Streptomyces toxytricini a Gram-positive bacterium belonging to the genus Streptomyces is producer of the pancreatic lipase inhibitor lipstatin,of which the antiobesity drug orlistat is a derivative.The study has been done for optimization of seed media and fermentation media using Plackett-Burman method for increasing the yield of the strain selected.Purification was done on silica gel column and identification of lipstatin molecule using sophisticated techniques such as IR, NMR, and mass spectroscopy. The presence and identification was also confirmed using TLC and HPLC method. This study also compares the impurity profile and the percentage purity of the lipstatin. The optimized media gave a titer of Lipstatin (1980 mg/ml) which is comparable to highest producing benchmarks (2400 mg/ml), but it is quite economical in a way that this work eliminates the use of L-Leucine as media component. The further increase in yield can be tried by different UV, chemical mutation methods in future.