Enzymatic synthesis of nucleoside 5′‐mono and ‐triphosphates

Abstract

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.

A comparison of enzymatic phosphorylation and phosphatidylation of β-l- and β-d-nucleosides

“…Nucleoside phosphotransferase (NPT) was isolated from Erwinia herbicola 47/3 obtained as reported earlier (Zinchenko et al 1990). For NPT purification (all operations were carried out at 4°C), bacterial cells (18 g wet wt; 3.6 g calculated as dry wt) were suspended in 100 ml buffer A (0.15% NaCl, 50 mM Tris/HCl, pH 7.0) and disrupted by sonication.…”

“…The mixture was incubated at 37°C. Aliquots (20 ll) of reaction medium were analyzed by TLC as described (Zinchenko et al 1990). NPT activity was measured till the product yield not exceeded 15-20% of maximal nucleoside conversion.…”

“…Earlier, we have reported on the efficient enzymatic transformation of natural as well as sugar and base modified b-D-nucleosides into their respective 5¢-monophosphates by using the whole cells of Erwinia herbicola 47/3 (Zinchenko et al 1990;Zaitseva et al 1994;Barai et al 2004). In the present study, we have used partially purified NPT from the Erwinia herbicola 47/3 cells that were cultivated as described (Zinchenko et al 1990). …”

An improved method for the enzymatic transformation of nucleosides into 5?-monophosphates

“…NMP are obtained by microbial synthesis or by isolation from hydrolysates of nucleic acids, which are the main sources of these compounds (Kuninaka & Shoyu 1976). Methods for the production of modified NMP are based either on chemical (Scheit 1980, Burgess & Cook 2000 or on an enzymatic phosphorylation of corresponding nucleosides (Humble et al 1984, Zinchenko et al 1990, Rutkowski & Draminski 1991, Zaitseva et al 1994. The yields of NMPs by chemical phosphorylation of modified nucleosides vary widely depending on the method employed, the structure of initial compounds, and from assay to assay.…”

Enzymatic synthesis of ATP from RNA and adenine

“…Moreover, it is well known that enzymes of alcohol fermentation and the kinases extracted from permeabilized yeast cells catalyze ATP regeneration from adenosine (Ado), using glucose as an energy source (Asada et al. 1978;Zinchenko et al, 1990c). With this background, in the present work we have studied an analogous approach for the synthesis of Ado from enzymatic RNA hydrolysate and Ade in order to convert it then to ATP.…”