Abstract: Full factorial design of experiment for medium optimization was employed for lipstatin production by Streptomyces toxytricini in shake flask study. The full factorial DOE was very much effective in screening of nutritional parameters within the stipulated time frame in a limited number of experiments. A maximum lipstatin production was achieved 3.290 g/l with the following optimized factors: soya flour 35g/l and soya oil 25g/l. Validation experiments were also carried out to verify the adequacy and the accuracy of the model. The results also give a scope for large scale fermentation of lipstatin production. [Luthra, U., Kumar, H., Kulshreshtha, N., Tripathi, A., Trivedi, A., Khadpekar, S., Chaturvedi, A. and Dubey, R.C. Medium optimization for the production of lipstatin by Streptomyces toxytricini using full factorial design of experiment. Nat Sci 2013;1