2026-06-27T15:15:43Zhttps://docta.ucm.es/rest/oai/request
oai:docta.ucm.es:20.500.14352/963322025-05-09T23:46:33Zcom_20.500.14352_14col_20.500.14352_15
Docta Complutense advisor Hernáiz Gómez-Degano, María Josefa author Hoyos Vidal, María Pilar author Bavaro, Teodora author Perona Requena, Almudena author Rumbero Sánchez, Ángel author Tengattini, Sara author Terreni, Marco author Hernáiz Gómez-Degano, María Josefa 2024-01-30T08:55:49Z 2024-01-30T08:55:49Z 2020 Hoyos Vidal, M. P., Bavaro, T., Perona Requena, A. et al. «Highly Efficient and Sustainable Synthesis of Neoglycoproteins Using Galactosidases». ACS Sustainable Chemistry & Engineering, vol. 8, n.o 16, abril de 2020, pp. 6282-92. https://doi.org/10.1021/acssuschemeng.9b07785. 10.1021/acssuschemeng.9b07785 https://hdl.handle.net/20.500.14352/96332 2168-0485 https://doi.org/10.1021/acssuschemeng.9b07785 Oligosaccharides are a key component on glycoconjugates with biomedical applications. However, the investigation of more efficient and environmental protocols for their preparation carrying defined oligosaccharides remains a big challenge due to lack of access to structurally well-defined and functionalized oligosaccharides. In this work a highly sustainable and efficient chemoenzymatic synthesis of neo-glycoproteins in green solvents was studied. In particular, we described the preparation of new activated disaccharides useful for direct protein binding under mild reaction conditions, reducing the protection/activation steps commonly needed in the traditional synthetic routes. Thus, N-acetylglucosamine (GlcNAc) was functionalized with a thiocyanomethyl group at C-1 and employed for the first time as acceptor in the enzymatic synthesis of the disaccharide of interest using β-Gal-3 from Bacillus circulans and β-galactosidase from Escherichia coli. The use of biosolvents promotes a very important increase of the enzymatic activity in the synthesis of functionalized Gal-β-(1 → 6)-GlcNAc (95%) preventing hydrolytic activity with full regioselectivity. Furthermore, reaction scaling up and biosolvent recycling are feasible without losing catalytic action. The enzyme–substrate recognition and the effect of biosolvent over the galactosidases activity has been investigated and explained by computational modeling studies. The presence of C-1 thiocyanomethyl moiety strongly hindered the normal course of transglycosylation for β-Gal-3 afforded β(1 → 6) glyosidic linkage instead of β(1 → 3). Then, this functionalized disaccharide has been activated with imino methoxyethyl (IME) linker and conjugated to ribonuclease A (RNase A). The glycosylation of RNase A, with IME-disaccharide provided the corresponding neo-glycoprotein with 85% of yield. This work opens to a new, simple, and green way to synthesize innovative glycoconjugate vaccines. eng Highly Efficient and Sustainable Synthesis of Neoglycoproteins Using Galactosidases journal article 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URL https://docta.ucm.es/bitstreams/ce388cb8-5480-49f1-889a-183baae3be11/download File MD5 857adafaa71d1f0aee0e728032b30e91 4729430 application/pdf Synthesis_of_Neoglycoproteins.pdf URL https://docta.ucm.es/bitstreams/10205e6c-a983-456d-9e2c-96303b8fb434/download File MD5 f2dc36793c39b6d7f27b92181a5d4618 69048 text/plain ACS Sustain. Chem. Eng. 2202.pdf.txt