The strains with strong ability to produce γ-aminobutyric acid (GABA) were preliminarily screened by thin-layer chromatography and identified by 16S rDNA. The high-yield GABA strains were further screened by amino acid automatic analyzer and inoculated into rice bran for fermentation experiments. The results showed that stale rice bran was conducive to the isolation of high yield GABA strains. The 16S rDNA sequences of strains with higher yields had the highest similarity to Enterococcus faecium, Enterococcus raffinosus, Escherichia coli, Lactobacillus brevis and Shigella sonnei. GABA content in rice bran which was just fermented by a single species did not increase obviously, however, it was 184.6 mg/100 g (dry mass) when rice bran was simultaneously fermented by Bacillus lincheniformis DY and lactic acid bacteria L-44A. Meanwhile, GABA content could reach 245.8 mg/100 g (dry mass) when rice bran was treated by protease then fermented by lactic acid bacteria L-44A.