A laboratory experiment was conducted to optimize the level of carbon and nitrogen sources in the LGI growthmedium to achieve a higher population of native isolate of Gluconacetobacter G1. The basal LGI medium was amended withfour levels of sucrose i.e., 5, 10, 15 and 20 per cent as carbon source in combination with three levels of ammonium sulphatei.e. 1, 2 and 3 g per liter of medium as a nitrogen source. It was observed that the highest population (118 × 109 CFU ml-l)was recorded in T10 (Basal LGI medium with 10 % sucrose and 3 g l-1 ammonium sulphate) at 96th h after inoculation whichwas on par with T6 (Basal LGI medium with 10 % sucrose and 2 g l-1 ammonium sulphate) with a population of 69.18 × 109CFU ml-1 at 96th h after inoculation. However, the treatment T6 recorded the shortest mean generation time of 2.99 h whichwas on par with T10 with a mean generation time of 3.06 h. The pH of the medium in all the treatments decreasedcontinuously with incubation time and low values of pH in all the treatments were recorded at 120th h after inoculation. ThepH dropped very quickly during exponential growth phase of the bacterium.