Improved synthesis of 2′-deoxyadenosine and 5-methyluridine by Escherichia coli using an auto-induction system

Abstract

Nucleoside analogues are used widely for the treatment of viral diseases and cancer, however the preparation of some important intermediates of these nucleoside analogues, including 2'-deoxyadenosine (dAR) and 5-methyluridine (5-MU), remains inconvenient. To optimize the synthesis of dAR and 5-MU, recombinant strains and auto-induction medium were employed in this study. E. coli BL21(DE3) strains overexpressing purine nucleoside phosphorylase (PNP), uridine phosphorylase (UP) and thymidine phosphorylase (TP) were constructed and cultured in auto-induction ZYM-Fe-5052 medium for 8 h. The cultures of these strains were then used directly to synthesize dAR and 5-MU. Under optimized conditions, 30 mM adenine was converted to 29 mM dAR in 1 h, and 32 mM 5-MU was obtained from 60 mM thymine, using 6% (v/v) cell solutions as biocatalysts. These results indicate that our convenient and efficient method is ideal for the preparation of dAR and 5-MU, and has potential for the preparation of other nucleoside analogue intermediates.

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].…”

Deoxyadenosine family: improved synthesis, DNA damage and repair, analogs as drugs

“…Hence, the development of convenient and environmentally friendly methods for 2 ′ -deoxyadenosine synthesis is important. Xiong et al (1) described the improved synthesis of 2 ′ -deoxyadenosine and 5-methyluridine by cloning three different nucleoside phosphorylases -purine nucleoside phosphorylase, uridine phosphorylase, and thymidine phosphorylase -into a recombinant E. coli strain, one at a time, using whole cells, thus avoiding isolation and purification of those enzymes. Instead of the more expensive IPTG method of induction, they have employed the autoinduction ZYM Fe 5052 medium successfully.…”

“…Xiong et al (1) have cloned three different nucleoside phosphorylases into a recombinant Escherichia coli strain and used thymidine and adenine as the more inexpensive starting materials to make 2 ′ -deoxyadenosine without using isopropyl β -d -thiogalactoside (IPTG). Horinouchi et al (2) used glucose, acetaldehyde, and nucleobase to produce the four deoxynucleosides using the coexpression of three enzymes in one E. coli strain.…”