CAL-B-Catalyzed Acylation of Nucleosides and Role of the Sugar Conformation: An Improved Understanding of the Enzyme-Substrate Recognition

• Published in: European journal of organic chemistry Vol. 2012; no. 28; pp. 5483 - 5490
• Main Authors: Martínez-Montero, Saúl, Fernández, Susana, Sanghvi, Yogesh S., Gotor, Vicente, Ferrero, Miguel
• Format: Journal Article
• Language: English
• Published: Weinheim WILEY-VCH Verlag 01.10.2012; WILEY‐VCH Verlag; Wiley; Wiley-VCH; Wiley Subscription Services, Inc
• Subjects: Acylation; Bioconversions. Hemisynthesis; Biological and medical sciences; Biotechnology; Carbohydrates with 4, 5, 6, ... C atoms, dissacharides and oligosaccharides; Carbohydrates. Nucleosides and nucleotides; Chemistry; Chemistry, Organic; Enzymes; Exact sciences and technology; Fundamental and applied biological sciences. Psychology; Methods. Procedures. Technologies; Molecular modeling; NMR; Nuclear magnetic resonance; Nucleosides; Nucleosides, nucleotides and oligonucleotides; Organic chemistry; Physical Sciences; Preparations and properties; Science & Technology; Sugar conformation; Enzymes; ENANTIOSELECTIVITY; BUILDING-BLOCKS; RESOLUTION; HYDROLYSIS; SOLUTION-PHASE SYNTHESIS; OLIGONUCLEOTIDES; NUCLEOTIDES; ANTARCTICA LIPASE-B; Sugar conformation; Molecular modeling; Nucleosides; LOCKED NUCLEIC-ACIDS; STEREOSPECIFICITY POCKET; Acylation; Catalytic reaction; candida antarctica; Enzyme; Triacylglycerol lipase; Molecular interaction; Oside; Esterases; NMR spectrometry; Selectivity; Carboxylic ester hydrolases; Fungi; Biotransformation; Force field method; Molecular recognition; Hydrolases; Nucleoside; Catalytic site; Fungi Imperfecti; Conformation
• ISSN: 1434-193X; 1099-0690
• DOI: 10.1002/ejoc.201200609

We demonstrate that the sugar ring conformation of nucleosides plays a critical role during Candida antarctica lipase B (CAL‐B) catalyzed acylation. Specifically, the North (N), but not the South (S) nucleoside sugar ring conformation is preferred for efficient binding at the catalytic site. In this study, we used nuclear magnetic resonance (NMR) spectroscopy experiments to establish the sugar ring conformation of nucleosides and performed molecular modeling studies to support the observations. The ribo‐ and 2′‐substituted (OMe, F) nucleosides displaying the N‐conformation undergo rapid and facile acylation compared to the 2′‐deoxynucleosides with the S‐conformation. This study improves our understanding of the critical role that sugar conformation plays in enzyme–substrate recognition during biotransformations using CAL‐B. To the best of our knowledge, this is the first experimental report offering a rationale for the observed selectivity during acylation of nucleosides containing the N‐sugar conformation. The sugar conformation of nucleosides plays a critical role in Candida antarctica lipase B catalyzed acylation. Specifically, the North nucleoside sugar ring conformation is preferred for efficient binding at the catalytic site. Nuclear magnetic resonance spectroscopy was used to establish the sugar conformation, which was confirmed by molecular modeling studies.