A review of innovative tools for creative nucleic acid chemists that open the door to novel probes and therapeutic agents

Nucleic acids continue to gain importance as novel diagnostic and therapeutic agents. With contributions from noted scientists and scholars, Enzymatic and Chemical Synthesis of Nucleic Acid Derivatives is a practical reference that includes a wide range of approaches for the synthesis of designer nucleic acids and their derivatives.

The book covers enzymatic (including chemo-enzymatic) methods, with a focus on the synthesis and incorporation of modified nucleosides. The authors also offer a review of innovative approaches for the non-enzymatic chemical synthesis of nucleic acids and their analogs and derivatives, highlighting especially challenging species. The book offers a concise review of the methods that prepare novel and heavily modified polynucleotides in sufficient amount and purity for most clinical and research applications. This important book:

-Presents a timely and topical guide to the synthesis of designer nucleic acids and their derivatives
-Addresses the growing market for nucleotide-derived pharmaceuticals used as anti-infectives and chemotherapeutic agents, as well as fungicides and other agrochemicals.
-Covers novel methods and the most recent trends in the field
-Contains contributions from an international panel of noted scientistics

Written for biochemists, medicinal chemists, natural products chemists, organic chemists, and biotechnologists, Enzymatic and Chemical Synthesis of Nucleic Acid Derivatives is a practice-oriented guide that reviews innovative methods for the enzymatic as well as non-enzymatic synthesis of nucleic acid species.

Jesús Fernández-Lucas, PhD, is a tenured professor at the European University of Madrid, where he leads the Applied Biotechnology research group. His research is focused on the design and application of novel biocatalysts for the enzymatic synthesis of nucleosides.

María José Camarasa Rius, PhD, is Full Professor at the Medicinal Chemistry Institute of the Spanish National Research Council (IQM-CSIC) in Madrid, working on the design of novel enzymatic inhibitors with potential anticancer, antifungal and antiviral activity.

Inhalt

Preface xi

1 Enzymatic Synthesis of Nucleoside Analogues by Nucleoside Phosphorylases 1
Sarah Kamel, Heba Yehia, Peter Neubauer, and AnkeWagner

1.1 Introduction 1

1.1.1 Nucleosides and Nucleoside Analogues 1

1.1.2 Enzymes Involved in the Enzymatic Synthesis of Nucleoside Analogues 3

1.2 Nucleoside Phosphorylases 3

1.2.1 Classification and Substrate Spectra of Nucleoside Phosphorylases 3

1.2.1.1 Nucleoside Phosphorylase-I Family 4

1.2.1.2 Nucleoside Phosphorylase-II Family 6

1.2.2 Limitations in the Current Classification 7

1.2.3 Reaction Mechanism 8

1.2.4 Domain Structure and Active Site Residues of Nucleoside Phosphorylases 9

1.2.4.1 NP-I Family Members 9

1.2.4.2 NP-II FamilyMembers 10

1.3 Enzymatic Approaches to Produce Nucleoside Analogues Using Nucleoside Phosphorylases 11

1.3.1 One-pot Two-Step Transglycosylation Reaction 11

1.3.2 Pentofuranose-1-phosphate as Universal Glycosylating Substrate for Nucleoside Phosphorylase (NP) 12

1.3.2.1 Nucleoside Synthesis from Chemically Synthesized Pentose-1P 12

1.3.2.2 Nucleosides Synthesis from d-Glyceraldehyde-3-phosphate 13

1.3.2.3 Nucleoside Synthesis from d-Pentose 13

1.3.2.4 Nucleoside Synthesis from Enzymatically Produced Pentose-1P 13

1.4 Approaches to Produce Nucleoside Analogues 14

1.4.1 Whole Cell Catalysis 14

1.4.2 Crude Enzyme Extract 15

1.4.3 Application of Purified Enzymes 15

1.4.3.1 Immobilized Enzymes 16

1.4.3.2 Enzyme Reactors 17

1.5 Upscaling Approaches for the Production of Nucleoside Analogues 18

1.6 Production of Pharmaceutically Active Compounds by Nucleoside Phosphorylases 18

1.7 Outlook for the Application of Nucleoside Phosphorylase in the Production of Nucleoside Analogues 19

References 20

2 Enzymatic Phosphorylation of Nucleosides 29
Daniela Ubiali and Giovanna Speranza

2.1 Introduction 29

2.2 Nonspecific Acid Phosphatases (NSAPs) 30

2.3 Deoxyribonucleoside Kinases (dNKs) 33

2.4 Conclusion 37

References 37

3 Enzymatic Synthesis of Nucleic Acid Derivatives UsingWhole Cells 43
Elizabeth S. Lewkowicz and AdolfoM. Iribarren

3.1 Introduction 43

3.2 Nucleoside Synthesis Mediated by Microbial Nucleoside Phosphorylases 45

3.3 Nucleoside Analogues Synthesis by the Combined Action of Microbial Nucleoside Phosphorylases and Other Enzymes 48

3.3.1 Nucleoside Phosphorylases Coupled to Deaminases 48

3.3.2 Nucleoside Phosphorylases Coupled to Phosphopentomutase 48

3.3.3 Nucleoside Phosphorylases Coupled to Phosphopentomutase and Other Enzymes 49

3.3.4 Nucleoside Phosphorylases Coupled to Other Enzymes 51

3.4 Chemoenzymatic Preparation of Nonconventional Nucleoside Analogues Involving Whole Cell Biocatalyzed Key Steps 51

3.4.1 l-Nucleosides 52

3.4.2 Carbocyclic Nucleosides 55

3.4.3 C-Nucleosides 56

3.5 Nucleoside Prodrugs Preparation by Whole Cell Systems 57

3.5.1 Acylnucleosides 57

3.5.2 Nucleoside Phosphates 59

3.6 Other Nucleoside Derivatives 61

3.6.1 NDP 61

3.6.2 NDP-sugar 61

3.7 Perspectives 65

References 65

4 Enzymatic Synthesis of Nucleic Acid Derivatives by Immobilized Cells 79
Jorge A. Trelles, CintiaW. Rivero, Claudia N. Britos, and María J. Lapponi

4.1 Introduction 79

4.2 Nucleic Acid Derivatives 81

4.3 Whole Cell Immobilization: Generalities 85

4.4 Synthesis of Nucleosides by Immobilized Cells 86

4.4.1 Natural Nucleoside Synthesis 87

4.4.2 Nucleoside Analogues Synthesis 88

4.4.3 Nucleoside Analogues Derivatives Synthesis 92

4.5 Conclusion 98

References 98

5 Enzymatic Synthesis of Nucleic Acid Derivatives b...