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Adding 0.15% nicotinic acid to medium significantly increased the γ-aminobutyric acid（GABA）production of Lactobacillus brevis. In order to clarify the role of nicotinic acid，L. brevis was inoculated in the medium with（experimental group）and without nicotinic acid（control group）for fermentation. The differential metabolites were studied by liquid chromatography-mass spectrometry（LC-MS）， and the integrity of cell membrane was analyzed using propidium iodide（PI）single staining method. The results showed that 33 metabolites were significantly different between the experimental group and the control group. The unsaturated fatty acids and some amino acids including glutamate，proline，aspartate，serine，histidine，and ornithine，significantly increased in the experimental group，whereas nicotinic acid nucleotide，2-phosphoglycerate，adenosine monophosphate and tyramine significantly decreased. PI single staining experiments demonstrated that the cells with red fluorescence in the experimental group were significantly more than those in the control group. Therefore，the addition of nicotinic acid has a broad impact on the physiological metabolism of L. brevis，and its promoting effect on GABA biosynthesis may be related to increasing the permeability of the cell membrane of L. brevis，resulting in easier substrates entry into the cells and easier products discharge from the cells. [ABSTRACT FROM AUTHOR]