R-mandelic acid production with immobilized recombinant Escherichia coli cells in a recirculating packed bed reactor

Abstract

A recirculating packed bed reactor (RPBR) was used for efficient production of R-mandelic acid (R-MA) by kinetic resolution of racemic R,S-mandelonitrile (R,S-MN) using the recombinant E. coli cells crosslinked with diatomite (DA)/glutaraldehyde (GA)/polyethyleneimine (PEI). The performance and productivity of RPBR were evaluated by several parameters, including cell load, substrate feeding rate, height diameter (H/D) ratio, reactor structures, and operation stability. The kinetic resolution process showed higher initial reaction rate (1.52 mM/min) and yield (100%) by recycling 100 mL of substrate solution (70 mM) through RPBR packed with 6.0 g immobilized cells at a substrate-feeding rate of 19 mL/min while the H/D ratio was 2.8. The immobilized cells were successfully applied into kinetic resolution of R,S-MN in the RPBR for 50 batches with an average productivity of 4.12 g/L/h for R-MA with >99% of enantiomeric excess.