The invention relates to a fermentation culture medium for increasing leucinostatin produced from purpureocillium lilacinum. The fermentation culture medium comprises, by weight, 45-55 parts of glucose, 4.5-5.5 parts of peptone, 4.5-5.5 parts of yeast extract, 0.5-2.0 parts of K2HPO4, 8.5-12.5 parts of Na2CO3, 0.1-0.5 part of MgSO4.7H2O, 1.5-10 parts of corn flour, 0.005-0.05 part of FeSO4.7H2O and 1.5-10.5 parts of urea. A culture method for increasing the leucinostatin produced from the purpureocillium lilacinum by the aid of the fermentation culture medium includes steps of carrying out fermentation culture on culture bacterium solution by the aid of purpureocillium lilacinum shake flasks; carrying out culture on the culture bacterium solution in seed tanks by the aid of the fermentation culture medium; inoculating the culture bacterium solution in the seed tanks into fermentation tanks and carrying out fermentation. The fermentation culture medium and the culture method have the advantages that the purpureocillium lilacinum cultured by the aid of the fermentation culture medium is low in cost, short in fermentation cycle and high in leucinostatin yield, and the like.