Development of an immobilized biocatalyst based on Bacillus psychrosaccharolyticus NDT for the preparative synthesis of trifluridine and decytabine

• The optimization of BpNDT immobilization is here reported.
• Crosslinking of enzyme adsorbed to functionalized supports increased its stability.
• Addition of 20% glycerol to the immobilization medium improved final activity.
• Selected BpNDT biocatalyst was stable in presence of 20% DMF or high pH.
• Immobilized BpNDT was used to synthesize decytabine and trifluridine.

The immobilization of Bacillus psychrosaccharolyticus nucleoside 2′-deoxyribosyltransferase was deeply investigated and finally optimized. The best immobilization procedure resulted to be ionic adsorption on PEI 600 Da agarose followed by crosslinking with 70% oxidized dextran (20 kDa). The percentage of recovered activity was further improved (from 21% to 33%) by the addition of 20% glycerol to the immobilization mixture. The resulting biocatalyst showed a stability profile similar to that of the soluble enzyme and it was used for the preparative synthesis of trifluridine and decytabine obtaining conversions ranging from 50% to 76%.

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