Methods to optimize the production of gamma-aminobutyric acid(GABA) by Lactobacillus brevis CGMCC 1306 were investigated. Results indicated that cell growth was maximal at pH 5.0, while pH 4.5 was preferable to GABA formation. The optimal temperature for cell growth(35 °C) was lower than that for GABA formation(40 °C). In a two-stage pH and temperature control fermentation, cultures were maintained at pH 5.0 and 35 °C for 32 h, then adjusted to pH 4.5 and 40 °C, GABA production increased remarkably and reached 474.79 mmol·L 1at 72 h, while it was 398.63 mmol·L 1with one stage pH and temperature control process, in which cultivation conditions were constantly controlled at pH 5.0 and 35 °C. In order to avoid the inhibition of cell growth at higher L-monosodium glutamate(L-MSG) concentrations, the two-stage control fermentation with substrate feeding strategy was applied to GABA production, with 106.87 mmol(20 g) L-MSG supplemented into the shaking-flask at 32h and 56 h post-inoculation separately. The GABA concentration reached 526.33 mmol·L 1at 72 h with the fermentation volume increased by 38%. These results will provide primary data to realize large-scale production of GABA by L. brevis CGMCC 1306.