransferases require UDP-glucose as a cofactor to glycosylate a hydroxyl of the polyphenol, which is expensive and unstable. To simplify these processes for industrial implementation, the preparation of self-sufficient heterogeneous biocatalysts is needed. In this study, a glycosyltransferase and a sucrose synthase (as an UDP-regenerating enzyme) were co-immobilized onto porous agarose-based supports coated with polycationic polymers: polyethylenimine and polyallylamine. In addition, the UDP cofactor was strongly ionically adsorbed and co-immobilized with the enzymes, eliminating the need to add it separately. Thus, the optimal self-sufficient heterogeneous biocatalyst was able to catalyze the glycosylation of three polyphenolic compounds (piceid, phloretin and quercetin) with in situ regeneration of the UDP-glucose, allowing multiple consecutive reaction cycles without the addition of exogenous cofactor. A TTN value of 50 (theoretical maximum) was obtained in the reaction of piceid glycosylation, after 5 reaction cycles, using the self-sufficient biocatalyst based on an improved sucrose synthase variant. This result was 5-fold higher than the obtained using soluble cofactor and the co-immobilized enzymes, and much higher than those reported in the literature for similar processes.engAttribution 4.0 Internationalhttp://creativecommons.org/licenses/by/4.0/Glycosylation of polyphenolic compounds: design of a self-sufficient biocatalyst by co-immobilization of a glycosyltransferase, a sucrose synthase and the cofactor UDPjournal article1879-0003https://doi.org/10.1016/j.ijbiomac.2023.126009open access544.478577.15Sucrose synthaseGlycosyltransferaseCo-immobilizationColocalizationGlycobiotechnologySelf-sufficient biocatalystBioquímica (Biología)Bioquímica (Química)Biología molecular (Química)Biología molecular (Biología)2403 Bioquímica2415 Biología Molecular