Defining a process operating window for the synthesis of 5-methyluridine by transglycosylation of guanosine and thymine

Immobilization of thermostable nucleoside phosphorylases on MagReSyn® epoxide microspheres and their application for the synthesis of 2,6-dihalogenated purine nucleosides

“…The bench scale reaction productivities of 1.5 and 2.0 g L -1 h -1 are comparable to other reactions with the natural substrates in literature (e.g. 5-methyluridine [32][33][34], 2'-deoxyadenosine [35] and 2'-deoxyruridine [36]), which implies that further optimization of the reaction conditions may improve the productivity in line with larger scale production as illustrated by Gordon et al [32,37].…”

“…We found that PNP coupled with PyNP worked more efficiently than PNP alone with-D-Rib-1P and the purine base. Concerning the effect of the phosphate ion concentration on the biocatalytic reactions, an unexpected low phosphate concentration (1.4 mM instead of the typical 10-50 mM reported in the literature [30][31][32]) was found to be the optimum for the product formation in the one-pot reaction. It is obvious that the phosphate is a co-substrate consumed in the phosphorolysis of the pentofuranose donor and regenerating in the synthetic reaction.…”

Comparative Analysis of Enzymatic Transglycosylation Using E. coli Nucleoside Phosphorylases: A Synthetic Concept for the Preparation of Purine Modified 2′-Deoxyribonucleosides from Ribonucleosides

“…Therefore, we performed a comparative analysis of transglycosylation conditions for the synthesis of 2′-deoxyribonucleosides, involving E. coli PNP and TP ( Table 1 ). The choice of enzymes of bacterial origin as catalysts was due to their broad substrate specificity, optimal pH in neutral/slightly alkaline media and a sufficiently wide operating temperature range, which allows to perform transglycosylation reactions under mild conditions without noticeable nonspecific cleavage of N -glycosidic bond [ 18 , 19 , 20 , 21 , 22 , 23 , 24 ]. Then, we analyzed the transglycosylation conditions for enzymatic synthesis of 2′-deoxynucleosides using different sources of sugar moiety and varying nucleoside-donor:Base-acceptor:Phospate-anion (D:B:P i ) ratio ( Table 1 ).…”

Enzyme‐mediated synthesis of Molnupiravir: paving the way for the application of biocatalysis in pharmaceutical industry

“…To this effect, several synthetic strategies have been previously described, such as the irreversible phoshorolysis of 7-methylguanosine iodide to 7-methylguanine [23] or the use of guanosine as nucleoside donor at neutral pH values. [24] In both cases, due to the nature of released nucleobase (7-methylguanine can not be recognized by NPs; guanine precipitates as a result of low water solubility), the equilibrium is shifted away from the reverse reaction. Additionally, both nucleobases are poorly water-soluble and easily separate the reaction, remaining 5 as single species in solution.…”

“…However, this undesired constraint can be used to favor the transglycosylation reaction in the desired way. To this effect, several synthetic strategies have been previously described, such as the irreversible phoshorolysis of 7‐methylguanosine iodide to 7‐methylguanine [23] or the use of guanosine as nucleoside donor at neutral pH values [24] . In both cases, due to the nature of released nucleobase (7‐methylguanine can not be recognized by NPs; guanine precipitates as a result of low water solubility), the equilibrium is shifted away from the reverse reaction.…”