Enzymatic production of ribavirin from pyrimidine nucleosides by Enterobacter aerogenes AJ 11125.

Abstract

Microorganisms that produce ribavirin(l-^-D-ribofuranosyl-l ,2,4-triazole-3-carboxamide; virazole®) directly from pyrimidine nucleosides and TCA(l,2,4-triazole-3-carboxamide) were screened from our stock cultures. Of the 400 strains tested, 16 were isolated as ribavirin-producers from uridine or cytidine. In particular, Enterobacter aerogenes AJ 1 1 125, Bacillus brevis AJ 1282 and Sarcina lutea AJ 1212 were found to possess potent activities of ribavirin production from them. In the presence of intact cells of Enterobacter aerogenes AJ 1 1 125, which was selected as the best strain, 1 10.2mMand 67.6mMribavirin were produced from uridine and cytidine, respectively, on 96hr reaction at 60°C. In addition, this strain could also produce ribavirin from guanosine, but could not produce it from orotidine, which is also a pyrimidine nucleoside.

Characterization of a recombinant cold-adapted purine nucleoside phosphorylase and its application in ribavirin bioconversion

“…Like other mesophilic enzymes, the optimum temperature of these PNPs is almost above 65°C (Shirae et al 1988;Shirae and Yokozeki 1991). Many articles reported that cold-adapted enzymes from psychrophilic bacteria have higher catalytic activity and efficiency than their mesophilic counterparts at lower temperature (Lorentzen et al 2006;Li et al 2008;Zhang et al 2007).…”

“…It has broad spectrum and potent antiviral activity against DNA and RNA viruses. Currently, ribavirin can be produced by three methods: chemical synthesis, fermentation synthesis and enzymatic synthesis (Shirae et al 1988). However, chemical synthesis usually takes long time, requires harsh conditions and brings many by-products during the coupling reaction in industrial production.…”

“…This enzyme has been used to produce ribavirin, doxifluridine, vidarabine and other nucleoside medications. Many microorganisms with PNP catalytic activity can be employed in ribavirin enzymatic synthesis (Shirae et al 1988;Shirae and Yokozeki 1991). Although enzymatic synthesis has been applied in industrial production of ribavirin, it still has some problems.…”

Bioproduction of ribavirin by green microbial biotransformation

“…Ribavirin biotransformation was carried out using 1 × 10 9 colony-forming units (CFU), in 1 mL of reaction media containing equimolar concentrations (2.5 mM) of uridine (Urd) and TCA in sodium phosphate buffer (30 mM, pH 7). Reactions were performed during 16 h at 60 • C and 200 rpm as previously reported [13].…”

“…It is well known that there are some biocatalysts able to biosynthesize several nucleoside analogues [13,16]. Thus, 63 strains corresponding to 20 microbial genera were tested for ribavirin biotransformation (Scheme 1).…”