Regioselective acylation of nucleosides and their analogs catalyzed by Pseudomonas cepacia lipase: enzyme substrate recognition

The substrate recognition of Pseudomonas cepacia lipase in the acylation of nucleosides was investigated by means of rational substrate engineering for the first time. P. cepacia lipase displayed excellent 3′-regioselectivities (96 to >99%) in the lauroylation of 2′-deoxynucleosides 1a-1e, while low to good 3′-regioselectivities (59-89%) in the lauroylation of ribonucleosides 1f-1j. It might be due to the unfavorable hydrogen bond interaction between 2′-hydroxyl group of 1f-1j and phenolic hydroxyl group of tyrosine residue present in the alternate hydrophobic pocket of the enzyme, which stabilizes the conformation of 5′-acylation transition state and thus increases the amount of the minor regioisomer. In addition, various ester derivatives of floxuridine were synthesized successfully by the lipase with high conversions (99%) and good to excellent 3′-regioselectivities under mild conditions. The recognition of various acyl donors by the enzyme was examined. The enzymatic recognition of acyl groups was rationalized in terms of the structure of the active site of the lipase, especially the size, shape, and physicochemical properties.

Graphical abstractThe substrate recognition of PSL-C in the acylation of nucleosides was revealed through rational substrate engineering for the first time. A deep understanding on the interactions between the lipase and the substrates on a molecular level was attempted. These findings would help us in controlling the regioselectivity of the synthetically useful enzyme via chemical modification and protein engineering.Download full-size image