Nucleoside Synthesis Mediated by Glycosyl Transferring Enzymes

Prasad, Ashok K.; Trikha, S. K.; Parmar, Virinder S.

doi:10.1006/bioo.1998.1127

Cited by 52 publications

(23 citation statements)

References 76 publications

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“…Indeed, chemical methods usually increase the difficulty of obtaining products with correct stereo-and regioselectivity, generating secondary products (13,34,56). As an alternative to the synthesis approach, enzymatic synthesis performed under very mild conditions in a stereoand regiospecific manner (42,55) is a valuable option. Nowadays, 2Ј-deoxyribosyltransferases are quite interesting since these enzymes can be used to carry out the synthesis of natural and nonnatural nucleosides with antiviral and antitumor activity in a one-pot reaction.…”

Section: Discussionmentioning

confidence: 99%

Lactobacillus reuteri 2′-Deoxyribosyltransferase, a Novel Biocatalyst for Tailoring of Nucleosides

Fernández‐Lucas

Acebal

Sinisterra³

et al. 2010

Appl Environ Microbiol

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A novel type II nucleoside 2-deoxyribosyltransferase from Lactobacillus reuteri (LrNDT) has been cloned and overexpressed in Escherichia coli. The recombinant LrNDT has been structural and functionally characterized. Sedimentation equilibrium analysis revealed a homohexameric molecule of 114 kDa. Circular dichroism studies have showed a secondary structure containing 55% ␣-helix, 10% ␤-strand, 16% ␤-sheet, and 19% random coil. LrNDT was thermostable with a melting temperature (T m ) of 64°C determined by fluorescence, circular dichroism, and differential scanning calorimetric studies. The enzyme showed high activity in a broad pH range (4.6 to 7.9) and was also very stable between pH 4 and 7.9. The optimal temperature for activity was 40°C. The recombinant LrNDT was able to synthesize natural and nonnatural nucleoside analogues, improving activities described in the literature, and remarkably, exhibited unexpected new arabinosyltransferase activity, which had not been described so far in this kind of enzyme. Furthermore, synthesis of new arabinonucleosides and 2-fluorodeoxyribonucleosides was carried out.Nucleoside 2Ј-deoxyribosyltransferases (NDTs) (EC 2.4.2.6) catalyze the exchange between the purine or pyrimidine base of 2Ј-deoxyribonucleosides and free pyrimidine or purine bases (10, 25). These enzymes are specific for 2Ј-deoxyribonucleosides, regioselective (N-1 glycosylation in pyrimidine and N-9 in purine), and stereoselective (␤-anomers are exclusively formed) (26) (Fig. 1).Deoxyribosyltransferases are classified into two classes depending on their substrate specificity: type I (NDT I), specific for purines (Pur 7 Pur), and type II (NDT II), which catalyzes the transfer between purines and/or pyrimidines (Pur 7 Pur, Pur 7 Pyr, Pyr 7 Pyr) (10, 25). These enzymes were initially described for lactobacilli (27,28), and they are involved in the nucleoside salvage pathway for DNA synthesis (23), although this remains unclear in Lactococcus lactis subsp. lactis (36). NDTs have been also found in some species of Streptococcus (11), in parasitic unicellular eukaryotic organisms such as Crithidia luciliae (49, 50), in Trypanosoma brucei (6), and in Borrelia burgdorferi (33). NDTs from Lactobacillus helveticus and Lactobacillus leichmannii have been well studied (2,25,26,28,29), and their kinetic mechanisms as well as their catalytic and substrate binding sites have been characterized. The transferase reaction proceeds via a ping-pong bi-bi mechanism by formation of a covalent deoxyribosyl enzyme intermediate (3,15,16). Likewise, a glutamyl residue (Glu98) has been proven essential for activity (40,41,46). Enzymatic natural and nonnatural nucleoside synthesis in a one-pot reaction by NDTs provides an interesting alternative to traditional multistep chemical methods (13,34). Indeed, chemical glycosylation includes several protection-deprotection steps and the use of chemical reagents and organic solvents that are expensive and environmentally harmful. Whereas previously described NDTs accept different nucleosides fr...

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Section: Discussionmentioning

confidence: 99%

Lactobacillus reuteri 2′-Deoxyribosyltransferase, a Novel Biocatalyst for Tailoring of Nucleosides

Fernández‐Lucas

Acebal

Sinisterra³

et al. 2010

Appl Environ Microbiol

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“…[3] Several procedures based on the "transglycosylation" reaction (Scheme 1) have been employed (using both isolated enzymes and whole cells) to prepare a wide variety of nucleosides. [4][5][6][7][8][9][10][11] Structural analysis revealed that there are two distinct families of NPs: NPI and NPII. Members of the NPI family show either a trimeric or a hexameric quaternary structure.…”

Section: Introductionmentioning

confidence: 99%

Production, Characterization and Synthetic Application of a Purine Nucleoside Phosphorylase from Aeromonas hydrophila

Ubiali

Serra

et al. 2012

Adv Synth Catal

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Purine nucleoside phosphorylase (PNP) from Aeromonas hydrophila encoded by the deoD gene has been over-expressed in Escherichia coli, purified, characterized about its substrate specificity and used for the preparative synthesis of some 6-substituted purine-9-ribosides. Substrate specificity towards natural nucleosides showed that this PNP catalyzes the phosphorolysis of both 6-oxo-and 6-aminopurine (deoxy)ribonucleosides. A library of nucleoside analogues was synthesized and then submitted to enzymatic phosphorolysis as well. This assay revealed that 1-, 2-, 6-and 7-modified nucleosides are accepted as substrates, whereas 8-substituted nucleosides are not. A few transglycosylation reactions were carried out using 7-methylguanosine iodide (4) as a d-ribose donor and 6-substituted purines as acceptor. In particular, following this approach, 2-amino-6-chloropurine-9-riboside (2c), 6-methoxypurine-9-riboside (2d) and 2-amino-6-(methylthio)purine-9-riboside (2g) were synthesized in very high yield and purity.

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“…Therefore, this enzyme provides an alternative to the synthesis of nucleosides using nucleoside phosphorylases. Nucleoside 2 0 -deoxyribosyl transferases are specific for 2 0 -deoxyribosyl nucleosides, regioselective (N-1 glycosylation in pyrimidine and N-9 in purine), as well as stereoselective (b-anomers are exclusively formed; Prasad et al, 1999). In nature, these enzymes have been described in various Lactobacilli (Cardinaud and Holguin, 1979;Holguin and Cardinaud, 1975;Huang et al, 1983;Kaminski, 2002) and in E. coli .…”

Section: Introductionmentioning

confidence: 99%

Low‐temperature synthesis of 2′‐deoxyadenosine using immobilized psychrotrophic microorganisms

Fernández‐Lucas

Condezo

Quezada

et al. 2007

Biotech & Bioengineering

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Selective biocatalyzed synthesis of 2'-deoxyadenosine from 2'-deoxypyrimidine nucleosides was carried out using free or immobilized whole cells. The reaction was performed at 57 degrees C without secondary reactions. Two psychrotrophic microorganisms, Bacillus psychrosaccharolyticus and Psychrobacter immobilis, are described for the first time as active and specific strains for the synthesis of 2'-deoxyadenosine. Adenosine deaminase activity was not detected. Whole cells were immobilized in different matrixes. Calcium alginate and calcium pectate gave the best biocatalysts. The synthesis of 2'-deoxyadenosine follows an apparent first order kinetic expression. External mass transfer control was negligible as deduced from k(s), N(A), and Omega values. Internal mass transfer was the rate controlling step according to eta(T) and phi values.

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Nucleoside Synthesis Mediated by Glycosyl Transferring Enzymes

Cited by 52 publications

References 76 publications

Lactobacillus reuteri 2′-Deoxyribosyltransferase, a Novel Biocatalyst for Tailoring of Nucleosides

Lactobacillus reuteri 2′-Deoxyribosyltransferase, a Novel Biocatalyst for Tailoring of Nucleosides

Production, Characterization and Synthetic Application of a Purine Nucleoside Phosphorylase from Aeromonas hydrophila

Low‐temperature synthesis of 2′‐deoxyadenosine using immobilized psychrotrophic microorganisms

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