Systematic investigations on the effect of varying dissolved oxygen (DO) concentrations and agitation profiles on glutaminase yield from Zygosaccharomyces rouxii NRRL Y-2547 were conducted in a batch stirred tank reactor. The agitation rate was varied in the range of 100-400 rpm, while the DO values studied were 20, 40 and 60 %, respectively. Furthermore, a stepwise reduction in DO was investigated wherein the DO concentration was decreased from 40 % for the first 12 h to 20 % for the following 24 h, and finally decreased to 5 % DO up to 54 h (end of batch period). The maximum enzyme yield in all cases was observed after 48 h at 37°C. An agitation rate of 200 rpm and a three-stage DO maintenance strategy (40-20-5 % DO) gave maximum glutaminase productivity. Yeast extract and glucose feeding increased the enzyme production from 370.75 ± 4.97 units/L to 437.14 ± 4.44 and 417.99 ± 5.82 units/L, respectively.