In the present paper, attempts have been made to control the production of pullulan by supplementing commercial source of carbons and protein, timely. Pullulan production was regulated by supplying full fat soya flour and hydrolyzed soya extract, individually and in combination. Pullulan quantification was assayed for sensitivity to pullulanase. Aureobasidium pullulans was found to produce 125.7 gL−1 of pullulan. The rotation speed of shake flask, the pH of broth, and the supply of air were maintained at 180 rpm, 5.9, and 1.5 vvm air, respectively. The effect of carbons and lipids on pullulan production was noticed to be substrate specific. However, after the lapse of 36 h, addition of full fat soya floor and hydrolyzed soya extract in combination enhanced the pullulan production 125.7 gL−1. Besides this, pH of broth was also noticed as a critical factor in monitoring pullulan biosynthesis. The newly isolated mutant Aureobasidium pullulans, having high potential for pullulan production as compared to existing data, can be well used for commercialization of pullulan.