Abstract: γ-aminobutyric acid（GABA）, a crucial inhibitory neurotransmitter in the central nervous system, plays an important role in alleviating insomnia, depression, and nervous disorders. To increase the content of GABA content in steamed bread and optimize their flavor and texture, a strain of Levilactobacillus sp. LB-2（with high GABA production ability）, Saccharomyces cerevisiae Y5, and Acetobacter sp. A3 was selected to make steamed bread by single strain and mixed-strain fermentation. The effects of mixed-strain fermentation on the edible qualities of the steamed bread, such as specific volume, color, and textural characteristics, were investigated. The results showed that the highest GABA content in the steamed bread fermented by the mixed fermentation of Saccharomyces cerevisiae Y5, Levilactobacillus sp. LB-2 and Acetobacter sp. A3 was 6.16 mg/g. Compared with the single-yeast fermentation（3.82 mg/g）, the GABA content in steamed bread increased by approximately 61.3%. However, adding corn flour and using the scalded-dough process were not conducive to increasing the GABA content in steamed bread. When the cell ratio of Saccharomyces cerevisiae Y5, Levilactobacillus sp. LB-2 and Acetobacter sp. A3 was 1∶ 5∶ 1 in the dough starter, the specific volume, height-to-diameter ratio, and L^* value of the steamed bread were significantly higher than the rest of the treatment groups（P＜0.05）, while the hardness was significantly lower（P＜0.05）. Moreover, the hardness of the steamed bread was the lowest after the scalded-dough process, indicating that the scalded-dough process contributes to the improvement of the quality of the steamed bread. This study provides a foundation for the development of GABA-rich steamed bread or other kinds of food products made by fermented dough.