Development of an oil-sealed anaerobic fermentation process for high production of γ-aminobutyric acid with Lactobacillus brevis isolated by directional colorimetric screening

γ-aminobutyric acid (GABA) is an important inhibitory neurotransmitter in the brain with many physiological functions. Therefore, this work establishes an efficient biosynthesis process to achieve the high production of food-grade GABA. Firstly, we adopted a directional colorimetric screening method and then obtained the strain Lactobacillus brevis ( Lb. brevis) CGMCC 24975 with 22.61 g/L GABA in shake flasks, much higher than Lb. brevis ATCC 367 (3.07 g/L) due to its more efficient and stable expression of its glutamate decarboxylase (GAD) system. After exploring the effects of optimal fermentation conditions and oxygen on GABA production, we found that anaerobic fermentation could increase GABA production by up-regulating gad operon expression and activating GADase. Finally, we developed an oil-sealed anaerobic fermentation process with 157.90 g/L GABA in a 5 L bioreactor, which was 6.98-fold of that in shake flasks. This work includes a complete process from screening GABA-producing strains to developing efficient fermentation processes, laying the foundation for producing food-safe GABA.