Nucleoside recovery in DNA and RNA synthesis (original) (raw)
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New trends in nucleoside biotechnology
Acta naturae, 2010
This review focuses on new trends in nucleoside biotechnology, which have emerged during the last decade. Continuously growing interest in the study of this class of compounds is fueled by a number of factors: ( i ) a growing need for large-scale production of natural 2 ' -deoxy- β -D-ribonucleosides as well as their analogs with modifications in the carbohydrate and base fragments, which can then be used for the synthesis and study of oligonucleotides, including short-interfering RNA (siRNA), microRNA (miRNA), etc.; ( ii ) a necessity for the development of efficient practical technologies for the production of biologically important analogs of natural nucleosides, including a number of anticancer and antiviral drugs; ( iii ) a need for further study of known and novel enzymatic transformations and their use as tools for the efficient synthesis of new nucloside analogs and derivates with biomedical potential. This article will review all of these aspects and also include a brie...
Some recent findings in the biotechnology of biologically important nucleosides
Some recent findings in the biotechnology of biologically important nucleosides will be discussed, viz., (i) a new strategy of the cascade one-pot transformation of D-pentoses into nucleosides based on the extension and deepening of the knowledge of the mechanism of functioning of the ribokinase (RK), phosphopentomutase (PPM), and uridine (UP), thymidine (TP) & purine nucleoside (PNP) phosphorylases, and the role of different factors (structural, electronic, stereochemical) in the glycoside bond formation, (ii) the modern chemistries of the chemo-enzymatic syntheses of nucleosides, (iii) the transglycosylation reaction using natural and sugar modified nucleosides as donors of carbohydrate residues and heterocyclic bases as acceptors catalyzed by nucleoside phosphorylases (NP).
Enzymatic synthesis of nucleosides by nucleoside phosphorylase co-expressed in Escherichia coli
Journal of Zhejiang University SCIENCE B, 2010
Nucleoside phosphorylase is an important enzyme involved in the biosynthesis of nucleosides. In this study, purine nucleoside phosphorylase and pyrimidine nucleoside phosphorylase were co-expressed in Escherichia coli and the intact cells were used as a catalyst for the biosynthesis of nucleosides. For protein induction, lactose was used in place of isopropyl β-D-1-thiogalactopyranoside (IPTG). When the concentration of lactose was above 0.5 mmol/L, the ability to induce protein expression was similar to that of IPTG. We determined that the reaction conditions of four bacterial strains co-expressing these genes (TUD, TAD, DUD, and DAD) were similar for the biosyntheses of 2,6-diaminopurine nucleoside and 2,6-diaminopurine deoxynucleoside. When the substrate concentration was 30 mmol/L and 0.5% of the recombinant bacterial cell volume was used as the catalyst (pH 7.5), a greater than 90% conversion yield was reached after a 2-h incubation at 50 °C. In addition, several other nucleosides and nucleoside derivatives were efficiently synthesized using bacterial strains co-expressing these recombinant enzymes.
New developments in nucleoside analogues biosynthesis: A review
Journal of Molecular Catalysis B: Enzymatic, 2016
Highlights Nucleoside analogues (NAs) are commonly used as antitumoral and antiviral compounds Chemoenzymatic synthesis of biologically important NAs is described The use of enzymes as biocatalysts improves chemo-, regio-and stereoselectivity Enzymatic modification of NAs improve the pharmacological properties of the existing drugs Biocatalyst immobilization favors separation, reuse, and bioprocess scale up
Enzymatic synthesis of nucleoside 5′-mono and -triphosphates
FEBS Letters, 1990
The transformation of fluorodeoxy-nucleosides into 5'-monophosphates with the use of whole bacterial cells of Erwinia herbicola as a biocatalyst and PNP as a phosphate donor and next into S-triphosphates by using an extract of the Saccharomyces cerevisiae cells has been demonstrated. Nucleoside 5'-monophosphate; Nucleoside phosphotransferase; Nucleoside 5'-triphosphate; Nucleotide kinase; (Saccharomyces cerevisiae, Erwinia herbicola)
Nucleic Acids Research, 1976
Nucleoside 3'-phosphotriesters as key intermediates for the oligoribonucleotide synthesis. IV. New method for removal of 2,2,2-trichloroethyl group and 31P NM R as a new tool for analysis of deblocking of intemucleotide phosphate protecting groups1 ABSTRACT Zinc/acetylacetone/pyridine treatment has been designed as a very efficient method for removal of 2,2,2-trichloroethyl group from phosphoesters. Internucleotide and terminal 2,2,2-trichloroethylphosphotriesters were transformed to corresponding diesters quantitatively. Much less reactive 2,2,2-trichloroethylphosphodiesters produced monoesters with ca. 909 yield. 31 NMR spectroscopy has been proposed as a new tool for analysis of removal of internucleotide phosphate protecting groupsa crucial step in oligonucleotides synthesis via phosphotriester approach.
Selective O-phosphitilation with nucleoside phosphoramidite reagents
Nucleic Acids Research, 1992
In contrast to tetrazole, pyridine hydrochloride/ imidazole converts nucleoside phosphoramidltes to intermediates that show a high preference for phosphltllating hydroxyl groups relative to nucleoside amino groups. Use of this activating agent and incorporation of a pyridine hydrochloride/aniline wash step in the synthetic cycles permit synthesis of mixed base twenty-mer oligonucleotides from nucleoside reagents containing unprotected amino groups. This approach should be useful for the synthesis of oligonucleotide analogues containing substltuents sensitive to reagents used in conventional deblocking steps. Pyridine hydrochloride itself is an effective reagent for activating nucleoside methylphosphonoamidites and ribonucleoside phosphoramidltes, as well as deoxyribonucleoside phosphoramldites, when high O/N selectively is not needed.