Fig. 3: Chemo-enzymatic cascade reactions in batch and flow systems.

a Cascade reaction for the synthesis of chiral O-acylated cyanohydrin by cascading Ti(Salen) and CALB. b Kinetic profiles for different batch cascade reactions. System A: using Ti(Salen) and CALB as catalysts; System B: Ti/SCs and CALB/SCs; System C: co-localization of Ti/SCs and CALB/SCs within a single microreactor (IL-in-oil emulsion). c Benzaldehyde conversion and e.e. of O-acylated cyanohydrin with time in the flow system with an IL-in-oil microreactor. d Catalysis efficiency in the batch and flow reactions. e Specific activity of CALB in different systems. For batch reactions, CE and specific activity were calculated according to the conversion within the first 10 h; for the flow reactions, CE was calculated after the conversion leveled off. f Calculated benzaldehyde concentration in IL as a function of the partition coefficient in the batch and flow systems. Batch reaction conditions: System A: 2.1 mL [BMIM]PF₆, 3 mL PEG-300, 0.9 mL PBS (pH 7.4, 0.05 M), 0.08 g Ti(Salen), 0.03 g 4-dimethylaminopyridine (DMAP, as co-catalyst)⁴³, 0.9 mL CALB (8.0 mg mL⁻¹ of protein), 3 mL n-octane containing benzaldehyde (0.05 M) and acetyl cyanide (0.2 M), 25 °C, 900 rpm; System B: using Ti/SCs and CALB/SCs;System C: Ti/SCs and CALB/SCs co-localized within a single microreactor. Flow reaction conditions: the system consists of 2.1 mL [BMIM]PF₆, 3 mL PEG-300, 0.9 mL PBS (pH 7.4, 0.05 M), 0.2 g Ti/SCs, 0.1 g CALB/SCs, 3 mL n-octane, 0.18 g emulsifier. Benzaldehyde (0.05 M) and acetyl cyanide (0. 2 M) in n-octane as mobile phase, 25 °C, flow rate = 1 mL h⁻¹. All the reactions were repeated twice to obtain an average.