Abstract
(R)-Mandelic acid (R-MA) is a key precursor for the synthesis of semi-synthetic penicillin, cephalosporin, anti-obesity drugs, antitumor agents, and chiral resolving agents for the resolution of racemic alcohols and amines. In this study, an enzymatic method for the large-scale production of R-MA by a stereospecific nitrilase in an aqueous system was developed. The nitrilase activity ofthe Escherichia coli BL21(DE3)/pET-Nit whole cells reached 138.6U/g in a 20,000-L fermentor. Using recombinant E. coli cells as catalyst, 500mM R,S-mandelonitrile (R,S-MN) was resolved into 426mM (64.85g/L) R-MA within 8h, and the enantiomeric excess (ee) value of R-MA reached 99%. During the purification process, pure R-MA with a recovery rate of 78.8% was obtained after concentration and crystallization. This study paved the foundation for the upscale production of R-MA using E. coli whole cells as biocatalyst.
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