Taking the lipase activity in the fermentation broth as the target， through single factor， Plackett-Burman， steepest climbing and response surface tests， the optimal conditions for the production of lipase by the Moesziomyces aphidis were optimized. The results showed that culture conditions such as glucose， NaNO3， yeast extract powder， MgSO4， peanut oil， and Tween-80 medium components， the amount of liquid filling， and the fermentation temperature all affected the lipase activity. Among them， the 3 most significant factors affecting the lipase activity are the addition amount of NaNO3， the addition amount of yeast extract and the culture temperature. The optimal conditions after optimization are: the addition amount of NaNO3 is 3.1 g/L， the addition amount of yeast extract is 2.24 g/L， and culture temperature is 25.8 ℃. On this condition， the lipase activity in the fermentation broth is 83.14 U/mL， which is close to the predicted value of the regression equation obtained by the response surface， and is 2.6 times higher than the enzyme activity in the initial medium， indicating that the regression model can better optimize the lipase fermentation conditions. This study explored the industrial application potential of the Moesziomyces aphidis， providing a reference for broadening the source of lipase.