Biotransformation of halogenated nucleosides by immobilized Lactobacillus animalis 2′-N-deoxyribosyltransferase

•This work describes a fast, simple and economical immobilized biocatalyst.•Lactobacillus animalis NDT was successfully stabilized in DEAE-Sepharose.•The developed immobilized biocatalyst was able to sinthetize floxuridine.•Halogenated nucleoside analogues of clinical interest were also obtained.

An immobilized biocatalyst with 2′-N-deoxyribosyltransferase (NDT) activity, Lactobacillus animalis NDT (LaNDT), was developed from cell free extracts. LaNDT was purified, characterized and then immobilized by ionic interaction. Different process parameters were optimized, resulting in an active derivative (2.6 U/g) able to obtain 1.75 mg/g of 5-fluorouracil-2′-deoxyriboside, an antimetabolite known as floxuridine, used in gastrointestinal cancer treatment. Furthermore, immobilized LaNDT was satisfactorily used to obtain at short reaction times other halogenated pyrimidine and purine 2′-deoxynucleosides such as 6-chloropurine-2′-deoxyriboside (4.9 U/g), 6-bromopurine-2′-deoxyriboside (4.3 U/g), 6-chloro-2-fluoropurine-2′-deoxyriboside (5.4 U/g), 5-bromo-2′-deoxyuridine (2.8 U/g) and 5-chloro-2′-deoxyuridine (1.8 U/g) compounds of pharmaceutical interest in antiviral or antitumor treatments. Besides, increasing the biocatalyst amount 8 times per volume unit allowed obtaining a 5-fold improvement in floxuridine biotransformation. The developed biocatalyst proved to be effective for the biosynthesis of a wide spectrum of nucleoside analogues by employing an economical, simple and environmentally friendly methodology.

Graphical abstractAn immobilized biocatalyst with 2′-N-deoxyribosyltransferase activity (NDT) was developed from cell free extracts, resulting in a derivative with an activity of 2.6 U/g for the biosynthesis of the 5-fluorouracil-2′-deoxyriboside, an antimetabolite known as floxuridine, used in gastrointestinal cancer treatment. Furthermore, immobilized NDT was satisfactorily used to obtain other halogenated pyrimidine and purine 2′-deoxynucleosides of pharmaceutical interest as antiviral or antitumor compounds at short reaction times. Besides, increasing the biocatalyst amount 8 times per volume unit allowed obtaining a 5-fold improvement in floxuridine biosynthesis. The developed biocatalyst proved to be effective for the biosynthesis of a wide spectrum of halogenated nucleosides by employing an economical, simple and environmentally friendly methodology.Figure optionsDownload full-size imageDownload as PowerPoint slide