Fig. 2: Development of the integrated chemo- and biocatalytic synthesis of amides.
From: Merging enzymes with chemocatalysis for amide bond synthesis
A Optimisation of the integrated reaction. Reaction conditions: 1. Nitrile (1) (50 mM), Equi_NHase (see table) in 0.1 M KPi buffer (pH = 7.8)/10% v/v iPrOH (2 mL) at RT, 24 h; 2. CuBr2 (10 mol%), L1 (20 mol%), d-glu (20 mol%), 3 (150 mM) at 50 °C, 24 h under N2 atmosphere (headspace purge). [a] Conversion determined in triplicates by HPLC/UV using benzophenone as external standard. [b] Yield of isolated product after column chromatography. [c] E. coli (Equi_NHase) whole cells from ca. 20 mL cell culture (OD600 = ~0.5) were used. B Activity screening of E. coli (NHase) whole cells. Relative conversion was determined in triplicates by HPLC/UV analysis (n = 3). Blue bars represent mean values and error bars represent ± SEM of benzamide (2) conversion. n.d. not detected; w/o without, OD600 Optical density measured at 600 nm.
