This protocol describes the regioselective deprotection of single hydroxyls in peracetylated monosaccharides and disaccharides by enzymatic or chemoenzymatic strategies. The introduction of a one-pot enzymatic step by using immobilized biocatalysts obviates the requirement to carry out tedious workups and time-consuming purifications. By using this straightforward protocol, different per-O-acetylated glycopyranosides (mono- or disaccharides, 1-substituted or glycals) can be transformed into a whole set of differentially monodeprotected 1-alcohols, 3-alcohols, 4-alcohols and 6-alcohols in high yields. These tailor-made glycosyl acceptors can then be used for stereoselective glycosylation for oligosaccharide and glycoderivative synthesis. They have been successfully used as building blocks to synthesize tailor-made di- and trisaccharides involved in the structure of lacto-N-neo-tetraose and precursors of the tumor-associated carbohydrate antigen T and the antitumoral drug peracetylated β-naphtyl-lactosamine. We are able to prepare a purified monoprotected carbohydrate in between 1 and 4 d. With this protocol, the small library of monodeprotected products can be synthesized in 1–2 weeks.
Subscribe to Journal
Get full journal access for 1 year
only $9.92 per issue
All prices are NET prices.
VAT will be added later in the checkout.
Tax calculation will be finalised during checkout.
Rent or Buy article
Get time limited or full article access on ReadCube.
All prices are NET prices.
Chen, S. & Fukuda, M. Cell-type-specific roles of carbohydrates in tumor metastasis. Methods Enzymol. 416, 371–380 (2006).
Rabinovich, G.A., Toscano, M.A., Jackson, S.S. & Vasta, G.R. Functions of cell surface galectin-glycoprotein lattices. Curr. Opin. Struct. Biol. 17, 513–520 (2007).
Seeberger, P.H. & Werz, D.B. Synthesis and medical applications of oligosaccharides. Nature 446, 1046–1051 (2007).
Walker-Nasir, E., Kaleem, A., Hoessli, D.C., Khurshid, A. & Nasir-ud-Din . Galactose: a specifically recognized, terminal carbohydrate moiety in biological processes. Curr. Org. Chem. 12, 940–956 (2008).
Murrey, H.E. & Hsieh-Wilson, L.C. The chemical neurobiology of carbohydrates. Chem. Rev. 108, 1708–1731 (2008).
Weymouth-Wilson, A.C. The role of carbohydrates in biologically active natural products. Nat. Prod. Rep. 14, 99–110 (1997).
Kren, V. & Rezanka, T. Sweet antibiotics—the role of glycosidic residues in antibiotic and antitumor activity and their randomization. FEMS Microbiol. Rev. 32, 858–889 (2008).
Vyas, A.A. et al. Gangliosides are functional nerve cell ligands for myelin-associated glycoprotein (MAG), an inhibitor of nerve regeneration. Proc. Natl. Acad. Sci. USA 99, 8412–8417 (2002).
Sidransky, E. Gaucher disease: complexity in a 'simple' disorder. Mol. Genet. Metab. 83, 6–15 (2004).
Campbell, C.T. & Yarema, K.J. Large-scale approaches for glycobiology. Genome Biol. 6, 236–244 (2005).
Palomo, J.M., Filice, M., Fernandez-Lafuente, R., Terreni, M. & Guisan, J.M. Regioselective hydrolysis of peracetylated β-monosaccharides by immobilized lipases. Key role of the immobilization protocol. Adv. Synth. Cat. 349, 1969–1976 (2007).
Filice, M., Fernandez-Lafuente, R., Terreni, M., Guisan, J.M. & Palomo, J.M. Screening of lipases for regioselective hydrolysis of peracetylated β-monosaccharides. J. Mol. Cat. B: Enzym. 49, 12–17 (2007).
Fernandez-Lorente, G. et al. Lecitase ultra as regioselective biocatalyst in the hydrolysis of fully protected carbohydrates. Strong modulation by using different immobilization protocols. J. Mol. Cat. B: Enzym. 51, 110–117 (2008).
Filice, M. et al. Preparation of linear oligosaccharides by a simple monoprotective chemoenzymatic approach. Tetrahedron 64, 9286–9292 (2008).
Filice, M. et al. A Chemo-biocatalytic approach in the synthesis of β-O-naphtylmethyl-N-peracetylated lactosamine. J. Mol. Cat. B: Enzym. 52–53, 106–112 (2008).
Mendes, A.A. et al. Regioselective monohydrolysis of per-O-acetylated 1-O-substituted-β-glucopyranosides catalyzed by immobilized lipases. Tetrahedron 64, 10721–10727 (2008).
Filice, M. et al. Chemo-biocatalytic regioselective synthesis of different deprotected monosaccharides. Catal. Today 140, 11–18 (2009).
Rodrigues, D.S. et al. Different derivatives of a lipase display different regioselectivity in the monohydrolysis of per-O-acetylated 1-O-substituted-β-galactopyranosides. J. Mol. Cat. B: Enzym. 58, 36–40 (2009).
Filice, M., Vanna, R., Terreni, M., Guisan, J.M. & Palomo, J.M. Lipase-catalyzed regioselective one-step synthesis of Penta-O-acetyl-3-hydroxy-lactal. Eur. J. Org. Chem. 20, 3327–3329 (2009).
Filice, M., Guisan, J.M. & Palomo, J.M. Recent trends in regioselective protection and deprotection of monosaccharides. Curr. Org. Chem. 14, 516–532 (2010).
Filice, M., Guisan, J.M. & Palomo, J.M. Effect of ionic liquids as additives in the catalytic properties of different immobilized preparations of Rhizomucor miehei lipase in the hydrolysis of peracetylated lactal. Green Chem. 12, 1365–1369 (2010).
Wang, P.G. Sugars synthesized in a snap. Nat. Chem. Biol. 3, 309–310 (2007).
Hung, S.-C. et al. Synthesis of heparin oligosaccharides and their interaction with eosinophil-derived neurotoxin. Org. Biomol. Chem. 10, 760–772 (2012).
Hsu, C.-H., Hung, S.-C., Wu, C.-Y. & Wong, C.-H. Toward automated oligosaccharide synthesis. Angew. Chem. Int. Ed. 50, 11872–11923 (2011).
Galan, M.C., Benito-Alifonso, D. & Watt, G.M. Carbohydrate chemistry in drug discovery. Org. Biomol. Chem. 9, 3598–3610 (2011).
Wang, W. et al. Preparation of oligosaccharides by homogenous enzymatic synthesis and solid-phase extraction. Chem. Commun. 47, 11240–11242 (2011).
Fujikawa, K., Ganesh, N.V., Tan, Y.H., Stine, K.J. & Demchenko, A.V. Reverse orthogonal strategy for oligosaccharide synthesis. Chem. Commun. 47, 10602–10604 (2011).
Chu, K.-C. et al. Efficient and stereoselective synthesis of α(2→9) oligosialic acids: from monomers to dodecamers. Angew. Chem. Int. Ed. 50, 9391–9395 (2011).
Wu, C.-Y. & Wong, C.-H. Chemistry and glycobiology. Chem. Commun. 47, 6201–6207 (2011).
Flitsch, S.L. Glycosylation with a twist. Nature 437, 201–202 (2005).
Ernst, B., Hart, G.W. & Sinay, P. (eds). Carbohydrates in Chemistry and Biology, Vol. 1 (Wiley-VCH, 2000).
Wang, C.C. et al. Regioselective one-pot protection of carbohydrates. Nature 446, 896–899 (2007).
Antoine, F., Urban, D. & Beau, J.-M . Tandem catalysis for a one-pot regioselective protection of carbohydrates. The example of glucose. Angew. Chem. Int. Ed. 46, 8662–8665 (2007).
Wang, C.-C., Kulkarni, S.S., Lee, J.-C., Luo, S.-Y. & Hung, S.-C. Regioselective one-pot protection of glucose. Nat. Protoc. 3, 97–113 (2008).
Pastore, A., Valerio, S., Adinolfi, M. & Iadonisi, A. An easy and versatile approach for the regioselective De-O-benzylation of protected sugars based on the I2/Et3SiH combined system. Chem. Eur. J. 17, 5881–5889 (2011).
Wuts, P.G.M. Greene's Protective Groups in Organic Synthesis, 4th edn. (John Wiley & Sons, 2007).
Greene, T.W. & Wuts, P.G.M. Protective Groups in Organic Synthesis, 3th edn. (John Wiley & Sons, 1999).
Jaeger, K.-E. & Eggert, T. Lipases for biotechnology. Curr. Opin. Biotechnol. 13, 390–397 (2002).
Reetz, M.T. Lipases as practical biocatalysts. Curr. Opin. Biotechnol. 6, 145–150 (2002).
Brady, L. et al. A serine protease triad forms the catalytic centre of a triacylglycerol lipase. Nature 343, 767–770 (1990).
Palomo, J.M. Modulation of enzymes selectivity via immobilization. Curr. Org. Synth. 6, 1–14 (2009).
Palomo, J.M. Lipases enantioselectivity alteration by immobilization techniques. Curr. Bio. Comp. 4, 126–138 (2008).
Cabrera, Z. & Palomo, J.M. Enantioselective desymmetrization of prochiral diesters catalyzed by immobilized Rhizopus oryzae lipase. Tetrahedron: Asymmetry 22, 2080–2084 (2011).
Bastida, A. et al. A single step purification, immobilization and hyperactivation of lipases via interfacial adsorption on strongly hydrophobic supports. Biotechnol. Bioeng. 58, 486–493 (1998).
Mateo, C. et al. Some special features of glyoxyl supports to immobilize proteins. Enzyme Microb. Technol. 37, 456–462 (2005).
Mong, T.K.K., Lee, L.V., Brown, J.R., Esko, J.D. & Wong, C.H. Synthesis of N-acetyllactosamine derivatives with variation in the aglycon moiety for the study of inhibition of sialyl Lewis x expression. ChemBioChem 4, 835–840 (2003).
Bradford, M.M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72, 248–254 (1976).
Palomo, J.M. et al. General trend of lipase to self-assemble giving bimolecular aggregates greatly modifies the enzyme functionality. Biomacromolecules 4, 1–6 (2003).
This work was supported by The Spanish National Research Council (CSIC) and the Spanish Ministry of Science. We acknowledge Á. Berenguer (Instituto Universitario de Materiales, Universidad de Alicante) for his help during the writing of this paper.
The authors declare no competing financial interests.
Per-O-acetylation of carbohydrates (PDF 125 kb)
Regioselective enzymatic C-6 monodeprotection of per-O-acetylated glycopyranosides (PDF 173 kb)
Regioselective enzymatic monodeprotection of per-O-acetylated glycopyranosides (PDF 171 kb)
Synthesis of 4-hydroxy-tetraacetylated monosaccharides by acyl-chemical migration from the 6-OH monodeprotected tetraacetylated products (PDF 136 kb)
Synthesis of 3-hydroxy-tetraacetylated monosaccharides by acyl-chemical migration from the 6-OH monodeprotected tetraacetylated products (PDF 137 kb)
HPLC trace for the regioselctive enzymatic hydrolysis of 1,2,3,4,6-Penta-O-acetyl-α-D-glucopyranose (20) (PDF 551 kb)
HPLC traces for the chemical acyl-migration of 39 to form 64 and 73 (PDF 528 kb)
TLC of the chemo-enzymatic process starting from 1,2,3,4,6-Penta-O-acetyl-α-D-glucopyranose (20) (PDF 427 kb)
About this article
Cite this article
Filice, M., Guisan, J., Terreni, M. et al. Regioselective monodeprotection of peracetylated carbohydrates. Nat Protoc 7, 1783–1796 (2012). https://doi.org/10.1038/nprot.2012.098
Nature Communications (2015)
Nature Protocols (2013)