Production of cyclodextrin glucanotransferase (CGTase) from Bacillus sp. G1 was investigated. Two level Full Factorial Design has been employed to study the effect of different experimental variables on the production of CGTase. Three variables of pH (8, 9, and 10), temperature (32 C, 34.5 C, and 37 C) and agitation (100 rpm, 200 rpm, and 300 rpm) were used to identify the significant effects and interactions in the batch studies. A polynomial regression model has been developed using the experimental data. The results show that production of CGTase was strongly affected by the variations in pH. The maximum CGTase activity was achieved when the production was carried out at 32C, pH 10 with 100 rpm of agitation. The predicted value produced 27.85 U/ml is in close agreement with CGTase activity produce from experiment, which is 27.82 U/ml.