Production of γ-aminobutyric acid in Escherichia coli by engineering MSG pathway

The compound γ-aminobutyric acid (GABA) has many important physiological functions. The effect of glutamate decarboxylases and the glutamate/GABA antiporter on GABA production was investigated in Escherichia coli. Three genes, gadA, gadB, and gadC were cloned and ligated alone or in combination into the plasmid pET32a. The constructed plasmids were transformed into Escherichia coli BL21(DE3). Three strains, E. coli BL21(DE3)/pET32a-gadA, E. coli BL21(DE3)/pET32a-gadAB and E. coli BL21(DE3)/pET32a-gadABC were selected and identified. The respective titers of GABA from the three strains grown in shake flasks were 1.25, 2.31, and 3.98 g/L. The optimal titer of the substrate and the optimal pH for GABA production were 40 g/L and 4.2, respectively. The highest titer of GABA was 23.6 g/L at 36 h in batch fermentation and was 31.3 g/L at 57 h in fed-batch fermentation. This study lays a foundation for the development and use of GABA.