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Enzymatic Synthesis of Therapeutic Nucleosides using a Highly Versatile Purine Nucleoside 2’‐DeoxyribosylTransferase from Trypanosoma brucei
Author(s) -
Pérez Elena,
SánchezMurcia Pedro A.,
Jordaan Justin,
Blanco María Dolores,
Mancheño José Miguel,
Gago Federico,
FernándezLucas Jesús
Publication year - 2018
Publication title -
chemcatchem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.497
H-Index - 106
eISSN - 1867-3899
pISSN - 1867-3880
DOI - 10.1002/cctc.201800775
Subject(s) - chemistry , trypanosoma brucei , purine nucleoside phosphorylase , nucleoside analogue , nucleoside , cladribine , combinatorial chemistry , biocatalysis , purine , deoxyadenosine , enzyme , stereochemistry , organic chemistry , biochemistry , ionic liquid , catalysis , biology , genetics , gene
The use of enzymes for the synthesis of nucleoside analogues offers several advantages over multistep chemical methods, including chemo‐, regio‐ and stereoselectivity as well as milder reaction conditions. Herein, the production, characterization and utilization of a purine nucleoside 2’‐deoxyribosyltransferase (PDT) from Trypanosoma brucei are reported. Tb PDT is a dimer which displays not only excellent activity and stability over a broad range of temperatures (50–70 °C), pH (4–7) and ionic strength (0–500 mM NaCl) but also an unusual high stability under alkaline conditions (pH 8–10). Tb PDT is shown to be proficient in the biosynthesis of numerous therapeutic nucleosides, including didanosine, vidarabine, cladribine, fludarabine and nelarabine. The structure‐guided replacement of Val11 with either Ala or Ser resulted in variants with 2.8‐fold greater activity. Tb PDT was also covalently immobilized on glutaraldehyde‐activated magnetic microspheres. M Tb PDT3 was selected as the best derivative (4200 IU/g, activity recovery of 22 %), and could be easily recaptured and recycled for >25 reactions with negligible loss of activity. Finally, M Tb PDT3 was successfully employed in the expedient synthesis of several nucleoside analogues. Taken together, our results support the notion that Tb PDT has good potential as an industrial biocatalyst for the synthesis of a wide range of therapeutic nucleosides through an efficient and environmentally friendly methodology.