Abstract: OBJECTIVE To find a best production process by studying Lipstatin fermentation formula and culture conditions. METHODS During the aerobic fermentation, pH, carbon source, nitrogen source, inorganic salt, inoculate age, inoculate quantity, temperature and fermentation time were studied to optimize the fermentation media and culture conditions. RESULTS The most suitable pH for original culture was 6.5-7.0; the best carbon source was glycerol; the best nitrogen source was skimmed bean flour; and zinc ion was mostly impact the fermentation process among the inorganic salt; the best culture temperature was 30 ℃; the suitable fermentation time was 25 h and the best inoculate quantity was 4%-6%. CONCLUSION Fermentation activity unit can be increased through optimized fermentation formula and culture condition.