Flow bioprocessing of citrus glycosides for high-value aglycone preparation

An efficient one-pot, 2-step flow bioprocess for the hydrolysis of hesperidin (HES) and rutin (RT), citrus rutinosyl flavonoids, has been developed for the obtainment of the corresponding aglycones. A commercially available α-rhamnosidase (RN) and an extremophilic home-made β-glycosidase (HOR) have been co-immobilized on glyoxyl-agarose beads to prepare a high-performing multi-active biocatalyst (imm-RN–HOR). After the optimization of the reaction conditions in batch mode, a “flow switch” was applied, increasing the productivity (>99% m.c., 5 min), cost-efficiency and sustainability of the overall process. Due to the recovery and reuse of all the materials involved in the flow biotransformation, this strategy is effectively a zero-waste process.