The invention relates to the biotechnology field, and particularly relates to penicillium capsulatum, and an isolated culture method and an application thereof. The invention provides a strain of penicillium capsulatum with the preservation number of CGMCC No.6020. Further researches on bacterial strain morphological characteristics, culture characteristics and carbon source utilization are carried out. The invention also provides the isolated culture and purification method of the bacterial strain, and the application in screening antifungal drugs.