One-Pot Multi-Enzymatic Production of Purine Derivatives with Application in Pharmaceutical and Food Industry

Abstract

Abstract:Biocatalysis reproduce nature's synthetic strategies in order to synthesize different organic compounds. Natural metabolic pathways usually involve complex networks to support cellular growth and survival. In this regard, multi-enzymatic systems are valuable tools for the production of a wide variety of organic compounds. Methods: The production of different purine nucleosides and nucleoside-5 -monophosphates has been performed for first time, catalyzed by the sequential action of 2 -deoxyribosyltransferase from Lactobacillus delbrueckii (LdNDT) and hypoxanthine-guanine-xanthine phosphoribosyltransferase from Thermus themophilus HB8 (TtHGXPRT). Results: The biochemical characterization of LdNDT reveals that the enzyme is active and stable in a broad range of pH, temperature, and ionic strength. Substrate specificity studies showed a high promiscuity in the recognition of purine analogues. Finally, the enzymatic production of different purine derivatives was performed to evaluate the efficiency of multi-enzymatic system LdNDT/TtHGXPRT. Conclusions: The production of different therapeutic purine nucleosides was efficiently catalyzed by LdNDT/TtHGXPRT. In addition, the resulting by-products were converted to IMP and GMP. Taking all of these features, this bioprocess entails an efficient, sustainable, and economical alternative to chemical synthetic methods.

Enzymatic Synthesis of Therapeutic Nucleosides using a Highly Versatile Purine Nucleoside 2’‐DeoxyribosylTransferase from Trypanosoma brucei

“…Because type II NDTs offer several advantages over PDTs, such as higher activity values under the same experimental conditions or the possibility of using a broad range of substrates, type II NDTs usually represent the best option for the enzymatic synthesis of nucleoside analogues. Nevertheless, the specific activities of Tb PDT are higher than those reported for other PDTs from Lactobacillus and Borrelia species under similar enzymatic assay conditions (Table ) and comparable to those previously described for several type II NDTs from Lactobacillus reuteri and Lactobacillus delbrueckii . Tb PDT can also use as substrates the manifold of purine derivatives and nucleoside analogues displayed in Tables 2 and 3, and it is to be preferred, in principle, over Lm PDT because it can transglycosylate on a larger number of purine analogues when dIno is used as the donor nucleoside (data not shown).…”

Synthesis of Ribavirin, Tecadenoson, and Cladribine by Enzymatic Transglycosylation

“…All the HPLC peaks were assigned on the basis of the pure reference compounds, either purchased or synthesized (see Materials and Methods). As recalled in the Introduction, the synthesis of nucleoside-based APIs by an enzymatic transglycosylation reaction has been investigated by many authors (for a comprehensive review, see References [3,51]; for a recent example see Reference [52]). However, the enzymatic synthesis of Tecadenoson has been reported herein for the first time (see Supplementary Materials, Figure S7).…”

Biocatalysis and Biotransformations

“…In more detail, Fresco-Taboada and coworkers investigated the synthesis of modified nucleosides by using novel 2 -deoxyribosyltransferases [13]. Acosta and coworkers reported the synthesis of purine derivatives by the sequential action of 2 -deoxyribosyltransferases and hypoxanthine-guanine-xanthine phosphoribosyltransferases [14]. The utilization of renewable free fatty acids as a source of α,ω-diols (C 8 -C 16 ) by the action of CYP153A monooxygenase, carboxylic acid reductase, and aldehyde reductases was demonstrated by Ahsan and coworkers [15].…”