The probiotic fermentation of the bioactive substance gamma-aminobutyric acid (GABA) is an attractive research topic. There is still room for further improvement in reported GABA fermentation methods based on a single substrate ( l -glutamic acid or l -monosodium glutamate). Here, we devised a pH auto-buffering strategy to facilitate the fermentation of GABA by Levilactobacillus brevis CD0817. This strategy features a mixture of neutral monosodium l -glutamate plus acidic l -glutamic acid as the substrate. This mixture provides a mild initial pH; moreover, the newly dissolved l -glutamic acid automatically offsets the pH increase caused by substrate decarboxylation, maintaining the acidity essential for GABA fermentation. In this study, a flask trial was first performed to optimize the GABA fermentation parameters of Levilactobacillus brevis CD0817. The optimized parameters were further validated in a 10 L fermenter. The flask trial results revealed that the appropriate fermentation medium was composed of powdery l -glutamic acid (750 g/L), monosodium l -glutamate (34 g/L [0.2 mol/L]), glucose (5 g/L), yeast extract (35 g/L), MnSO 4 ·H 2 O (50 mg/L [0.3 mmol/L]), and Tween 80 (1.0 g/L). The appropriate fermentation temperature was 30 °C. The fermenter trial results revealed that GABA was slowly synthesized from 0–4 h, rapidly synthesized until 32 h, and finally reached 353.1 ± 8.3 g/L at 48 h, with the pH increasing from the initial value of 4.56 to the ultimate value of 6.10. The proposed pH auto-buffering strategy may be popular for other GABA fermentations.
Haixing Li,Tianyi Sun,Mengya Jia,Lingqin Wang,Cheng Wei,Jinfeng Pei,Zhiyu Lin,Shuixing Wang
There is a recent trend of using lactic acid bacteria for the production of gamma-aminobutyric acid (GABA). This study described a method that combines fermentation and self-buffered whole-cell catalysis for the efficient production of GABA usingLevilactobacillus brevisCD0817. Upon the completion of GABA fermentation, cells were recovered to conduct whole-cell catalysis by which the substrate L-glutamic acid was catalytically decarboxylated to GABA. L-glutamic acid itself maintained the acidity essential for decarboxylation. To maximize the whole-cell catalysis ability, the effects of the cell culture method, catalysis temperature, catalysis time, cell concentration, and L-glutamic acid dosage were investigated. The results illustrate that the cells that were cultivated for 16 h in a fermentation medium supplemented with 20.0 g/L of glucose were the most suitable for the whole-cell catalytic production of GABA. At 16 h, the fermentative GABA content reached 204.2 g/L. Under optimized whole-cell catalytic conditions (temperature 45.0 °C, time 12.0 h, wet cells 25.0 g/L, and L-glutamic acid 120.0 g/L), 85.1 g/L of GABA was obtained, with 3.7 ± 0.9 g/L of substrate residue. GABA was recovered from the system by sequentially performing rotary vacuum evaporation, precipitation with ethanol, filtration with filter paper, and drying. The purity of the GABA product reached 97.1%, with a recovery rate of 87.0%. These data suggest that the proposed method has potential applications in the production of GABA.Keywords:gamma-aminobutyric acid;Levilactobacillus brevisCD0817;fermentation;whole-cell catalysis
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