Optimization of the medium composition and product extraction for R-mandelic acid using recombinant Escherichia coli expressing Alcaligenes sp. nitrilase

Abstract

The aim of this study is to investigate the effects of the components of the medium on nitrilase expression in recombinant Escherichia coli (E. coli), which is applied in the hydrolysis of racemic mandelonitrile (MN) to R-mandelic acid (MA). In addition, the separation of R-MA from the reaction mixture is studied. The glycerol medium is screened, and compositions are optimized in single-factor experiments. The nitrilase activity is 1.96-fold higher than before using the optimal medium containing peptone 15 g/L, yeast extract 12 g/L, NaCl 10 g/L, glycerol 15 g/L, (NH4)2SO4 5 g/L, KH2PO4 10 mM, K2HPO4·3H2O 10 mM, and MgSO4·7H2O 15 mM. During the acid-base extraction of R-MA, the optimal alkaline pH, the alkaline extractant ratio, the optimal acidic pH, the acidic extractant ratio and the amount of activated carbon are 10.0, 1:0.5 (once), 2.50, 1:2 (twice) and 1%, respectively. This study provides a basis for the industrial production of R-MA.