Efficient Resolution of cis-(±)-Dimethyl 1-Acetylpiperidine-2,3-dicarboxylate by Covalently Immobilized Mutant Candida antarctica Lipase B in Batch and Semicontinuous Modes

Abstract

Candida antarctica lipase B (CALB) is a robust biocatalyst for production of chemicals. In this study, an engineered lipase B (CALB-I189K) was covalently immobilized onto a glutaraldehyde activated amino resin (ESR-3) and used for the preparation of chiral moxifloxacin precursor though enzymatic resolution of cis-(±)-dimethyl 1-acetylpiperidine-2,3-dicarboxylate (cis-(±)-1). The immobilization conditions were optimized, and the immobilized CALB-I189K exhibited improved thermal stability, with a half-life of 247.5 h at 30 °C. It enables the resolution of 100 g L–1 cis-(±)-1 for 50 cycles in a stirred tank reactor, with an average productivity of 49.6 g L–1 h–1. The resolution reaction was also performed in a recirculating packed bed reactor (RPBR) in a semicontinuous mode, and the average productivity reached 59.4 g L–1 h–1 under the optimal operating condition. Moreover, the RPBR bioconversion system has high operational stability and conserved over 85.7% of the initial conversion capacity after 50 cycles...