Batch and Flow Nitroaldol Synthesis Catalysed by Granulicella tundricola Hydroxynitrile Lyase Immobilised on Celite R‐633

Granulicella tundricola hydroxynitrile lyase (GtHNL) catalyses the synthesis of chiral (R)‐ cyanohydrins and (R)‐β‐nitro alcohols. The triple variant GtHNL‐A40H/V42T/Q110H (GtHNL‐3V) was immobilised on Celite R‐633 and used in monophasic MTBE saturated with 100 mM KPi buffer pH 7 for the synthesis of (R)‐2‐nitro‐1‐phenylethanol (NPE) in batch and continuous flow systems. Nitromethane was used as a nucleophile. A total of 82% of (R)‐NPE and excellent enantioselectivity (>99%) were achieved in the batch system after 24 hours of reaction time. GtHNL‐3V on Celite R‐ 633 was successfully recycled five times. During more recycling steps a significant decrease in yield was observed while the enantioselectivity remained excellent over eight cycles. The use of a flow system enabled the continuous synthesis of (R)‐NPE. A total of 15% formation of (R)‐NPE was reached using a flow rate of 0.1 mL min⁻¹; unfortunately, the enzyme was not stable, and the yield decreased to 4% after 4 hours on stream. A similar yield was observed during 15 hours at a rate of 0.01 mL min⁻¹. Surprisingly the use of a continuous flow system did not facilitate the process intensification. In fact, the batch system displayed a space‐time‐yield (STY/mgenzyme) of 0.10 g L⁻¹ h⁻¹ mgenzyme⁻¹ whereas the flow system displayed 0.02 and 0.003 g L⁻¹ h⁻¹ mgenzyme⁻¹ at 0.1 and 0.01 mL min⁻¹, respectively. In general, the addition of 1 M nitromethane potentially changed the polarity of the reaction mixture affecting the stability of Celite‐GtHNL‐3V. The nature of the batch system maintained the reaction conditions better than the flow system. The higher yield and productivity observed for the batch system show that it is a superior system for the synthesis of (R)‐NPE compared with the flow approach.