The cultural conditions of Bullera singularis were optimized for the efficient production of galactooligosaccharide (GOS), Optimum temperature was 25$\circ$C, pH was 6.0, inoculum size was over 5% (v/v), initial lactose concentration was over 5% (w/v). The GOS production increased with microbial growth. Maximum amount of 72% (w/w) GOS was obtained from the optimized medium (5% lactose and 0.75% yeast extract) in 70 hours. Seven types of GOS (3 of dimer, 2 of trimer, 1 of tetramer, and 1 of pentamer) were identified by two-dimensional TLC. A new mechanism of GOS production is proposed based on the metabolism of carbon source.